Abstract
Objective(s):
Despite of wide range applications of polymeric nanoparticles in protein delivery, there are some problems for the field of protein entrapment, initial burst and controlled release profile.
Materials and Methods:
In this study, we investigated the influence of some changes in PLGA nanoparticles formulation to improve the initial and controlled release profile. Selected parameters were: pluronic F127, polysorbate 80 as surfactant, pH of inner aqueous phase, L/G ratio of PLGA polymer, volume of inner aqueous phase and addition of polyvinylpyrrolidone as an excipient. FITC-HSA was used as a model hydrophilic drug. The nanoparticles were prepared by nanoprecipitation.
Results:
Initial release of FITC-HSA from PLGA-tween 80 nanoparticles (opt-4, 61%) was faster than control (PLGA-pluronic) after 2.30 h of incubation. Results showed that decrease in pH of inner aqueous phase to pI of protein can decrease IBR but the release profile of protein is the same as control. Release profile with three phases including a) initial burst b) plateau and c) final release phase was observed when we changed volume of inner aqueous phase and L/G ratio in formulation. Co-entrapment of HSA with PVP and pluronic reduced the IBR and controlled release profile in opt-19. Encapsulation efficiency was more than 97% and nanoparticles size and zeta potentials were mono-modal and -18.99 mV, respectively.
Conclusion:
In this research, we optimized a process for preparation of PLGA-PVP-pluronic nanoparticles of diameter less than 300 nm using nanoprecipitation method. This formulation showed a decreased initial burst and long lasting controlled release profile for FITC-HSA as a model drug for proteins.
Combined effects of PEGylation and particle size on uptake of PLGA particles ...Nanomedicine Journal (NMJ)
Abstract
Objective:
At the present study, relationship between phagocytosis of PLGA particles and combined effects of particle size and surface PEGylation was investigated.
Materials and Methods:
Microspheres and nanospheres (3500 nm and 700 nm) were prepared from three types of PLGA polymers (non-PEGylated and PEGylation percents of 9% and 15%). These particles were prepared by solvent evaporation method. All particles were labeled with FITC-Albumin. Interaction of particles with J744.A.1 mouse macrophage cells, was evaluated in the absence or presence of 7% of the serum by flowcytometry method.
Results:
The study revealed more phagocytosis of nanospheres. In the presence of the serum, PEGylated particles were phagocytosed less than non-PEGylated particles. For nanospheres, this difference was significant (P<0/05) and their uptake was affected by PEGylation degree. In the case of microsphere formulation, PEGylation did not affect the cell uptake. In the serum-free medium, the bigger particles had more cell uptake rate than smaller ones but the cell uptake rate was not influenced by PEGylation.
Conclusion:
The results indicated that in nanosized particles both size and PEgylation degree could affect the phagocytosis, but in micron sized particles just size, and not the PEGylation degree, could affect this.
The present article deals with design of antibacterial and antifungal pH-responsive hydrogels based on Quaternary Ammonium Functionalized-Tragacanth Gum (QTG) biopolymer as drug delivery systems. The antimicrobial effects of the graft-copolymer hydrogels QTG/ polyacrylic acid (QTG-AA) and QTG/polyacrylamide (QTG-AM) were investigated against fi ve standard microorganisms including Candida albicans, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa. The results of the in-vitro release of quercetin as a drug from the functionalized copolymer hydrogels exhibited dependence on the pH, immersion time, medium, and temperature. All copolymer hydrogels demonstrated antibacterial and antifungal properties and moreover, QTG-AM copolymers presented higher antimicrobial activity than QTG-AA copolymers.
Long acting injectable microparticle formulation - a new dimension for peptid...Merck Life Sciences
Explore the clinical benefits and applications of sustained release drug delivery with this presentation. Access the findings from a technical feasibility study as well as a case study on sustained release microparticle formulation for a sensitive peptide.
Combined effects of PEGylation and particle size on uptake of PLGA particles ...Nanomedicine Journal (NMJ)
Abstract
Objective:
At the present study, relationship between phagocytosis of PLGA particles and combined effects of particle size and surface PEGylation was investigated.
Materials and Methods:
Microspheres and nanospheres (3500 nm and 700 nm) were prepared from three types of PLGA polymers (non-PEGylated and PEGylation percents of 9% and 15%). These particles were prepared by solvent evaporation method. All particles were labeled with FITC-Albumin. Interaction of particles with J744.A.1 mouse macrophage cells, was evaluated in the absence or presence of 7% of the serum by flowcytometry method.
Results:
The study revealed more phagocytosis of nanospheres. In the presence of the serum, PEGylated particles were phagocytosed less than non-PEGylated particles. For nanospheres, this difference was significant (P<0/05) and their uptake was affected by PEGylation degree. In the case of microsphere formulation, PEGylation did not affect the cell uptake. In the serum-free medium, the bigger particles had more cell uptake rate than smaller ones but the cell uptake rate was not influenced by PEGylation.
Conclusion:
The results indicated that in nanosized particles both size and PEgylation degree could affect the phagocytosis, but in micron sized particles just size, and not the PEGylation degree, could affect this.
The present article deals with design of antibacterial and antifungal pH-responsive hydrogels based on Quaternary Ammonium Functionalized-Tragacanth Gum (QTG) biopolymer as drug delivery systems. The antimicrobial effects of the graft-copolymer hydrogels QTG/ polyacrylic acid (QTG-AA) and QTG/polyacrylamide (QTG-AM) were investigated against fi ve standard microorganisms including Candida albicans, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa. The results of the in-vitro release of quercetin as a drug from the functionalized copolymer hydrogels exhibited dependence on the pH, immersion time, medium, and temperature. All copolymer hydrogels demonstrated antibacterial and antifungal properties and moreover, QTG-AM copolymers presented higher antimicrobial activity than QTG-AA copolymers.
Long acting injectable microparticle formulation - a new dimension for peptid...Merck Life Sciences
Explore the clinical benefits and applications of sustained release drug delivery with this presentation. Access the findings from a technical feasibility study as well as a case study on sustained release microparticle formulation for a sensitive peptide.
Improving Downstream Processing: Application of Excipients in DSPMilliporeSigma
This study investigates the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Applications of Poly (lactic acid) in Tissue Engineering and Delivery SystemsAna Rita Ramos
Applications of Poly (lactic acid) in Tissue Engineering and Delivery Systems
Poly (lactic acid) is a thermoplastic derived from renewable resources and is at present, one of the most promising biodegradable and nontoxic biopolymers. In addition to its versatility and consequent large-scale production, PLA can be processed with a large number of techniques.
Due to its excellent mechanical properties and biocompatibility, this polymer is becoming largely applied in the biomedical field such as in tissue engineering for scaffolds and in delivery systems in the form of micro and nanoparticles. Furthermore, because it’s relatively cheap and an eco-friend, it has been considered as one of the solutions to lessen the dependence on petroleum-based plastics and solid waste problems.
In order to maximize the knowledge and development of this polymer, it is necessary to understand the material synthesis, proprieties, manufacturing processes, main applications, commercialization and its market state, which will be presented in this review.
A presentation discussing amino acids, peptide bonds and peptide synthesis. The Merrifield Synthesis of Peptides is further discussed covering principle, methodology, isolation and purification, its advantages and disadvantages. A brief note on protecting groups is mentioned. A few practical applications of synthetic peptides are mentioned and discussed in brief as well.
Industrial fermentation-Does Fermentation Really Increase the Phenolic Conten...ShreyaMandal4
Decortication leads to a reduction in minerals, fibers, and antioxidants as phenolic compounds located in the peripheral parts of the grains. Fermentation can be applied to treat the whole nondecorticated flour to prevent functional loss. Bringing out the nutritional benefits of millets upon fermentation will serve us to include millets at a proportion in our meals along with traditional cereals.
Optimization of Corynebacterium glutamicum Immobilization on Alginate and In...IJMER
The parameters of the immobilized process of Corynebacterium glutamicum VTCC – B – 0632
on alginate were identified by Plackett-Burman matrix, and the experiments were designed by response
surface methodology having the central composite designs (RSM-CCD). The maximum yield of cell
immobilization on alginate carrier reached at 92.6%. Optimal parameters were the cell density of 89.3
million cells/mL in the 4% sterile alginate with ratio 1:1. This mixture went through the syringe system of
the 2M CaCl2 solution at 200C with the shaking speed of 75 rpm until the gels get in shape. Then, these
gels were soaked in the CaCl2 liquor and shaken for 41 minutes (150 rpm). At last, the particle size of
final products was 4mm and the average cell density was 14.75 million cells/gram. This immobile product
is maintained under the suitable condition in the CaCl2 liquor (w/v), pH=7. The cell survival percentage
after 72 hours were 98% when it was stored in 4
0C, 0.5% CaCl2 and pH of 7
THE SOLID PHASE PEPTIDE SYNTHESIS IS SLIGHTLY DIFFRENT FROM PEPTIDE SYNTHESIS WHICH IS DISCUSSED HERE, ITS SYNTHESIS WITH STRUCTURE ANS BASICS ARE DISCUSSED WHICH WILL BE VERY USEFUL FOR READERS.
Gelatin-grafted N- proflavine acryl amide was synthesized through two steps; firstly the Gelatin was grafted with
acrylic acid free radically using Ammonium per-sulfate at 60℃, Then it was modified to its corresponding acyl
chloride derivation, second step included the substitution with amino group of proflavine, in this research Gelatin
was used as a natural nontoxic, water soluble polymer as a drug carrier.
The prepared pro drug polymer was characterized by FTIR and 1H-NMR spectroscopies, Controlled drug release
was studied in different pH values at 37℃. Many advantages were obtained comparing with other known
methods.
Preparation and characterization of nimesulide loaded cellulose acetate hydro...Jing Zang
The aim of this study is to prepare nimesulide loaded cellulose acetate hydrogen phthalate nanoparticles by salting out technique. In this study Cellulose acetate Hydrogen phthalate was taken as polymer. Nimesulide was selected as a model drug. This technique is suitable for drugs and polymers that are soluble in polar solvents such as acetone or ethanol. The effect of drug concentration and polymer concentration on nanoparticle size, shape, uniform size distribution and stability was studied. Nanoparticles were evaluated for particle size, zetapotential and particle size distribution. Size of the particle was measured by SEM.(Scanning electron microscope).Surface charge and stability of the resultant nanoparticles was determined by Zetasizer. Particle size distribution was determined by Photon Correlation Spectroscopy (PCS) with a Malvern Zetasizer Nano-ZS. The cellulose acetate hydrogen phthalate concentration and nimesulide concentration was varied from 5mg/ml to 10 mg/ml. The effect of drug and polymer concentrations on nanoparticle size, shape, particle size distribution was studied. Increased drug concentration has no impact on the particle size. The size of the particle was found to be decreased with increased polymer concentration. Increased polymer concentration has resulted in uniform particle size distribution. Higher the polymer concentrations and lower the drug concentrations resulted in uniform particle size distribution.
Improving Downstream Processing: Application of Excipients in DSPMilliporeSigma
This study investigates the benefits of adding excipients during downstream processing on protein stability, chromatographic performance and viral inactivation.
Applications of Poly (lactic acid) in Tissue Engineering and Delivery SystemsAna Rita Ramos
Applications of Poly (lactic acid) in Tissue Engineering and Delivery Systems
Poly (lactic acid) is a thermoplastic derived from renewable resources and is at present, one of the most promising biodegradable and nontoxic biopolymers. In addition to its versatility and consequent large-scale production, PLA can be processed with a large number of techniques.
Due to its excellent mechanical properties and biocompatibility, this polymer is becoming largely applied in the biomedical field such as in tissue engineering for scaffolds and in delivery systems in the form of micro and nanoparticles. Furthermore, because it’s relatively cheap and an eco-friend, it has been considered as one of the solutions to lessen the dependence on petroleum-based plastics and solid waste problems.
In order to maximize the knowledge and development of this polymer, it is necessary to understand the material synthesis, proprieties, manufacturing processes, main applications, commercialization and its market state, which will be presented in this review.
A presentation discussing amino acids, peptide bonds and peptide synthesis. The Merrifield Synthesis of Peptides is further discussed covering principle, methodology, isolation and purification, its advantages and disadvantages. A brief note on protecting groups is mentioned. A few practical applications of synthetic peptides are mentioned and discussed in brief as well.
Industrial fermentation-Does Fermentation Really Increase the Phenolic Conten...ShreyaMandal4
Decortication leads to a reduction in minerals, fibers, and antioxidants as phenolic compounds located in the peripheral parts of the grains. Fermentation can be applied to treat the whole nondecorticated flour to prevent functional loss. Bringing out the nutritional benefits of millets upon fermentation will serve us to include millets at a proportion in our meals along with traditional cereals.
Optimization of Corynebacterium glutamicum Immobilization on Alginate and In...IJMER
The parameters of the immobilized process of Corynebacterium glutamicum VTCC – B – 0632
on alginate were identified by Plackett-Burman matrix, and the experiments were designed by response
surface methodology having the central composite designs (RSM-CCD). The maximum yield of cell
immobilization on alginate carrier reached at 92.6%. Optimal parameters were the cell density of 89.3
million cells/mL in the 4% sterile alginate with ratio 1:1. This mixture went through the syringe system of
the 2M CaCl2 solution at 200C with the shaking speed of 75 rpm until the gels get in shape. Then, these
gels were soaked in the CaCl2 liquor and shaken for 41 minutes (150 rpm). At last, the particle size of
final products was 4mm and the average cell density was 14.75 million cells/gram. This immobile product
is maintained under the suitable condition in the CaCl2 liquor (w/v), pH=7. The cell survival percentage
after 72 hours were 98% when it was stored in 4
0C, 0.5% CaCl2 and pH of 7
THE SOLID PHASE PEPTIDE SYNTHESIS IS SLIGHTLY DIFFRENT FROM PEPTIDE SYNTHESIS WHICH IS DISCUSSED HERE, ITS SYNTHESIS WITH STRUCTURE ANS BASICS ARE DISCUSSED WHICH WILL BE VERY USEFUL FOR READERS.
Gelatin-grafted N- proflavine acryl amide was synthesized through two steps; firstly the Gelatin was grafted with
acrylic acid free radically using Ammonium per-sulfate at 60℃, Then it was modified to its corresponding acyl
chloride derivation, second step included the substitution with amino group of proflavine, in this research Gelatin
was used as a natural nontoxic, water soluble polymer as a drug carrier.
The prepared pro drug polymer was characterized by FTIR and 1H-NMR spectroscopies, Controlled drug release
was studied in different pH values at 37℃. Many advantages were obtained comparing with other known
methods.
Preparation and characterization of nimesulide loaded cellulose acetate hydro...Jing Zang
The aim of this study is to prepare nimesulide loaded cellulose acetate hydrogen phthalate nanoparticles by salting out technique. In this study Cellulose acetate Hydrogen phthalate was taken as polymer. Nimesulide was selected as a model drug. This technique is suitable for drugs and polymers that are soluble in polar solvents such as acetone or ethanol. The effect of drug concentration and polymer concentration on nanoparticle size, shape, uniform size distribution and stability was studied. Nanoparticles were evaluated for particle size, zetapotential and particle size distribution. Size of the particle was measured by SEM.(Scanning electron microscope).Surface charge and stability of the resultant nanoparticles was determined by Zetasizer. Particle size distribution was determined by Photon Correlation Spectroscopy (PCS) with a Malvern Zetasizer Nano-ZS. The cellulose acetate hydrogen phthalate concentration and nimesulide concentration was varied from 5mg/ml to 10 mg/ml. The effect of drug and polymer concentrations on nanoparticle size, shape, particle size distribution was studied. Increased drug concentration has no impact on the particle size. The size of the particle was found to be decreased with increased polymer concentration. Increased polymer concentration has resulted in uniform particle size distribution. Higher the polymer concentrations and lower the drug concentrations resulted in uniform particle size distribution.
Está es la primera parte de una serie de presentaciones sobre ARRITMIAS, posteriormente subiré las demás arritmias activas y luego las pasivas. Espero que les sirvan.
Dejen sus comentarios !!
Power Point Presentation on GREEN CHEMISTRY
(info on pollution, causes and its prevention)
Friends if you found this helpful please click the like button. and share it :)
Similar to Preparation of protein-loaded PLGA-PVP blend nanoparticles by nanoprecipitation method: entrapment, Initial burst and drug release kinetic studies
Intranasal delivery of drug loaded thiolated co-polymeric microparticles for...Gaurav Patil
E-Presentation at Two days 15th Indo-US virtual International Conference
on “Global advances in Pharmaceutical and Allied Science”
In collaboration with
APP Gujarat State branch, AAP American International branch,
AAP Pharmedu Healthcare Manag Division
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Colloids and Surfaces B: Biointerfaces 101 (2013) 353– 360
Contents lists available at SciVerse ScienceDirect
Colloids and Surfaces B: Biointerfaces
j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / c o l s u r f b
harmacokinetics of curcumin-loaded PLGA and PLGA–PEG blend nanoparticles
fter oral administration in rats
ajeh Maissar Khalil , Thuane Castro Frabel do Nascimento , Diani Meza Casa , Luciana Facco Dalmolin ,
na Cristina de Mattos, Ivonete Hoss, Marco Aurélio Romano, Rubiana Mara Mainardes ∗
epartment of Pharmacy, Universidade Estadual do Centro-Oeste/UNICENTRO, Rua Simeão Camargo Varela de Sá 03, 85040-080 Guarapuava, PR, Brazil
r t i c l e i n f o
rticle history:
eceived 2 March 2012
eceived in revised form 10 June 2012
ccepted 12 June 2012
vailable online 28 June 2012
eywords:
urcumin
C–MS/MS
ioavailability
anoparticles
a b s t r a c t
The aim of this study was to assess the potential of nanoparticles to improve the pharmacokinetics of
curcumin, with a primary goal of enhancing its bioavailability. Polylactic-co-glycolic acid (PLGA) and
PLGA–polyethylene glycol (PEG) (PLGA–PEG) blend nanoparticles containing curcumin were obtained
by a single-emulsion solvent-evaporation technique, resulting in particles size smaller than 200 nm. The
encapsulation efficiency was over 70% for both formulations. The in vitro release study showed that cur-
cumin was released more slowly from the PLGA nanoparticles than from the PLGA–PEG nanoparticles. A
LC–MS/MS method was developed and validated to quantify curcumin in rat plasma. The nanoparticles
were orally administered at a single dose in rats, and the pharmacokinetic parameters were evaluated
and compared with the curcumin aqueous suspension. It was observed that both nanoparticles formu-
lations were able to sustain the curcumin delivery over time, but greater efficiency was obtained with
the PLGA–PEG nanoparticles, which showed better results in all of the pharmacokinetic parameters ana-
lyzed. The PLGA and PLGA–PEG nanoparticles increased the curcumin mean half-life in approximately 4
and 6 h, respectively, and the Cmax of curcumin increased 2.9- and 7.4-fold, respectively. The distribution
and metabolism of curcumin decreased when it was carried by nanoparticles, particularly PLGA–PEG
nanoparticles. The bioavailability of curcumin-loaded PLGA–PEG nanoparticles was 3.5-fold greater than
the curcumin from PLGA nanoparticles. Compared to the curcumin aqueous suspension, the PLGA and
PLGA–PEG nanoparticles increased the curcumin bioavailability by 15.6- and 55.4-fold, respectively.
These results suggest that PLGA and, in particular, P.
ABSTRACT Human platelet-derived growth factor-BB (hPDGF-BB), a proliferation factor, has been successfully manufactured and approved by FDA as the treatment for diabetic foot ulcer and self bone grafting. There have been no reports on the storage of the recombinant hPDGF-BB (rhPDGF-BB) in the Pichia pastoris fermentation broth although during the research of process development and the production manufacture it needs to be stored at low temperature. The concentration of rhPDGF-BB protein in the fed-batch fermentation broth of P. pastoris was stable during 3-week storage at -20oC, but its bioactivity was reduced by 20%. The addition of a mixture of 50% glycerol with either 1 mM EDTA or 1 mM PMSF into the fermentation broth could fully preserve the bioactivity of rhPDGF-BB until 3 weeks at -20oC. The addition of 50% glycerol with either 1 mM EDTA or 1 mM PMSF was found no affection on the protein purification process.
Key-words: rhPDGF-BB, Pichia pastoris, Fed-batch fermentation, Protein stability, Bioactivity, Storage
Method Development, Validation and Forced Degradation Studies of Dapagliflozi...ijtsrd
A simple, sensitive, robust, precise, and efficient RP HPLC approach for the simultaneous determination of Dapagliflozin and Pioglitazone Hydrochloride in Synthetic Mixture. As per ICH Q2 R1 guidelines, the final chromatographic conditions were Optimized with a mobile phase ratio of 25 75 v v in ACN Potassium Dihydrogen Phosphate Buffer pH 4 was adjusted by adding OPA at a flow rate of 1 mL min, column temperature of 30 °C, injection volume of 20 µL, Kromstar Vertex C18 analytical column, and UV detection at 228 nm wavelength. Dapagliflozin and Pioglitazone Hydrochloride reported retention times of 3 min and 6.5 min, respectively. Validation of a method was found to be linear in the range of 2 10 µg ml for Dapagliflozin and 3–15 µg mL for Pioglitazone Hydrochloride. The Recovery for Dapagliflozin was discovered to be 98.52 99.90 , while for Pioglitazone Hydrochloride, it was found to be 99.67 99.94 . The Precision results for both drugs were within the limits while expressed Intraday and Interday. For Dapagliflozin, the LOD and LOQ were reported to be 0.041 µg mL and 0.13 µg mL, respectively, and for Pioglitazone Hydrochloride, 0.105 µg mL and 0.32 µg mL. As per ICH Q1A R2 guidelines, the synthetic mixture was subjected to acid, base, oxidation, thermal, and photolysis stress conditions. Mr. Tarang Patel | Ronak Parikh "Method Development, Validation and Forced Degradation Studies of Dapagliflozin and Pioglitazone Hydrochlorides in Synthetic Mixtures by RP-HPLC" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-6 | Issue-6 , October 2022, URL: https://www.ijtsrd.com/papers/ijtsrd52165.pdf Paper URL: https://www.ijtsrd.com/pharmacy/analytical-chemistry/52165/method-development-validation-and-forced-degradation-studies-of-dapagliflozin-and-pioglitazone-hydrochlorides-in-synthetic-mixtures-by-rphplc/mr-tarang-patel
Poly caprolactone loaded with antibiotics may be used to treat infections of the respiratory tract, like tuberculosis . this is important for biodegradation of rubber compound.
This presentation deals with the brief study of functionalized graphene oxide nanoparticles for the delivery of model anticancer drug. It represents benefits of targeted drug delivery over the conventional drug delivery,
It is one of the remingtons journal club seminar which is part of masters degree progam in pharmacy.
1. bioconversion of progesterone using aqueous two phase system by comamonas ...Darshan Rudakiya
Strains of Comamonas acidovoras have been
reported to have vital role in degradation of natural
as well as complex organic compounds.
Comamonas acidovoras MTCC 3364 has been
routinely reported for steroid bioconversion by
activated beads in aqueous system. Previously
studies observed that progesterone was converted
into AD and ADD steroids. These compounds were
higher valuable steroids which were mainly used in
different types of drugs. Novel system was used for
the bioconversion of progesterone in to AD and
ADD. Aqueous two-phase system was initially
optimized for the progesterone solubility, separation
of two phases and extraction of product steroids
from the system. Even 1% of PEG can easily
dissolved progesterone but 5% of PEG needed for
separating phases and for extraction purposes.
Beads was easily tolerate up to 20% of PEG system
even for 120 hours. Hexane used as better
extraction solvent among different type of solvent
system. Products were shown in PEG 6000 system
after 96 hours in fewer amounts. PEG 8000 system
showed products 48 hours in minor amount but
increased in 96 hours. Advantage of aqueous twophase
system in progesterone bioconversion was
better stability of progesterone, extraction of steroid
products and stability of the system.
In vitro and in vivo evaluation of positively charged liposaccharide derivati...Adel Abdelrahim, PhD
Similar to Preparation of protein-loaded PLGA-PVP blend nanoparticles by nanoprecipitation method: entrapment, Initial burst and drug release kinetic studies (20)
Evaluation of the effect of crocetin on antitumor activity of doxorubicin enc...Nanomedicine Journal (NMJ)
Objective(s): The current study reports investigation of codelivery by PLGA nanoparticles (NPs) loaded with crocetin (Cro), a natural carotenoid dicarboxylicHYPERLINK “http://en.wikipedia.org/wiki/Carboxylic_acid” acid that is found in the crocus flower, and Doxorubicin (DOX).
Materials and Methods: Double emulsion/solvent evaporation method was used for preparation of PLGA nanoparticles containing Dox and Cro. Characterizations of prepared NPs were investigated by atomic force microscopy (AFM) and dynamic light scattering analysis. In vitro Cytotoxicity of DOX and Cro loaded PLGA NPs (PLGA-DOX-Cro) on MCF-7 cell line was evaluated using MTT test. Flow cytometry experiments were implemented to distinguish cells undergoing apoptosis from those undergoing necrosis. Furthermore the expression of caspase 3 was examined by western blot analysis.
Results: The prepared formulations had size of 150- 300 nm. Furthermore, PLGA-DOX-Cro nanoparticles inhibited MCF-7 tumor cells growth more efficiently than either DOX or Cro alone at the same concentrations, as quantified by MTT assay and flow cytometry. Studies on cellular uptake of DOX-Cro-NPs demonstrated that NPs were effectively taken up by MCF-7 tumor cells.
Conclusion: This study suggested that DOX-Cro-NPs may have promising applications in breast cancer therapy.
Effects of combination of magnesium and zinc oxide nanoparticles and heat on ...Nanomedicine Journal (NMJ)
Objective: The objective of this study was to investigate the antibacterial activities of combination of MgO and ZnO nanoparticles in the presence of heat against Escherichia coli and Staphylococcus aureus.
Materials and Methods:Bacteria were grown on either agar or broth media followed by the addition of ZnO and MgO nanoparticles. Then the combined effect of ZnO and MgO nanoparticles was investigated. Furthermore, the media containing nanoparticles were treated with mild heat and their synergistic antibacterial activity was investigated against E. coli and S. aureus in milk.
Results: The data showed that the nanoparticles used in this study had no effect on the bacteria in the agar medium. However, the results showed that ZnO and MgO nanoparticles resulted in a significant decrease in the number of E. coli (P<0.000) and S. aureus (Pd”0.05) in the broth medium. The combination of nanoparticles and mild heat exhibited a significant decrease in the number of E. coli and S. aureus indicating the synergistic effects of nanoparticles and heat.
Conclusion: Using a combination of mild heat, ZnO and MgO nanoparticles, E. coli and S. aureus can be controlled successfully in the milk. Mild heating plus ZnO and MgO nanoparticles has a synergistic effect which would reduce the need for high temperature and also the concentrations of ZnO and MgO nanoparticles required for pathogen control in minimally processed milk during maintaining.
Preparation and evaluation of electrospun nanofibers containing pectin and ti...Nanomedicine Journal (NMJ)
Objective(s):The aim of this study was to prepare electrospun nanofibers of celecoxib using combination of time-dependent polymers with pectin to achieve a colon-specific drug delivery system for celecoxib.
Materials and Methods:Formulations were produced based on two multilevel 22 full factorial designs. The independent variables were the ratio of drug:time-dependent polymer (X1) and the amount of pectin in formulations (X2). Electrospinning process was used for preparation of nanofibers. The spinning solutions were loaded in 5 mL syringes. The feeding rate was fixed by a syringe pump at 2.0 mL/h and a high voltage supply at range 10-18 kV was applied for electrospinning. Electrospun nanofibers were collected and evaluated by scanning electron microscopy and drug release in the acid and buffer with pH 6.8 with and without pectinase.
Results:Electrospun nanofibers of celecoxib with appropriate morphological properties were produced via electrospinning process. Drug release from electrospun nanofibers was very low in the acidic media; while, drug release in the simulated colonic media was the highest from formulations containing pectin.
Conclusion: Formulation F2 (containing drug:ERS with the ratio of 1:2 and 10% pectin) exhibited acceptable morphological characteristics and protection of drug in the upper GI tract and could be a good candidate as a colonic drug delivery system for celecoxib.
The combined effects of Aloe vera gel and silver nanoparticles on wound heali...Nanomedicine Journal (NMJ)
Objective(s): This study was aimed at investigating the synergy effects of Aloe vera gel and silver nanoparticles on the healing rate of the cutting wounds.
Materials and Methods: In order to determine the concentration of silver nanoparticles in Aloe vera gel, the MBC methods were applied on the most common bacteria infecting wounds, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa. The cutting wounds with Full-thickness skin were dorsally created on rats; then the rats were divided into 4 groups. The treatments groups included: mixture of Aloe vera gel and silver nanoparticles, Aloe vera gel alone and silver nanoparticles alone in addition to control groups. The treatment was carried out for 2 weeks and the size of the wound closures were measured by an image software analysis.
Results:There was no significant difference (p<0.05) in healing rate between the control and mixture group. However, there were significant differences between the silver nanoparticles and Aloe vera groups using Tukey’s analysis on the 6th, 8th and 10th days.
Conclusion:The Aloe vera gel increased the rate of wound healing whereas the silver nanoparticles had a delay effect; and when they were mixed, it was similar to the average effect of both Aloe vera gel and silver nanoparticles.
Simultaneous loading of 5-florouracil and SPIONs in HSA nanoparticles: Optimi...Nanomedicine Journal (NMJ)
Objective(s): Over the past two decades, considerable interest has been focused on utilizing biocompatible magnetic nanoparticles (MNPs) for biomedical applications. In this study, production of human serum albumin (HSA) nanoparticles using desolvation technique that were simultaneous loaded with high amounts of superparamagnetic iron oxide nanoparticles (SPIONs) and 5-flourouracil (5-FU) was investigated.
Materials and Methods: 5-FU loading (%) and SPIONs entrapment efficiency (%) were optimized using response surface methodology (RSM). The design expert software used to analyse the interactive effects of pH, 5-FU and SPIONs concentrations.
Results:The optimum conditions found to be pH of 8.2, drug concentration of 1.5 mg/ml and SPIONs concentration of 2.79 mg/ml. Under the mentioned optimum conditions, particles with the size of 111.8 nm, zeta potential of -37.1 mV, 5-FU loading of 15.8% and SPIONs entrapment efficiency of 41.1% were obtained. In vitro cumulative release of 5-FU from the nanoparticles was evaluated in phosphate buffer saline (pH 7.4, 37 °C). Results indicated that 85% of the 5-FU released during 95 h, which revealed a sustained release profile. In addition, Vibrating Sample Magnetometer (VSM) analyses confirmed the superparamagnetic properties of magnetic albumin nanoparticles manufactured under the optimum conditions.
Conclusion: According to the findings,SPIONs and 5-FU loaded HAS nanoparticles arepromising for use as novel targeted delivery system due to proper magnetic and drug release behaviours.
Antimicrobial and cytotoxicity effect of silver nanoparticle synthesized by C...Nanomedicine Journal (NMJ)
Objective(s): For the development of reliable, ecofriendly, less expensive process for the synthesis of silver nanoparticles and to evaluate the bactericidal, and cytotoxicity properties of silver nanoparticles synthesized from root extract of Croton bonplandianum, Baill.
Materials and Methods: The synthesis of silver nanoparticles by plant part of Croton bonplandianum was carried out. The formation of nanoparticles was confirmed by Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), XRD and UV-Vis spectrophotometric analysis. The biochemical properties were assayed by antibacterial study, cytotoxicity assay using cancer cell line.
Results: The formation of silver nanoparticles was confirmed by UV-VIS spectroscopic analysis which showed absorbance peak at 425 nm. X-ray diffraction photograph indicated the face centered cubic structure of the synthesized AgNPs. TEM has displayed the different dimensional images of biogenic silver nanoparticles with particle size distribution ranging from 15-40 nm with an average size of 32 nm. Silver particles are spherical in shape, clustered. The EDX analysis was used to identify the elemental composition of synthesized AgNPs. Antibacterial activity of the synthesized AgNPs against three Gram positive and Gram negative bacteria strains like Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa carried out showed significant zones of inhibition. The cytotoxicity study by AgNPS also showed cytotoxicity on ovarian cancer cell line PA-1 and lung epithelial cancer cell line A549.
Conclusion: The present study confirms that the AgNPs have great promise as antibacterial, and anticancer agent.
Investigation of the effect of different parameters on the phase inversion te...Nanomedicine Journal (NMJ)
Objective(s): Nanoemulsions are a kind of emulsions that can be transparent, translucent (size range 50-200 nm) or “milky” (up to 500 nm). Nanoemulsions are adequatly effective for transfer of active component through skin which facilitate the entrance of the active component . The transparent nature of the system and lack of the thickener and fluidity are among advantages of nanoemulsion.
Materials and Methods: In this study, a nanoemulsion of lemon oil in water was prepared by the phase inversion temperature (PIT) emulsification method in which the tween 40 was used as surfactant. The effect of concentration of NaCl in aqueous phase, pH and weight percent of surfactant and aqueous on the PIT and droplet size were investigated. Results: The results showed that with increasing of concentration of NaCl from 0.05 M to 1 M, PIT decrease from 72 to 50. The average droplet sizes, for 0.1, 0.5 and 1 M of NaCl in 25 ºC are 497.3, 308.1 and 189.9 nm, respectively and the polydispersity indexes are 0.348, 0.334 and 0.307, respectively.
Conclusion: Considering the characteristics of nanoemulsions such as being transparent, endurance of solution and droplet size can provide suitable reaction environment for polymerization process used in making hygienic and medical materials.
Mechanism of oxidative stress involved in the toxicity of ZnO nanoparticles a...Nanomedicine Journal (NMJ)
ZnO NPs (zinc oxide nanoparticles) has generated significant scientific interest as a novel antibacterial and anticancer agent. Since oxidative stress is a critical determinant of ZnO NPs-induced damage, it is necessary to characterize their underlying mode of action. Different structural and physicochemical properties of ZnO NPs such as particle surface, size, shape, crystal structure, chemical position, and presence of metals can lead to changes in biological activities including ROS (reactive oxygen species) production. However, there are some inconsistencies in the literature on the relation between the physicochemical features of ZnO NPs and their plausible oxidative stress mechanism. Herein, the possible oxidative stress mechanism of ZnO NPs was reviewed. This is worthy of further detailed evaluations in order to improve our understanding of vital NPs characteristics governing their toxicity. Therefore, this study focuses on the different reported oxidative stress paradigms induced by ZnO NPs including ROS generated by NPs, oxidative stress due to the NPs-cell interaction, and role of the particle dissolution in the oxidative damage. Also, this study tries to characterize and understand the multiple pathways involved in oxidative stress induced by ZnO NPs. Knowledge about different cellular signaling cascades stimulated by ZnO NPs lead to the better interpretation of the toxic influences induced by the cellular and acellular parameters. Regarding the potential benefits of toxic effects of ZnO NPs, in-depth evaluation of their toxicity mechanism and various effects of these nanoparticles would facilitate their implementation for biomedical applications.
Synthesis of silver nanoparticles and its synergistic effects in combination ...Nanomedicine Journal (NMJ)
Abstract
Objectives:
Biofilms are communities of bacteria attached to surfaces through an external polymeric substances matrix. In the meantime, Acinetobacterbaumannii is the predominant species related to nosocomial infections. In the present study, the effect of silver nanoparticles alone and in combination with biocides and imipenem against planktonic and biofilms of A. baumannii was assessed.
Materials and Methods:
Minimum inhibitory concentrations (MICs) of 75 planktonic isolates of A. baumannii were determined by using the microdilution method as described via clinical and laboratory standards institute (CLSI). Among all strains, 10 isolates which formed strong biofilms were selected and exposed to silver nanoparticles alone and in combination with imipenem, bismuth ethandithiol (BisEDT) and bismuth propanedithiol (BisPDT) to determine minimum biofilm inhibitory concentrations (MBIC). Subsequently, minimum biofilm eradication concentrations (MBECs) of silver nanoparticles alone and in combination with imipenem against mature biofilm of the isolates were evaluated.
Results:
Results showed that 29.3% of isolates were susceptible to silver nanoparticles and could inhibit the growth and eradicate biofilms produced by the isolates. For this reason, ∑FIC, ∑FBIC and ∑FBEC ≤ 0.05 were reported which shows synergism between silver nanoparticles and imipenem against not only planktonic cells but also inhibition and eradication of biofilms. The results of ∑FBIC >2 indicated to antagonistic impacts between silver nanoparticles and BisEDT/BisPDT against biofilms.
Conclusion:
It can be concluded that silver nanoparticles alone can inhibit biofilm formation but in combination with imipenem are more effective against A. baumannii in planktonic and biofilm forms.
Abstract
Objective(s):
Zinc oxide nanoparticles (ZNP) are increasingly used in sunscreens, biosensors, food additives and pigments. In this study the effects of ZNP on liver of rats was investigated.
Materials and Methods:
Experimental groups received 5, 50 and 300 mg/kg ZNP respectively for 14 days. Control group received only distilled water. ALT, AST and ALP were considered as biomarkers to indicate hepatotoxicity. Lipid peroxidation (MDA), SOD and GPx were detected for assessment of oxidative stress in liver tissue. Histological studies and TUNEL assay were also done.
Results:
Plasma concentration of zinc (Zn) was significantly increased in 5 mg/kg ZNP-treated rats. Liver concentration of Zn was significantly increased in the 300 mg/kg ZNP-treated animals. Weight of liver was markedly increased in both 5 and 300 mg/kg doses of ZNP. ZNP at the doses of 5 mg/kg induced a significant increase in oxidative stress through the increase in MDA content and a significant decrease in SOD and GPx enzymes activity in the liver tissue. Administration of ZNP at 5 mg/kg induced a significant elevation in plasma AST, ALT and ALP. Histological studies showed that treatment with 5 mg/kg of ZNP caused hepatocytes swelling, which was accompanied by congestion of RBC and accumulation of inflammatory cells. Apoptotic index was also significantly increased in this group. ZNP at the dose of 300 mg/kg had poor hepatotoxicity effect.
Conclusion:
It is concluded that lower doses of ZNP has more hepatotoxic effects on rats, and recommended to use it with caution if there is a hepatological problem.
Synthesis of graphene oxide-TiO2 nanocomposite as an adsorbent for the enrich...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
In our study, graphene oxide-TiO2 nanocomposite (GO/TiO2) was prepared and used for the enrichment of rutin from real samples for the first time.
Materials and Methods:
The synthesized GO/TiO2 was characterized by X-ray diffraction, scanning electron microscopy, and FT-IR spectra. The enrichment process is fast and highly efficient. The factors including contact time, pH, and amount of GO/TiO2 affecting the adsorption process were studied.
Results:
The maximum adsorption capacity for ciprofloxacin was calculated to be 59.5 mg/g according to the Langmuir adsorption isotherm. The method yielded a linear calibration curve in the concentration ranges from 15 to 200 μg/L for the rutin with regression coefficients (r2) of 0.9990. The limits of detection (LODs, S/N=3) and limits of quantification (LOQs, S/N=10) were found to be 8 μg/Land 28 μg/L, respectively. Both the intra-day and inter-day precisions (RSDs) were < 10% .
Conclusion:
The developed approach offered wide linear range, and good reproducibility. Owing to the diverse structures and unique characteristic, GO/TiO2 possesses great potential in the enrichment and analysis of trace rutin in real aqueous samples.
Preparation and evaluation of vitamin A nanosuspension as a novel ocular drug...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
The aim of this study was to prepare a nanosuspension formulation as a new vehicle for the improvement of the ocular delivery of vitamin A.
Material and Methods:
Formulations were designed based on full factorial design. A high pressure homogenization technique was used to produce nanosuspensions. Fifteen formulations were prepared by the use of different combinations of surfactants Tween 80, benzalkonium chloride and Pluronic and evaluated for pH, particle size, entrapment efficiency, differential scanning calorimetry (DSC), stability and drug release. Also, Draize test was used to evaluate the irritation of rabbit eye by formulations.
Results:
All formulations showed a small mean size that is well suited for ocular application. Also it was observed that the particle size decreased with increase in the amount of surfactant. Drug entrapment increased with increasing amount of surfactant. It was shown that initial and final drug release can be controlled by the ratio and the total amount of surfactants, respectively.
Conclusion:
It was concluded that the use of Tween 80 and Pluronic in the formualtions with a proper ratio does not show eye irritation and could be useful to achieve a suitable nanosuspension of vitamin A as a novel ocular delivery system.
A comparative study about toxicity of CdSe quantum dots on reproductive syste...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Medicinal benefits of quantum dots have been proved in recent years but there is little known about their toxicity especially in vivo toxicity. In order to use quantum dots in medical applications, studies ontheir in vivo toxicity is important.
Materials and Methods:
CdSe:ZnS quantum dots were injected in 10, 20, and 40 mg/kg doses to male mice10 days later, mice were sacrificed and five micron slides were prepared structural and optical properties of quantum dots were evaluated using XRD.
Results:
Histological studies of testis tissue showed high toxic effect of CdSe:ZnS in 40 mg/kg group. Histological studies of epididymis did not show any effect of quantum dots in terms of morphology and tube structure. Mean concentration of LH and testosterone and testis weight showed considerable changes in mice injected with 40 mg/kg dose of CdSe:ZnS compared to control group. However, FSH and body weight did not show any difference with control group.
Conclusion:
Although it has been reported that CdSe is highly protected from the environment by its shell, but this study showed high toxicity for CdSe:ZnS when it is used in vivo which could be suggested that shell could contribute to increased toxicity of quantum dots. Considering lack of any previous study on this subject, our study could potentially be used as an basis for further extensive studies investigating the effects of quantum dots toxicity on development of male sexual system.
Functionalization of carbon nanotubes and its application in nanomedicine: A ...Nanomedicine Journal (NMJ)
Abstract
This review focuses on the latest developments in applications of carbon nanotubes (CNTs) in medicine. A brief history of CNTs and a general introduction to the field are presented.
Then, surface modification of CNTs that makes them ideal for use in medical applications is highlighted. Examples of common applications, including cell penetration, drug delivery, gene delivery and imaging, are given. At the same time, there are concerns about their possible adverse effects on human health, since there is evidence that exposure to CNTs induces toxic effects in experimental models. However, CNTs are not a single substance but a growing family of different materials possibly eliciting different biological responses. As a consequence, the hazards associated with the exposure of humans to the different forms of CNTs may be different. Understanding the structure–toxicity relationships would help towards the assessment of the risk related to these materials. Finally, toxicity of CNTs, are discussed. This review article overviews the most recent applications of CNTs in Nanomedicine, covering the period from 1991 to early 2015.
The role of surface charge of ISCOMATRIX nanoparticles on the type of immune ...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
ISCOMATRIX vaccines have now been shown to induce strong antigen-specific cellular or humoral immune responses to a broad range of antigens of viral, bacterial, parasite or tumor. In the present study, we investigated the role of ISCOMATRIX charge in induction of a Th1 type of immune response and protection against Leishmania major infection in BALB/c mice.
Materials and Methods:
Positively and negatively charged ISCOMATRIX were prepared. BALB/C mice were immunized subcutaneously, three times with 2-week intervals, with different ISCOMATRIX formulations. Soluble Leishmania antigens (SLA) were mixed with ISCOMATRIX right before injection. The extent of protection and type of immune response were studied in different groups of mice.
Results:
The group of mice immunized with negatively charged ISCOMATRIX showed smaller footpad swelling upon challenge with L. major and the highest IgG2a production compared with positively charged one. The mice immunized with positively charged ISCOMATRIX showed the lowest splenic parasite burden compared to the other groups. Cytokine assay results indicated that the highest level of IFN- γ and IL-4 secretion was observed in the splenocytes of mice immunized with negatively charged ISCOMATRIX as compared to other groups.
Conclusion:
The results indicated that ISCOMATRIX formulations generate an immune response with mixed Th1/Th2 response that was not protective against challenge against L. major.
Abstract
In the last decade, developments in nanotechnology have provided a new field in medicine called “Nanomedicine”. Nanomedicine has provided new tools for photodynamic therapy. Quantum dots (QDs) are approximately spherical nanoparticles that have attracted broad attention and have been used in nanomedicine applications. QDs have high molar extinction coefficients and photoluminescence quantum yield, narrow emission spectra, broad absorption, large effective stokes shifts. QDs are more photostable and resistant to metabolic degradation. These photosensitizing properties can be used as photosensitizers for Photodynamic Therapy (PDT). PDT has been recommended for its unique characteristic, such as low side effect and more efficiency. Therefore, nanomedicine leads a promising future for targeted therapy in cancer tumor. Furthermore, QDs have recently been applied in PDT, which will be addressed in this review letter. Also this review letter evaluates key aspects of nano-particulate design and engineering, including the advantage of the nanometer scale size range, biological behavior, and safety profile.
Abstract
Objective(s):
Abdominal adhesions are one of the most important problems, occurring after intra-abdominal surgery in more than 90% of cases. This condition is the leading cause of bowel obstruction, infertility, and abdominal/pelvic pain. Gold nanoparticles (GNPs) have been shown to be non-toxic and exhibit anti-inflammatory, anti-angiogenic and antioxidant activities. The purpose of this study was to determine the effect of intraperitoneal lavage with GNP solutions on the development of postoperative peritoneal adhesion (PPA).
Materials and Methods:
In the current experimental study, thirty-five male Wistar rats were randomly assigned to seven groups of five rats. After a standardized peritoneal injury, GNP solutions in different concentrations (1, 2.5, 5, 10, 50 and 100 ng/ml) were locally administered through nebulization; normal saline (NS) was administered to the control group. Two weeks later, the rats were sacrificed and cecum and peritoneal samples were harvested for histopathological assessment. Blood samples were obtained to determine serum concentrations of inflammatory biomarkers including tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β) and vascular endothelial growth factor (VEGF).
Results:
The rats treated with GNPs had significantly lower microscopic and macroscopic peritoneal adhesion scores, compared to the control group (P<0.05). Score 5 of macroscopic adhesions was reported in all the rats of the control group, unlike the GNP groups. Furthermore, microscopic adhesions were reported with all rats in the control group, unlike the GNP groups (reported in 0 out of 5 rats in all GNP groups). In addition, serum levels of IL-1β, TNF-α and VEGF underwent no significant changes.
Conclusion:
Compared to the control group, GNPs decreased the severity of peritoneal adhesions, although they did not alter TNF-α, IL-1β or VEGF serum levels.
Preparation and characterization of Sr-Ti-hardystonite (Sr-Ti-HT) nanocomposi...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Hardystonite (HT) is Zn-modified silicate bioceramics with promising results for bone tissue regeneration. However, HT possesses no obvious apatite formation. Thus, in this study we incorporated Sr and Ti into HT to prepare Sr-Ti-hardystonite (Sr-Ti-HT) nanocomposite and evaluated its in vitro bioactivity with the purpose of developing a more bioactive bone substitute material.
Materials and methods:
The HT and Sr-Ti-HT were prepared by mechanical milling and subsequent heat treatment. Calcium oxide (CaO), zinc oxide (ZnO) and silicon dioxide (SiO2) (all from Merck) were mixed with molar ratio of 2:1:2. The mixture of powders mixture was then milled in a planetary ball mill for 20 h. In the milling run, the ball-to-powder weight ratio was 10:1 and the rotational speed was 200 rpm. After synthesis of HT, 3% nanotitanium dioxide (TiO2, Degussa) and 3% strontium carbonate (SrCO3, Merck) were added to HT and then the mixture was ball milled and calcined at 1150°C for 6 h. Simultaneous thermal analysis (STA), X-ray diffraction (XRD), Transmission electron microscopy (TEM) and Fourier transform infra-red spectroscopy (FT-IR) performed to characterize the powders.
Results:
XRD and FT-IR confirmed the crystal phase and silicate structure of HT and TEM images demonstrated the nanostructure of powders. Further, Sr-Ti-HT induced apatite formation and showed a higher human mesenchymal stem cell (hMSCs) adhesion and proliferation compared to HT.
Conclusion:
Our study revealed that Sr-Ti-HT with a nanostructured crystal structure of 50 nm, can be prepared by mechanical activation to use as biomaterials for orthopedic applications.
How to Split Bills in the Odoo 17 POS ModuleCeline George
Bills have a main role in point of sale procedure. It will help to track sales, handling payments and giving receipts to customers. Bill splitting also has an important role in POS. For example, If some friends come together for dinner and if they want to divide the bill then it is possible by POS bill splitting. This slide will show how to split bills in odoo 17 POS.
How to Create Map Views in the Odoo 17 ERPCeline George
The map views are useful for providing a geographical representation of data. They allow users to visualize and analyze the data in a more intuitive manner.
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
Ethnobotany and Ethnopharmacology:
Ethnobotany in herbal drug evaluation,
Impact of Ethnobotany in traditional medicine,
New development in herbals,
Bio-prospecting tools for drug discovery,
Role of Ethnopharmacology in drug evaluation,
Reverse Pharmacology.
Preparation of protein-loaded PLGA-PVP blend nanoparticles by nanoprecipitation method: entrapment, Initial burst and drug release kinetic studies
1. Nanomed. J., 2(3):175-186, Summer 2015
175
Preparation of protein-loaded PLGA-PVP blend nanoparticles by
nanoprecipitation method: entrapment, Initial burst and drug release
kinetic studies
1
S. Shakeri; 2*
R. Roghanian; 2
G. Emtiazi; 3
C. Errico; 3
F. Chiellini; 3*
E. Chiellini
1
Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences,
Graduate University of Advanced Technology, Kerman, Iran
2
Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran
3
Department of Chemistry and Industrial Chemistry, University of Pisa, Via Risorgimento 35, 56126 Pisa, Italy
ABSTRACT:
Objective(s): Despite of wide range applications of polymeric nanoparticles in protein delivery, there are some problems
for the ûeld of protein entrapment, initial burst and controlled release profile.
Materials and Methods: In this study, we investigated the influence of some changes in PLGAnanoparticles formulation
to improve the initial and controlled release profile. Selected parameters were: pluronic F127, polysorbate 80 as
surfactant, pH of inner aqueous phase, L/G ratio of PLGA polymer, volume of inner aqueous phase and addition of
polyvinylpyrrolidone as an excipient. FITC-HSA was used as a model hydrophilic drug. The nanoparticles were
prepared by nanoprecipitation.
Results: Initial release of FITC-HSA from PLGA-tween 80 nanoparticles (opt-4, 61%) was faster than control (PLGA-
pluronic) after 2.30 h of incubation. Results showed that decrease in pH of inner aqueous phase to pI of protein can
decrease IBR but the release profile of protein is the same as control. Release profile with three phases including a)
initial burst b) plateau and c) final release phase was observed when we changed volume of inner aqueous phase and L/
G ratio in formulation. Co-entrapment of HSA with PVP and pluronic reduced the IBR and controlled release profile in
opt-19. Encapsulation efficiency was more than 97% and nanoparticles size and zeta potentials were mono-modal and
-18.99 mV, respectively.
Conclusion: In this research, we optimized a process for preparation of PLGA-PVP-pluronic nanoparticles of diameter
less than 300 nm using nanoprecipitation method. This formulation showed a decreased initial burst and long lasting
controlled release profile for FITC-HSA as a model drug for proteins.
Keywords: Controlled releaseprofile, FITC-HSA, Initial burst release, PLGA nanoparticles, Protein delivery
Nanomed. J., 2(3):175-186, Summer 2015
ISSN 2322 - 5904
*Corresponding Author Email: rasoul_roghanian
@yahoo.co.uk; emochie@doci.unipi.it
Tel.: (+98) 3556126;
Note. This manuscript was submitted on April 3, 2015;
approved on June 12, 2015;
Received; 3 April 2015 Accepted; 12 June 2015
INTRODUCTION
Nanoparticles have wide range applications in
medicine and pharmacy, because of their specific
properties (1). In particular; biopolymeric nanoparticles
have become increasingly attractive due to
biocompatibility and biodegradability (2). The design of
nano-sized biodegradable polymeric nanoparticles to
target tissues has certain unique advantages in drug
delivery, pharmacology and tissue engineering (3, 4).
These novel formulations reduce therapeutic dosage,
related toxicity and side effects (5-7). Polymeric
nanoparticles have wide applications in biomedicine (8)
and especially for the ûeld of nanobiotechnology,
nanoparticlescomposedofpoly(D,L-lactide-co-glycolide)
(PLGA) have shown several technological advantages
such as sustained and controlled release and safety (9,
10).PLGAisacopolymerofpoly(D,L-lactide-co-glycolide)
and is an ideal candidate of biodegradable polymers for
2. Nanomed. J., 2(3):175-186, Summer 2015
176
S. Shakeri et al.
formulation into nanoparticles (11-13). The PLGAbased
colloidal systems are stable in biological fluid and are
suitable for protein encapsulation due to their long
half-life (14). In the recent years, proteins have been
proved to be very effective candidates for therapeutic
use in a large number of human diseases (15, 16).
Proteins are hydrophilic drugs and thereby are unstable
and degradable in the body system and therefore need
effective drug delivery devices to complete protection
from enzymes degradation and loss of their activity
(17). Polymeric nanoparticulated systems such as PLGA
nanoparticles can control protein release kinetics and
ease of administration in addition of increasing protein
stability and activity in the biological systems (18).
However, the bulk core of PLGA nanoparticles is
hydrophobic and can dissolve only hydrophobic drugs
and hence nanoparticles formulated using PLGA
polymers, demonstrate low proteintaceus drug
encapsulation efûciency and rapid release (19). In this
research we demonstrated a new strategy by using of
polyvinylpyrrolidone (PVP) to enhance encapsulation
of FITC- HSA as a model hydrophilic drug into the
hydrophobic core of PLGA nanoparticles. PVP or
povidone, is soluble in water and polar solvents and
made from monomeric unite of vinylpyrrolidone (20).
PVP was synthesized in 1939 after development of
acetylene chemistry at BASF in the 1920 by Walter
Reppe (21). The U.S. Food and Drug Administration
(FDA) has approved PVP.PVP was first as a
substitution and expander for blood plasma. Its wide
range solubility in conventional solvents,
biocompatibility and non-toxicity characteristics gave
it numerous applications in medicine, pharmacy and
cosmetics. Some of advantageous characteristics of
PVP are shown in Table 1. PVP has a long, proven
history of pharmaceutical applications as delivery
systems of poorly soluble drugs (22). It also possesses
a high degree of biocompatibility with the possibility
to control the rate of drug release so as to improve the
in vivo pharmacokinetics (23). PVP has also been
shown to inhibit drug crystallization in solution as well
as in the solid state and protects against degradation
in solution (24).The aim of this work was to solubilize
water-soluble drug molecules in polymeric organic
solution and make the homogeneous dispersion of
protein with PLGA chains for nanoparticle formation
during nanoprecipitation methods.We have
investigated the effects of various formulation
parameters on encapsulation efficiency and initial burst
release (IBR) of FITC-HSA in comparison with PLGA-
PVP blend nanoparticles.
MATERIALS AND METHODS
PLGA (poly(D,L-lactide-co-glycolide)) with 50:50
monomer ratio and inherent viscosity midpoint of 0.4
dl/g was purchased from purac-Hollandunder the
commercial nameofPurasorb®
PDLG5004.Thepluronic
F-127, polyvinylpyrrolidone (PVP) with molecular
weights of K10 and fluorescein isothiocyanate
conjugated to human serum albumin (FITC-HSA) were
obtained from Sigma Aldrich. PVP with molecular
weights of K15 and K25 were purchased from Fluka.
Other chemicals and solvents used were of the highest
grade commercially available.
Preparation of FITC-HSAloaded PLGA nanoparticles
Nanoparticles were formulated by a nano-
precipitation technique (25). FITC-HSA solution in
water (1 mg/200 µl and pH=7.0) was emulsified into
PLGA-Pluronic solution in organic solution (ratio of
50/50 mg in 2 ml dichloromethane). The primary
emulsion was mixed by vortexing (3000/min, Vibromix)
for 30s and then was poured onto a polar phase (25ml
ethanol) under moderate magnetic stirring leading to
immediate polymer nanoprecipitation in the form of
nanoparticles. This nanoparticle suspension was
immediately diluted with 25ml water and stirred using a
magnetic stirrer for 10 min. Solvent was evaporated
under vacume at 30°C (BUCHI, Rotavapor R-114,
Germany). Nanoparticles were reco-vered by
centrifugation (1 h, 8,000 × g, 15°C, model) and
resuspended in water or PBS.
Preparation of different formulations for protein
entrapment and release kinetic studies
In order to evaluate the effect of the different
formulations on the entrapment efficiency and kinetic
of drug release, some variables or modification
in formulation were investigated (the effect of pH, L/G
ratio, volume of internal aqueous phase, tween 80
instead of pluronic and PVP as excipient with/without
pluronic. Individual exper ments for preparation of
PLGAnano-particles are summarized in Table 2.
Preparation of PLGA-PVP and PLGA-PVP-pluronic
blend nanoparticles
PLGA-PVP with/without pluronic blend nanoparticle
was prepared by nano-precipitation method. Briefly,
3. Nanomed. J., 2(3):175-186, Summer 2015
177
an amount ofPLGA (50/50 ratio) and PVP (K10, 15 and
25) with/without pluronic were dissolved in 2 ml of
dichloromethane. This organic solution was mixed by
magnet stirrer and poured in polar phase (25 ml ethanol).
Nanoparticles solution was diluted with distilled water
and solvent was evaporated under vacuum at 30°C.
Nanoparticles were collected by centrifugation and
resuspended in water or PBS. The method for
preparation of FITC-HSA loaded PLGA-PVP nano-
particles was the same as method for preparation of
HSA loaded PLGA nano-particle.
Characterization of nanoparticles
Size and zeta potential determination
Particle size analyzingfor determination ofdimension
was performed by means of a Coulter LS230 Laser
Diffraction Particle SizeAnalyzer equipped with small
volume module plus. 30% obscuration ofPIDS detector
was used for nanoparticle suspension size analysis
with three runs on each sample. Deionized water was
used as background. Zeta potential of the nanoparticles
was determined by Beckman coulter delsa™ nano zeta
potential and submicron particle size analyzer.
Nanoparticles were dispersed in deionized water and
were subjected to zeta potential analysis at room
temperature.
Scanning electron microscopy (SEM)
SEM experiments were carried out on a JEOL
LSM5600LV scanning electron microscopy. The
nanoparticles samples were puriûed by centrifugation.
The pellets were frozen at -20°C, freeze-dried in a
5Pascal Lio 5P lyophilizator. These powders were used
for gold sputtering and SEM analysis.
Determination of FITC-HSA entrapment efficiency
and yield of nanoparticle production
Protein entrapment efficiency was calculated from
total FITC-HSAadded in formulation and the amount of
FITC-HSA present in the supernatant samples
recollected after centrifugation of nanoparticle
suspension. Total HSA content was calculated as the
sum of FITC-HSA amount both in the supernatant and
entrapped in nanoparticles. The amount of FITC-HSA
present in the supernatant was determined by
fluorescence spectroscopy (excitation/emission wave-
lengths of 495/518 nm) using spectro-photometer. Drug
entrapment efficiency was calculated as the percent of
the total HSA added in formulation. Standard curve was
obtained by plotting the fluorescence intensity of FITC-
HSA versus the concentration of HSA (mg/ml). To
measure yield of nanoparticle production, nano-particle
suspension in pre-weight falcon tube was centrifuged
and the pellet was frozen at -20°C, freeze-dried in a
5Pascal Lio 5Plyophilizator.Yield ofnanoparticle (%w/
w) was calculated as the percent of the total polymers
(PLGA, PVP and pluronic) were used in formulation.
In vitro release kinetic studies
In vitro release kinetic of nanoparticle encapsulated
drug was determined during one month. Nanoparticles
pellet was re-dispersed in 25 ml phosphate buffered
saline (PBS, pH=7.4) after centrifugation. The
nanoparticle suspension was incubated at 37°C with
horizontal shaking (100 rpm). At determined time
intervals, 1 mlofmediumwascentrifuged and the amount
of FITC-HSApresent in the supernatant was determined
by fluorescence spectroscopy. Drug concentration in
the release medium and cumulative release profiles were
calculated using calibration curve.
Cell culture and in vitro cell cytotoxicity assessment
The human fibroblast cell line was obtained from
national cell bank of Iran (Pasteur institute of Iran,
Tehran) and cultured in DMEM medium supplemented
with 10% heat-inactivated FBS and 100U/mL pen-icillin
and 100 µg/mL streptomycin at 37ºC in humidiûed
incubator with an atmosphere of 5% CO2
. The cell
cytotoxicity of PLGA-PVP blend nanoparticles was
evaluated by MTT assay. Briefly 2×104
cells/mL were
seeded in 96-well culture plates containing 200 µl of
medium in triplicate, allowed to attach, grow for 24 h
and subsequently the culture medium was replaced by
fresh medium containing nanoparticles (0.5-10 mg/mL),
then incubated for 48 h. The wells without drug and
nanoparticle were used as control. Negative control
cells were treated with DMSO. After treatment, media
were carefully removed and 20 µl of MTT (5 mg/ml) is
added to each well. Microplates were incubated for 4 h
at 37ºC and then the medium was removed; formazan
was dissolved in 200 µl of acidic isopropanol (0.04-0.1
N HCl in absolute isopropanol). Formazan absorbance
was assessed at 490 nm using a microplate reader. A
wavelength of 630 nm was used as the reference. The
results are given as relative values to the negative control
in percentage, whereas the untreated (positive) control is
set to be 100% viable. The data were analyzed using
Student’s t-test with significant difference at P < .05.
4. Nanomed. J., 2(3):175-186, Summer 2015
178
Protein-loaded PLGA-PVP blend nanoparticles
RESULTS
Entrapment efficiency and in-vitro release kinetic
studies
All of entrapment efficiencies for different
formulations were higher than 90%. The entrapment
efficiency was increased more than 97% by adding of
PVP in nanoparticles formulation (Table 3, opt-6 and
opt-17, 18, 19). A nanoprecipitation method was used
for preparation of PLGA nanoparticles with different
formulations.
The release ofFITC-HSA from PLGA nanoparticles
was monitored in PBS (pH=7.4), 100 rpm at 37°C for
more than one month. Percentage of protein that had
been released was shown in Figs. 1, 2 and 3. FITC-HSA
loaded PLGA-pluronic blend nanoparticles were
considered as control formulation in release kinetic
studies. Nanoparticles were prepared as noted in
materials and methods and monitored for protein
release in PBS. In this research, our aims were
decreasing ofIBR and increasing duration of the release
profile. For these resions, we changed some of factors
in control formulation (e.g tween 80 as surfactant, pH
of inner aqueous phase, L/G ratio of PLGA polymer,
volume of inner aqueous phase and addition of PVP
Table 1. Some advantageous characteristics of polyvinylpyrrolidone.
Broad specterum solubility in hydrophilic and hydrophobic solvents
Good affinity to different kinds of polymers and substrates
High hygroscopicity
Good adhesiveness to various substrates and formation of stable complex with active substances
Formation of chelate or complex
Improve the release rate, bioavailability and solubility of drugs
Reduction of drugs toxicity
Stabilization of suspension
Homogeneous distribution of substances in solution or in the coating
Table 2. Individual experiments with different formulations for protein encapsulation and release.
Experiments
PLGA (50/50)
ratio (mg)
PLGA (75/25)
ratio (mg)
Pluronic
(mg)
PVP (mg)
Tween 80
(µl)
aqueous phase
volume (µl)
aqueous
phase pH
Control a
50 - 50 - - 200 7.0
Opt-1 50 - 50 - - 100 7.0
Opt-2 - 50 50 - - 200 7.0
Opt-4 b
50 - - - 50 200 7.0
Opt-5 50 - 50 - - 200 3.0
Opt-6 c
50 - - 50 (K25) 250 200 7.0
Opt-7 50 - - 50 (K10) 250 200 7.0
Opt-8 50 - - 50 (K15) 250 200 7.0
Opt-11 50 - 25 25 (K25) - 200 7.0
Opt-17 d
25 - 41 13 (K15) - 200 7.0
Opt-18 e
25 - 41 13 (K10) - 200 7.0
Opt-19f
25 41 13 (K25) - 200 7.0
with different molecular weight as excipient). IBR was
measured after 2.30 h of incubation. IBR in control
formulation (Fig. 1) is about 58% of the nanoparticle
bounded FITC-HSA after 2.30 h. The release profile
was increased and continued with constant slope
during 10 days and more than 80% of total entraped
protein, released to the PBS solution.Addition of tween
80 instead of pluronic in organic phase (opt-4) and
change of pH of inner aqueous phase (opt-5) increased
and decreased IBR of protein, respectively (Fig. 1). IBR
in opt-4 is the same as control formulation, but release
was dramatically increased and more than 85% of
entraped protein released into the PBS after 24 h of
incubation. These data indicate that the rate of release
of HSA is enhanced due to use of tween-80. Change in
pH of inner aquoes phase (opt-5) considerably
decreased IBR. A slow IBR was observed in PLGA-
pluronic nanoparticles when pH of aquoes phase
containing of HSA was decreased below pH of
isoelecteric point of HSA protein (pH=3.0<4.5). In this
formulation, 37% of total proteins were released during
2.30 h of incubation in PBS (pH=7.4) at 37°C. But the
release profile was the same as control and opt-4 during
5. Nanomed. J., 2(3):175-186, Summer 2015
179
10 days. Release profiles from formulations with change
in volume of inner aquoes phase (opt-1), L/G ratio (opt-
2) andaddition of PVP asexcipient with/withoutpluronic
in formulation (opt-6, opt-19) were completely different
from release profile of control formulation (Figs. 2 and
3).IBRis50%foropt-1after2.30 hofincubation(Fig.2).
An slow release profile was observed to 10% of total
drug loaded in nanoparticles between 1-13 days and
continued by a final release up to 85% between 13 to 18
days of incubation. The same profile was observed for
opt-2 formulation. IBR is 46% after 2.30 h. This phase
was continued and 66% of proteins were released during
2 days of incubation. The release profile showed a
constant slop with slow increasing of protein release till
18 days of incubation. The significant influence of
addition ofPVP instead of pluronic onIBR was observed
in PLGA-PVP nanoparticles. Lower IBR was observed
in opt-6 and there was 37% of HSA released during the
2.30 h ofincubation and 58% of total drugs was released
during 2 days of incuation. The plateau phase was
observed between 2-10 days of incubation and
continued by a final release of 20% of total drug during
10-18 days.Addition of PVPinformulation withpluronic
(opt-19) resulted in change in IBR and duration (Fig. 3).
IBR and duration were decreased and increased in opt-
19 formulation (withpluronicas surfactant), respectively.
35% of total drugs were released during 2.30 h of
incubation and there is a plateau phase between 1-16
days of incubation. No significant release was not
observed during this phase. Final release was observed
after 16 days and reached to 75% of total protein. In this
formulation, thequantityofPLGA, PVP and pluronic are
optimized andnanoparticlesizeis 297±52nmwithmono-
modal distribution (Table 3).
Nanoparticles characterization: SEM, nanoparticle
size and distribution
Nanoparticles morphology was investigated by SEM.
All nanoparticles showed a swell-defined spherical shape
with nano-size (Figs. 5 and 6). The size of nanoparticles
were depended on the formulation conditions. Decreases
of volume and pH of inner aquoeus phase (opt-1 and 5
respectively) and use of tween 80 instead of pluronic in
organicphase(opt-4)resulted indecreasesofnanoparticle
sizeto132,124and149nmwithmono-modaldistribution,
respectively(Table3).Thenanoparticlesizewasincreased
and rangedbetween250-297nm,whenformulationswere
changedinamountofpluronicandPVP.Thenanoparticles
size was increased when 50 mg of PVP was used instead
ofpluronicinopt-6formulation.TheaveragesizeofPLGA-
PVP nanoparticles was 250±32 nm with tri modal-
distribution (Fig. 4). The other two populations (3 and
200 µm) were observed with very low number. Average
size of nanoparticles ranged 169±34 nm in control
formulation (Fig.5). Opt-19 is the optimized formulation
(Fig.6). Intheseformulation,PVPwasusedincombination
with pluronic to improve of nanoparticles size and
distribution, initial burst release and stability. All of
nanoparticles were mono-odal and very stable.
Nanoparticles size is 297±52 nm with mono-modal
distribution.
Fig. 1. The influence of the tween-80 (²%), pH (×) and
pluronic (control, Ï%) on the FITC-HSA in vitro release
from PLGA nanoparticles incubated in PBS (pH=7.4) at 37
°C and 100 rpm.
Fig. 2. The influence of the inner water volume (f&), L/G
ratio ( %) and PVP instead of pluronic (×) on the FITC-HSA
in vitro release from PLGA nanoparticles incubated in PBS
(pH=7.4) at 37 °C and 100 rpm.
6. Nanomed. J., 2(3):175-186, Summer 2015
180
S. Shakeri et al.
Fig. 3. The influence of the inner water volume (f&), L/G
ratio ( %) and PVP instead of pluronic (×) on the FITC-HSA
in vitro release from PLGA nanoparticles incubated in PBS
(pH=7.4) at 37 °C and 100 rpm
Fig. 4. Particle size distribution of FITC-HSA-loaded PLGA-
PVP nanoparticles (Opt 6) prepared by nanoprecipitation
method.
Homogeneous distribution of FITC-HAS
We observed two different regions with different
colors in nanoparticles pellet after nanoprecipitation.
Aqueous suspension of FITC-HSAis in yellow color. In
general, hydrophilic properties of HSA prevent it to
dissolve homogeneously in organic phase (DCM phase).
Heterogeneous dispersion of HSA was observed in PLGA
nanoparticle in control formulation after addition of first
emulsion (water phase in organic solution) into the
ethanol phase. Two regions were observed in
nanoparticles pellet (Fig. 7-A). The region R1
is in white
color and region R2
is in yellow color. Homogeneous
distribution of FITC-HSAmolecules was observed within
the nanoparticles pellet when PVP was used without/
with of pluronic in fromulations of opt-6 and opt-19
respectively (Fig. 7-C, B). The united yellow color
was observed in whole nanoparticles pellet in both
formulations.
Yield of PLGA nanoparticles
The effect of concentration of PVP on the yield of
lyophilized PLGAnanoparticles is shown in Table 4.
The yield of control formulation based on polymers
and surfactant used in formulation is 39%. The yield
of PLGA nanoparticles is increased and reached a
value of about 60%, when 50 mg of PVP was used
instead of pluronic (opt-6). We used tween 80 as
surfactant in ethanolic phase (1% v/v) in this
formulation. The percentage of yield decreased in opt-
19 to a value of 33% in comparision with control
formulation, when w/w ratio of Pluronic/PLGA
increased from 1 to 1.65 and w/w ratio of PVP/PLGA
decreased from 1 to 0.5. As a result, the yield of
nanoparticle formations was increased as the
concentration of PVP increased. The yild in PLGA-
Pluronic nanoparticle (control formulation) was low
in comparision of formulation with PVP addition (opt-
6). The zeta potential and mobility of opt-19
nanoparticles were -18.99 mV and -1.483e-004 Cm2
/
Vs, respectively and nanoparticles were stable in
suspension.
In vitro cytocompatibility assay
Cytocompatibility of prepared PLGA-PVP
nanoparticles were assessd by MTT test. The
viability of fibroblast cells treated with 0.5, 1, 2, 4 and
6 mg /mL of nanoparticles was 98%, 87%, 63%, 11%
and 1.8%, respectively. Results showed PLGA-PVP
nanoparticles have good cytocompatibility (IC50
=2.4
mg/ml).
DISCUSSION
Low protein encapsulation efficiency and rapid
initial burst release (IBR) are major obstacles for
application of nanoparticles in nanomedicine (26).
Thereby, hydrophilic molecules of proteins with short
physiological half-life need to the delivery systems
to enhance entrapment, their half-life and controlled
release after administration (27, 28). The main objects
of this research were to investigate different
formulations of PLGA nanoparticles for enhanced
entrapment efficiency, controlled initial burst and in
vitro release of FITC-HSA as a model proteinaceous
drug. It had been shown that PLGA:pluronic
7. Nanomed. J., 2(3):175-186, Summer 2015
181
Fig.6. Scanning electron micrograph and particle size distribution of FITC-HSA-loaded PLGA-PVP-pluronic nanoparticles prepared
by nanoprecipitation method (Opt-19). Nanoparticles size is 297±52 nm with mono-modal distribution
Fig. 5. Scanning electron micrograph and particle size distribution of FITC-HSA-loaded PLGA-pluronic nanoparticles prepared by
nanoprecipitation method. Average size of nanoparticles ranged 169±34 nm (control formulation)
8. Nanomed. J., 2(3):175-186, Summer 2015
182
S. Shakeri et al.
Table 3. Entrapment efficiency (%) and initial burst release of PLGA nanoparticles: effect of volume of aqueuse phase, L/G ratio,
surfactant (pluronic and tween 80), polyvinylpyrrolidon and effect of pH of aqueouse phase.
Experiments Entrapment
efficiency (%)
Size (nm) Initial burst release (%) after 2.30 h Size distribution
Control 92 169±34 58% Mono-modal
Opt-1 95 132±52 50% Mono-modal
Opt-2 91 174±47 46% Mono-modal
Opt-4 93 149±33 61% Mono-modal
Opt-5 95 124±25 37% Mono-modal
Opt-6 96 250 nm,3 µm, 200 µm 37% Tri-modal
Opt-7 ND 600 nm,2 µm, 5 µm, 15 µm ND Tetra-modal
Opt-8 ND 2 µm ND Mono-modal
Opt-11 >97% 155 nm, 3 µm 45% Bi-modal
Opt-17 >97% 271±42 34% Mono-modal
Opt-18 >97% 263±58 38% Mono-modal
Opt-19 >97% 297±52 35% Mono-modal
Table 4. Yield of nanoparticle production with various formulations via nanoprecipitation method.
Factors (mg) Control Opt-6 Opt-19
PLGA (mg) 50 50 25
PVP (mg) --- 50 13
Pluronic (mg) 50 --- 41
PLGA+PVP+pluronic (mg) 100 100 79
PVP/PLGA ratio (w/w) --- 1 0.52
Pluronic/PLGA ratio (w/w) 1 --- 1.65
Nanoparticle (mg) 39±2 60±3 26±1
Yield (based on PLGA+PVP +pluronic) 39±2% 60±3% 33±2%
Fig. 7. The effect of different concentrations of polyvinylpyrrolidon on dispersion of FITC-HSA in nanoparticles pellet. A)
Heterogeneous dispersion of HSA in control formulation. Two regions were observed with white and yellow colors. B)
Homogeneous dispersion of HSA in PLGA nanoparticle (opt-6) with formulation of of PLGA 50 mg, polyvinylpyrrolidon 50
mg and HSA-FITC 1 mg/200 µl water. Tween 80 (1%) was used as surfactant in ethanol phase, only one region was observed
in nanoparticle pellet in yellow color. C) Homogeneous dispersion of HSA in PLGA nanoparticle (opt-19) with formulation
of PLGA 25 mg, pluronic 41 mg, polyvinylpyrolidon 13 mg and HSA-FITC 1 mg/200 µl water, only one region was
observed in nanoparticle pellet in yellow color
A B C
R1
R2
Homogeneous
dispersion of
HSA in PLGA
ananoparticle
nanoparticles encapsulate hydrophilic proteins with
minimum loss in their biological activity (29, 30) and
protect them against degradation by acidic condition
(10). But, IBR is one of the main problems that still
associated to PLGA nanoparticles as protein delivery
system (31). Initial drug release and long time release
during incubation time are dependent to composition
of nanoparticles formulation. There were two separate
release phases based on release kinetic for opt-4, opt-
5 and control formulations. The release profiles were
9. Nanomed. J., 2(3):175-186, Summer 2015
183
biphasic. The first phase is acceleration or initial burst
release during 2.30h of incubation. The main reasons
for IBR are deficiently protein entrapment and
weakly attachment on the surface of the PLGA
nanoparticles. This release profile was continued by
a constant release with increasing slop during 10
days in opt-4, 5 and control. This phase named as
final release phase or secound phase. This phase is
related to protein release from nanoparticles matrics.
The release of HSA from PLGA-tween 80
nanoparticle formulation (opt-4) was faster than
control (PLGA-pluronic). The results showed that
PLGA nanoparticles prepared in opt-4 formulation
have a higher initial burst (61%) than those prepared
by control formulation reached up to 85% after 1
day. Tween 80 instead of pluronic not only increased
the IBR, but also final release was ocurred during 1
day of incubation. The most of HSA was released
during acceleration phase and only less than 10%
was release during final release phase. The weak
interaction between HSA and PLGA polymer or
inhibitory effect of tween 80 on adsorbtion of protein
to the polymer chains are probable resions for fast
release of drug in the first phase. The results showed
that the most of protein molecules are attached
weakly on the surface of PLGA nanoparticles not
inside of core of nanoparticles. The use of tween-80
as a surfactant instead of pluronic reduced the
affinity of HSA to entrap inside of PLGA
nanoparticles. The results suggested that tween 80
has an inhibitory role in protein entrapment. These
results could be explained by the reduced HSA–
PLGA interaction caused by the presence of tween
80, as a surfactant. Results showed that decreases
of pH of inner aqueous phase to pI of protein can
decrease IBR but not duration of protein release.
Proteins are stable in pH 6.5-7.0, but protein
refolding, aggregation and formation of proteins
fibrils take place at a pH close to pI of proteins (32).
Change of pH in opt-5 formulation can affect physic-
chemical stabilities of proteins. There is a new phase
in addition of IBR and final release phases in opt-1,
2, 6 and 19. Initial burst release or acceleration phase
was continued by a deceleration phase and then
there is a plateau phase before final release phase.
The slow release or plateau phase observed when
we changed volume of inner aqueous phase, L/G
ratio and when we used PVP without or with pluronic
in formulation. PLGA with L/G ratio of 75:25 is less
hydrophilic than copolymer with a 50:50 ratio of L/G
(33). This hydrophobic nature is due to lactic acid
monomers. So, this kind of PLGA degrades in a
longer time in comparison of more hydrophilic PLGA
(34, 35). Results indicate that, in the absence of
pluronic, HSA was very efficiently encapsulated in
the opt-6 formulation. The results showed that the
FITC-HSA initial burst release from opt-6 and 19 is
lower than nanoparticles prepared by formulations
without PVP. This could be interpreted to the
enhanced entrapment of the protein molecules inside
of the nanoparticles matrix by application of PVP in
formulations. Opt-6 wasn’t the best formulation for
nanoparticle preparation and some works were
needed to find the optimized formulation for
nanoparticles size, mono-modal distribution and long
time release. However, the best formulation for
protein encapsulation and release was opt-19 (PLGA-
PVP-pluronic). On the other hand, the mono-
dispersed nanoparticles size and long time release
of protein were observed in opt-19 formulation by
the incorporation of the PVP and pluronic together.
PVP has wide range solubility in conventional
solvents and has a long, proven history of
applications as delivery systems for poorly soluble
drugs (20). PVP inhibits and protects drug
crystallization and degradation in solution (24). We
did not analyze inner structure of nanoparticles in
opt-19 which could affect the release of the
encapsulated Protein, but the size of nanoparticles
is bigger than other formulations. As shown in Table
3, the size of the nanoparticles in opt-6 and opt-19
formulations was highly affected by introduction of
PVP in formulation with or without pluronic. The
increase in EE% and decrease in IBR maybe due to
the stabilizing property of PVP. Stabilizers and their
concentration are important factors in nanoparticles
size, homogeneous or heterogeneous distribution
of nanoparticles, entrapment efficiency and release
rate profile (33). Coombes et al (36) used PVP as
stabilizer for ovalbumin (OVA) loaded microparticles.
Their results showed that PVP enhanced entrapment
efficiency and release rate profile. Nanoparticles
based on PLGA-pluronic (control formulation)
showed faster onset of initial and final release than
nanoparticles based on PLGA-PVP-pluronic.
Hydration property of nanoparticles and
formation of water channels inside of nanoparticles
structure increase by pluronic and thereby resulted
10. Nanomed. J., 2(3):175-186, Summer 2015
184
Protein-loaded PLGA-PVP blend nanoparticles
in initial burst and faster final release (10, 29, 30).
Amphiphilic property of PVP make this polymer
soluble in water and many organic solvents (37).
Addition of PVP in formulation changes hydrophobic
or hydrophilic properties of nanoparticles. Its
potential as a hydrophilizing agent for biomaterials,
reduce hydrophobic interactions in the surface and
matrix of PLGA nanoparticles (21, 38). In the other
hand, hydrophilic property of protein and its interaction
with polymer chains and nanoparticles matrix determine
protein entrapment and IBR (31, 39). The best
interaction between proteins molecules and PLGA
matrix resulted in entrapment of protein inside of
nanoparticles with high efficacy. It has been reported
that conjugation of PVP with proteins prevents them
from autolysis, degradation and enhances half life of
proteins (40-43). This conjugation can control initial
burst and release profile as tailored made nanoparticle
(44).Applications of PVPnanoparticles in drug delivery
have been shown.This kind of nanoparticles can evade
fro reticoloendothelial system and have long time
circulation in blood stream (22, 23). These results
showed that PVP as a nanoparticle, conjugate with
proteins or as a polymeric modifier in combination with
other biomaterials (e.g., PLGA and pluronic) is a good
candidate for delivery of protein and other hydrophilic
therapeutic agents like as plasmid DNA (45). In this
paper we optimized a process for preparation of PLGA-
PVP-pluronic nanoparticles ofdiameter less than 300nm
using nanoprecipitation method. This formulation
showed an decreased initial burst and long time
controlled release profile for FITC-HSA as amodel drug
for proteins. These results showed that the release of
HSA or proteinaceus hydrophilic drugs from different
formulation based nanoparticles is not dependent only
to polymer degradation. There are effective factors
such as protein properties, surfactants, inhibitors (for
inhibition of protein interaction with polymer chains)
and stabilizing agants that can affect protein
entrapment and release from nanoparticles.
CONCLUSION
In this research, optimized formulation of PLGA-
PVP-pluronic nanoparticles were prepared by nano-
precipitation method for improvement of initial
release and kinetic of hydrophilic model drug. Initial
burst release was decreased in this formulation and
controlled release profile was observed in a long
time for FITC-HSA as a model drug for proteins. In
conclusion, PLGA-PVP nanoparticles have good
cytocompatibility and can be used as a nanocarrier
for controlled release of protein and peptide drugs.
ACKNOWLEDGEMENTS
The financial support from National Elite Foundation
(NEF: IRAN) is gratefully acknowledged.
CONFLICTSOFINTERESTS
The authors declare that they have no competing
interests.
AUTHORS’CONTRIBUTIONS
SS and CE performed the experiments and collected
data. RR, GE and EC: Supervisors and designed the
experiment and interpreted results. FC and CE:Advisors
and interpreted results. SS prepared the manuscript.
All authors read and approved the final manuscript.
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How to cite this article:
Shakeri S, Roghanian R, Emtiazi G, Errico C, Chiellini F, Chiellini E. Preparation of protein-loaded PLGA-PVP blend nanoparticles by
nanoprecipitation method: Entrapment, Initial burst and drug release kinetic studies. Nanomed. J., 2015; 2(3): 175-186.
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