Sishant Rao Divya, M.Pharm thesis ppt

1. A
synopsis on
Development and evaluation of N-
Acetylcysteine loaded nanostructure lipid
carrier to treat ischemic stroke
Submitted to: Submitted by
Dr. Rajinikanth Siddalingam Sishant Rav Divya
(Department of Pharmaceutical science) M.Pharm III sem.(Pharmaceutics
BBAU Lucknow
Session: 2017-2019

2. Index
Introduction
Ischemic Strokes
Types of Stroke
Ischemic Stroke Treatment
Problem with brain stroke

3. INTRODUCTION
 Ischemic Strokes: is a second most leading cause of death,
The 62000 stroke occurs every year in Canada effect all age
group.
 Thrombotic stroke ( block arteries)
 Hemorrhagic stroke ( Blood vessel ruptured)

4. Types of Stroke

5. Ischemic Stroke Treatment
tPA, the Gold Standard
 The gold standard, tissue Plasminogen activator (or
Alteplase IV r-tPA) is an FDA-approved treatment for
ischemic stroke.
 Plasminogen activator dissolve the clot and improve the
blood flow to the part of brain.
 430 potentially medicine use to treat ischemic stroke over the
period 1995-2015 such as-
1. Adenosine diphosphate
2. Butylphthalide/3n-butylphthalide
3. α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid or more.

6. Problem with brain stroke
The blood Brain Barrier (BBB) is major problem with
hydrophilic drug.

7. • Application of nanostructured lipid carriers: the prolonged protective
effects for sesamol in in vitro and in vivo models of ischemic stroke
via activation of PI3K signalling pathway

8. Background:
• Treatment of the ischemic stroke has remained a major healthcare
challenge.
• The phenolic compound, sesamol, has shown antioxidant and
neuroprotective effects, however, fast clearance may negatively
affect its efficiency.
• This, prompted us to incorporate sesamol into the nanostructured
lipid carriers (S-NLCs) and evaluate its therapeutic potential in in vitro
and in vivo models of ischemic stroke

9. Methods
Materials
• Cell culture materials were all purchased from GIBCO/ Invitrogen,
Germany. Cetyl palmitate and Tween 80 were provided by Merck
(Darmstadt, Germany) and other chemicals or kits were purchased
from Sigma Al- drich, Germany.

10. Preparation of sesamol-loaded NLCs (S-NLCs)
 The nanostructure lipid carrier (NLCs) is a type oil in water type of
emulsion
 where the oil phase place with the solid lipid.
Indridients
Solid Lipid cetyl palmitate
Liquid Lipid oleic acid
(cetyl palmitate and oleic acid; 85:15 or 70:30
poloxamer 188 (0.5 or 1%, w/ v) and Tween 80 (1 or 2%, w/v)
Double distilled water

11. Methods of preparation of NLCs
1. Hot homogenization
2. cold homogenization
3. micro emulsion technique
4. solvent emulsification evaporation
5. emulsification solvent diffusion
6. solvent injection ( or solvent displacement)
7. phase inversion technique
8. Melt emulsification Sonication
9. multiple emulsion method and membrane contractor technique
10. high-pressure homogenization
high-pressure homogenization is use to formulate fabrication of NLCs.

12. Characterization of S-NLCs
• Particle size, polydispersity index (PDI), and zeta potential (ZP) NLCs
dispersions were diluted by deionized water and the mean particle
size, PDI, and ZP were analyzed at 25 °C by photon correlation
spectroscopy (Zetasizer, Malvern Instruments, UK) (n=6).

13. Morphological assessment
Morphology
Scanning electron microscope (KYKY-EM3200, China) was used to
evaluate the shape of nanoparticles.
• Entrapment efficiency (EE) and drug loading capacity (DL)

14. • Differential scanning calorimetry (DSC)
• In-vitro release
• Storage stability

15. Evaluation of the bioactivity of S-NLCs In vitro
experiments
• Cell culture
• Rat pheochromocytoma-derived PC12 cells were grown in Dulbecco’s
modified Eagle’s medium supple- mented with fetal bovine serum (10%),
horse serum
• In vitro model of ischemic stroke and treatments
• Oxygen-glucose deprivation (OGD) is a well-recognized in vitro model of
stroke and may also be used to develop novel neuroprotective agents

16. Cell viability
• Cell viability assay
• The viability of PC12 cells was assessed using MTT (3- [4,5-
dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium brom- ide) colorimetric
assay

17. In vivo experiments
•Animals Male Wistar rats (300–350 g
•Visualization of S-NLCs in rat brain
•Brain distribution study
•Animal groups, induction of ischemic stroke, and
treatments
•Behavioural assessments- Spatial learning and memory was
assessed by Morris water-maze (MWM)
• Assessment of the brain infarcted area
• Histological assessments
• Biochemical analysis

18. In vivo experiments
• The effects of sesamol solution or S-NLCs on I/R-induced
neurobehavioral deficits

19. The effects of sesamol or S-NLCs on I/R-
induced neuronal damage in the
hippocampal CA1 region

20. Representative photographs of brain coronal
sections stained with TTC

21. Conclusions
•NLCs serve as promising carriers for sesamol, a
phenolic compound with a wide spectrum of
pharmacological ac- tivities. S-NLCs by improving
the pharmacological pro- file of sesamol provide
longer-lasting therapeutic effects in both in vitro
and in vivo models of stroke. This nano- formulation
through the activation of PI3K pathway might be a
suitable controlled release drug carrier sys- tem
against the ischemic injuries or other
neurodegener- ative pathologies.