This is my first YEAR Second-semester presentation in NIPER GUWAHATI

1. Presented By:
Harshad Arvind Takate
Second Semester
Dept: Pharmaceutical Technology (Formulation)
Nose to brain delivery of rotigotine loaded chitosan
nanoparticles in Parkinson's disease
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FLOW OF SEMINAR
ARTICLE CASE STUDY
MATERIALS AND METHODS
PREPARATION OF NANOPARTICLES
DETERMINATION OF ENTRAPMENT EFFICIENCY
CHARACTERIZATION OF NANOPARTICLES
CELL CULTURE AND CELL UPTAKE STUDY
RESULT AND DISCUSSION
CONCLUSION
REFERENCES
INTRODUCTION

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 Tremor,
 Rigidity,
 Bradykinesia
 Postural
instability
SYMPTOMS
Management of therapy
Conventional therapies
STATISTICS
Advanced targeted therapies
INTRODUCTION
PD

4. IMPACT FACTOR: 6.321
PUBLICATION: 2020
JOURNAL NAME: INTERNATIONAL
JOURNAL OF PHARMACEUTICS
ARTICLE CASE STUDY
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Preparation of Nanoparticles
Determination of entrapment efficiency
Characterization of Nanoparticles
Cell Culture
Experimental model
Fig: Chitosan nanoparticle forming via ionic
gelation method.
Ref: Yanat M, Schroën K. Preparation methods and applications of chitosan nanoparticles; with an outlook toward reinforcement of biodegradable packaging. Reactive and
Functional Polymers, 2021;161(104849)
MATERIALS AND METHODS
 Rotigotine
 Chitosan
 Sodium tripolyphosphate
 Coumarin-6

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CONCENTRATION WAS DETERMINE BY CALIBRATION
CURVE OF ROTIGOTINE
SUPERNATANT WAS DILUTED AND ANALYSED BY UV
SPECTROMETRY
RNPs WAS CONCENTRATED BY ULTRACENTRIFUGATION.
%EE = INITIAL WT OF DRUG –
FREE DRUG IN SUPERNATANT.
INITIAL WT OF DRUG
DETERMINATION OF ENTRAPMENT EFFICIENCY

7. Ref: Bhattamisra SK, Shak AT, Xi LW, Safian NH, Choudhury H, Lim WM, et al. Nose to brain delivery of rotigotine loaded chitosan nanoparticles in human
SH-SY5Y neuroblastoma cells and animal model of Parkinson’s disease. International Journal of Pharmaceutics. 2020;579(119148):119148 7
CHARACTERIZATION OF NANOPARTICLES
1. Zetasizer
• particle size
• size distribution
• zeta potential
2. TEM
3. FESEM (SEM)

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CELL CULTURE AND CELL UPTAKE STUDIES
SH-SY5Y CELL
(NEUROBLAST
OMA CELLS)
COUMARIN 6
SOLUTION
AND
COUMARIN 6
LABELED RNPs
UPTAKE OF NPs
BY SH-SY5Y
CELLS WAS
DETERMINE
BY
FLUORESCEN
CE
MICROSCOPY
SH-SY5Y CELLS
HUMAN
NEUROBLASTOMA
CELLS
DULBECCO’S
MODIFICATION OF
EAGLE’S MEDIUM
CELL CULTURE
CELL UPTAKE STUDIES

9. Ref: Bhattamisra SK, Shak AT, Xi LW, Safian NH, Choudhury H, Lim WM, et al. Nose to brain delivery of rotigotine loaded chitosan nanoparticles in human
SH-SY5Y neuroblastoma cells and animal model of Parkinson’s disease. International Journal of Pharmaceutics. 2020;579(119148):119148 9
RESULT AND DISCUSSION
CELLULAR UPTAKE OF COUMARIN-6 AND COUMARIN-6-NPS

10. Ref: Bhattamisra SK, Shak AT, Xi LW, Safian NH, Choudhury H, Lim WM, et al. Nose to brain delivery of rotigotine loaded chitosan nanoparticles in human
SH-SY5Y neuroblastoma cells and animal model of Parkinson’s disease. International Journal of Pharmaceutics. 2020;579(119148):119148 10
Fig.3. Cytotoxic effects of rotigotine and RNPs on SH-SY5Y cells. (A & B) SH-SY5Y cells were treated with
Rotigotine, RNPs and placebo NPs (2.5–25 μg/mL) for 24 and 48 h.
CELL VIABILITY

11. Ref: Bhattamisra SK, Shak AT, Xi LW, Safian NH, Choudhury H, Lim WM, et al. Nose to brain delivery of rotigotine loaded chitosan nanoparticles in human SH-
SY5Y neuroblastoma cells and animal model of Parkinson’s disease. International Journal of Pharmaceutics. 2020;579(119148):119148 11
NEUROPROTECTIVE ACTIVITY OF RNPS
Fig.4.(C) Neuroprotective activity of RNPs and rotigotine solution was measured in SH-SY5Y cells through MTT assay on 6-OHDA (45 µm)
induced neurotoxicity for 24 h followed by treatment with 5 µg/mL of rotigotine solution, equivalent RNPs and placebo NPs individually for
24 h. Data was represented as % cell viability in a bar graph. Viability of control cells was 100%.

12. Ref: Bhattamisra SK, Shak AT, Xi LW, Safian NH, Choudhury H, Lim WM, et al. Nose to brain delivery of rotigotine loaded chitosan nanoparticles in human
SH-SY5Y neuroblastoma cells and animal model of Parkinson’s disease. International Journal of Pharmaceutics. 2020;579(119148):119148
12
Fig.5. (A) Plasma concentration versus time profile, (B) Brain concentration versus time profile and (c) Brain/blood concentration ratio in
rat at 0.5 h following RNPs (IN), Rotigotine solution (IN) and RNPs (IV) administrations
PHARMACOKINETIC STUDIES

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CONCLUSION
Rotigotine loaded chitosan nanoparticles were successfully
prepared and evaluated for nose to brain delivery.
Neuroprotective effect of RNPs was demonstrated in human
neuroblastoma cell line.
Intranasal route provide a therapeutic advantages compared to
conventional .
Analysis of pharmacokinetic data suggest that for intranasal route
is best route for direct transport of rotigotin to the brain.
Lower dose required for intranasal formulation

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1. Bhattamisra SK, Shak AT, Xi LW, Safian NH, Choudhury H, Lim WM, et al. Nose to brain
delivery of rotigotine loaded chitosan nanoparticles in human SH-SY5Y neuroblastoma cells and
animal model of Parkinson’s disease. International Journal of Pharmaceutics.
2020;579(119148):119148.
2. Yanat M, Schroën K. Preparation methods and applications of chitosan nanoparticles; with an
outlook toward reinforcement of biodegradable packaging. Reactive and Functional Polymers
2021;161(104849)
3. Formica ML, Real DA, Picchio ML, Catlin E, Donnelly RF, Paredes AJ. On a highway to the brain: A
review on nose-to-brain drug delivery using nanoparticles. Applied Materials Today
2022;29(101631):101631.
4. Liu M, Zhang J, Shan W, Huang Y. Developments of mucus penetrating nanoparticles. Asian J
Pharm Sci. 2015;10(4):275–82.
REFERENCES

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