Transcript Objective - Conferences

Chitosan Nanoparticles - Challenges
for Antibiotic Drug Delivery using
Penicillin as a Model Drug
5th International Conference and Exhibition on
Pharmaceutics & Novel Drug Delivery Systems
March 16-18, 2015 Crowne Plaza, Dubai, UAE
Dr. B. B.Barik
Professor, Dept. of Pharmaceutics
Mohammed M. Safhi, S.M.Sivakumar,
Aamena Jabeen, Foziyah Zakir & Farah Islam
College of Pharmacy, Jazan University,
Kingdom of Saudi Arabia.
E-mail: [email protected]
INTRODUCTION
Antibiotic resistance is one of the major health and economic
problems worldwide eroding the discovery of antibiotics and their
application to clinical medicine.
It has been estimated that each year in US, approximately 2
million people get infected with antibiotic resistance bacteria and
23,000 die as a result of these infections .
Resistance to penicillin is well recognized in Staphylococcus
aureus and Staphylococcus epidermidis .
Use of nanoparticles for antibiotic delivery is increasing due to
increased prevalence of antibiotic resistant bacterial strains.
Antibiotic Resistance
WHY NANOPARTICLES AND WHY CHITOSAN?
Nanoparticles can easily bypass efflux mechanism, particularly
chitosan nanopartilcles because chitosan is highly poly cationic
can easily bind with anionic microbial cells.
Efflux mechanism: Membrane proteins that export antibiotics
from the cell and maintain their low intracellular concentrations.
Outer membrane permeability: An asymmetric bilayer: the
phospholipid form the inner leaflet and the lipopolysaccharides
(LPS) form the outer leaflet, provides a formidable barrier that
must be overcome by drugs.
Objective
The objective of this study was to formulate
and characterize chitosan nanoparticles of
penicillin G to improve the bioavailability and
prevent drug resistance.
Method
Ionic gelation technique was used to encapsulate penicillin G in
chitosan nanoparticles.
 The anti bacterial efficacy of penicillin-loaded chitosan
nanoparticles were evaluated against five bacterial strains namely:
• Staphylococcus aureus
• Streptococcus pyogenes
• Bacillus subtilis
• Escherichia coli and
• Klebsiella planticola
 The antibacterial effect of penicillin nanoparticles was compared
with conventional penicillin drug.
Preparation of chitosan nanoparticle of penicillin G
1% w/v chitosan solution was prepared with 1% v/v glacial acetic acid
having pH 4.
The pH was increased to 6 by adding 1ml of 1N NaOH.
The chitosan gel was homogenized at 3600 rpm for 30 minutes
During homogenization 1 ml of 1% w/v of penicillin G was added.
Sonication was done for 30 seconds at pre determined time interval.
500 μl of formaldehyde was added drop by drop and homogenization
was continued for one hour.
The penicillin loaded nanoparticles were washed & dried.
Prepartion of pencillin – chitosan nanoparticles
IONIC GELLATION TECHNIQUE
Chitosan
Chitosan cross linking
Characterisation of nanoparticles
• Morphology
The nanoparticles were observed under TEM with an accelerating
voltage of 100 kv
• Zeta potential, Particle size and PDI
The zeta potential, size and PDI of penicillin loaded chitosan
nanoparticles were measured by using Nano – ZS zeta sizer,
Malvern Instruments, UK.
• In vitro release profile
2 ml of penicillin loaded chitosan nanoparticles were taken in 50 ml
of phosphate buffer of pH 6.0 placed in a beaker and stirred in a
magnetic stirrer at 50 rpm for 48 hrs at a temperature of 25ºC. The
samples were withdrawn, centrifused and assayed
spectrophotometrically at 208 nm.
Assessment of antibacterial activity
Following five bacterial strains were used:
• Staphylococcus aureus,
• Streptococcus pyogenes,
• Bacillus subtilis,
• Escheriachia coli and
• Klebsiella planticola
The MIC was determined in Nutrient Broth using two-fold serial
dilutions of chitosan nanoparticles with initial bacterial inoculums
of 10-6 CFU and the time and temperature of incubation being 24
hours and 370C, respectively.
Morphology and Size
TEM analysis
Zeta potential, Particle size and pdi:
Polymer Cross
linker
Before sonication
After sonication
Size
PdI
Zeta
Sonication Size
PdI
(nm)
potntial
time
(nm)
(Sec)
Zeta
potntial
0.1%
250 µl
644.2
1.00
35.4 mV
10
497.2 0.59
55.4
0.1%
500 µl
585.9
0.63
42.4
15
401.1 0.48
62.8
0.1%
1000 µl 1057
0.7
37.3
20
3
0.3
52.3
0.1%
1500 µl 1167
0.8
33.7
25
661
0.51
46.8
In vitro release
Anti bacterial studies
CONCLUSION
This work confirms that ionic gelation technique is a successful
method to encapsulate penicillin in chitosan nanoparticles.
Penicillin is released in sustained manner from chitosan
nanoparticle.
It showed better microbial action than conventional penicillin.
References
1. CDC.Active Bacterial Core Surveillance Methodology (2012).http://www.cdc.gov/abcs/
index.html
2. Kimberlin, C., Khalid, F., Hariri, A. Resistance of staphylococci to penicillin-G and cloxacillin.
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nanoparticles. Chem. Mater. 12, 2000, 2268-2274
4. Pertha, S., Amit, K.G., Goutam, R. Formulation and evaluation of chitosan based ampicillin
trihydrate nanoparticles. Trop J Pharm Res. 9(5), 2010 : 483 – 488.
5. Sarmento, B., Riberio A, Veiga F, Sampaio P, Neufeld R, Ferreira D. Alginate/Chitosan
nanoparticles are effective for oral insulin delivery. Pharmaceutical Research. 2007; 24(12):
2198 – 2206