Transcript In-situ - Conferences

Hydrogels of Solid lipid Nanoparticles of Curcumin
Presented by
RUCHI CHAWLA
ASSISTANT PROFESSOR
IIT(BHU), VARANASI
5th International Conference and Exhibition on Pharmaceutics
& Novel Drug Delivery Systems
March 16-18, 2015 Crowne Plaza, Dubai, UAE
Contents

Introduction

Experimental work
 Preparation of SLNs
 Analytical Method for Curcumin
 Formulation characterization

Summary and conclusion

References
Introduction
Hydrogels:
Advantages of Hydrogels (Kopecek, 2009):
Shape stability and softness similar to that of the soft surrounding tissues
Chemical and biochemical stability
Absence of extractables
High permeability for water-soluble nutrients and metabolites across the biomaterial tissue-interface
Convenient handling
Easy application
Excellent tissue biocompatibility due to their high water content
Curcumin
 Yellow spice derived from the roots, rhizome of Curcuma longa
 Adaptogen, bio-protectant, anti-bacterial, antioxidant and anti-inflammatory
 Chemically it is a mixture of three principal compounds (Strimpakos and Sharma, 2008):
 Curcumin (sometimes referred to as curcumin I),
 Demethoxycurcumin (curcumin II), and
 Bisdemethoxycurcumin (curcumin III)
 Limitations:
 Poor systemic bioavailability because of poor absorption and rapid systemic
elimination via glucuronidation (Aggarwal and Sung, 2009).
 Hydrophobic compound with a high partition coefficient of 3.2 and water solubility
around 0.6 μg/ml (Kurien et al.,2007 and Patel et al., 2009).
Why Solid lipid Nanoparticles?
 Small size
 Possibility of controlled rug release
 Increased drug stability
 High drug pay load
 Incorporation of lipophilic and hydrophilic drugs
 Biocompatibility
 Enhancement of bioavailability of the incorporated drugs (Mehnert and Mader, 2001)
 Enhanced penetration of drug into the skin from lipid nanoparticles, occlusion effect and film
formation of lipid nanoparticles on the skin
 Larger surface area and contact point to adhere to the skin layer than that of microparticle.
 SLN can provide benefits of accumulation of drug in the skin strata to various skin diseases
such as acne and eczema (Korting et al, 2007).
Experimental Work
Preparation of SLNs of Curcumin by nanoprecipitation method (Chorny et
al., 2002)
Organic Phase (58.5 ml acetone and 1.5 ml
dichloromethane) consisting of 600 mg of stearic acid and
9.0 mg Curcumin
Injected slowly with stirring at 1000 rpm Aqueous Phase (120 mL) containing 0.5, 1.0 & 2.0% w/v
PVA
Solvent removal by evaporation room temperature
for 24hours
Evaporation of organic phase
SLNs suspended in aqueous medium
Ultracentrifugation at 10,000 x g for 5 min
Collection of SLNs
UV spectrophotometric method for Curcumin
Calibration curve in water @λmax 405 nm
(n=3)
1
Calibration curve at pH 6.0 @λmax 415 nm
(n=3)
1.0000
0.9
0.9000
0.8
y = 0.0164x + 0.0048
R² = 0.9995
0.8000
y = 0.0171x - 0.002
R² = 0.9999
0.7000
Absorbance
Absorbance
0.7
0.6
0.5
0.4
0.6000
0.5000
0.4000
0.3
0.3000
0.2
0.2000
0.1
0.1000
0
0
10
20
30
40
50
Concentration (µg/ml)
60
0.0000
0
10
20
30
40
Concentration (µg /ml)
50
60
Characterization of SLNs
Compatibility studies
40
45
%T
%T
35
40
FT-IR Spectrum of Curcumin
FT-IR Spectrum of Stearic acid
30
35
25
547.80
30
20
466.79
25
1464.02
719.47
686.68
1433.16
2661.85
1705.13
2916.47
1508.38
0
2848.96
1205.55
1153.47
1280.78
1429.30
5
1298.14
10
5
1627.97
10
1026.16
1114.89
15
937.44
856.42
808.20
15
962.51
3508.63
20
0
2500
2250
2000
1750
1500
1250
1000
750
500
1/ c m
4000
3750
s t earic ac id
3500
3250
3000
2750
2500
2250
2000
27. 5
%T
25
FT-IR Spectrum of Curcumin + Stearic acid
22. 5
543.94
17. 5
466.79
20
1026.16
1153.47
1205.55
7. 5
1278.85
1508.38
1464.02
1429.30
10
962.51
12. 5
719.47
808.20
15
1627.97
2750
1701.27
3000
5
2. 5
4000
3750
3500
s tearic ac id + c urc um in
3250
3000
2848.96
3250
2918.40
3500
3510.56
4000
3750
c urc um in
2750
2500
2250
2000
1750
1500
1250
1000
750
500
1/ c m
1750
1500
1250
1000
750
500
1/ c m
Particle size and polydispersity of C-SLNs
Batches
Particle-Size
Poly Dispersity index
Batch-1 (0.5 % PVA)
697.7 + 0.06 nm
-0.9375 + 0.0004
Batch -2 (0.75 % PVA)
939.8 + 0.01 nm
0.383+ 0.001
Batch 3 (1 % PVA)
527.6 + 0.04 nm
-1.597+ 0.001
Entrapment Efficiency of C-SLNs
Batch (n=3)
Total Drug content (mg)
Free Drug Content (mg)
Entrapment efficiency
(%EE)
(TDC –FDC)/TDC * 100)
Batch 1
5.448+ 0.02
1.006+ 0.07
81.53 + 0.02%
Batch 2
5.880+ 0.09
1.150+ 0.06
80.44 + 0.09%
Batch 3
5.860+ 0.10
1.012+ 0.07
82.73 + 0.01%
Hydrogel preparation
S. No.
Batches
Carbopol concentration
(% w/v)
Hydrogel type
1
B1
0.5
Blank gel
2
B2
1.0
Blank gel
3
B3
2.0
Blank gel
4
Y1
0.5
In-situ hydrogels*
5
Y2
1.0
In-situ hydrogels*
6
Y3
2.0
In-situ hydrogels*
7
D1
0.5
Enriched hydrogels^ (1:1
ratio of B2 and C-SLNs#)
8
D2
0.66
Enriched hydrogels (1:1
ratio of B2 and C-SLNs#)
*In situ hydrogels were prepared by adding carbopol (0.5, 1.0 and 2.0 %) to fixed volume of C-SLNs suspension containing 1% PVA
^Enriched hydrogels were prepared by adding carbopol hydrogel (1%w/v) to # C-SLNs prepared using 1% PVA
Texture profile analysis
Force (g)
1
2
3
60
F
1
50

40


30


20

































10



 Y5

0
0
2
4
6
8
10


-10

-20

-30


-40
-50
Typical force-time plot of Texture analysis
F
2
12





14
Time (sec)
Texture analysis of blank hydrogels
Firmness(g)
Firmness(g)
70
59.879
60
50
40
30
17.214
20
19.643
10
0
0
0.5
1
1.5
2
2.5
2
2.5
Concentration ( %w/v)
Cohesiveness(g)
Cohesiveness(g)
0
-5 0
0.5
1
1.5
-10
-15
-20
-25
-30
-35
-40
-45
-31.787
-35.012
-41.926
R² = 0.9997
Concentration ( %w/v)
Consistency (g-sec)
Consistency (g-sec)
500
434.176
400
300
138.496
200
79.688
100
0
0
0.5
1
1.5
Concentration ( %w/v)
2
2.5
Texture analysis of In-situ hydrogels
Texture analysis of In-situ hydrogels
Texture analysis of Enriched hydrogels
160
138.496
140
120
96.858
100
79.042
80
60
40
20
19.643
14.257 12.673
0
-20
-40
-60
Control(L2)
D1
D2
-35.012
Firmness(g)
19.643
14.257
12.673
Consistency (g-sec)
138.496
96.858
79.042
-29.886 -28.936
Cohesiveness(g)
-35.012
-29.886
-28.936
-19.93
-12.109
-5.179
Index of Viscosity(g-sec)
-19.93
-12.109
-5.179
Comparative Texture analysis
In-vitro release studies
 Conducted using Franz diffusion cells and dialysis membrane
 Dissolution media: phosphate buffer pH 6.0 solution containing 1% v/v Methanol maintained at 37 ±
1 ◦C on a magnetic hot plate with moderate stirring
CUMULATIVE PERCENTAGE RELEASE
SLN Release Profile
Hydrogel Release Profile
70
60
50
40
30
20
10
0
0
200
400
600
800
1000
TIME(IN MIUTES)
1200
1400
1600
In-vitro occlusion test
120
100
80
60
40
20
0
% water loss (relative to Control )
Occlusion Factor, F
Control
100
0
Occlusion Factor, F
0.5 % Carbopol Gel
61.33333333
38.66666667
% water loss (relative to Control )
0.5 % SLN-Gel
28.83333333
71.16666667
Summary & Conclusion
 Curcumin loaded SLNs were prepared by nano-precipitation technique using stearic acid as lipid and PVA as surfactant.
 Batch-3 (1% PVA) exhibited particle size of 527.6 nm, PDI of -1.597 and % Entrapment efficiency 82.73%
 In vitro release from C-SLN enriched hydrogel showed controlled release of drug in comparison to C-SLNs upto
72 hr with approx.58 % release within 24 hours.
 Occlusion test showed reduced water loss from carbopol and even lesser from enriched hydrogel.
 Stability studies over a period of 90 days, showed increase in firmness, cohesiveness and index of viscosity and
decrease in consistency.
 In-situ hydrogels exhibited a concentration (carbopol) dependent increase in firmness, consistency, cohesiveness and
viscosity, however, presence of C-SLNs significantly decreased (p < 0.05) these values in comparison to blank
hydrogels.
 Similar observations were made in enriched hydrogels
 Also, a significant difference (p < 0.05) in hydrogel properties was observed between in-situ and enriched hydrogels
indicating effect of SLNs on the swelling properties of Carbopol.
 Occlusive properties of in-situ hydrogels were better than enriched and blank hydrogels.
 In-situ hydrogels also exhibited uniform and extended release of curcumin, alongwith higher permeation characteristics.
 Better formulation characteristics of in-situ hydrogels might be because of homogenous deposition and gelling of
carbopol around curcumin nanoparticles.
References:

Strimpakos, A. S., Sharma, R. A. (2008). Comprehensive invited review curcumin: preventive and therapeutic properties in
laboratory studies and clinical trials. Antioxidants & Redox Signaling 10, 511-546.

Kurien, B. T., Singh, A., Matsumoto, H., Scofield, R. H. (2007). Improving the solubility and pharmacological efficacy of
curcumin by heat treatment. Assay and Drug Development Technologies 5, 567-576.

Patel, R., Singh, S. K., Singh, S., Sheth, N.R., Gendle, R. (2009). Development and characterization of curcumin loaded
transfersome for transdermal delivery. Journal of Pharmaceutical Sciences and Research 1, 71-80.

Aggarwal, B. B., Sung, B. (2009). Pharmacological basis for the role of curcumin in chronic diseases: an age-old spice with
modern targets. Trends in Pharmacological Sciences 30, 85-94.

Mehnert, W. and Mader, K. (2001) Solid lipid nanoparticles: Production, characterization and applications. Adv. Drug Deliv. Rev.,
47, 165-196.

Yang, S., Zhu, J., Lu, Y., Liang, B., Yang C. (1999) Body distribution of camptothecin solid lipid nanoparticles after oral
administration. Pharm. Res., 16 , pp. 751–757.

Korting, M.S., Mehnert, W. and Korting, H.C. (2007) lipid nanoparticles for improved topical applicationof drugs for skin
disease, Adv Drug Del Review, 59, 427-443.

Chorny, M., Fishbein, I., Danenberg, H.D., Golomb, G. (2002) Lipophilic drug loaded nanospheres prepared by
nanoprecipitation: effect of formulation variables on size, drug recovery and release kinetics Journal of Controlled Release 83
389–400.
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Thank You