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  • Bi-layer composite dressing of gelatin nanofibrous mat and poly vinyl alcohol hydrogel for drug delivery and wound healing application: in-vitro and in-vivo studies.

Bi-layer composite dressing of gelatin nanofibrous mat and poly vinyl alcohol hydrogel for drug delivery and wound healing application: in-vitro and in-vivo studies.

Journal of biomedical nanotechnology (2013-08-29)
Maneesh Jaiswal, Asheesh Gupta, Ashwini K Agrawal, Manjeet Jassal, Amit Kr Dinda, Veena Koul
ABSTRACT

Present investigation involves the development of a bi-layer dressing of gelatin nanofibrous mat loaded with epigallocatechin gallate (EGCG)/poly vinyl alcohol (PVA) hydrogel and its in-vivo evaluation on full-thickness excision wounds in experimental Wistar rats. Nanomorphological observation, porosity, effect of crosslinking on tensile strength, physical stability and drug release profile in phosphate buffer and biocompatibility aspects of electrospun nanomat were investigated by various physico-chemical tools. EGCGa release profile was found to increase from 2-4 days with decreasing crosslinking time from 15 to 5 min. PVA hydrogels were prepared by freeze-thaw method and has been utilized as a protective and hydrating outer layer of the bi-layer dressing. Topical application of bi-layer composite dressing loaded with EGCG improve the healing rate in experimental rats as acute wounds model which was evidenced by significant increase in DNA (approximately 42%), total protein (approximately 32%), hydroxyproline (approximately 26%) and hexosamine approximately 24%) contents. A faster wound contraction was observed in wounds treated with composite dressing from approximately 14% to 47%. Histopathological examination revealed significant improvement in angiogenesis, re-epithelialization and less inflammatory response in comparison to control. Van-Gieson's collagen stains revealed matured, compact and parallel deposition of collagen fibrils on day 12. These results were supported by up-regulated expressions of matrix metalloproteinase (MMPs-2 and 9) by gelatin zymography. Control release of EGCG, 3D porous architecture of nanofibrous scaffolds as well as moist microenvironment provides ideal conditions for uninterrupted wound healing.

MATERIALS
Product Number
Brand
Product Description

Supelco
(−)-Epigallocatechin gallate, analytical standard
Sigma-Aldrich
Kuraray Poval ®56-98, Mw ~195,000
Sigma-Aldrich
Mowiol® 28-99, Mw ~145,000
Sigma-Aldrich
Gelatin from cold water fish skin, solid
Sigma-Aldrich
Mowiol® 18-88, Mw ~130,000
Millipore
Gelatin from porcine skin, medium gel strength, suitable for microbiology
Millipore
Gelatin from porcine skin, suitable for microbiology, high gel strength
Sigma-Aldrich
(−)-Epigallocatechin gallate, ≥80% (HPLC), from green tea
Sigma-Aldrich
Gelatin from bovine skin, Type B, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
(−)-Epigallocatechin gallate, ≥95%
Sigma-Aldrich
Gelatin from porcine skin, gel strength 300, Type A
Sigma-Aldrich
Gelatin from porcine skin, gel strength ~175 g Bloom, Type A
Sigma-Aldrich
Gelatin from porcine skin, gel strength 80-120 g Bloom, Type A
Sigma-Aldrich
Prionex® Highly purified Type A, aqueous solution
Supelco
(+)-Catechin, analytical standard
Epigallocatechin gallate, primary reference standard
Sigma-Aldrich
Mowiol® 40-88, average Mw ~205,000 g/mol
Millipore
Gelatin from porcine skin, suitable for microbiology, low gel strength
Sigma-Aldrich
Gelatin solution, Type B, 2% in H2O, tissue culture grade, BioReagent, suitable for cell culture
Millipore
Gelatin from porcine skin, suitable for microbiology, ultrahigh gel strength
Sigma-Aldrich
(−)-Epicatechin, ≥90% (HPLC)
Sigma-Aldrich
(−)-Epicatechin, ≥98% (HPLC), from green tea
Sigma-Aldrich
Poly(vinyl alcohol), Fully hydrolyzed
Sigma-Aldrich
Gelatin from bovine skin, Type B
Sigma-Aldrich
Gelatin from bovine skin, gel strength ~225 g Bloom, Type B
Sigma-Aldrich
Gelatin from porcine skin, powder, gel strength ~300 g Bloom, Type A, BioReagent, suitable for electrophoresis, suitable for cell culture
Epicatechin, primary reference standard
Sigma-Aldrich
Mowiol® 10-98, Mw ~61,000
Sigma-Aldrich
Poly(vinyl alcohol), Mw 8,000-10,000, 80% hydrolyzed
Sigma-Aldrich
Mowiol® 6-98, Mw ~47,000