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Biocomposite nanofibrous strategies for the controlled release of biomolecules for skin tissue regeneration.

International journal of nanomedicine (2014-10-23)
Chinnasamy Gandhimathi, Jayarama Reddy Venugopal, Velmurugan Bhaarathy, Seeram Ramakrishna, Srinivasan Dinesh Kumar
ABSTRACT

Nanotechnology and tissue engineering have enabled engineering of nanostructured strategies to meet the current challenges in skin tissue regeneration. Electrospinning technology creates porous nanofibrous scaffolds to mimic extracellular matrix of the native tissues. The present study was performed to gain some insights into the applications of poly(l-lactic acid)-co-poly-(ε-caprolactone) (PLACL)/silk fibroin (SF)/vitamin E (VE)/curcumin (Cur) nanofibrous scaffolds and to assess their potential for being used as substrates for the culture of human dermal fibroblasts for skin tissue engineering. PLACL/SF/VE/Cur nanofibrous scaffolds were fabricated by electrospinning and characterized by fiber morphology, membrane porosity, wettability, mechanical strength, and chemical properties by Fourier transform infrared (FTIR) analysis. Human dermal fibroblasts were cultured on these scaffolds, and the cell scaffold interactions were analyzed by cell proliferation, cell morphology, secretion of collagen, expression of F-actin, and 5-chloromethylfluorescein diacetate (CMFDA) dye. The electrospun nanofiber diameter was obtained between 198±4 nm and 332±13 nm for PLACL, PLACL/SF, PLACL/SF/VE, and PLACL/SF/VE/Cur nanofibrous scaffolds. FTIR analysis showed the presence of the amide groups I, II, and III, and a porosity of up to 92% obtained on these nanofibrous scaffolds. The results showed that the fibroblast proliferation, cell morphology, F-actin, CMFDA dye expression, and secretion of collagen were significantly increased in PLACL/SF/VE/Cur when compared to PLACL nanofibrous scaffolds. The accessibility of human dermal fibroblasts cultured on PLACL/SF/VE/Cur nanofibrous scaffolds proved to be a potential scaffold for skin tissue regeneration.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trypan Blue solution, 0.4%, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
1,1,1,3,3,3-Hexafluoro-2-propanol, ≥99%
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Curcumin, from Curcuma longa (Turmeric), powder
Sigma-Aldrich
Hexamethyldisilazane, reagent grade, ≥99%
Sigma-Aldrich
Curcumin, ≥94% (curcuminoid content), ≥80% (Curcumin)
Sigma-Aldrich
Trypan Blue, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Trypan Blue solution, 0.4%, for microscopy
Supelco
Curcumin, analytical standard
Supelco
1,1,1,3,3,3-Hexafluoro-2-propanol, for GC derivatization, LiChropur, ≥99.8%
Sigma-Aldrich
Hexamethyldisilazane, ReagentPlus®, 99.9%
Supelco
Hexamethyldisilazane, for GC derivatization, LiChropur, ≥99.0% (GC)
Sigma-Aldrich
Trypan Blue, Dye content 60 %
Supelco
Curcumin, matrix substance for MALDI-MS, ≥99.5% (HPLC)
Sigma-Aldrich
Hexamethyldisilazane, produced by Wacker Chemie AG, Burghausen, Germany, ≥97.0% (GC)
USP
Curcumin, United States Pharmacopeia (USP) Reference Standard
Curcumin, primary reference standard