Study of fibroblast adhesion on RGD-modified electrospun Thai silk fibroin nanofiber for scaffold material in Dentistry: A preliminary study
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2010-11-03
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Khon Kaen University Dental Journal
Abstract
Silk fibroin is gaining interest as a novel scaffold for regeneration. The fabrication of nanofibers using an electrospinning technique is one of the most promising means of fabricating for Tissue engineering. RGD (arg-gly-asp) sequence, a novel cell adhesion molecule, is a candidate for silk surface modification to improve cell adhesion. The purposes of this study were: 1 to optimize the concentration of silk fibroin solution in order to fabricate the electrospun Thai silk fibroin nanofibers, 2 to fabricate RGD-modified electrospun Thai silk fibroin nanofibers and 3 to compare adhesion of human gingival fibroblast cell line between RGD- and non-RGD modified electrospun nanofibers. Materials and methods: fibroin protein was extracted from Thai (Bombyx mori) Nangnoi srisaket-I domestic silk cocoons. It was dialyzed and lyophilized, and 98% formic acid was used to dissolve the fibroin powder to yield 22.5, 23.5, 25, 27.5 and 30% w/v solutions. Electrospinning was performed under 9-12 kV, 10 mA with a syringe tip to collector distance of 12 cm, and loading rate of 0.5 mL/hr. Then RGD was incorporated onto the nanofiber surfaces using covalent coupling method. Scanning electron microscope was used to observe the morphology of the electrospun nanofibers. Finally, a cell adhesion assay was performed. Results: Silk fibroin solution concentrations of 22.5%, 23.5%, 25%, 27.5% and 30% w/v produced the average fiber diameters of 284.20±165.92 nm, 298.91±71.78 nm, 395.34±140.26 nm, 494.7±113.39 and 601.5±157.37 nm respectively. The optimal concentration, which produced continuous and uniform fibers, was 23.5% w/v. High performance liquid chromatography assay demonstrated that electrospun Thai silk fibroin nanofibers could be successfully modified with RGD. The RGD-modified nanofibers had a significantly higher cell number compared with conventional ones. Thus after optimum modification, we may use Thai silk fibroin-derived scaffold as a candidate for Tissue engineering in Dentistry.
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Khon Kaen University Dental Journal; Vo.13 No. 1 Jan - June 2010