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Abstract

ABSTRACT:The aim of this study was to determine the properties of β-glycerol phosphate (GP)/collagen/chitosan blended films for the potential application to skin tissue engineering. Various ratios of collagen to chitosan (8:2 and 7:3) and amounts of GP (0.5, 1, and 1.5% w/w) of total polymers were blended in solution form and cast into films. According to SEM images, the casted films showed a non-porous surface. For the mechanical properties, the prepared scaffolds exhibited the maximum elongation ranging from 15-23%, which is lower than those found by other researchers. However, the maximum tensile strength values of the scaffolds made from the collagen/chitosan (ratios 7:3) crosslinked with 0.5 or 1% w/w GP were in the range of 8-10 MPa which achieve the recommended values for application in skin tissue engineering. The scaffolds showed ability to retain their structure after immersion in phosphate buffer saline solution (pH 7.4) for 1 h, and their volume increased about 20%. After incubation in collagenase solution (200 U of collagenase/5 g of collagen) at 37°C, the scaffolds were degraded within 24 to 26 days which coincides very well with the healing time of acute wounds (about 25 days). FT-IR studies revealed the possibility of an interaction of GP with collagen/chitosan via ionic interaction that enhances the strength and stability of the prepared scaffold. The results from an in vitro culture study showed that the keratinocyte HaCaT culture could adhere well and grow on the selected scaffold with a typical morphology at 98.1 ± 1.8% of the control (cells growth on tissue culture plate) after cultivation for 5 days. The results suggest the potential of the GP/collagen/chitosan blended films for use as skin scaffolds.

Efficacy evaluation of the fibroblast-seeded collagen/chitosan scaffold on application in skin tissue engineering

“…Type I collagen was isolated from bovine tendon, according to our previous study 17 . Chitosan used in the present study was derived from crab shell (crab chitosan, molecular weight of 100 000-1 000 000 and more than 90% degree of deacetylation, Bannawach Bio-line Co. Ltd., Chonburi, Thailand).…”

“…The total amount of polymer used was fixed at 3% w/v and the proportion of collagen to chitosan was 7:3 17 . Type I collagen and chitosan were separately dissolved in 0.5 M acetic acid solution.…”

“…Its swelling ratio was defined as the ratio of weight increase (w − w 0 ) to the initial weight 9 . To determine its enzymatic degradation, the scaffold film was incubated in collagenase (200 U/5 g of collagen) in 10 ml phosphate buffered saline (PBS, pH 7.4) at 37°C for 1 month 17,19 and the enzyme was refreshed every 2 days. The percentage of remaining weight was calculated from w/w 0 .…”

“…The number of cells remaining on the scaffolds was quantified by XTT assay as used previously 17,21 . Briefly, the required amount of serum-free DMEM and XTT labelling mixture was added to wells containing the cell-adherent scaffolds.…”

“…In our previous study 17 , we used type I collagen for testing potential scaffolds for wound healing. The scaffolds were intended as temporary dressings to facilitate wound repair because of its abundance in skin tissue.…”