Human Dermal Fibroblast Viability Against Time and Calcium Alginate Density in Vitro

Han, Seung Kyu; Kim, Hyun Peel; Kim, Woo Kyung

Abstract

One approach for soft tissue augmentation is tissue engineering technique which uses autogenous fibroblasts localized within a biocompatible polymer. Many reports of chondrocytes within calcium alginate for cartilage transplantation in vitro have been presented, but no report of fibroblasts within calcium alginate has been presented. This study is related to the conditions of polymerization depending on alginate density and culture time in the viability of human fibroblasts encapsulated in calcium alginate matrix. Human dermal fibroblasts were obtained from STSG patients, enzymatically dissociated, and cultured in DMEM/ Ham's F-12, then the fibroblasts were collected by centrifugation. Alginate discs containig fibroblasts were made from 1 or 2% sodium alginate mixed in 150 mM CaCl2 solution. Viability of fibroblasts was measured by quantification of the DNA content per alginate disc at six different time intervals from 1 to 8 weeks. Significant initial cell loss was observed in the first two weeks after which the survival rate remained stationary. According to the alginate density, fibroblasts seeded at 1% alginate showed higher survival rate than at 2% alginate in early periods(by 6 weeks), then inversed in late periods(p<0.05). Our study provides a significant information in manufacturing alginate-fibroblast induced soft tissue by tissue engineering.