Cellular biocompatibility of shengji solution-gelatin-chitosan drug-loading skin scaffold / 中国组织工程研究

ABSTRACT

BACKGROUND: Tissue-engineered skin plays an important role in the repair and reconstruction of large-area skin lesion. Little is known about the effects of Chinese medicine-loading tissue-engineered skin scaffold on cell adhesion and wound surface infection. OBJECTIVE: To screen the application concentration of shengji solution, which can promote tissue regeneration, using keratinocytes and fibroblasts, and to observe the effects of shengji solution-gelatin-chitosan drug-loading skin scaffold on cellular adhesion and biocompatibility. DESIGN, TIME AND SETTING: Taking keratinocytes and fibroblasts as subjects, the present randomized and controlled experiment was performed at the Laboratory of Cell Engineering, Orthopedic Institute, Tianjin Hospital between February and August 2005. MATERIALS One healthy big-ear rabbit was included for harvesting skin seed cells. Shengji solution, Danggui (Radix Angelicee Sinensis) and Shengdi (rehmannia dride rhizome) extract (self-extracted, 1.5 g/mL), gelatin-chitosan scaffold and shengji solution-gelatin-chitosan scaffold were provided by Tianjin University, China. METHODS: Passage 3 keratinocytes and fibroblasts were harvested by dispase Ⅱ- trypsase- ethylenediamine tetraacetic acid(EDTA) digestion. The prepared cells were divided into 5 groups control, shengji solution, Danggui, Shengdi, and epidermal growth factor (EGF). Common culture solution containing 0.1 volume fraction of fetal bovine serum, shengjisolution (5, 6, 8, and 12 g/L), Danggui extract (8 g/L), Shengdi extract (8 g/L), and EGF culture medium (10 μ g/L) were used in corresponding groups for cell culture. At 1, 3, 5, and 7 days, cellular proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. Keratinocytes and fibroblasts were cultured in different concentrations of shengji solution (50, 60, 80, and 120 g/L) and grouped according to different concentrations. At 7 and 14 days, cellular adhesion was observed by semi-quantitative method. At 4, 7, and 14 days, keratinocytes cultured by 60 and 80 g/L shengji solution were harvested for hematoxylin-eosin staining and subsequent scanning electron microscope observation. MAIN OUTCOME MEASURES: Effects of shengji solution on skin seed cell proliferation; effects of drug-loading scaffold on cellular adhesion; and cell-carrier biocompatibility. RESULTS: MIF detection results demonstrated that cells significantly proliferated after treatment of 5, 6, and 8 g/L shengji solution compared to the control group (P < 0.05). Obvious proliferation of passage 3 keratinocytes and fibroblasts was found on 60 and 80 g/L drug-loading scaffold. Keratinocytes on the drug-loading scaffold exhibited good cellular morphology and closely adhered to the scaffold. Shengji solution had no apparent toxic effects on cells. CONCLUSION: Shengji solution (60 and 80 g/L)-gelatin-chitosan scaffold can effectively promote the adhesion and proliferation of keratinocytes and fibroblasts.