Study On The Biocompatibility Of Polylactic Acid-Polyglycolic Acid/Type I Collagen Composite Scaffold
Keywords
Tissue Engineered Vagina, Polylactic Acid-Polyglycolic Acid, Type I Collagen, Vaginal Epithelial Cells, Biocompatibility
Abstract
Objective To observe the biocompatibility of poly-lactide-co-glycolide (PLGA)/type I collagen composite scaffold and explore its feasibility as a tissue engineering vaginal scaffold. Methods Polylysine-coated PLGA was placed in an acetic acid aqueous solution containing 0.25% type I collagen to prepare a PLGA/type I collagen composite scaffold. The vaginal tissue of 10- to 12-week-old female SD rats was taken, vaginal epithelial cells were isolated and cultured using enzyme digestion method, and the second-generation cells were taken for experiments. The composite scaffold extract was taken to culture vaginal epithelial cells and the cytotoxicity of the material was observed. Vaginal epithelial cells were co-cultured with the composite scaffold for 48 hours (experimental group), and the cell adhesion rate was detected; cells were seeded on the simple PLGA scaffold as the control group. The cell-scaffold complex was implanted under the skin of SD rats, and the cells were harvested at 2, 4, and 8 weeks for HE staining and immunohistochemical staining to observe the growth of cells on the scaffold. The cell-scaffold complex was transplanted into the vagina of 6 SD rats whose vaginal tissue was removed. The vaginal growth was observed 3 and 6 months after surgery. The vaginal tissue was taken for histological observation 6 months later. Results Rat vaginal epithelial cells grew and proliferated well in the leaching solution of PLGA/type I collagen composite scaffold material, and the cytotoxicity was level 1. The cell adhesion rate of the experimental group was 71.8%¡À9.2%, which was significantly higher than the 63.4%¡À5.7% of the control group (t=2.195, P=0.005). Vaginal epithelial cells can adhere and grow on the PLGA/¢ñ collagen composite scaffold material. After 2 weeks of subcutaneous implantation in rats, cells grew and proliferated in the pores of the scaffold, and fibroblasts grew; 1 to 3 layers of epithelium were formed on the surface of the scaffold material after 4 weeks. ; After 8 weeks, the scaffold material was partially degraded, the epithelial layer increased and was arranged in a polar manner, and keratin immunohistochemical staining was positive. Three months after the orthotopic transplantation of the cell-scaffold complex, the rat vaginal mucosa was pink and shiny, and most of the scaffold material was degraded; at 6 months, the vagina was approximately 1.2 cm deep, with no obvious stenosis, and the appearance of the vaginal mucosa was similar to that of a normal vagina. The mucosa has fewer folds. Histological observation shows that the epithelial layer is not significantly different from that of the normal vagina. The number of nail-like protrusions is visible in the basal layer, which is less than that of the normal vagina. Keratin immunohistochemical staining is positive. Conclusion The PLGA/type I collagen composite scaffold has good biocompatibility and can be used as a scaffold material for constructing tissue-engineered vaginas.
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Original research was done by Zhang Mingle, Li Yanan, Jia Jingfei, Chen Yexing, Huang Xianghua
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