Experimental Study On Constructing Neural Tissue Engineering Scaffolds Using Collagen Gel
Keywords
Neural Tissue Engineering, Collagen Gel, Scaffold Material, Biocompatibility, Bmscs, Rat
Abstract
Objective To study the effect of collagen gel scaffold on the proliferation and differentiation of rat BMSCs and explore its feasibility as a neural tissue engineering scaffold. Methods Type I collagen was used to prepare collagen gel scaffolds. Rat BMSCs were isolated and cultured using density gradient centrifugation, and collagen gel-BMSCs complexes were prepared from fifth-generation cells. Scanning electron microscopy and HE staining were used to observe the morphological structure of the collagen gel scaffold and the cell morphology after composite culture; MTT was used to detect the effect of the scaffold on the proliferation of BMSCs. Green fluorescent protein (GFP)-positive (GFP+) BMSCs were cultured in collagen gel scaffolds for 24 hours, and cell growth and adhesion to materials were observed through laser confocal microscopy and live cell workstation. Results: Laser confocal microscopy and live cell workstation observation showed that GFP+BMSCs were evenly distributed in the collagen gel scaffold. Most of the GFP+BMSCs were spindle-shaped, with some cells extending out protrusions and some forming connections between cells, suggesting that BMSCs were in a three-dimensional space. Grows well in medium. Scanning electron microscopy showed that the collagen gel scaffold had a porous fibrous network structure, and BMSCs could adhere to the scaffold material with good cell morphology. MTT assay showed that the absorbance (A) values of BMSCs cultured in collagen gel scaffolds at 3, 5, and 7 d were all higher than those of BMSCs alone, and the difference between groups at 5 and 7 d was statistically significant (t=4.472, P=0.011; t=4.819, P=0.009). HE staining showed that the collagen gel scaffold was a homogeneous light pink-stained filament-like substance. The BMSCs were evenly distributed in the collagen gel after being cultured for 24 hours; at 7 days, the BMSCs had various shapes, with some cells extending out slender processes and having a neuron-like morphology cultured in vitro. Conclusion Collagen gel scaffold is easy to prepare, has good biocompatibility, and can be used as a scaffold material for neural tissue engineering.
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Original research was done by Zhu Liang, Li Shen, Wang Suping, Qin Huamin, Lan Xiaoyan
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