hIGF-1促进软骨细胞在脱细胞软骨基质材料中的增殖及黏附

hIGF-1 promotes proliferation and adhesion of chondrocytes on the acellular cartilaginous matrix

目的探究脱细胞软骨基质(ACM)作为天然细胞支架的可行性及人胰岛素样生长因子1(hIGF-1)对软骨细胞在ACM上的增殖及黏附作用的影响。方法兔耳软骨细胞原代培养,取第3代细胞备用。将脱细胞处理获得的ACM进行HE染色、Masson染色及电镜下观察,用CCK-8法检测支架浸提液的毒性。将软骨细胞接种至ACM中,实验组加入57.14μg/L的hIGF-1,培养7 d后于电镜下观察细胞在ACM上的增殖及黏附。结果1)ACM疏松多孔,电镜下孔径(3.48±1.39)μm,孔隙率(90.13±2.52)%。2)实验组软骨细胞增殖旺盛,细胞占据绝大部分支架空穴。而对照组软骨细胞增殖较缓慢,且仅有少量细胞进入材料空穴内。结论实验制备的ACM是一种可靠的天然细胞支架。且hIGF-1能促进软骨细胞在ACM上的增殖及黏附。

Objective To explore the feasibility of acelluar cartilaginous matrix as a natural material and the effect of human insulin-like growth factor 1 on the proliferation and adhesiveness of chondrocytes on the acelluar cartilaginous matrix. Methods The chondrocytes were isolated from rabbit’s aural cartilage.The third passage chondrocytes were identified with toluidine blue staining and collagen Ⅱ immunofluorescence staining.The cartilage of rabbit’s knee joint was taken for decellularization. The acelluar cartilaginous matrix was microsopied by HE staining and Masson staining. Using CCK-8(cell counting kit) to detect the acelluar cartilaginous matrix extracts’ toxicity of 25%, 50%, 100% concentration. Chondrocytes were inoculated into the acelluar cartilaginous matrix. In the experimental group, DMEM medium containing 57.14 μg/L human insulin-like growth factor 1 was added.After 7 days,observing chondrocytes with scanning electron microscopy. Results 1)The acelluar cartilaginous matrix was loose and porous.The pore size was(3.48±1.39)μm and the porosity was(90.13±2.52)% under electron microscope. 2)The experimental group showed that the chondrocytes were growing quickly and they were entered the matrix and connected with each other.They occupied most of the cavity.The chondrocytes in control group was growing slower than the experimental group.And a large amount of cells clustered onthe surface of the material,only a small amount of cells entered the cavity. Conclusions The acelluar cartilaginous matrix is a reliable natural material for repairing after articular cartilage injury. Moreover, human insulin-like growth factor 1 can stimulate the proliferation and adhesiveness of chondrocytes on the acelluar cartilaginous matrix.