聚氨酯材料复合软骨细胞构建组织工程化软骨

Tissue engineered cartilage constructed by polyurethane composite chondrocytes

背景:聚氨酯材料作为一种非降解高分子化合物具有良好的稳定性和力学特性,但作为组织工程材料的软骨细胞生物相容性研究未见报道。目的:观察聚氨酯材料与兔膝关节软骨细胞复合培养的生物相容性。方法:采用酶消化法提取兔膝关节软骨细胞,体外扩增至第3代时与聚氨酯材料复合培养,采用MTT法检测复合培养与单纯培养的细胞增殖活性;复合培养2,4周后,倒置显微镜、苏木精-伊红染色与扫描电镜观察软骨细胞生长情况,免疫组化染色观察Ⅱ型胶原表达。结果与结论:(1)MTT检测显示,复合培养6,8,10 d的细胞增殖活性高于单纯培养的细胞(P<0.05)。(2)倒置显微镜显示,2周后细胞基本长满材料表面,部分细胞融入材料内部;4周后材料表面长满细胞,成团细胞填塞材料孔隙。(3)苏木精-伊红染色显示,软骨细胞在聚氨酯材料内部稳定贴壁生长;扫描电镜显示,软骨细胞成团贴附于聚氨酯材料表面。(4)免疫组化染色显示,支架中的软骨细胞表达Ⅱ型胶原。(5)结果表明,聚氨酯材料与软骨细胞的生物相容性良好,有望成为理想的组织工程软骨材料。

BACKGROUND:Polyurethane material,as an excellent non-degradable polymer compound,has good stability and excellent mechanical properties,but the biocompatibility of chondrocytes as a tissue engineering material has not been reported.OBJECTIVE:To observe the biocompatibility of compound culture of rabbit knee cartilage with polyurethane.METHODS:Rabbit knee chondrocytes were extracted by enzyme digestion method,and were cultured with polyurethane composite material when expanded to the third generation in vitro.The proliferation activity of the composite and simple cultured cells was detected by MTT assay.After 2 and 4 weeks of compound culture,inverted microscope,hematoxylin-eosin staining and scanning electron microscope were utilized to observe the chondrocyte growth.Immunohistochemical staining was applied to observe expression of typeⅡcollagen.RESULTS AND CONCLUSION:(1)MTT assay showed that the proliferation activity of cells cultured for 6,8 and 10 days was higher than that of cells cultured only(P<0.05).(2)The inverted microscope showed that after 2 weeks,the cells were almost all over the surface of the material,and some of the cells melted into the material.After 4 weeks,the surface of the material was covered with cells,and clusters of cells filled the pores of the material.(3)Hematoxylin-eosin staining showed that chondrocytes grew stably and adhered to the wall inside the polyurethane.Scanning electron microscopy showed that chondrocytes clustered inside the polyurethane material.(4)Immunohistochemical staining showed that chondrocyte in the scaffold presented the expression of type II collagen.(5)It is concluded that polyurethane biological scaffolds have good biocompatibility with chondrocytes and are expected to be ideal tissue-engineered cartilage materials.