中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (22): 3455-3459.doi: 10.12307/2022.270

• 组织工程软骨材料Tissue-engineered cartilage • 上一篇    下一篇

聚氨酯材料复合软骨细胞构建组织工程化软骨

韩  志1,王志苗2,嘎西斯甲2,卢庆玲1,郭  涛3   

  1. 1贵州中医药大学第二附属医院骨一科,贵州省贵阳市  550003;2贵州中医药大学,贵州省贵阳市  550002;3贵州省人民医院骨一科,贵州省贵阳市  550001
  • 收稿日期:2021-01-05 修回日期:2021-02-22 接受日期:2021-05-23 出版日期:2022-08-08 发布日期:2022-01-10
  • 通讯作者: 郭涛,博士,主任医师,贵州省人民医院骨一科,贵州省贵阳市 550001
  • 作者简介:韩志,男,1984年生,山东省枣庄市人,汉族,2013年贵州中医药大学毕业,硕士,主治医师,主要从事骨损伤修复及组织工程的研究。
  • 基金资助:
    2019年贵州省卫健委科技基金(GZWJKJ 20191137号),项目负责人:郭涛

Tissue engineered cartilage constructed by polyurethane composite chondrocytes

Han Zhi1, Wang Zhimiao2, Gaxi Sijia2, Lu Qingling1, Guo Tao3   

  1. 1First Department of Orthopedics, Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550003, Guizhou Province, China; 2Guizhou University of Traditional Chinese Medicine, Guiyang 550002, Guizhou Province, China; 3First Department of Orthopedics, Guizhou Provincial People’s Hospital, Guiyang 550001, Guizhou Province, China
  • Received:2021-01-05 Revised:2021-02-22 Accepted:2021-05-23 Online:2022-08-08 Published:2022-01-10
  • Contact: Guo Tao, MD, Chief physician, First Department of Orthopedics, Guizhou Provincial People’s Hospital, Guiyang 550001, Guizhou Province, China
  • About author:Han Zhi, Master, Attending physician, First Department of Orthopedics, Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550003, Guizhou Province, China
  • Supported by:
    the Science and Technology Fund of Health Commission of Guizhou Province, No. GZWJKJ 20191137 (to GT) 

摘要:

文题释义:
聚氨酯:是有氨脂结合基团的一类聚合物,既有橡胶的弹性又有塑料的强度,在耐久性和弹性方面有其他材料无法比拟的优势,是继聚乙烯、聚丙烯、聚苯乙烯后的新兴材料。
软骨细胞:是组成软骨组织的主要细胞,幼稚的软骨细胞位于软骨表层,单个分布,成熟的软骨细胞多成群分布,由同源母细胞分裂增殖形成。软骨细胞分泌Ⅱ型胶原蛋白,呈网状结构,具有一定的机械强度。

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:①MTT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。

https://orcid.org/0000-0002-1419-2272 (韩志) 

中国组织工程研究杂志出版内容重点:生物材料;骨生物材料口腔生物材料纳米材料缓释材料材料相容性;组织工程

关键词: 聚氨酯, 软骨细胞, 支架, 复合材料, 生物相容性, Ⅱ型胶原

Abstract: 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 type II collagen.
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.

Key words: polyurethane, chondrocytes, scaffold, composite materials, biocompatibility, type II collagen

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