PhaP-RGD融合蛋白修饰的PHBHHx与人鼻中隔软骨细胞的生物相容性研究  

Biocompatibility of Human Nasal Septal Chondrocytes on PHBHHx Coated with PhaP-RGD Fusion Peptide

作　　者：常会敏[1] 高天喜[2] 王正辉[2] 卢晓云[3] 吴宝俊[2] 张向红[2] 

机构地区：[1]西安医学院附属医院耳鼻喉科,陕西省西安市710077 [2]西安交通大学第二附属医院耳鼻咽喉-头颈外科,陕西省西安市710004 [3]西安交通大学生命科学与技术学院,陕西省西安市710049

出　　处：《组织工程与重建外科杂志》2013年第6期319-323,共5页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81000416);中央高校基本科研业务费专项资金(西安交通大学国际合作类;交叉学科类);西安交通大学第二附属医院人才培养基金[RC(XM)201102];西安交通大学第二附属医院重点项目基金[YJ(ZD)201103]

摘　　要：目的观察经PhaP-RGD融合蛋白修饰的3-羟基丁酸和3-羟基己酸的共聚酯(PHBHHx)生物材料与人鼻中隔软骨细胞的生物相容性。方法体外培养人鼻中隔软骨细胞,接种于经PhaP-RGD融合蛋白修饰的PHBHHx膜性材料上进行体外培养,通过MTT比色法、DAPI染色、扫描电镜、甲苯胺蓝染色等实验方法观察在体外条件下生物修饰后的PHBHHx膜与鼻中隔软骨细胞的生物相容性。结果经PhaP-RGD融合蛋白修饰的PHBHHx膜表面接触角变小,亲水性增加;人鼻中隔软骨细胞在蛋白修饰后的生物膜材料表面能保持较高的增殖率;经蛋白修饰的PHBHHx膜表面的软骨细胞活力明显高于细胞培养板及未经蛋白修饰的PHBHHx膜上的软骨细胞;甲苯氨蓝染色发现经PhaP-RGD修饰的PHBHHx膜能促进软骨细胞分泌胞外基质糖胺多糖。结论经PhaP-RGD融合蛋白修饰的PHBHHx与软骨细胞的生物相容性良好,是一种较理想的软骨组织工程支架材料。Objective To explore the biocompatibility of PHBHHx biomaterial coated with PhaP-RGD fusion protein on human nasal septal chondrocytes. Methods The human nasal septal chondrocytes were cultured and seeded onto the PHBHHx biomaterials modified with PhaP-RGD fusion protein in vitro. The biocompatibility of modified PHBHHx on human nasal septal chondrocytes was detected by MTT assay, DAPI staining, scanning electron microscopy and toluidine blue staining. Results The contact angle of modified PHBHHx was smaller and the surface hydrophilicity increased; The human nasal septal chondrocytes on modified maintained a high proliferation rate. The viability of chondrocytes on modified PHBHHx film was significantly higher than that on PHBHHx film and cell culture plate. It is discovered that PHBHHx film coated with PhaP-RGD can promote the secretion of extracellular matrix of cartilage--glycosaminoglycan by toluidine blue staining. Conclusion PHBHHx coated with PhaP-RGD fusion protein has better biocompatibility on chondrocytes, and is an ideal scaffold material for cartilage tissue engineering.

关 键 词：PhaP—RGD融合蛋白 3-羟基丁酸与3-羟基己酸共聚酯 人鼻中隔软骨细胞 组织工程 

分 类 号：Q813.11[生物学—生物工程]