三维多孔丝素蛋白羟基磷灰石支架细胞相容性的实验研究  

Experimental study on the cytocompatibility of three-dimensional porous silk fibroin hydroxyapatite scaffolds

作　　者：陈仲春 苗变梁 谢幸巧 姚晋荣[2] 赵霞 CHEN Zhongchun;MIAO Bianliang;XIE Xingqiao;YAO Jinrong;ZHAO Xia(Department of Otorhinolaryngology-Head and Neck Surgery,Huashan Hospital of Fudan University,Shanghai 200040,China;Department of Macromolecular Science,Fudan University,Shanghai 200433,China)

机构地区：[1]复旦大学附属华山医院耳鼻咽喉头颈外科,上海200040 [2]复旦大学高分子科学系,上海200438

出　　处：《中国眼耳鼻喉科杂志》2023年第5期406-410,共5页Chinese Journal of Ophthalmology and Otorhinolaryngology

基　　金：上海市自然科学基金(19ZR1407700);上海科委科技发展基金(17441901600)。

摘　　要：目的 探讨丝素羟基磷灰石(HA)复合支架的细胞相容性及其作为组织工程化软骨支架的潜力。方法 以丝素蛋白(SF)和HA为主要成分,通过乳液冷冻法研制出三维多孔丝素蛋白HA复合支架。在支架上接种人鼻中隔原代软骨细胞,通过扫描电镜和活细胞染色倒置相差显微镜观察细胞形态和活性,细胞计数试剂8(CCK8)检测细胞增殖情况。结果 制作的支架具有类似细胞外基质的孔隙相互连通的三维多孔结构,能为细胞的黏附和生长提供充分的三维空间。支架能促进人原代软骨细胞在其上粘附、生长、增殖,并保持良好的细胞活性。扫描电镜观察:培养14 d的细胞覆盖99%支架表面较培养3 d的细胞覆盖面积(74%)增大(P<0.01)。CCK8检测培养3 d和7 d较1 d时细胞明显增多(P<0.01)。结论 丝素HA复合支架具有良好的细胞相容性,是一种潜在的组织工程软骨修复材料。Objective To investigate the cytocompatibility of silk fibroin-hydroxyapatite composite scaffolds and their potential as tissue-engineered chondrocyte scaffolds.Methods Three-dimensional porous silk fibroin-hydroxyapatite composite scaffolds were prepared by emulsion freezing method using silk fibroin and hydroxyapatite.Primary human nasal septum chondrocytes were seeded on the scaffolds,and cell morphology and activity were observed by scanning electron microscope,inverted phase contrast microscope after live cell staining,and cell proliferation detected by CCK8.Results The scaffolds had a three-dimensional porous structure similar to extracellular matrix.The scaffold could promote the adhesion,growth and proliferation of human primary chondrocytes.Scanning electron microscope observed that the cells covered more areas(99%)of the scaffold surface on 14 days of culture than on 3 days(74%)(P<0.01).CCK8 detection showed that the cells proliferated at 3 days and 7 days compared with 1 day(P<0.01).Conclusions The 3D porous silk fibroin-hydroxyapatite composite scaffold has good cytocompatibility and is a promising biomaterial for cartilage repair.

关 键 词：丝素蛋白 羟基磷灰石 软骨重建 软骨细胞 细胞培养 组织工程 

分 类 号：R762[医药卫生—耳鼻咽喉科]