明胶-硫酸软骨素-透明质酸钠作为组织工程软骨支架的实验研究  

EXPERIMENTAL STUDY ON GELATIN-CHONDROITIN SULFATE-SODIUM HYALURONATE TRI-COPOLYMER AS NOVEL SCAFFOLDS FOR CARTILAGE TISSUE ENGINEERING

作　　者：范宏斌[1] 胡蕴玉[1] 李旭升[1] 吕荣[1] 白建萍[1] 王军[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,西安710032

出　　处：《中国修复重建外科杂志》2005年第6期473-477,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家"九五"高技术计划新材料领域基金资助项目(009-0160)~~

摘　　要：目的探索明胶-硫酸软骨素-透明质酸钠多孔支架的制备方法和其作为组织工程软骨支架的可行性。方法光镜和扫描电镜观察-20℃、-80℃及液氮条件下冷冻抽真空干燥制备的明胶-硫酸软骨素-透明质酸钠支架的孔径、孔隙率、交通孔和密度;测定不同条件制备的支架材料与正常软骨的压缩载荷-形变曲线;分离、扩增兔骨髓基质干细胞(mesenchymalstemcells,MSCs),接种于不同条件制备的支架材料上培养,MTT检测MSCs在支架上的黏附率及多孔支架对细胞增殖功能的影响。结果在-20℃、-80℃及液氮条件下制备的支架材料具有疏松多孔结构,孔径依次为300±45μm、230±30μm和45±10μm,孔隙率为81%、79%和56%,密度为9.41±0.25μg/mm3、11.50±0.36μg/mm3和29.50±0.61μg/mm3;-80℃和液氮条件下制备的支架材料,力学强度接近于正常软骨;MTT测得细胞在-20℃、-80℃及液氮条件下制备的支架材料,黏附率分别为85.0%、87.5%和56.3%;可轻度促进MSCs增殖。结论-80℃条件下抽真空干燥制备的明胶-硫酸软骨素-透明质酸钠多孔支架,具有良好的孔径、孔隙率和抗压缩载荷能力,与MSCs具有较好的相容性,是软骨组织工程中的一种新型仿生支架材料。Objective To fabricate a novel gelatin-chondroitin sulfate-sodium hyaluronate tri-copolymer scaffold and to confirm the feasibility of serving as a scaffold for cartilage tissue engineering. Methods Different scaffolds was prepared with gelatin-chondroitin sulfate-sodium hyaluronate tri-copolymer by varying the freezing temperatures (-20℃,-80℃ and liquid nitrogen). Pore size, porosity, inter pores and density were observed with light microscopy and scanning electron microscopy (SEM). The load-stiffness curves were compared between different scaffolds and normal cartilage. The number of MSCs attaching to different scaffolds and the function of cells were also detected with MTT colorimetric microassay. Results The pore size was 300±45, 230±30 and 45±10 μm; the porosity was 81%, 79% and 56%; the density was 9.41±0.25, 11.50±0.36 and 29.50±0.61 μg/mm^3 respectively in different scaffolds fabricated at -20℃,-80℃ and liquid nitrogen; the latter two scaffolds had nearly the same mechanical property with normal cartilage; the cell adhesion rates were 85.0%, 87.5% and 56.3% respectively in different scaffolds and the scaffolds can mildly promote the proliferation of MSCs. Conclusion Gelatin-chondroitin sulfate-sodium hyaluronate tri-copolymer scaffold fabricated at -80℃ had proper pore size, porosity and mechanical property. It is a novel potential scaffold for cartilage tissue engineering.

关 键 词：组织工程 骨髓基质干细胞 软骨支架材料 实验研究 明胶-硫酸软骨素-透明质酸钠 

分 类 号：R318.08[医药卫生—生物医学工程]