摘要
目的观察单抗CD44-生物素（Biotin）-亲和素（Avidin）绑定系统在构建组织工程软骨中能否提高软骨细胞与支架的黏附能力。方法分别制备软骨细胞二维和三维培养体系，分3组：A：壳聚糖＋软骨细胞；B：生物素＋亲和素化壳聚糖＋软骨细胞；C：生物素化单抗CD44＋亲和素化壳聚糖＋软骨细胞。分别测定细胞展平面积、细胞接种脱落率、细胞增殖率，逆转录-聚合酶链反应（1iT-PCR）法检测Ⅱ型胶原（ColII）、聚集蛋白聚糖（aggrecan）、sox9的mRNA表达，组织学切片观察壳聚糖支架内细胞黏附生长情况。结果细胞展平面积、细胞增殖率及ColII、aggrecan、sox9的mRNA表达量皆为：C组〉B组〉A组（P〈0．05）。细胞接种脱落率：A组为c组的3．71倍、为B组的2．17倍，而B组为C组的1．71倍，A组〉B组〉C组（P〈0．05）。组织切片显示细胞在支架内C组增殖旺盛、胞外基质表达较丰富，A组增殖较弱、胞外基质表达较弱，B组介于C与A组间。结论单抗CD44．Biotin．Avidin绑定系统能比Biotin．Avidin绑定系统更显著的提高软骨细胞与支架的黏附能力，促进组织工程软骨种子细胞的增殖和软骨细胞表型的表达。
Objective To oberserve the efficiency of CD44 monoclonal antibody-biotin-avidin binding system for the improvement of cells adhesion to scaffolds in the cartilage tissue engineering. Meth- otis The chondrocytes were cultured in two-dimensional and three-dimensional culture system respective- ly. And in each system the cells were divided into 3 groups： Group A, the chondrocytes were seeded rou- finely on the chitosan membrane or scaffolds; Group B, the chondroeytes were seeded on the chitosan mem- brane or scaffolds with biotin-avidin binding system; Group C, the chondrocytes were seeded on the chi- tosan membrane or scaffolds with CD44 monoclonal antibody-biotin-avidin binding system. The spreading area in the two-dimensional culture system, cell exfoliation rate and cell proliferation rate in the three-di- mensional culture system were investigated. The histologic analysis was used to evaluate the cell growth in the scaffolds. The mRNA expression level of type H collagen, aggrecan and sox9 was detected by using re- verse transcription-polymerase chain reaction （RT-PCR）. Results Group C showed better results in the cell spreading area, the cell proliferation rate and the mRNA expression level of type II collagen, aggrecan and sox9 than group B, and group B more than group A, group C 〉 group B 〉 group A （ P 〈 0.05 ）. The results showed that the cell exfoliation rate in group A was 2. 17 times of that in group B which was 1.71 times of that in group C, group A 〉 group B 〉 group C （ P 〈 0. 05 ）. The histologic sections showed the pro- liferation of cells was exuberant and expression of extracellutar matrix was stronger in group C, that in group B was between group C and group A in which they were weaker. Conclusion The CD44 monoclonal anti- body-biotin-avidin binding system could improve the chondrocytes adhesion, proliferation and phenotypic expression in cartilage tissue engineering.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第2期295-298,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30872623、81071468）
上海市科委基础重点科研项目（09JCl403600）
