PGA支架上人与猪皮肤成纤维细胞增殖和基质分泌的比较  

Comparison of Growth and Matrix Secretion in Human and Pig Dermal Fibroblasts Seeded on PGA Scaffolds

作　　者：徐福[1] 李鹏程[2] 刘伟[1] 王玉明[2] 魏娴[1] 周广东[1] 崔磊[1] 曹谊林[1] 

机构地区：[1]上海交通大学医学院第九人民医院整复外科上海市组织工程研究重点实验室,上海200011 [2]贵州遵义医学院

出　　处：《上海交通大学学报（医学版）》2006年第10期1138-1142,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家重点基础研究发展规划基金("973"项目)(2002AA20503)资助项目

摘　　要：目的对人与猪皮肤成纤维细胞作为肌腱组织工程种子细胞,进行可行性比较。方法酶消化法获取人与猪皮肤成纤维细胞,培养扩增,取第2代细胞,以1×106/mL的细胞密度接种至聚羟基乙酸(PGA)圆柱形支架上体外培养。通过大体、组织学及增殖曲线观察,羟脯氨酸定量测定以及免疫组化检测,比较两种细胞在PGA支架的黏附、增殖及基质分泌能力。结果组织学及三维增殖曲线观察结果显示,与猪皮肤成纤维细胞比较,人皮肤成纤维细胞与支架的黏附能力及细胞增殖能力良好;接种后第6天和第12天,HE染色可见人皮肤成纤维细胞-PGA复合物有连续的基质形成,两者羟脯氨酸含量分别为(2.23±0.25)μg/mgprotvs(1.07±0.18)μg/mgprot和(3.72±0.39)μg/mgprotvs(0.27±0.04)μg/mgprot,差异具有统计学意义(P<0.05);免疫组化结果显示,接种后第12天,两种复合物的Ⅰ、Ⅲ型胶原表达均呈阳性。结论与猪皮肤成纤维细胞相比,人皮肤成纤维细胞更有希望作为肌腱组织工程的种子细胞。Objective To compare the possibility of human and pig dermal fibroblasts used in tendon engineering as seed cells. Methods Human and pig dermal fibroblasts were isolated from the skin by enzymation and cultured in vitro. The second passage of human and pig dermal fibroblasts were collected and seeded at a density of 1.0 × 10^6 cells/mL onto cylinder-shaped polyglycolic acid （PGA）scaffolds. Changes in growth curves of the second passage cells were observed, and cell-scaffold constructs were harvested for histological and hydroxyproline （HPR） production assessment. Growth and matrix secretion of the two types of cells on PGA scaffolds were compared. Results In the three-dimensional PGA scaffolds, it was found that human dermal fibroblasts had better adhesion and extracellular matix secretion than pig fibroblasts, and the growth curves showed that human dermal fibroblasts had much better proliferation potential than pig cells. HE staining showed that tissue formed in human dermal fibroblasts-PGA complex on d 6 and d 12 in vitro. HPR production of human and pig dermal fibroblasts on PGA scaffolds was （2.23 ±0.25） μg/mgprot vs （ 1.07 ± 0.18）μg/mgprot, and （3.72 ± 0.39） μg/mgprot vs （0.27 ± 0.04 ） μg/mgprot on d 6 and d 12, respectively, with significant statistical differences （P 〈 0.05）. Positive staining of type Ⅰ and Ⅲ collagen could be detected by immunohistochemical staining in both types of cell-scaffolds on d12. Conclusion It is supposed that human dermal fibroblasts may be used in tendon engineering as seed cells.

关 键 词：组织工程 肌腱 皮肤成纤维细胞 聚羟基乙酸 

分 类 号：R318[医药卫生—生物医学工程]