摘要
目的 探讨利用皮肤成纤维细胞作为肌腱构建种子细胞的可行性。方法 取外伤病人术中修剪的多余肌腱和皮肤组织,一部分用于石蜡包埋做组织学检测,一部分用酶消化法培养细胞,取第2代细胞进行细胞学检测。本实验采用偏光显微镜、免疫组织化学技术对人正常肌腱组织和皮肤组织中Ⅰ、Ⅲ型胶原的表达进行比较;采用免疫细胞化学、免疫细胞荧光、RT-PCR技术从蛋白水平、RNA水平比较第2代人肌腱细胞和皮肤成纤维细胞合成Ⅰ、Ⅲ型胶原的情况。结果 偏光显微镜下可见肌腱组织中以粗大的Ⅰ型胶原为主,而皮肤组织中Ⅰ、Ⅲ型胶原交错排列。免疫组织化学和显示石蜡包埋的肌腱组织和皮肤组织的Ⅰ型胶原染色均为强阳性,肌腱组织Ⅲ型胶原染色阴性,皮肤组织Ⅲ型胶原染色阳性。免疫细胞化学和免疫细胞荧光显示第2代肌腱细胞和皮肤成纤维细胞Ⅰ、Ⅲ型胶原染色均为阳性。RT-PCR灰度分析结果示肌腱组织和皮肤组织Ⅰ型胶原灰度比值为1.25±0.03,Ⅲ型胶原肌腱组织不表达。第2代肌腱细胞与皮肤成纤维细胞Ⅰ型胶原灰度比值为1.09±0.05,Ⅲ型胶原灰度比值为0.93±0.08。结论 人正常肌腱组织和皮肤组织中在胶原组成上均以Ⅰ型胶原为主。第2代人肌腱细胞和皮肤成纤维细胞的生物学特性相近,均合成Ⅰ型胶原和Ⅲ型胶原。皮肤成纤维细胞很有?
Objective The purpose of this study was to compare types Ⅰ and Ⅲ collagen expression in human tendon and skin as well as tenocytes and dermal fibroblasts. Methods The samples of tendon and skin were obtained from discarded tissues of traumatic patients. Part of the samples were embedded in paraffin, others were digested by enzyme to obtain tenocytes and dermal fibroblasts. Polarimicroscopy and immunohistochemistry were employed to compare the expression of type Ⅰ and type Ⅲ collagens in human tendon and skin. We also studied the synthesis of type Ⅰ and type Ⅲ collagens in tenocytes and dermal fibroblasts using immunocytochemistry, immunofluores-cence, and RT-PCR. Results Polarimicroscopy revealed that tendon tissue was mainly composed of bulky type Ⅰcollagen bundles, while type Ⅰ collagen and type Ⅲ collagen were arrayed crosswise in skin tissue. Immunohistochemistry showed that both tendon and skin expressed type Ⅰ collagen, but only skin expressed type Ⅲ collagen. In both tenocytes and dermal fibroblasts of second passage, both types Ⅰ and Ⅲ collagen staining were positive in immunohistochemistry and immunofluorescence. RT-PCR showed that grey density ratio of procollagen I between ten-don tissue and skin tissue was 1.25±0.03 and no type Ⅲprocollagen expressed in tendon. In passage two tenocytes and dermal fibroblasts, grey density ratio was 1. 09±0. 05 for type Ⅰprocollagen, and 0. 93±0. 08 for type Ⅲ pro-collagen. Conclusion Human normal tendon and skin expressedmainly type Ⅰ collagen. The second passage of human tenocytes and dermal fibroblasts showed similar biological features, both synthesizing type Ⅰand Ⅲ collagen. Therefore, dermal fibroblasts may serve as a new source of seed cells for tendon engineering.
出处
《上海第二医科大学学报》
CSCD
2004年第4期275-278,283,共5页
Acta Universitatis Medicinalis Secondae Shanghai
基金
国家高技术研究发展计划资助项目("863"计划)(2002AA205031)
上海市"重中之重"重点学科基金资助.
