Gelberman R H, Amiel D, Harwood F
Department of Orthopaedic Surgery, Massachusetts General Hospital, Boston.
J Hand Surg Am. 1992 May;17(3):551-8. doi: 10.1016/0363-5023(92)90370-5.
To determine the precise mechanism by which contact tendon healing occurs at the cellular level, the production of pro alpha (I) collagen messenger RNA (mRNA) produced by fibroblasts of healing intrasynovial flexor tendons was determined by an in situ hybridization technique. The repair site and the proximal and distal tendon stumps of repaired tendons treated with early controlled passive mobilization were fixed and buffered in formalin, 3, 7, 10, and 17 days after repair. A complimentary DNA (cDNA) probe corresponding to alpha (I) procollagen mRNA was labeled with [32P]d-CTP. After hybridization, autoradiography, and staining of the sections, the level of procollagen mRNA was assessed by microscopic examination. Rising levels of procollagen mRNA, indicating progressively increasing levels of synthetic collagen activity, were detected in the healing tendons through 10 days. A moderate decrease in procollagen mRNA was seen at 17 days. Genetic expression for procollagen mRNA was localized specifically to the epitenon cells on the tendon surface overlying the repair site and to cells in the gap between the tendon stumps. No detectable expression was noted in endotenon fibroblasts. The finding of high levels of expression for procollagen type I mRNA in the surface layer of healing tendons demonstrates that cells intrinsic to tendon epitenon contribute the greatest quantity of native tendon collagen to the repair site during these important early intervals after tendon suture.
为了确定滑膜内屈肌腱愈合在细胞水平上发生的精确机制,采用原位杂交技术测定愈合中的滑膜内屈肌腱成纤维细胞产生的前α(I)型胶原信使核糖核酸(mRNA)。对早期接受控制性被动活动的修复肌腱的修复部位以及修复肌腱的近端和远端残端在修复后3天、7天、10天和17天进行固定并用福尔马林缓冲。用[32P]d-CTP标记与α(I)型前胶原mRNA相对应的互补DNA(cDNA)探针。杂交、放射自显影和切片染色后,通过显微镜检查评估前胶原mRNA的水平。在愈合肌腱中直至10天均检测到前胶原mRNA水平升高,表明合成胶原活性水平逐渐增加。在17天时观察到前胶原mRNA有适度下降。前胶原mRNA的基因表达特异性定位于修复部位上方肌腱表面的腱外膜细胞以及肌腱残端之间间隙中的细胞。在内腱成纤维细胞中未观察到可检测到的表达。在愈合肌腱表层中I型前胶原mRNA高水平表达的发现表明,在肌腱缝合后的这些重要早期阶段,肌腱腱外膜固有的细胞为修复部位贡献了最大量的天然肌腱胶原。