骨骼肌对前交叉韧带重建腱性移植物关节内愈合效果的影响

摘要

目的探讨骨骼肌对前交叉韧带(ACL)重建腱性移植物关节内愈合效果的影响.方法取新西兰兔跟腱制备半腱半肌移植物(SSG)与全腱移植物(TTG),切断其双侧ACL后,一侧采用SSG(SSG侧)、另一侧采用TTG(TTG侧)重建ACL,于术后2、4、8周将造模成功的30只新西兰兔处死(每个时相点10只),于移植物关节内中央部取材制成5μm纵向切片,行苏木精—伊红(HE)染色和甲苯胺蓝(TB)染色,观察组织结构,高倍镜视野下对切片中央部进行细胞计数.结果术后2周,SSG骨骼肌占总面积的10%~40%,骨骼肌变性、萎缩,腱性组织胶原纤维排列规整,内部有少量成纤维细胞；TTG胶原纤维排列规整,内部偶见成纤维细胞.术后4周, SSG内已无大片骨骼肌残留,中央部为致密胶原纤维,排列不规整,可见大量新生成纤维细胞；TTG胶原纤维排列不规整,中央部仅见少量成纤维细胞.术后8周,SSG为致密结缔组织,胶原纤维排列较规整,中央部仍有较多成纤维细胞；TTG亦为致密结缔组织,胶原纤维排列较规整,移植物中央部细胞数较少.术后2、4、8周,SSG侧中央部成纤维细胞数均多于TTG侧,两侧比较,差异有统计学意义(t =6.194、16.738、9.613,P<0.05).2×3析因方差分析结果显示,不同种类的移植物中央部成纤维细胞计数存在显著差异(F =277.67,P<0.05)；术后不同观察时相点,移植物中央部成纤维细胞计数存在显著差异(F=262.72,P<0.05)；移植物处理方式与观察时相点之间存在显著交互作用(F =69.68,P<0.05).结论腱性移植物上残留骨骼肌有利于关节内段移植物内组织细胞的增生,有可能加快移植物的愈合与改建.%　　Objective To explore the effect of residual skeletal muscle on intra-articular healing of tendon graft in anterior cruciate ligament (ACL) reconstruction. Methods After ablation of ACL from bilateral of 30 New Zealand rabbits, the semi-tendon and semi-muscle graft (SSG) as well as the total tendon graft (TTG) prepared with their own achilles tendon were used separately for ACL reconstruction at each side of knee. After animals' sacrifice, specimens of the intra-articular grafts were collected at 2, 4 and 8 weeks after operation, with 10 animals in each time point. Five micron thickness slices at the central part of the graft in longitudinal plane, stained with HE and TB staining methods, were used for histological evaluation and fibroblast count under high power fields in the center of the sample. Results At 2 weeks, in SSG group, skeletal muscle became degenerative and atrophic, accounted for 10%-40% of total area of the SSG sample. In the tendon part of SSG, collagen fibers arranged regularly with few fibroblasts. Similarly, collagen fibers of the tendon in TTG group also organized in order with very few fibroblasts. At 4 weeks, no large sheet of residual skeletal muscle was seen in SSG, dense connective tissue with poor oriented collagen fibers and vigorous proliferation of fibroblast were found in the center of SSG sample. In contrast, the collagen fibers of TTG arranged irregularly with slack proliferation of fibroblast. At 8 weeks, there was still vigorous proliferation of fibroblast in the center of SSG, and alignment of collagen fibers tended to orientation. However, TTG was still dense connective tissue with oriented collagen fibers and slight proliferation of fibroblast in the center. The numbers of fibroblast in the center of the sample were much higher in SSG group than those in TTG group at 2, 4 and 8 weeks, the differences between two groups were statistically significant (t =6.194, 16.738, 9.613, P<0.05). 2 × 3 factorial variance analysis showed that among the different types of grafts, as well as from different observative time points, central compartmental fibroblast counts showed significant difference (F = 277.67, 262.72, P <0.05); between graft treatment methods and observative time points, there existed significant interaction (F = 69.68, P <0.05). Conclusion The residual skeletal muscle in the tendon graft is favorable to the proliferation of tissue cell within the intra-articular grafts, which may improve incorporation, healing and remodeling of the graft.