自体肌腱加富血小板血浆重建比格犬前交叉韧带的实验研究

Reconstruction of Anterior Cruciate Ligament of Beagle Dogs by Autologous Tendon with Plateletrich Plasma

目的研究应用自体肌腱添加富血小板血浆重建比格犬前交叉韧带的早期组织学转归及生物力学变化。方法选用健康成熟的雄性比格犬36只,随机分为两组(模型组和实验组),将每组比格犬36个膝关节分别取同侧趾长屈肌腱进行单束双股重建,实验组添加自体富血小板血浆。术后第4、8、12周每组随机抽取6只比格犬处死,分别取12个膝关节(包括股骨端与前交叉韧带),每组随机取8个膝关节做生物力学拉脱试验,另4个膝关节标本进行组织学检查,并对实验数据进行统计学分析。结果组织学观察模型组术后4周时,腱骨界面有纤维血管形成,实验组腱骨界面纤维血管更加密集,排列较有序并有少量Ⅱ型胶原纤维。术后8周模型组有肌腱周围的新骨形成,骨隧道壁增厚。但在此期间,胶原纤维机化不充分,腱骨的连续性偶尔可见;实验组胶原纤维生长成熟,可见腱骨的连续性,并在腱骨界面中心出现少量垂直排列的Sharpey样纤维。术后12周模型组可见少量垂直排列的Sharpey样纤维;实验组腱骨界面可见大量的垂直排列的Sharpey样纤维,此时腱骨愈合已逐步趋于稳定。生物力学测试显示术后第4、8周,两组肌腱都能从骨隧道里拉脱出来。在第12周时,模型组仍可以从骨髓道里被拉脱;实验组肌腱均不能从隧道中被拉出,而是在肌腱中间断裂。结论应用自体肌腱可以重建比格犬前交叉韧带并达到腱骨愈合,添加富血小板血浆可以加快腱骨愈合的速度。

Objective To research early histological and biomechanical outcome of reconstruction anterior cruciate ligament of beagle dogs by autologous tendon adding platelet-rich plasma. Methods We selected 36 healthy adult male beagle dogs and divided them into model group and research group. The same side tendons of the flexor digitorum longus（ 8 cm in length） were harvested in all dogs and folded in half to create a 4-cm-long double-stranded graft in two groups,but we added platelet-rich plasma when reconstruction of autologous tendon in research group. At the four,eight and twelve-week,6 beagle dogs drawing randomly from two groups were sacrificed and 12 knee joints were obtained including the femoral components and the anterior cruciate ligament. Eight of them were to study the biomechanics and the rest were for histological examination. At last,statistical analysis was carried out on the experimental data. Results Four-weeks after opertiaon,histological findings showed that there were the formation of fibrovascular in the tendon-bone interface in model group and there were more intensive fibrovascular and arranged in order associated with type Ⅱcollagen deposition of chondriod cells from bone growth to the tendon in the tendon-bone interface in research group. Eight-week after operation,there were formation of the new bone around tendons and the thickening of the bone tunnel wall. However,during this period,the collagen fibers machine were not sufficient and tendon continuity of bone could only occasionally be seen in model group. In research group,the collagen fibers grow to maturity and the continuity of the tendon bone and the small amounts of vertical arrangement of Sharpey-like fibers in the tendon bone interface center visible could be seen. At the twelve-week,the small amounts of vertical arrangement of Sharpey-like fibers could be seen in model group. In research group,the tendon bone healing process to enter a period of chronic inflammatory reaction and the tendon bone healing had gradually in a stable. The biomechanics research showed that the tendons of two groups could be pulled out from the bone tunne at the four-week and eight-week. At the twelve-week,the tendons in model group could be gradually pulled out of the tendon from the clamp. Whether,the tendons in research group could not be pulled out from the bone tunne and ruptured in the middle. Conclusion The reconstruction of anterior cruciate ligament can be realized by applying autologous tendon and achieve tendon bone healing. In addition,autologous tendon adding platelet-rich plasma can promote tendon-bone healing.