Chinese Journal of Tissue Engineering Research ›› 2025, Vol. 29 ›› Issue (4): 713-720.doi: 10.12307/2025.222
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Xiong Bohan1, Wang Guoliang1, Yu Yang1, Xue Wenqiang2, Yu Hong1, Liu Jinrui1, Ruan Zhaohui1, Li Yajuan1, Liu Haolong1, Dong Kaiyan1, Long Dan1, Chen Zhao3
Received:
2023-12-14
Accepted:
2024-02-07
Online:
2025-02-08
Published:
2024-05-30
Contact:
Wang Guoliang, PhD, Associate chief physician, Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China
About author:
Xiong Bohan, Master candidate, Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China
Supported by:
CLC Number:
Xiong Bohan, Wang Guoliang, Yu Yang, Xue Wenqiang, Yu Hong, Liu Jinrui, Ruan Zhaohui, Li Yajuan, Liu Haolong, Dong Kaiyan, Long Dan, Chen Zhao. Internal tension relieving technique assisted anterior cruciate ligament reconstruction to promote ligamentization of Achilles tendon grafts in small ear pigs in southern Yunnan province[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(4): 713-720.
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2.1 实验动物数量分析 实验过程中30只雌性滇南小耳猪均无发生死亡,全部进入结果分析。 2.2 实验动物一般情况观察 见图1。雌性滇南小耳猪30只术前动物精神、食欲、运动状态均良好。造模后,两组中各有1只动物右膝关节出现轻微感染及关节腔积液现象,术后予以常规抗感染治疗,连续3 d肌肉注射苄星青霉素控制感染,伤口每日消毒换药后,感染症状消除。其余动物生命体征及健康状态均良好,手术前后无实验动物的丢失。各组动物术中出血及体液量的丢失较少,手术前后实验动物的体质量均无明显变化。术后14 d,各组实验动物膝关节手术切口均良好愈合,关节功能及活动逐渐恢复正常,双后肢能伸直并支撑身体躯干,可以在饲养笼内自由走动。麻醉后查体:膝关节重建的交叉韧带连续性完整,膝关节无僵硬,活动度正常,前抽屉试验及Lachman实验均阴性。"
2.3 重建韧带形态学观察 分别打开各组滇南小耳猪膝关节暴露前交叉韧带观察,见图2。 2.3.1 减张组 12周时,减张组前交叉韧带移植物呈现灰白色较致密纤维束状结构,完整性尚可,有细小韧带纤维束膨出及断裂,韧带表面有部分滑膜覆盖,张力可,胫骨隧道入口与股骨隧道入口移植物呈扁束状,中段移植物呈现较规则圆柱状,韧带表面有极少数红色血管覆盖;24周时,移植物呈现白色较致密纤维束状结构,完整性有所提高,有极少韧带纤维束膨出,未见纤维束断裂,韧带表面见较致密滑膜组织覆盖,移植物色泽较正常前交叉韧带苍白,韧带表面血管覆盖较12周时增多;48周时,移植物完整性较好,张力优,滑膜组织将跟腱移植物完全覆盖,韧带纤维束整合紧密,未见断裂及膨出,色泽接近正常前交叉韧带组织,胫骨隧道入口、股骨隧道入口、韧带中段移植物均呈现较为规则的圆柱状,形态均一、重塑良好。 2.3.2 普通组 12周时,普通组关节腔内移植物呈现苍白色较疏松纤维束状结构,完整性较差,有较多韧带纤维束膨出及断裂,韧带表面有较少滑膜组织覆盖,张力较弱,胫骨隧道入口、股骨隧道入口及韧带中段移植物均呈现扁平束状结构;24周时,关节腔内移植物呈现灰白色较致密纤维束状结构,张力稍欠佳,完整性提高,可见少部分韧带纤维束膨出及紊乱,韧带表面有部分滑膜组织覆盖,移植物表面见少量滑膜血管翳生成,但色泽仍较正常前交叉韧带苍白;48周时,移植物完整性好,在关节腔内呈现白色致密纤维束状结构,张力尚可但差于跟腱组,完整性较好,未见韧带纤维束的膨出及紊乱,韧带表面有大量滑膜组织覆盖,移植物可见部分滑膜血管翳在其间,移植物色泽接近正常前交叉韧带。 2.3.3 对照组 在12,24,48周,对照组自体右前肢正常前交叉韧带表面光滑、胫骨隧道入口、股骨隧道入口及韧带中段移植物均呈现规则圆柱状结构,滑膜组织覆盖完全,具有明显光泽度,两端牵拉时可感受到明显的张力及弹性。"
2.5 韧带苏木精-伊红染色结果和血管化评分 见图4,5,表2。术后12周,减张组与普通组血管分布均较少,血管内皮细胞排列较为疏松,重建韧带内血管分散,表明移植物内血管分布较少,血管化程度尚不成熟。对照组韧带内血管分布密集均匀,血管内皮细胞排列紧密有序。术后24周,减张组与普通组在同一视野下血管数量均增多,但减张组血管再生优于普通组,排列较12周时更加均匀,且出现部分肥厚炎性血管。术后48周,减张组韧带内血管数量进一步增多,血管化进程快于普通组,出现较大量肥厚炎性血管,但普通组视野下血管增生数量仍较少,分布较不均匀。术后12,24,48周减张组血管化评分均低于普通组,组间、组内比较差异均有显著性意义 (P < 0.05)。"
2.6 扫描电镜结果 见图6。在扫描电镜10 μm的放大倍数下,对照组在12,24,48周均可见平行或横向交错排列的胶原纤维,几乎未见断裂的胶原纤维束,平行对称排列的胶原纤维中间还可见部分纤维横向或交错排列(见红色箭头),这种多方向、类网状的排列结构提示韧带具有良好的强度和塑形程度。 还可以看到减张组、普通组与对照组韧带均由直行或扭曲的胶原束和纤维束构成,大部分胶原纤维主要呈现单向平行排列,呈现一定的排列周期性与规律性。12周时,减张组及普通组均可见多量断裂的胶原纤维束,排列紊乱无规则,被覆在正常塑形的深层胶原纤维束表面。但减张组在同一视野下胶原纤维断裂的数目及程度均小于普通组,表明此时移植物的超微结构重塑缓慢,韧带化成熟度较低。 24周时,减张组与普通组致密排列的胶原纤维表面已出现部分纤维的横向连接,减张组多于普通组。两组同一视野下仍可见部分断裂的胶原纤维束,但断裂纤维的数量及程度均少于12周时,组间比较减张组断裂的胶原纤维数量少于普通组(见黑色箭头)。 48周时,减张组胶原纤维重塑程度已经接近对照组,出现多量胶原纤维的横向连接,提示移植物的刚度及塑形程度达到了较为成熟的水平。普通组胶原纤维排列较规则,但仍偶见断裂的胶原纤维断端在其中散在分布,视野中可见部分纤维出现了较明显的横向连接。这提示48周时,减张组不良纤维的数量少于普通组,韧带化重塑已经接近对照组正常前交叉韧带。"
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