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骨-前交叉韧带-骨同种异体移植后的细胞存活:犬模型中多种标志物的DNA指纹图谱、分离及胶原形态分析

Cell survival following bone-anterior cruciate ligament-bone allograft transplantation: DNA fingerprints, segregation, and collagen morphological analysis of multiple markers in the canine model.

作者信息

Goertzen M J, Buitkamp J, Clahsen H, Möllmann M

机构信息

Department of Orthopaedic Surgery, Heinrich Heine University, Düsseldorf, Germany.

出版信息

Arch Orthop Trauma Surg. 1998;117(4-5):208-14. doi: 10.1007/s004020050231.

Abstract

Bone-anterior cruciate ligament-bone allograft transplantation has become recognized as a potential solution to reconstruction of the anterior cruciate ligament (ACL). The purpose of this study was to determine the time-dependent fibrocyte donor cell survival rate after cryopreserved bone-ACL-bone allograft transplantation. Additionally, bony incorporation of the pediculated bone plugs was examined. The ability to successfully transplant allogenous ACL fibrocytes and have them survive has not previously been documented. In this study, DNA fingerprints identified and documented the survival rate of the cellular DNA in transplanted ACL allografts for ACL re-construction in the knee joints of 10 skeletally mature dogs. At 4, 8, 26 and 52 weeks after ACL allograft transplantation, DNA probes, H & E, Giemsa, Goldner, PAS and polarized light staining was done to demonstrate the time-dependent changes in the allografts after transplantation. At 4 weeks host fibrocytes began to grow into the graft; however, histologically the cells could not be distinguished as to host or donor origin. After 4 weeks the DNA pattern reflected only the band pattern of the host. This reveals the early cellular infiltration activity of the host into the ACL allograft, also demonstrated in the light microscopy stainings. The survival rate of transplanted allogenous ACL fibrocytes had not been documented before this study. There is no evidence that ACL allograft cells survive in the intra-articular environment of the host's knee. Within 4 weeks ACL allografts became completely repopulated with host cells. The cells that migrate early into the ACL allografts are probably of synovial origin because they are present before revascularization and collagen reorganization occur. We conclude from this study that viable cells in transplanted ACL allografts did not survive longer than 4 weeks after intra-articular transplantation. Advances in molecular biology may offer new approaches to alter or stimulate fibrocyte population and function in the transplanted ACL allograft used for ACL reconstruction. New methods to maintain the viability of donor cells may be necessary to improve the biomechanical and histological properties of autografts or allografts for ACL reconstruction.

摘要

骨-前交叉韧带-骨同种异体移植已被公认为是重建前交叉韧带(ACL)的一种潜在解决方案。本研究的目的是确定冷冻保存的骨-ACL-骨同种异体移植后,随时间变化的纤维细胞供体细胞存活率。此外,还检查了带蒂骨栓的骨融合情况。此前尚未有成功移植异体ACL纤维细胞并使其存活的相关记录。在本研究中,通过DNA指纹识别并记录了10只骨骼成熟犬膝关节内用于ACL重建的移植ACL同种异体移植物中细胞DNA的存活率。在ACL同种异体移植后4周、8周、26周和52周,进行DNA探针、苏木精和伊红染色、吉姆萨染色、戈德纳染色、过碘酸希夫染色和偏振光染色,以显示移植后同种异体移植物随时间的变化。4周时,宿主纤维细胞开始长入移植物;然而,从组织学上无法区分这些细胞是宿主来源还是供体来源。4周后,DNA图谱仅反映宿主的条带模式。这揭示了宿主对ACL同种异体移植物的早期细胞浸润活性,在光学显微镜染色中也得到了证实。在本研究之前,移植异体ACL纤维细胞的存活率尚无记录。没有证据表明ACL同种异体移植细胞能在宿主膝关节的关节内环境中存活。4周内,ACL同种异体移植物完全被宿主细胞重新填充。早期迁移到ACL同种异体移植物中的细胞可能来自滑膜,因为它们在血管再生和胶原重组发生之前就已存在。我们从本研究得出结论,关节内移植后,移植的ACL同种异体移植物中的活细胞存活时间不超过4周。分子生物学的进展可能会为改变或刺激用于ACL重建的移植ACL同种异体移植物中的纤维细胞数量和功能提供新方法。可能需要新的方法来维持供体细胞的活力,以改善用于ACL重建的自体移植物或同种异体移植物的生物力学和组织学特性。

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