Ouyang Hong Wei, Goh James C H, Lee Eng Hin
Department of Orthopaedic Surgery, National University of Singapore, Singapore 119260.
Division of Bioengineering, National University of Singapore, Singapore 119260.
Cell Transplant. 2004 Sep;13(6):649-658. doi: 10.3727/000000004783983549.
Bone marrow stromal cells are potentially useful for tendon repair and regeneration. To provide lasting benefits, the seeded cells must survive implantation at local tendon sites. Our objective was to determine the in vivo fate of allogeneic bone marrow stromal cells (bMSCs) at different time points after implantation into patella tendon defects (i.e., at 2, 3, 5, and 8 weeks). The protocol involved the labeling of bMSCs with green fluorescent protein (GFP) or carboxyfluorescein diacetate (CFDA) before implantation. A window defect (5 × 5 mm) was created at the central portion of rabbit patella tendon and subsequently treated with GFP- or CFDA-marked bMSCs. The marked bMSCs were loaded into the window defect with fibrin glue. Upon sacrifice of the rabbits at the different time points, the implant site of the patellar tendon was immediately retrieved and the viability of the labeled cells was assessed under confocal microscopy. The results showed that the seeded bMSCs remained viable within the tendon wound site for at least 8 weeks after implantation. The cell morphology was changed from a round shape at 2 weeks to a spindle shape at 5 weeks after implantation. This study demonstrated that the bMSCs remained viable for prolonged periods after implantation and therefore have the potential to influence the formation and remodeling of neotendon tissue after tendon repair.
骨髓基质细胞对肌腱修复和再生具有潜在的应用价值。为了提供持久的益处，植入的细胞必须在局部肌腱部位存活。我们的目的是确定同种异体骨髓基质细胞（bMSCs）在植入髌腱缺损后不同时间点（即2周、3周、5周和8周）在体内的命运。实验方案包括在植入前用绿色荧光蛋白（GFP）或羧基荧光素二乙酸酯（CFDA）标记bMSCs。在兔髌腱中央部分制造一个窗口缺损（5×5毫米），随后用GFP或CFDA标记的bMSCs进行处理。将标记的bMSCs与纤维蛋白胶一起加载到窗口缺损处。在不同时间点处死兔子后，立即取出髌腱的植入部位，并在共聚焦显微镜下评估标记细胞的活力。结果表明，植入的bMSCs在肌腱伤口部位至少在植入后8周内保持存活。植入后2周细胞形态从圆形变为5周时的纺锤形。这项研究表明，bMSCs在植入后能长时间保持存活，因此有可能影响肌腱修复后新肌腱组织的形成和重塑。
