Morizaki Yutaka, Zhao Chunfeng, An Kai-Nan, Amadio Peter C
Department of Orthopedic Surgery, Mayo Clinic Rochester, 200 First Street SW, Rochester, MN55905, USA.
J Hand Surg Am. 2010 Nov;35(11):1833-41. doi: 10.1016/j.jhsa.2010.07.034. Epub 2010 Oct 15.
In this study, we investigated the effect of platelet-rich plasma (PRP) and bone marrow-derived stromal cell (BMSC)-seeded interposition in an in vitro canine tendon repair model.
Bone marrow, peripheral blood, and tendons were harvested from mixed-breed dogs. Bone marrow-derived stromal cells were cultured and passaged from adherent cells of bone marrow suspension. Platelet-rich plasma was purified from peripheral blood using a commercial kit. A total of 192 flexor digitorum profundus tendons were used for the study. Tendons repaired with a simple suture were used as a control group. In treatment groups, a collagen gel patch was interposed at the tendon repair site before suture. There were 3 treatment groups, according to the type of collagen patch: a patch with PRP, a patch with BMSC, and a patch with PRP and BMSC. The repaired tendons were evaluated by biomechanical testing and by histological survey after 2 and 4 weeks in tissue culture. To evaluate viability, cells were labeled with PKH26 red fluorescent cell linker (Sigma, St. Louis, MO) and surveyed under confocal microscopy after culture.
The maximum breaking strength and stiffness of the healing tendons with the BMSC-seeded PRP patch were significantly higher than those of the healing tendons without a patch or with a cell-seeded patch (p < .02). Viable BMSCs were present at both 2 and 4 weeks.
Platelet-rich plasma enhanced the effect of BMSC-seeded collagen gel interposition in this in vitro model. Based on these results, we now plan to investigate this effect in vivo.
在本研究中,我们在体外犬类肌腱修复模型中研究了富血小板血浆(PRP)和骨髓源性基质细胞(BMSC)植入的效果。
从杂种犬采集骨髓、外周血和肌腱。从骨髓悬液的贴壁细胞中培养并传代骨髓源性基质细胞。使用商业试剂盒从外周血中纯化富血小板血浆。总共192条指深屈肌腱用于该研究。用单纯缝合修复的肌腱用作对照组。在治疗组中,在缝合前于肌腱修复部位植入胶原凝胶贴片。根据胶原贴片类型有3个治疗组:含PRP的贴片、含BMSC的贴片以及含PRP和BMSC的贴片。在组织培养2周和4周后,通过生物力学测试和组织学检查评估修复的肌腱。为评估细胞活力,用PKH26红色荧光细胞连接剂(Sigma,圣路易斯,密苏里州)标记细胞,并在培养后在共聚焦显微镜下观察。
植入BMSC的PRP贴片修复的肌腱的最大断裂强度和刚度显著高于未贴片或植入细胞贴片的修复肌腱(p <.02)。在2周和4周时均存在有活力的BMSC。
在该体外模型中,富血小板血浆增强了植入BMSC的胶原凝胶的效果。基于这些结果,我们现在计划在体内研究这种效果。