Department of Orthopaedic Surgery, University of Connecticut, Farmington, Connecticut 06034, USA.
Arthroscopy. 2011 Nov;27(11):1459-71. doi: 10.1016/j.arthro.2011.06.029. Epub 2011 Oct 5.
The purpose of this study was to determine whether a one-time physiologic dose of insulin when compared with the growth factors insulin-like growth factor 1, β-fibroblastic growth factor, and growth differentiation factor 5 is capable of differentiating bone marrow-derived mesenchymal stem cells (MSCs) into tendon.
Eleven patients undergoing arthroscopic rotator cuff repair consented to undergo aspiration of bone marrow. A dose-response curve was calculated to determine the optimal dose of insulin needed to differentiate MSCs into tendon. After purification of bone marrow in the operating room, MSCs were exposed to either insulin or tendon-inducing growth factors or were left untreated to serve as a control. The potential for MSCs in each of these groups to differentiate into tendon was evaluated with a multistep process that included determination of the genetic upregulation for tendon-specific proteins, confirmation that the levels of these proteins were actually increased, staining of the MSCs with antibodies for these proteins to ensure that they were expressed on the cell surface, and finally, evaluation of cell morphology to verify the MSCs' tendon-like appearance.
MSCs treated with insulin showed increased gene expression of tendon-specific markers (P < .05), increased content of tendon-specific proteins (P < .05), and increased receptors on the cell surface (P < .05) compared with control cells. Histologic analysis showed a tendon-like appearance compared with the control cells.
Bone marrow-derived MSCs treated with a single physiologic dose of insulin differentiated into cells with characteristics consistent with tendon.
The potential for MSCs to differentiate into tendon after a 1-time dose of insulin may assist in developing practical biologic options for augmentation of rotator cuff repairs.
本研究旨在确定一次性生理剂量的胰岛素与生长因子胰岛素样生长因子 1、β-成纤维细胞生长因子和生长分化因子 5 相比，是否能够将骨髓间充质干细胞（MSCs）分化为肌腱。
11 例行关节镜下肩袖修复术的患者同意进行骨髓抽吸。计算剂量反应曲线以确定将 MSCs 分化为肌腱所需的最佳胰岛素剂量。在手术室对骨髓进行纯化后，将 MSCs 暴露于胰岛素或肌腱诱导生长因子中，或不进行处理作为对照。通过包括确定肌腱特异性蛋白的基因上调、确认这些蛋白的水平实际上增加、用这些蛋白的抗体对 MSCs 进行染色以确保它们在细胞表面表达以及最后评估细胞形态以验证 MSCs 的肌腱样外观在内的多步过程来评估这些组中 MSC 分化为肌腱的潜力。
用胰岛素处理的 MSC 表现出肌腱特异性标记物的基因表达增加（P <.05），肌腱特异性蛋白含量增加（P <.05），细胞表面受体增加（P <.05）与对照细胞相比。组织学分析显示与对照细胞相比具有肌腱样外观。
用单次生理剂量的胰岛素处理的骨髓来源的 MSCs 分化为具有与肌腱一致特征的细胞。
MSC 在接受一次性胰岛素剂量后分化为肌腱的潜力可能有助于开发用于增强肩袖修复的实用生物选择。
