Kreuz Peter Cornelius, Krüger Jan Philipp, Metzlaff Sebastian, Freymann Undine, Endres Michaela, Pruss Axel, Petersen Wolf, Kaps Christian
Department of Orthopaedic Surgery, University Medical Center Rostock, Rostock, Germany.
TransTissue Technologies, Berlin, Germany.
Arthroscopy. 2015 Oct;31(10):1951-61. doi: 10.1016/j.arthro.2015.03.033. Epub 2015 May 13.
To evaluate the chondrogenic potential of platelet concentrates on human subchondral mesenchymal progenitor cells (MPCs) as assessed by histomorphometric analysis of proteoglycans and type II collagen. Furthermore, the migratory and proliferative effect of platelet concentrates were assessed.
Platelet-rich plasma (PRP) was prepared using preparation kits (Autologous Conditioned Plasma [ACP] Kit [Arthrex, Naples, FL]; Regen ACR-C Kit [Regen Lab, Le Mont-Sur-Lausanne, Switzerland]; and Dr.PRP Kit [Rmedica, Seoul, Republic of Korea]) by apheresis (PRP-A) and by centrifugation (PRP-C). In contrast to clinical application, freeze-and-thaw cycles were subsequently performed to activate platelets and to prevent medium coagulation by residual fibrinogen in vitro. MPCs were harvested from the cortico-spongious bone of femoral heads. Chondrogenic differentiation of MPCs was induced in high-density pellet cultures and evaluated by histochemical staining of typical cartilage matrix components. Migration of MPCs was assessed using a chemotaxis assay, and proliferation activity was measured by DNA content.
MPCs cultured in the presence of 5% ACP, Regen, or Dr.PRP formed fibrous tissue, whereas MPCs stimulated with 5% PRP-A or PRP-C developed compact and dense cartilaginous tissue rich in type II collagen and proteoglycans. All platelet concentrates significantly (ACP, P = .00041; Regen, P = .00029; Dr.PRP, P = .00051; PRP-A, P < .0001; and PRP-C, P < .0001) stimulated migration of MPCs. All platelet concentrates but one (Dr.PRP, P = .63) showed a proliferative effect on MPCs, as shown by significant increases (ACP, P = .027; Regen, P = .0029; PRP-A, P = .00021; and PRP-C, P = .00069) in DNA content.
Platelet concentrates obtained by different preparation methods exhibit different potentials to stimulate chondrogenic differentiation, migration, and proliferation of MPCs. Platelet concentrates obtained by commercially available preparation kits failed to induce chondrogenic differentiation of MPCs, whereas highly standardized PRP preparations did induce such differentiation. These findings suggest differing outcomes with PRP treatment in stem cell-based cartilage repair.
Our findings may help to explain the variability of results in studies examining the use of PRP clinically.
通过对蛋白聚糖和II型胶原蛋白的组织形态计量学分析，评估血小板浓缩物对人软骨下间充质祖细胞（MPCs）的软骨形成潜能。此外，还评估了血小板浓缩物的迁移和增殖作用。
使用制备试剂盒（自体条件血浆[ACP]试剂盒[Arthrex，那不勒斯，佛罗里达州]；Regen ACR - C试剂盒[Regen Lab，瑞士洛桑湖畔勒蒙]；以及Dr.PRP试剂盒[Rmedica，韩国首尔]）通过单采术（PRP - A）和离心法（PRP - C）制备富血小板血浆（PRP）。与临床应用不同，随后进行冻融循环以激活血小板并防止体外培养基因残留纤维蛋白原而凝固。从股骨头的皮质 - 松质骨中采集MPCs。在高密度微团培养中诱导MPCs的软骨分化，并通过典型软骨基质成分的组织化学染色进行评估。使用趋化性分析评估MPCs的迁移，并通过DNA含量测量增殖活性。
在5% ACP、Regen或Dr.PRP存在下培养的MPCs形成纤维组织，而用5% PRP - A或PRP - C刺激的MPCs形成富含II型胶原蛋白和蛋白聚糖的致密软骨组织。所有血小板浓缩物均显著（ACP，P = 0.00041；Regen，P = 0.00029；Dr.PRP，P = 0.00051；PRP - A，P < 0.0001；PRP - C，P < 0.0001）刺激MPCs的迁移。除一种血小板浓缩物（Dr.PRP，P = 0.63）外，所有血小板浓缩物均对MPCs有增殖作用，DNA含量显著增加（ACP，P = 0.027；Regen，P = 0.0029；PRP - A，P = 0.00021；PRP - C，P = 0.00069）表明了这一点。
通过不同制备方法获得的血小板浓缩物在刺激MPCs的软骨分化、迁移和增殖方面表现出不同的潜能。通过市售制备试剂盒获得的血小板浓缩物未能诱导MPCs的软骨分化，而高度标准化的PRP制剂确实诱导了这种分化。这些发现表明在基于干细胞的软骨修复中PRP治疗的结果存在差异。
我们的发现可能有助于解释临床研究中检查PRP使用结果的变异性。
