Bone Regeneration Laboratory, Dipartimento di Patologie Ortopediche-Traumatologiche Specialistiche, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, I-40136 Bologna, Italy.
Eur Cell Mater. 2010 Jul 1;20:13-23. doi: 10.22203/ecm.v020a02.
Platelet-rich plasma (PRP) is used clinically in liquid or gel form to promote tissue repair. Because of the poor mechanical properties, conventional PRP is often difficult to handle when used in clinical settings and requires secure implantation in a specific site, otherwise when released growth factors could be washed out during an operation. In this study, we analyzed the end product of a recently developed commercially available system (FIBRINET), which is a dense pliable, platelet-rich fibrin matrix (PRFM). We characterized the mechanical properties of PRFM and tested whether PRFM releases growth factors and whether released factors induce the proliferation of mesenchymal stem cells (MSC). Mechanical properties as well as platelet distribution were evaluated in PRFM. PRFM demonstrated robust mechanical properties, with a tear elastic modulus of 937.3 +/- 314.6 kPa, stress at a break of 1476.0 +/- 526.3 kPa, and an elongation at break of 146.3 +/- 33.8 %. PRFM maintained its mechanical properties throughout the testing process. Microscopic observations showed that the platelets were localized on one side of the matrix. Elevated levels of PDGF-AA, PDGF-AB, EGF, VEGF, bFGF and TGF-beta1 were measured in the day 1-conditioned media (CM) of PRFM and growth factor levels decreased thereafter. BMP2 and BMP7 were not detectable. MSC culture media supplemented with 20% PRFM-CM stimulated MSC cell proliferation; at 24 and 48 hours the induction of the proliferation was significantly greater than the induction obtained with media supplemented with 20% foetal bovine serum. The present study shows that the production of a dense, physically robust PRFM made through high-speed centrifugation of intact platelets and fibrin in the absence of exogenous thrombin yields a potential tool for accelerating tissue repair.
富含血小板的血浆(PRP)以液体或凝胶形式用于临床,以促进组织修复。由于机械性能差,传统的 PRP 在临床应用中通常难以处理,需要在特定部位安全植入,否则在手术过程中释放的生长因子可能会被冲洗掉。在这项研究中,我们分析了最近开发的一种商业上可用的系统(FIBRINET)的最终产物,这是一种致密柔韧的富含血小板的纤维蛋白基质(PRFM)。我们对 PRFM 的机械性能进行了表征,并测试了 PRFM 是否释放生长因子以及释放的因子是否诱导间充质干细胞(MSC)的增殖。在 PRFM 中评估了机械性能和血小板分布。PRFM 表现出强大的机械性能,撕裂弹性模量为 937.3 ± 314.6 kPa,断裂时的应力为 1476.0 ± 526.3 kPa,断裂伸长率为 146.3 ± 33.8 %。PRFM 在整个测试过程中保持其机械性能。显微镜观察表明,血小板位于基质的一侧。在 PRFM 的第 1 天条件培养基(CM)中测量到 PDGF-AA、PDGF-AB、EGF、VEGF、bFGF 和 TGF-β1 的水平升高,此后生长因子水平下降。BMP2 和 BMP7 无法检测到。补充有 20% PRFM-CM 的 MSC 培养基刺激 MSC 细胞增殖;在 24 和 48 小时时,细胞增殖的诱导明显大于补充有 20%胎牛血清的培养基的诱导。本研究表明,通过在不存在外源性凝血酶的情况下高速离心完整血小板和纤维蛋白来制备致密、物理性能强的 PRFM,可以成为加速组织修复的潜在工具。