Jalowiec Jagoda M, D'Este Matteo, Bara Jennifer Jane, Denom Jessica, Menzel Ursula, Alini Mauro, Verrier Sophie, Herrmann Marietta
1 AO Research Institute Davos , Davos, Switzerland .
2 Biologie Fonctionnelle et Adaptative, CNRS UMR 8251, Université Paris Diderot Paris 7 , Paris, France .
Tissue Eng Part C Methods. 2016 Jan;22(1):49-58. doi: 10.1089/ten.TEC.2015.0223. Epub 2015 Dec 1.
Platelet-rich plasma (PRP) has been used for different applications in human and veterinary medicine. Many studies have shown promising therapeutic effects of PRP; however, there are still many controversies regarding its composition, properties, and clinical efficacy. The aim of this study was to evaluate the influence of different platelet concentrations on the rheological properties and growth factor (GF) release profile of PRP-gels. In addition, the viability of incorporated bone marrow-derived human mesenchymal stem cells (MSCs) was investigated. PRP (containing 1000 × 10(3), 2000 × 10(3), and 10,000 × 10(3) platelets/μL) was prepared from human platelet concentrates. Platelet activation and gelification were achieved by addition of human thrombin. Viscoelastic properties of PRP-gels were evaluated by rheological studies. The release of GFs and inflammatory proteins was measured using a membrane-based protein array and enzyme-linked immunosorbent assay. MSC viability and proliferation in PRP-gels were assessed over 7 days by cell viability staining. Cell proliferation was examined using DNA quantification. Regardless of the platelet content, all tested PRP-gels showed effective cross-linking. A positive correlation between protein release and the platelet concentration was observed at all time points. Among the detected proteins, the chemokine CCL5 was the most abundant. The greatest release appeared within the first 4 h after gelification. MSCs could be successfully cultured in PRP-gels over 7 days, with the highest cell viability and DNA content found in PRP-gels with 1000 × 10(3) platelets/μL. The results of this study suggest that PRP-gels represent a suitable carrier for both cell and GF delivery for tissue engineering. Notably, a platelet concentration of 1000 × 10(3) platelets/μL appeared to provide the most favorable environment for MSCs. Thus, the platelet concentration is an important consideration for the clinical application of PRP-gels.
富含血小板血浆(PRP)已被用于人类和兽医学的不同领域。许多研究表明PRP具有良好的治疗效果;然而,关于其组成、性质和临床疗效仍存在许多争议。本研究的目的是评估不同血小板浓度对PRP凝胶流变学性质和生长因子(GF)释放曲线的影响。此外,还研究了掺入的骨髓来源的人间充质干细胞(MSCs)的活力。从人血小板浓缩物中制备PRP(含有1000×10³、2000×10³和10000×10³个血小板/μL)。通过添加人凝血酶实现血小板活化和凝胶化。通过流变学研究评估PRP凝胶的粘弹性性质。使用基于膜的蛋白质阵列和酶联免疫吸附测定法测量GF和炎症蛋白的释放。通过细胞活力染色在7天内评估MSCs在PRP凝胶中的活力和增殖。使用DNA定量检查细胞增殖。无论血小板含量如何,所有测试的PRP凝胶均显示出有效的交联。在所有时间点均观察到蛋白质释放与血小板浓度之间呈正相关。在检测到的蛋白质中,趋化因子CCL5含量最高。最大释放量出现在凝胶化后的前4小时内。MSCs可以在PRP凝胶中成功培养7天,在含有1000×10³个血小板/μL的PRP凝胶中发现细胞活力和DNA含量最高。本研究结果表明,PRP凝胶是用于组织工程的细胞和GF递送的合适载体。值得注意的是,1000×10³个血小板/μL的血小板浓度似乎为MSCs提供了最有利的环境。因此,血小板浓度是PRP凝胶临床应用的重要考虑因素。