FORM, Frankfurt Oral Regenerative Medicine, Clinic for Maxillofacial and Plastic Surgery, Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt, Germany.
Osteopore International, Singapore 618305, Singapore.
Int J Mol Sci. 2021 Feb 22;22(4):2159. doi: 10.3390/ijms22042159.
Resorbable synthetic scaffolds are promising for different indications, especially in the context of bone regeneration. However, they require additional biological components to enhance their osteogenic potential. In addition to different cell types, autologous blood-derived matrices offer many advantages to enhance the regenerative capacity of biomaterials. The present study aimed to analyze whether biologization of a PCL-mesh coated using differently centrifuged Platelet rich fibrin (PRF) matrices will have a positive influence on primary human osteoblasts activity in vitro. A polymeric resorbable scaffold (Osteomesh, Osteopore (OP), Singapore) was combined with differently centrifuged PRF matrices to evaluate the additional influence of this biologization concept on bone regeneration in vitro. Peripheral blood of three healthy donors was used to gain PRF matrices centrifuged either at High (710× , 8 min) or Low (44× , 8 min) relative centrifugal force (RCF) according to the low speed centrifugation concept (LSCC). OP-PRF constructs were cultured with pOBs. POBs cultured on the uncoated OP served as a control. After three and seven days of cultivation, cell culture supernatants were collected to analyze the pOBs activity by determining the concentrations of VEGF, TGF-β1, PDGF, OPG, IL-8, and ALP- activity. Immunofluorescence staining was used to evaluate the Osteopontin expression of pOBs. After three days, the group of OP+PRF+pOBs showed significantly higher expression of IL-8, TGF-ß1, PDGF, and VEGF compared to the group of OP+PRF+pOBs and OP+pOBs. Similar results were observed on day 7. Moreover, OP+PRF+pOBs exhibited significantly higher activity of ALP compared to OP+PRF+pOBs and OP+pOBs. Immunofluorescence staining showed a higher number of pOBs adherent to OP+PRF+pOBs compared to the groups OP+PRF+pOBs and OP+pOBs. To the best of our knowledge, this study is the first to investigate the osteoblasts activity when cultured on a PRF-coated PCL-mesh in vitro. The presented results suggest that PRF centrifuged according to LSCC exhibits autologous blood cells and growth factors, seem to have a significant effect on osteogenesis. Thereby, the combination of OP with PRF showed promising results to support bone regeneration. Further in vivo studies are required to verify the results and carry out potential results for clinical translation.
可吸收合成支架在不同适应症中具有广阔的应用前景，特别是在骨再生方面。然而，它们需要额外的生物成分来增强其成骨潜力。除了不同的细胞类型外，自体血液衍生基质还具有许多优势，可以增强生物材料的再生能力。本研究旨在分析使用不同离心的富含血小板纤维蛋白（PRF）基质涂覆的 PCL 网是否会对原代人成骨细胞的体外活性产生积极影响。将一种聚合可吸收支架（Osteomesh、Osteopore（OP）、新加坡）与不同离心的 PRF 基质结合，以评估这种生物化概念对体外骨再生的额外影响。使用来自 3 名健康供体的外周血获得 PRF 基质，根据低速离心概念（LSCC）在高（710×，8 分钟）或低（44×，8 分钟）相对离心力（RCF）下离心。OP-PRF 构建体与 pOB 共培养。在未涂覆 OP 的 pOB 上培养的 pOB 作为对照。培养 3 天和 7 天后，收集细胞培养上清液以通过测定 VEGF、TGF-β1、PDGF、OPG、IL-8 和 ALP-活性来分析 pOB 的活性。免疫荧光染色用于评估 pOB 的骨桥蛋白表达。培养 3 天后，与 OP+PRF+pOBs 和 OP+pOBs 相比，OP+PRF+pOBs 组的 IL-8、TGF-ß1、PDGF 和 VEGF 表达显著升高。第 7 天观察到类似的结果。此外，OP+PRF+pOBs 的碱性磷酸酶活性明显高于 OP+PRF+pOBs 和 OP+pOBs。免疫荧光染色显示，与 OP+PRF+pOBs 和 OP+pOBs 相比，OP+PRF+pOBs 上附着的 pOBs 数量更多。据我们所知，这项研究是第一个研究在体外培养在 PRF 涂覆的 PCL 网上的成骨细胞活性的研究。所呈现的结果表明，根据 LSCC 离心的 PRF 显示出自体血细胞和生长因子，似乎对成骨有显著影响。因此，OP 与 PRF 的结合显示出支持骨再生的有希望的结果。需要进一步的体内研究来验证这些结果，并为临床转化进行潜在的研究。
