Dental Research Center, Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, 3010 Bern, Switzerland.
Department of Periodontology, School of Dental Medicine, University of Bern, 3010 Bern, Switzerland.
Int J Mol Sci. 2021 Apr 14;22(8):4051. doi: 10.3390/ijms22084051.
The aim of the present study was to investigate the influence of a novel volume-stable collagen matrix (vCM) on early wound healing events including cellular migration and adhesion, protein adsorption and release, and the dynamics of the hemostatic system. For this purpose, we utilized transwell migration and crystal violet adhesion assays, ELISAs for quantification of adsorbed and released from the matrix growth factors, and qRT-PCR for quantification of gene expression in cells grown on the matrix. Our results demonstrated that primary human oral fibroblasts, periodontal ligament, and endothelial cells exhibited increased migration toward vCM compared to control cells that migrated in the absence of the matrix. Cellular adhesive properties on vCM were significantly increased compared to controls. Growth factors TGF-β1, PDGF-BB, FGF-2, and GDF-5 were adsorbed on vCM with great efficiency and continuously delivered in the medium after an initial burst release within hours. We observed statistically significant upregulation of genes encoding the antifibrinolytic thrombomodulin, plasminogen activator inhibitor type 1, thrombospondin 1, and thromboplastin, as well as strong downregulation of genes encoding the profibrinolytic tissue plasminogen activator, urokinase-type plasminogen activator, its receptor, and the matrix metalloproteinase 14 in cells grown on vCM. As a general trend, the stimulatory effect of the vCM on the expression of antifibrinolytic genes was synergistically enhanced by TGF-β1, PDGF-BB, or FGF-2, whereas the strong inhibitory effect of the vCM on the expression of profibrinolytic genes was reversed by PDGF-BB, FGF-2, or GDF-5. Taken together, our data strongly support the effect of the novel vCM on fibrin clot stabilization and coagulation/fibrinolysis equilibrium, thus facilitating progression to the next stages of the soft tissue healing process.
本研究旨在探讨新型体积稳定胶原基质（vCM）对包括细胞迁移和黏附、蛋白吸附和释放以及止血系统动力学在内的早期伤口愈合事件的影响。为此，我们利用 Transwell 迁移和结晶紫黏附试验、ELISA 定量分析吸附和释放基质中的生长因子，以及 qRT-PCR 定量分析细胞在基质上生长的基因表达。结果表明，与缺乏基质的对照组相比，原代人口腔成纤维细胞、牙周韧带细胞和内皮细胞向 vCM 的迁移明显增加。与对照组相比，细胞在 vCM 上的黏附特性显著增加。TGF-β1、PDGF-BB、FGF-2 和 GDF-5 等生长因子能高效地吸附在 vCM 上，并在最初的几小时内爆发性释放后，持续地在培养基中释放。我们观察到，编码抗纤维蛋白溶解的血栓调节素、纤溶酶原激活物抑制剂 1、血小板反应蛋白 1 和凝血酶原的基因显著上调，而编码纤维蛋白溶解的组织型纤溶酶原激活物、尿激酶型纤溶酶原激活物及其受体和基质金属蛋白酶 14 的基因则显著下调。一般来说，TGF-β1、PDGF-BB 或 FGF-2 协同增强了 vCM 对抗纤维蛋白溶解基因表达的刺激作用，而 PDGF-BB、FGF-2 或 GDF-5 则逆转了 vCM 对纤维蛋白溶解基因表达的强烈抑制作用。总之，我们的数据强烈支持新型 vCM 对纤维蛋白凝块稳定和凝血/纤溶平衡的影响，从而促进软组织愈合过程的下一阶段的进展。
