Marconi Guya Diletta, Fonticoli Luigia, Della Rocca Ylenia, Rajan Thangavelu Soundara, Piattelli Adriano, Trubiani Oriana, Pizzicannella Jacopo, Diomede Francesca
Department of Medical, Oral and Biotechnological Sciences, University "G. d'Annunzio" Chieti-Pescara, 66100 Chieti, Italy.
Department of Innovative Technologies in Medicine & Dentistry, University "G. d'Annunzio" Chieti-Pescara, 66100 Chieti, Italy.
Biology (Basel). 2021 Sep 18;10(9):931. doi: 10.3390/biology10090931.
The major challenge for dentistry is to provide the patient an oral rehabilitation to maintain healthy bone conditions in order to reduce the time for loading protocols. Advancement in implant surface design is necessary to favour and promote the osseointegration process. The surface features of titanium dental implant can promote a relevant influence on the morphology and differentiation ability of mesenchymal stem cells, induction of the osteoblastic genes expression and the release of extracellular matrix (ECM) components. The present study aimed at evaluating the in vitro effects of two different dental implants with titanium surfaces, TEST and CTRL, to culture the human periodontal ligament stem cells (hPDLSCs). Expression of ECM components such as Vimentin, Fibronectin, N-cadherin, Laminin, Focal Adhesion Kinase (FAK) and Integrin beta-1 (ITGB1), and the osteogenic related markers, as runt related transcription factor 2 (RUNX2) and alkaline phosphatase (ALP), were investigated. Human PDLSCs cultured on the TEST implant surface demonstrated a better cell adhesion capability as observed by Scanning Electron Microscopy (SEM) and immunofluorescence analysis. Moreover, immunofluorescence and Western blot experiments showed an over expression of Fibronectin, Laminin, N-cadherin and RUNX2 in hPDLSCs seeded on TEST implant surface. The gene expression study by RT-PCR validated the results obtained in protein assays and exhibited the expression of RUNX2, ALP, Vimentin (VIM), Fibronectin (FN1), N-cadherin (CDH2), Laminin (LAMB1), FAK and ITGB1 in hPDLSCs seeded on TEST surface compared to the CTRL dental implant surface. Understanding the mechanisms of ECM components release and its regulation are essential for developing novel strategies in tissue engineering and regenerative medicine. Our results demonstrated that the impact of treated surfaces of titanium dental implants might increase and accelerate the ECM apposition and provide the starting point to initiate the osseointegration process.
牙科面临的主要挑战是为患者提供口腔修复,以维持健康的骨条件,从而缩短加载方案的时间。种植体表面设计的进步对于促进和推动骨整合过程是必要的。钛牙种植体的表面特征可对间充质干细胞的形态和分化能力、成骨细胞基因表达的诱导以及细胞外基质(ECM)成分的释放产生相关影响。本研究旨在评估两种具有钛表面的不同牙种植体TEST和CTRL对培养人牙周膜干细胞(hPDLSCs)的体外作用。研究了ECM成分如波形蛋白、纤连蛋白、N-钙黏着蛋白、层粘连蛋白、黏着斑激酶(FAK)和整合素β-1(ITGB1)的表达,以及成骨相关标志物,如与 runt 相关转录因子 2(RUNX2)和碱性磷酸酶(ALP)的表达。通过扫描电子显微镜(SEM)和免疫荧光分析观察到,在TEST种植体表面培养的人PDLSCs表现出更好的细胞黏附能力。此外,免疫荧光和蛋白质印迹实验表明,接种在TEST种植体表面的hPDLSCs中纤连蛋白、层粘连蛋白、N-钙黏着蛋白和RUNX2表达上调。通过RT-PCR进行的基因表达研究验证了在蛋白质测定中获得的结果,并显示与CTRL牙种植体表面相比,接种在TEST表面的hPDLSCs中RUNX2、ALP、波形蛋白(VIM)、纤连蛋白(FN1)、N-钙黏着蛋白(CDH2)、层粘连蛋白(LAMB1)、FAK和ITGB1的表达。了解ECM成分释放的机制及其调节对于开发组织工程和再生医学的新策略至关重要。我们的结果表明,钛牙种植体处理表面的影响可能会增加并加速ECM的沉积,并为启动骨整合过程提供起点。