Zheng Zheng, He Yanjin, Long Li, Gan Shuaiqi, Chen Shujiang, Zhang Min, Xu Jia, Fu Ruijie, Liao Yihan, Zhu Zhimin, Wang Hang, Chen Wenchuan
State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
Department of Oral Prosthodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.
Front Bioeng Biotechnol. 2022 Sep 16;10:975840. doi: 10.3389/fbioe.2022.975840. eCollection 2022.
Non-thermal atmospheric plasma (NTAP) modification to induce a hydrophilic titanium (Ti) surface with less carbon contamination, has been demonstrated to boost the osteogenic responses. In this study, we investigated the underlying bone formation mechanism of NTAP-Ti, and the involvement of PI3K/Akt signaling pathway in regulating osteogenic activities on NTAP-Ti surfaces. NTAP was employed for Ti activation, and PI3K inhibitor, LY294002, was applied to the suppression of PI3K/Akt pathway. We systematically and quantitatively detected the cell morphology, attachment, proliferation, osteogenic differentiation and mineralization of MC3T3-E1 mouse preosteoblasts, and molecular expressions involved in osteogenesis and PI3K/Akt signaling pathway and . A descent in osteoblast proliferation on Ti surfaces in relation to LY294002. Alkaline phosphatase (ALP) activity, as well as matrix mineralization, was mitigated by PI3K inhibitor in NTAP-Ti. Likewise, the expression levels of osteogenesis-related genes [ALP, osteocalcin (Ocn), osteopontin (Opn) and runt-related transcription factor 2 (Runx2)] on NTAP-Ti were notably attenuated by LY294002, as confirmed by the results of osteogenesis-related proteins (ALP, and Runx2) expression analysis. In addition, the expression of PI3K/Akt signal pathway proteins further verified the inhibition of LY294002 on Ti surfaces modified by NTAP. Collectively, the PI3K/Akt signal pathway was involved in the amelioration of osteogenesis induced by NTAP modification. NTAP treatment for Ti activation is promising in augmented osteogenic potential through the activation of PI3K/Akt signal pathway.
非热大气等离子体（NTAP）修饰可诱导产生碳污染较少的亲水性钛（Ti）表面，已被证明能促进成骨反应。在本研究中，我们探究了NTAP-Ti促进骨形成的潜在机制，以及PI3K/Akt信号通路在调节NTAP-Ti表面成骨活性中的作用。采用NTAP对Ti进行活化，并使用PI3K抑制剂LY294002抑制PI3K/Akt通路。我们系统且定量地检测了MC3T3-E1小鼠前成骨细胞的细胞形态、黏附、增殖、成骨分化和矿化情况，以及与成骨和PI3K/Akt信号通路相关的分子表达。LY294002使Ti表面的成骨细胞增殖减少。PI3K抑制剂使NTAP-Ti中的碱性磷酸酶（ALP）活性以及基质矿化减轻。同样，LY294002显著减弱了NTAP-Ti上成骨相关基因[ALP、骨钙素（Ocn）、骨桥蛋白（Opn）和 runt相关转录因子2（Runx2）]的表达水平，成骨相关蛋白（ALP和Runx2）表达分析结果证实了这一点。此外，PI3K/Akt信号通路蛋白的表达进一步验证了LY294002对NTAP修饰的Ti表面的抑制作用。总体而言，PI3K/Akt信号通路参与了NTAP修饰诱导的成骨改善过程。通过激活PI3K/Akt信号通路，NTAP处理Ti活化在增强成骨潜能方面具有前景。
