Lumetti S, Ferrillo S, Mazzotta S, Macaluso G M, Bonanini M, Passeri G, Galli C
Department of Biomedical, Biotechnological, and Translational Sciences, University of Parma, Italy.
Department of Clinical and Experimental Medicine, University of Parma, Italy.
J Biol Regul Homeost Agents. 2014 Jul-Sep;28(3):489-95.
Rough titanium surfaces enhance the activation of Wnt canonical signaling, a pathway required for osteoblast differentiation. The present study investigated the effects of GSK3b-inhibitor (2'Z,3'E)- 6-Bromoindirubin-3'-oxime (BIO) on osteoblastic differentiation on titanium surfaces with different topography and wettability. C2C12 cells were plated on pickled, acid-etched/sand-blasted (SLA), modified hydrophilic SLA titanium discs (modSLA) and stimulated with increasing doses of BIO. Activation of Wnt canonical signaling was measured with a reporter system. Gene expression was measured in the same cell system by Real Time PCR. Osteoblastic MC3T3 cells were then plated on discs with or without BIO and the expression of osteoblast specific genes was assessed by Real Time PCR. One mM BIO activated Wnt canonical signaling in C2C12 cells on all surfaces, and the highest effect was on rough surfaces. BIO markedly increased the expression of Osteoprotegerin and Osteocalcin in MC3T3 cells on rough surfaces at the concentration of 100 nM, and on all surfaces at the concentration of 1 mM. BIO enhances Wnt signaling activation and the expression of osteoblastic genes on rough surfaces and could be a viable approach to improve cell response to implant surfaces.
粗糙的钛表面可增强Wnt经典信号通路的激活，而该通路是成骨细胞分化所必需的。本研究调查了GSK3β抑制剂（2'Z,3'E）-6-溴靛玉红-3'-肟（BIO）对具有不同形貌和润湿性的钛表面上成骨细胞分化的影响。将C2C12细胞接种在酸洗、酸蚀/喷砂（SLA）、改性亲水性SLA钛盘（modSLA）上，并用递增剂量的BIO进行刺激。使用报告系统测量Wnt经典信号通路的激活情况。通过实时PCR在同一细胞系统中测量基因表达。然后将成骨MC3T3细胞接种在添加或未添加BIO的圆盘上，并通过实时PCR评估成骨细胞特异性基因的表达。1 mM的BIO可激活所有表面上C2C12细胞中的Wnt经典信号通路，且在粗糙表面上的效果最为显著。在浓度为100 nM时，BIO显著增加了粗糙表面上MC3T3细胞中骨保护素和骨钙素的表达，在浓度为1 mM时，在所有表面上均有此效果。BIO可增强粗糙表面上Wnt信号通路的激活和成骨细胞基因的表达，可能是改善细胞对植入物表面反应的一种可行方法。
