激光改性钛表面通过调节成骨细胞中DKK1的表达改善旁分泌骨生成。

Laser-Modified Ti Surface Improves Paracrine Osteogenesis by Modulating the Expression of DKK1 in Osteoblasts.

作者信息

Teixeira Jorge Felipe Lima, de Souza João Antônio Chaves, Magalhães Fernando Augusto Cintra, de Oliveira Guilherme José Pimentel Lopes, de Santis José Bernardo, de Souza Costa Carlos Alberto, de Souza Pedro Paulo Chaves

机构信息

Department of Physiology and Pathology, School of Dentistry, São Paulo State University, Araraquara 14801-385, Brazil.

School of Dentistry, Federal University of Goiás, Goiânia 74605-020, Brazil.

出版信息

J Funct Biomater. 2023 Apr 16;14(4):224. doi: 10.3390/jfb14040224.

DOI:10.3390/jfb14040224

PMID:37103314

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10145280/

Abstract

Titanium surface modifications are widely used to modulate cellular behavior by recognition of topographical cues. However, how those modifications affect the expression of mediators that will influence neighboring cells is still elusive. This study aimed to evaluate the effects of conditioned media from osteoblasts cultured on laser-modified titanium surfaces on the differentiation of bone marrow cells in a paracrine manner and to analyze the expression of Wnt pathway inhibitors. Mice calvarial osteoblasts were seeded on polished (P) and Yb:YAG laser-irradiated (L) Ti surfaces. Osteoblast culture media were collected and filtered on alternate days to stimulate mice BMCs. Resazurin assay was performed every other day for 20 days to check BMC viability and proliferation. After 7 and 14 days of BMCs maintained with osteoblasts P and L-conditioned media, alkaline phosphatase activity, Alizarin Red staining, and RT-qPCR were performed. ELISA of conditioned media was conducted to investigate the expression of Wnt inhibitors Dickkopf-1 (DKK1) and Sclerostin (SOST). BMCs showed increased mineralized nodule formation and alkaline phosphatase activity. The L-conditioned media enhanced the BMC mRNA expression of bone-related markers , , and . L-conditioned media decreased the expression of DKK1 compared with P-conditioned media. The contact of osteoblasts with Yb:YAG laser-modified Ti surfaces induces the regulation of the expression of mediators that affect the osteoblastic differentiation of neighboring cells. DKK1 is among these regulated mediators.

摘要

钛表面改性通过识别地形线索被广泛用于调节细胞行为。然而，这些改性如何影响将影响邻近细胞的介质的表达仍然不清楚。本研究旨在评估在激光改性钛表面培养的成骨细胞条件培养基以旁分泌方式对骨髓细胞分化的影响，并分析Wnt信号通路抑制剂的表达。将小鼠颅骨成骨细胞接种在抛光（P）和Yb:YAG激光照射（L）的钛表面上。每隔一天收集成骨细胞培养基并过滤以刺激小鼠骨髓间充质干细胞（BMCs）。每隔一天进行刃天青试验20天以检查BMCs的活力和增殖。在用成骨细胞P和L条件培养基培养BMCs 7天和14天后，进行碱性磷酸酶活性、茜素红染色和RT-qPCR检测。对条件培养基进行ELISA以研究Wnt抑制剂Dickkopf-1（DKK1）和硬化蛋白（SOST）的表达。BMCs显示矿化结节形成增加和碱性磷酸酶活性增强。L条件培养基增强了BMCs中骨相关标志物、和的mRNA表达。与P条件培养基相比，L条件培养基降低了DKK1的表达。成骨细胞与Yb:YAG激光改性钛表面的接触诱导了影响邻近细胞成骨细胞分化的介质表达的调节。DKK1是这些受调节的介质之一。