Mariscal-Muñoz Eduardo, Costa Carlos A S, Tavares Hewerson S, Bianchi Jonas, Hebling Josimeri, Machado João P B, Lerner Ulf H, Souza Pedro P C
Department of Physiology and Pathology, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil.
Department of Orthodontics and Pediatric Dentistry, Faculty of Dentistry at Araraquara, Univ. Estadual Paulista-UNESP, Araraquara, São Paulo, 14801-903, Brazil.
Clin Oral Investig. 2016 Apr;20(3):503-11. doi: 10.1007/s00784-015-1533-1. Epub 2015 Jul 30.
The aim of this study was to analyze the capacity of a new modified laser surface to stimulate calvarial osteoblasts isolated from neonatal mouse bones to differentiate and form mineralized nodules.
Titanium discs were subjectezd or not to laser irradiation according to specific parameters and characterized. Osteoblasts isolated from neonatal mouse calvaria were cultured over the discs, and the capacity of these cells to proliferate (MTT assay), form mineralized nodules (Alizarin red assay), and enhance alkaline phosphatase activity (ALPase activity) was analyzed. Real-time PCR was used for quantification of gene expression.
Laser-irradiated titanium discs (L) presented a rough nano-to-micrometric oxidized surface contrasting with the smooth pattern on polished discs (P). The Ra on the micrometric level increased from 0.32 ± 0.01 μm on P surfaces to 10.57 ± 0.39 μm on L surfaces. When compared with P, L promoted changes in osteoblast morphology, increased mineralized nodule formation in osteoblasts cultured on the surfaces for 14 days, and enhanced ALPase activity at days 7 and 14. Transcription factors triggering osteoblast differentiation (Runx2 and Sp7) and genes encoding the bone extracellular matrix proteins collagen type-1 (Col1a1), osteopontin (Spp1), and osteocalcin (Bglap) were upregulated in cells on L surfaces compared with those on P surfaces at days 1-14.
Laser treatment of titanium surfaces created a rough surface that stimulated osteoblast differentiation.
Laser treatment of titanium generates a reproducible and efficient surface triggering osteoblast differentiation that can be of importance for osteointegration.
本研究旨在分析一种新型改良激光表面刺激从新生小鼠骨骼分离的颅骨成骨细胞分化并形成矿化结节的能力。
根据特定参数对钛盘进行或不进行激光照射并进行表征。将从新生小鼠颅骨分离的成骨细胞培养在这些钛盘上，分析这些细胞的增殖能力（MTT 法）、形成矿化结节的能力（茜素红法）以及增强碱性磷酸酶活性（碱性磷酸酶活性）。使用实时 PCR 对基因表达进行定量。
激光照射的钛盘（L）呈现出粗糙的纳米至微米级氧化表面，与抛光盘（P）上的光滑表面形成对比。微米级水平上的 Ra 从 P 表面的 0.32±0.01μm 增加到 L 表面的 10.57±0.39μm。与 P 相比，L 促进了成骨细胞形态的变化，增加了在表面培养 14 天的成骨细胞中矿化结节的形成，并在第 7 天和第 14 天增强了碱性磷酸酶活性。与 P 表面上的细胞相比，在第 1 - 14 天，触发成骨细胞分化的转录因子（Runx2 和 Sp7）以及编码骨细胞外基质蛋白 I 型胶原蛋白（Col1a1）、骨桥蛋白（Spp1）和骨钙素（Bglap）的基因在 L 表面上的细胞中上调。
钛表面的激光处理产生了刺激成骨细胞分化的粗糙表面。
钛的激光处理产生了一种可重复且有效的表面，触发成骨细胞分化，这对于骨整合可能具有重要意义。
