Abstract

The purpose of this study is to explore a storage solution for titanium implants and investigate its osteogenic properties. The commercial pure titanium (cp-Ti) surface and double-etched (SLA) titanium surface specimens were preserved in air, saline, 10 mM Vitamin C (VitC)-containing saline and 100 mM VitC-containing saline storage solutions for 2 weeks. The surface microtopography of titanium was observed by scanning electron microscopy (SEM), the surface elemental compositions of the specimens were analyzed by Raman and X-ray photoelectron spectroscopy (XPS), and water contact angle and surface roughness of the specimens were tested. The protein adsorption capacity of two titanium surfaces after storage in different media was examined by BCA kit. The MC3T3-E1 osteoblasts were cultured on two titanium surfaces after storage in different media, and the proliferation, adhesion and osteogenic differentiation activity of osteoblasts were detected by CCK-8, laser confocal microscope (CLSM) and Western blot. The SEM results indicated that the titanium surfaces of the air group were relatively clean while scattered sodium chloride or VitC crystals were seen on the titanium surfaces of the other three groups. There were no significant differences in the micromorphology of the titanium surfaces among the four groups. Raman spectroscopy detected VitC crystals on the titanium surfaces of two experimental groups. The XPS, water contact angle and surface roughness results suggested that cp-Ti and SLA-Ti stored in 0.9% NaCl and two VitC-containing saline storage solutions possessed less carbon contamination and higher surface hydrophilicity. Moreover, the protein adsorption potentials of cp-Ti and SLA-Ti surfaces were significantly improved under preservation in two VitC-containing saline storage solutions. The results of in vitro study showed that the preservation of two titanium surfaces in 100 mM VitC-containing saline storage solution upregulated the cell adhesion, proliferation, osteogenic related protein expressions of MC3T3-E1 osteoblasts. In conclusion, preservation of cp-Ti and SLA-Ti in 100 mM VitC-containing saline storage solution could effectively reduce carbon contamination and enhance surface hydrophilicity, which was conducive to osteogenic differentiation of osteoblasts.

Graphical Abstract

中文翻译：

---

含维生素C盐储存液中保存的钛的表面特性和体外生物相容性

摘要

本研究的目的是探索钛种植体的存储解决方案并研究其成骨特性。将商业纯钛 (cp-Ti) 表面和双蚀刻 (SLA) 钛表面标本在空气、盐水、含 10 mM 维生素 C (VitC) 的盐水和含 100 mM VitC 的盐水储存溶液中保存 2 周。采用扫描电子显微镜（SEM）观察钛表面微观形貌，采用拉曼和X射线光电子能谱（XPS）分析试件的表面元素组成，并测试试件的水接触角和表面粗糙度。采用BCA试剂盒检测两种钛表面在不同介质中保存后的蛋白质吸附能力。将MC3T3-E1成骨细胞在不同培养基中保存后在两个钛表面培养，通过CCK-8、激光共聚焦显微镜（CLSM）和Western blot检测成骨细胞的增殖、粘附和成骨分化活性。SEM结果表明，空气组的钛表面相对干净，而其他三组的钛表面可见分散的氯化钠或VitC晶体。四组之间钛表面的微观形貌没有显着差异。拉曼光谱在两个实验组的钛表面上检测到了 VitC 晶体。XPS、水接触角和表面粗糙度结果表明，保存在 0.9% NaCl 和两种含 VitC 盐溶液中的 cp-Ti 和 SLA-Ti 具有较少的碳污染和较高的表面亲水性。此外，在两种含 VitC 的盐储存溶液中保存时，cp-Ti 和 SLA-Ti 表面的蛋白质吸附能力显着提高。体外研究结果表明，在含100 mM VitC的盐储存液中保存两个钛表面可上调MC3T3-E1成骨细胞的细胞粘附、增殖、成骨相关蛋白表达。总之，将cp-Ti和SLA-Ti保存在含100 mM VitC的盐储存液中可以有效减少碳污染并增强表面亲水性，有利于成骨细胞的成骨分化。

图形概要