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羟基磷灰石纳米晶体处理的钛表面对成骨细胞行为的评估:一项体外研究。

Evaluation of a Titanium Surface Treated with Hydroxyapatite Nanocrystals on Osteoblastic Cell Behavior: An In Vitro Study.

作者信息

Martinez Elizabeth Ferreira, Ishikawa Guilherme Junji, de Lemos Alexandre Barboza, Barbosa Bezerra Fábio José, Sperandio Marcelo, Napimoga Marcelo Henrique

出版信息

Int J Oral Maxillofac Implants. 2018 May/June;33(3):597–602. doi: 10.11607/jomi.5887. Epub 2017 Sep 22.

Abstract

PURPOSE

In the context of macrostructural and microstructural modifications to the design of dental implants, surface topography changes with different treatments have the purpose of accelerating bone formation. The aim of this study was to evaluate in vitro the influence of aggregated hydroxyapatite nanocrystals to surfaces treated with double acid etching (Nano) on osteoblastic cell behavior compared with a conventional double acid-etched surface (DE).

MATERIALS AND METHODS

Commercially pure Grade 4 titanium discs (6 × 2 mm) were selected, and both cell proliferation and viability were assessed at 24, 48, and 72 hours using Trypan blue vital dye and MTT, respectively. The expression of type I collagen and osteopontin on such surfaces was evaluated using ELISA. Immunostaining for fibronectin was also performed. Quantitative data were analyzed statistically using two-way analysis of variance (ANOVA) followed by Bonferroni post-test with a 5% significance level.

RESULTS

The results showed that in all evaluated time periods, cells expressed fibronectin on both surfaces. The cells presented greater morphologic spreading on the Nano surface when compared with the conventional DE surface in all assessed times. Increased cell proliferation and viability were detected in the Nano surface (P < .05) when compared with the conventional DE surface, especially after 72 hours. Osteopontin expression was higher after 24 hours in the Nano surface when compared with the conventional DE surface (P < .05). For type I collagen, a higher expression was observed with the Nano surface than with the DE surface, again after 72 hours (P < .05).

CONCLUSION

This in vitro study showed that the treated Nano surface tested promoted increased cell proliferation and viability when compared with the control surface. Additionally, increased cell spreading as well as type I collagen and osteopontin secretion were observed, favoring the early events of osseointegration.

摘要

目的

在对牙种植体设计进行宏观结构和微观结构修改的背景下,不同处理方式引起的表面形貌变化旨在加速骨形成。本研究的目的是在体外评估与传统双酸蚀刻表面(DE)相比,聚集的羟基磷灰石纳米晶体对经双酸蚀刻处理的表面(纳米表面)上成骨细胞行为的影响。

材料与方法

选用商业纯4级钛盘(6×2毫米),分别使用台盼蓝活细胞染料和MTT在24、48和72小时评估细胞增殖和活力。使用酶联免疫吸附测定法(ELISA)评估此类表面上I型胶原蛋白和骨桥蛋白的表达。还进行了纤连蛋白的免疫染色。定量数据采用双向方差分析(ANOVA)进行统计分析,随后进行Bonferroni事后检验,显著性水平为5%。

结果

结果表明,在所有评估时间段内,细胞在两种表面上均表达纤连蛋白。在所有评估时间点,与传统DE表面相比,细胞在纳米表面呈现出更大的形态铺展。与传统DE表面相比,纳米表面检测到细胞增殖和活力增加(P < 0.05),尤其是在72小时后。与传统DE表面相比,纳米表面在24小时后骨桥蛋白表达更高(P < 0.05)。对于I型胶原蛋白,同样在72小时后,纳米表面的表达高于DE表面(P < 0.05)。

结论

这项体外研究表明,与对照表面相比,所测试的经处理的纳米表面促进了细胞增殖和活力的增加。此外,观察到细胞铺展增加以及I型胶原蛋白和骨桥蛋白分泌增加,有利于骨结合的早期事件。

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