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载有辛伐他汀的聚乳酸-共-羟基乙酸酸涂层钛:牙科修复体基台的生物功能化。

Titanium coated with poly(lactic-co-glycolic) acid incorporating simvastatin: Biofunctionalization of dental prosthetic abutments.

机构信息

Dentistry Post Graduation, Federal University of Santa Catarina, Florianópolis, Brazil.

Biocentro, Florianópolis, Brazil.

出版信息

J Periodontal Res. 2020 Jan;55(1):116-124. doi: 10.1111/jre.12695. Epub 2019 Sep 7.

Abstract

OBJECTIVE

To propose a biofunctionalized prosthetic abutment by analyzing physico-chemical and morphological properties, simvastatin (SIM) release, and biocompatibility of titanium (Ti) disks coated with poly(lactic-co-glycolic) acid (PLGA) incorporating SIM.

METHODS

Titanium disks (8 × 3 mm) were distributed into four groups: Ti: pure Ti; Ti + PLGA: Ti coated with PLGA; Ti + PLGA + SIM6%: Ti + PLGA with 6% SIM; and Ti + PLGA + SIM0.6%: Ti + PLGA incorporating 0.6% SIM. PLGA was prepared through chloroform evaporation technique. After complete dissolution of PLGA, SIM was diluted in the solution. Ti + PLGA, Ti + PLGA + SIM6%, and Ti + PLGA + SIM0.6% were dip coated with PLGA and PLGA + SIM, respectively. Samples were sterilized by ethylene oxide. For SIM release assay, disks were submerged in PBS, pH 7.4, 37°C, 30 rpm up to 600 hours. At different time intervals, SIM was quantified by spectrophotometry (238 nm). For characterization of the biomaterial components, it was performed Fourier-transform infrared spectroscopy, differential scanning calorimetry, scanning electron microscopy (SEM), optical profilometry, and atomic force microscopy. Biocompatibility analyses were performed by MTS colorimetric assay on murine fibroblasts L929, human gingival fibroblasts (HGFs), and stem cells from human exfoliated deciduous teeth (SHEDs). Absorbance was measured at 490 nm, and percentages of viable cells were calculated in relation to positive control (Ti). SEM images were obtained to verify cell adhesion and morphology. One-way ANOVA followed by Tukey's post hoc test was applied (P < 0.05) for statistical analyses.

RESULTS

SIM release was slow and continuous, reaching about 21% of the incorporated SIM after 600 hours. Topographical analyses revealed success in coating Ti disks with PLGA incorporating SIM. Regarding biocompatibility test, Ti + PLGA + SIM0.6% showed the highest percentage of L929 viability at days 3 and 7. There was no significant difference for Ti, Ti + PLGA, and Ti + PLGA + SIM0.6% groups on cell viability of both SHEDs and HGFs at days 3 and 7. SEM corroborates that SHEDs and HGFs were able to adhere and proliferate on Ti, Ti + PLGA, and Ti + PLGA + SIM0.6% surfaces.

CONCLUSION

A slow and controlled release of SIM was achieved, attributed to a diffusional mass transfer mechanism. Moreover, a homogenous coating topography was obtained. Additionally, 0.6% SIM incorporated into PLGA coating improved fibroblasts L929 viability compared to titanium or PLGA. Also, 0.6% SIM incorporated into PLGA promoted cell viability of about 100% for HGFs and approximately 150% for human mesenchymal stem cells. Therefore, this study allows to consider the use of PLGA-coated titanium incorporating SIM as a biofunctionalized abutment for dental implants.

摘要

目的

通过分析物理化学和形态特性、辛伐他汀(SIM)释放和钛(Ti)盘的生物相容性,提出一种经过生物功能化处理的义齿基台。Ti 盘(8×3mm)分为四组:Ti:纯 Ti;Ti+PLGA:涂有 PLGA 的 Ti;Ti+PLGA+SIM6%:Ti+含有 6%SIM 的 PLGA;Ti+PLGA+SIM0.6%:Ti+含有 0.6%SIM 的 PLGA。PLGA 通过氯仿蒸发技术制备。PLGA 完全溶解后,将 SIM 稀释在溶液中。Ti+PLGA、Ti+PLGA+SIM6%和 Ti+PLGA+SIM0.6%分别用 PLGA 和 PLGA+SIM 进行浸涂。样品通过环氧乙烷进行灭菌。为了进行 SIM 释放试验,将圆盘浸入 PBS(pH7.4,37°C,30rpm)中,直至 600 小时。在不同的时间间隔内,通过分光光度法(238nm)定量 SIM。为了对生物材料成分进行表征,进行了傅里叶变换红外光谱、差示扫描量热法、扫描电子显微镜(SEM)、光学轮廓仪和原子力显微镜分析。通过 MTS 比色法分析对小鼠成纤维细胞 L929、人牙龈成纤维细胞(HGFs)和人脱落乳牙牙髓干细胞(SHEDs)进行了生物相容性分析。在 490nm 处测量吸光度,并计算与阳性对照(Ti)相关的存活细胞百分比。获得 SEM 图像以验证细胞黏附和形态。采用单因素方差分析(ANOVA)和 Tukey 事后检验(post hoc test)(P<0.05)进行统计学分析。

结果

SIM 释放缓慢且持续,600 小时后释放了约 21%的掺入 SIM。形貌分析表明成功地用含有 SIM 的 PLGA 涂覆了 Ti 盘。关于细胞相容性试验,Ti+PLGA+SIM0.6%在第 3 天和第 7 天的 L929 活力最高。在第 3 天和第 7 天,Ti、Ti+PLGA 和 Ti+PLGA+SIM0.6%组的 SHEDs 和 HGFs 的细胞活力均无显著差异。SEM 证实了 SHEDs 和 HGFs 能够在 Ti、Ti+PLGA 和 Ti+PLGA+SIM0.6%表面黏附和增殖。

结论

通过扩散质量传递机制实现了 SIM 的缓慢和控制释放。此外,还获得了均匀的涂层形貌。此外,与钛或 PLGA 相比,PLGA 涂层中掺入 0.6%SIM 提高了成纤维细胞 L929 的活力。此外,PLGA 中掺入 0.6%SIM 促进了 HGFs 约 100%和人间充质干细胞约 150%的细胞活力。因此,这项研究可以考虑将含有 SIM 的 PLGA 涂覆的钛用作牙科植入物的生物功能化基台。

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