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具有纳米形貌表面结构的阳极氧化-水热处理钛上培养的成纤维细胞样细胞的生物学行为。

Biological behavior of fibroblast-like cells cultured on anodized-hydrothermally treated titanium with a nanotopographic surface structure.

机构信息

Division of Fixed Prosthodontics, Department of Prosthodontics, School of Dentistry, Iwate Medical University, 1-3-27, Chuodori, Morioka, Iwate 020-8505, Japan.

出版信息

J Prosthodont Res. 2012 Jul;56(3):178-86. doi: 10.1016/j.jpor.2011.11.004. Epub 2012 Jan 20.

Abstract

PURPOSE

The interface between the transmucosal portion of endosseous implants surface and the connective tissue is characterized by fibroblast-rich barrier tissue, which is important for the long-term stability and maintenance of the implant. This study investigated the effect of cell adhesion on focal adhesion kinase (FAK) protein and on gene expression over a 72-h culture period. Fibroblast-like cells were cultured on anodized-hydrothermally treated commercially pure titanium with nanotopographic structure (SA-treated c.p.Ti) surfaces.

METHODS

Murine fibroblast-like NIH/3T3 cells were cultured for 10-72h on c.p.Ti, anodic oxide (AO) c.p.Ti, and SA-treated c.p.Ti disks. Cell morphology was analyzed using scanning electron microscopy (SEM). Cytoskeletal structure and FAK protein localization were analyzed using confocal laser scanning microscopy (CLSM). FAK mRNA levels were analyzed using real-time quantitative RT-PCR.

RESULTS

SEM and CLSM showed increased NIH/3T3 cell adhesion with time, and actin filaments oriented parallel with the filopodium-like extensions on all disks. Filopodium-like extensions were bound tightly to the nanotopographic structure surface of cultures on SA-treated c.p.Ti, and especially at 72h. FAK protein was localized along cellular extensions on SA-treated c.p.Ti and the expression of FAK mRNA was significantly higher on these disks than on c.p.Ti and AO c.p.Ti after 72h (P<0.05).

CONCLUSIONS

NIH/3T3 fibroblast-like cells have the capacity to adhere to SA-treated c.p.Ti as a transmucosal portion of implant surface material and express focal adhesion molecules, which may play a key role in the maintenance of a mucosal tissue barrier.

摘要

目的

骨内种植体表面粘膜部分与结缔组织之间的界面的特征是富含成纤维细胞的屏障组织,这对于种植体的长期稳定性和维持非常重要。本研究探讨了细胞黏附对粘着斑激酶(FAK)蛋白的影响,以及在 72 小时培养期间基因表达的影响。成纤维样细胞培养在具有纳米形貌结构的阳极氧化-水热处理商用纯钛(SA 处理的 c.p.Ti)表面。

方法

鼠类成纤维样 NIH/3T3 细胞在 c.p.Ti、阳极氧化 c.p.Ti 和 SA 处理的 c.p.Ti 盘上培养 10-72 小时。使用扫描电子显微镜(SEM)分析细胞形态。使用共聚焦激光扫描显微镜(CLSM)分析细胞骨架结构和 FAK 蛋白定位。使用实时定量 RT-PCR 分析 FAK mRNA 水平。

结果

SEM 和 CLSM 显示 NIH/3T3 细胞随时间的延长而增加黏附,细胞骨架纤维与所有盘上的类丝状伪足平行排列。类丝状伪足紧密结合在 SA 处理的 c.p.Ti 培养物的纳米形貌结构表面上,特别是在 72 小时。FAK 蛋白沿着 SA 处理的 c.p.Ti 上的细胞延伸定位,并且在这些盘上的 FAK mRNA 表达明显高于 c.p.Ti 和 AO c.p.Ti 72 小时后(P<0.05)。

结论

NIH/3T3 成纤维样细胞具有黏附于作为植入物表面材料的粘膜部分的 SA 处理的 c.p.Ti 的能力,并表达粘着斑分子,这可能在维持粘膜组织屏障中起关键作用。

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