TiO2纳米管表面聚合物辅助沉积Ta2O5涂层对MC3T3-E1细胞生物学活性的影响

doi:10.11958/20230648

天津医药 ›› 2023, Vol. 51 ›› Issue (8): 803-808.doi: 10.11958/20230648

TiO₂纳米管表面聚合物辅助沉积Ta₂O₅涂层对MC3T3-E1细胞生物学活性的影响

王非凡¹(), 王庆福¹, 张硕¹, 李广忠²^,^△()

1 天津市口腔医院种植科，南开大学医学院（邮编300041）
2 西北有色金属研究院多孔金属材料国家重点实验室

收稿日期:2023-04-28 修回日期:2023-06-02 出版日期:2023-08-15 发布日期:2023-08-10
通讯作者: ^△E-mail：shlgznin@126.com
作者简介:王非凡（1991），女，医师，主要从事种植体表面改性方面研究。E-mail：1257097144@qq.com
基金资助:
陕西省重点研发计划项目(2023—YBGY—187);天津市卫生健康科技项目(KJ20108)

Effects of polymer assisted deposition of Ta₂O₅ coating on the surface of TiO₂ nanotubes on improving the biological activity of MC3T3-E1 cells

WANG Feifan¹(), WANG Qingfu¹, ZHANG Shuo¹, LI Guangzhong²^,^△()

1 Department of Oral Implantology, Tianjin Stomatological Hospital, School of Medicine, Nankai University, Tianjin 300041, China
2 State Key Laboratory of Porous Metal Materials, Northwest Institute for Nonferrous Metal Research

Received:2023-04-28 Revised:2023-06-02 Published:2023-08-15 Online:2023-08-10
Contact: ^△E-mail: shlgznin@126.com

王非凡, 王庆福, 张硕, 李广忠. TiO₂纳米管表面聚合物辅助沉积Ta₂O₅涂层对MC3T3-E1细胞生物学活性的影响[J]. 天津医药, 2023, 51(8): 803-808.

WANG Feifan, WANG Qingfu, ZHANG Shuo, LI Guangzhong. Effects of polymer assisted deposition of Ta₂O₅ coating on the surface of TiO₂ nanotubes on improving the biological activity of MC3T3-E1 cells[J]. Tianjin Medical Journal, 2023, 51(8): 803-808.

摘要/Abstract

摘要：

目的探讨TiO₂纳米管表面聚合物辅助沉积五氧化二钽（Ta₂O₅）涂层对小鼠成骨前体细胞MC3T3-E1生物学活性的影响。方法制备出Ta/TiO₂纳米管（Ta/NT组）、Ta₂O₅/纯钛（Ta₂O₅/PT组）及Ta₂O₅/TiO₂纳米管（Ta₂O₅/NT组）3组样本，其中后2组的Ta₂O₅涂层通过聚合物辅助沉积法制备。对3组样品进行表征检测：扫描电镜（SEM）观察表面形貌，X射线光电子能谱（XPS）分析表面元素组成，X射线衍射（XRD）检测表面化合物。选用MC3T3-E1细胞探讨3组样品对细胞生物学活性影响；荧光显微镜观察细胞骨架和细胞核的黏附；CCK-8法测定细胞活性；检测细胞碱性磷酸酶（ALP）活性；茜素红染色后半定量分析钙沉积；Real-time PCR检测成骨标志基因重组人骨形态发生蛋白-2（BMP-2）、ALP、骨钙蛋白（OCN）和骨桥蛋白（OPN）的基因表达。结果 SEM图像显示Ta₂O₅/NT组表面可见TiO₂纳米管基底表面覆盖片状、带状交联的微-纳米级Ta₂O₅涂层结构；XRD显示Ta₂O₅/NT组的TiO₂纳米管呈现微量的脱矿钛及金红石晶体结构；与Ta/NT组比，Ta₂O₅/NT组和Ta₂O₅/PT组黏附细胞核密度较高，细胞骨架块状、丝状结构相对较大，交联、团簇状较多，细胞骨架张力较大，具有较高的细胞活力；茜素红染色样本半定量分析显示Ta₂O₅/NT组、Ta/NT组、Ta₂O₅/PT组钙沉积量依次降低（P<0.01）；Ta₂O₅/NT组、Ta/NT组和Ta₂O₅/PT组7 d和14 d 的ALP活性、钙沉积量和ALP、OCN、OPN mRNA含量依次降低，14 d Ta₂O₅/NT组的BMP-2 mRNA含量在3组中最高（P<0.05）。结论 Ta₂O₅/TiO₂纳米管复合涂层可促进MC3T3-E1细胞黏附、细胞活性、ALP活性、矿化及成骨相关基因的表达，有利于其生物学活性的发挥。

关键词: 钛, 纳米管, Ta, Ta₂O₅涂层, 聚合物辅助沉积, 表面改性, 生物活性

Abstract:

Objective To explore the effect of polymer assisted deposition of tantalum pentoxide (Ta₂O₅) coating on the surface of TiO₂ nanotubes on the biological activity of mouse bone forming precursor cells, MC3T3-E1 cells. Methods Three sets of samples were prepared, including Ta/TiO₂nanotubes (the Ta/NT group), Ta₂O₅/pure titanium (the Ta₂O₅/PT group) and Ta₂O₅/ TiO₂ nanotubes (the Ta₂O₅/NT group). The Ta₂O₅/PT group and the Ta₂O₅/NT group were prepared by polymer-assisted deposition. The three sets of samples were characterized and tested by scanning electron microscopy (SEM) to observe the surface morphology. X-ray photoelectron spectroscopy (XPS) was used to analyze surface element composition, and X-ray diffraction (XRD) was used to detect surface compounds. MC3T3-E1 cells were selected to explore the effect of three groups of samples on cell biological activity. Fluorescence microscope was used to observe the adhesion of cytoskeleton and nucleus. Cell activity was determined by CCK-8 method. Cell alkaline phosphatase (ALP) activity was detected. Calcium deposition was detected by alizarin red staining and semi-quantitative analysis. The expression levels of recombinant human bone morphogenetic protein-2 (BMP-2), ALP, osteocalcin (OCN) and osteopontin (OPN) were detected by real-time PCR. Results SEM images showed that on the surface of the Ta₂O₅/NT group, the TiO₂ nanotube substrate was covered with a sheet-like, banded cross-linked micro-nano Ta₂O₅ coating structure. XRD showed that the TiO₂ nanotubes in the Ta₂O₅/NT group presented a trace of demineralized titanium and rutile crystal structure. Compared with the Ta/NT group, the Ta₂O₅/NT group and the Ta₂O₅/PT group showed higher cell density, larger cytoskeletal mass and filamentous structure, more crosslinking and clusters, higher cytoskeletal tension and higher cell activity. The semi-quantitative analysis of alizarin red staining samples showed that calcium deposition decreased successively in the Ta₂O₅/NT group, the Ta/NT group and the Ta₂O₅/PT group (P<0.01). The ALP activity, calcium deposition and ALP, OCN and OPN mRNA contents on day 7 and 14 were decreased successively in the Ta₂O₅/NT group, the Ta/NT group and the Ta₂O₅/PT group, and the BMP-2 mRNA content on day 14 in the Ta₂O₅/NT group was the highest in the three groups (P<0.05). Conclusion Ta₂O₅/TiO₂ nanotube composite coating can promote the adhesion of MC3T3-E1 cells, the expression of mineralization and osteogenic genes, and enhance the activities of cell and ALP, which is conducive to the play of biological activities.

Key words: Titanium, nanotubes, Tantalum, Tantalum pentoxide coating, polymer-assisted deposition, surface characterization, biological activity

中图分类号:

R783.1

图/表 7

参考文献 21

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组别	1 d	4 d	7 d
Ta/NT组	0.267±0.015	0.403±0.021	0.633±0.015
Ta₂O₅/PT组	0.307±0.015^a	0.493±0.015^a	0.873±0.021^a
Ta₂O₅/NT组	0.330±0.020^a	0.487±0.015^a	0.860±0.017^a
F	10.654^＊	235.148^＊＊	169.379^＊＊

组别	1 d	4 d	7 d
Ta/NT组	0.267±0.015	0.403±0.021	0.633±0.015
Ta₂O₅/PT组	0.307±0.015^a	0.493±0.015^a	0.873±0.021^a
Ta₂O₅/NT组	0.330±0.020^a	0.487±0.015^a	0.860±0.017^a
F	10.654^＊	235.148^＊＊	169.379^＊＊

组别	7 d	14 d
Ta/NT组	2.993±0.051	2.420±0.072
Ta₂O₅/PT组	2.577±0.132^a	2.133±0.112^a
Ta₂O₅/NT组	3.197±0.083^ab	2.677±0.112^ab
F	33.298^＊＊	21.968^＊＊

组别	7 d	14 d
Ta/NT组	2.993±0.051	2.420±0.072
Ta₂O₅/PT组	2.577±0.132^a	2.133±0.112^a
Ta₂O₅/NT组	3.197±0.083^ab	2.677±0.112^ab
F	33.298^＊＊	21.968^＊＊

组别	ALP		OCN		OPN		BMP-2
组别	7 d	14 d	7 d	14 d	7 d	14 d	7 d	14 d
Ta/NT组	2.326±0.114	3.015±0.210	2.696±0.116	8.307±0.396	1.978±0.082	0.699±0.020	2.089±0.241	5.570±0.692
Ta₂O₅/PT组	1.690±0.108^a	2.590±0.154^a	0.840±0.070^a	1.013±0.121^a	1.003±0.106^a	0.553±0.078^a	0.913±0.060^a	4.353±0.235^a
Ta₂O₅/NT组	2.563±0.091^ab	3.727±0.078^ab	3.063±0.138^ab	10.283±0.286^ab	2.433±0.120^ab	0.810±0.044^ab	1.670±0.111^ab	6.727±0.250^ab
F	55.895^＊＊	40.233^＊＊	324.817^＊＊	847.407^＊＊	148.039^＊＊	17.871^＊＊	43.111^＊＊	21.236^＊＊

TiO₂纳米管表面聚合物辅助沉积Ta₂O₅涂层对MC3T3-E1细胞生物学活性的影响

Effects of polymer assisted deposition of Ta₂O₅ coating on the surface of TiO₂ nanotubes on improving the biological activity of MC3T3-E1 cells

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