中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (34): 5486-5491.doi: 10.12307/2022.460

• 组织工程骨材料 tissue-engineered bone • 上一篇    下一篇

碱热处理联合紫外线光催化纯钛对成骨细胞活性的影响

郑  锐1,2,孙  勇2   

  1. 1绵阳口腔医院种植科,四川省绵阳市  621000;2西南医科大学口腔医学院,四川省泸州市  646000
  • 收稿日期:2021-01-29 接受日期:2021-03-31 出版日期:2022-12-08 发布日期:2022-04-15
  • 通讯作者: 孙勇,主任医师,西南医科大学口腔医学院,四川省成都市 646000
  • 作者简介:郑锐,女,1991年生,四川省绵阳市人,汉族,2019年西南医科大学毕业,硕士,主治医师,主要从事口腔种植义齿的基础与临床研究。

Effects of alkali-and-heat treatment combined with ultraviolet photofunctionalization pure titanium on osteoblast activity

Zheng Rui1, 2, Sun Yong2   

  1. 1Department of Implantology, Mianyang Stomatological Hospital, Mianyang 621000, Sichuan Province, China; 2School of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2021-01-29 Accepted:2021-03-31 Online:2022-12-08 Published:2022-04-15
  • Contact: Sun Yong, Chief physician, School of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Zheng Rui, Master, Attending physician, Department of Implantology, Mianyang Stomatological Hospital, Mianyang 621000, Sichuan Province, China; School of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China

摘要:

文题释义:
碱热处理:是指纯钛经过强碱溶液碱蚀和热处理后会形成纳米粗糙表面,并且TiO2氧化膜与OH-结合形成钛酸根离子HTiO3-,然后HTiO3-水合作用形成水合钛酸根HTiO3-·nH2O,带负电的HTiO3-·nH2O进一步与NaOH溶液中的Na+反应形成一层多孔网状的钛酸钠凝胶层,能够促进类骨磷灰石的形成,明显增强钛的生物诱导性。
紫外线光催化:TiO2是半导体材料,短波紫外线可以产生大于TiO2禁带宽度的能量,催化TiO2表面形成氧空穴和自由电子,自由电子与TiO2中的Ti4+反应生成Ti3+,而氧空穴则吸附空气中解离的水产生活性羟基,其次光催化还能够降解钛表面的碳氢化合物污染,获得十分清洁的表面。

背景:钛表面经过碱热处理后会形成一层钛酸钠凝胶层促进类骨磷灰石的形成,从而与活骨发生结合;钛表面经过紫外线光催化后会形成亲水性表面,增加表面自由能,促进在体液中吸附蛋白质,进一步促进细胞与钛表面的黏附和反应。
目的:探索碱热处理联合紫外线光催化纯钛表面的的理化性质,以及其对小鼠胚胎成骨细胞前体细胞MC3T3-E1黏附、增殖及分化的影响。
方法:将TA2纯钛片分3组,A组进行酸蚀处理,B组进行酸蚀+碱热处理,C组进行酸蚀+碱热+紫外线光催化处理,表征3组钛片的微观形貌、元素组成及静态接触角。将对数生长期的MC3T3-E1细胞接种至3组钛片表面,采用CCK-8法检测细胞增殖情况,扫描电镜下观察细胞黏附情况;成骨诱导培养后,检测细胞的碱性磷酸酶活性、Ⅰ型胶原分泌与矿化结节形成情况。
结果与结论:①扫描电镜下可见,A组钛片表面粗糙不平呈蜂窝状,分布着较为均匀的孔隙;B组钛片表面疏松分布着纳米级孔隙与“脑回路”状的沟壑,沟壑孔隙相互交联;C组钛片具有纳米结构,孔隙的分布、大小与B组无明显区别;②3组钛片表面均有C、O、Ti、P元素,B、C组钛片表面还检测出Ca元素,A组表面几乎无Ca元素;A组钛片表面静态接触角最小,C组最大;③接种后24 h扫描电镜下可见,各组钛片表面的细胞较多,A、B组细胞呈长梭形、三角形或扁平形,C组细胞直径增大且具有数条伪足,呈扁平的多边形在钛片表面铺展成一薄层,相互连接,无法分清细胞边界;④C组钛片表面的细胞增殖快于A、B组,碱性磷酸酶活性、Ⅰ型胶原分泌与矿化结节数量多于A、B组;⑤结果表明,碱热处理联合紫外线光催化能有效改善纯钛表面的理化性质,促进成骨细胞的生物活性。

https://orcid.org/0000-0002-5292-2088 (郑锐) 

中国组织工程研究杂志出版内容重点:生物材料;骨生物材料口腔生物材料纳米材料缓释材料材料相容性;组织工程

关键词: 种植体, 钛片, 紫外线, 光催化, 碱热处理, 成骨细胞, 表面处理

Abstract: BACKGROUND: After alkali-and-heat treatment on the titanium surface, a layer of sodium titanate gel will be formed to promote the formation of bone-like apatite, thereby combining with living bone. The titanium surface will form a hydrophilic surface after ultraviolet photofunctionalization, which can increase the surface free energy, promote the adsorption of protein in body fluid, and further promote the adhesion and reaction between cells and titanium surface.  
OBJECTIVE: To explore the physical and chemical properties of alkali-and-heat treatment combined with ultraviolet photofunctionalization pure titanium surface, and its effects on adhesion, proliferation and differentiation of MC3T3-E1 cells.  
METHODS: TA2 pure titanium discs were randomly divided into three groups. The group A received acid etching treatment. The group B received acid etching + alkali-and-heat treatment. The group C received acid etching + alkali-and-heat + ultraviolet photofunctionalization treatment. The microscopic morphology, element composition, and static contact angle were characterized in the three groups of titanium discs. MC3T3-E1 cells in logarithmic growth phase were inoculated on the surface of three groups of titanium discs. CCK-8 assay was used to detect cell proliferation. Cell adhesion was observed under scanning electron microscope. After osteogenic induction and culture, the alkaline phosphatase activity, the secretion of type I collagen, and the formation of mineralized nodules were detected.
RESULTS AND CONCLUSION: (1) Under scanning electron microscope, the surface of group A was rough and honeycomb shaped, with relatively uniform pore. The surfaces of group B were distributed with loosely nano-level pores and brain-circuit like gullies. The gullies and pores were cross-linked with each other. The surfaces of group C also had nanoporous network structure, and the distribution and size of gullies and pores were not significantly different from that of group B. (2) C, O, Ti, and P elements were both found on the surfaces of the three groups, and Ca element was also detected on the surfaces of groups B and C, but Ca element was scarcely found on the surface of group A. The static contact angle of group A was the smallest, while that of group C was the largest. (3) Scanning electron microscope showed that there were more cells on the surface of the titanium plates in each group at 24 hours after injection. The cells in groups A and B were long fusiform, triangular or flat. The cells in group C were larger in diameter and had several pseudopods, and the flat polygons were spread into a thin layer on the surface of the titanium plates, interconnected, and unable to distinguish cell boundaries. (4) The cell proliferation on the surface of titanium plates in group C was faster than that in groups A and B, and the alkaline phosphatase activity, type I collagen secretion and the number of mineralized nodules in group C were higher than those in groups A and B. (5) The results showed that alkali-and-heat treatment combined with ultraviolet photofunctionalization could effectively improve the physical and chemical properties of pure titanium surface and promote the biological activity of osteoblasts.

Key words: implant, titanium disc, ultraviolet, photofunctionalization, alkali-and-heat treatment, osteoblast, surface treatment

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