氧化钛涂层润湿性对免疫成骨性能的影响规律

doi:10.15541/jim20230242

无机材料学报 ›› 2023, Vol. 38 ›› Issue (12): 1457-1565.DOI: 10.15541/jim20230242

所属专题：【生物材料】骨骼与齿类组织修复(202312)

氧化钛涂层润湿性对免疫成骨性能的影响规律

上官丽¹^,²(), 聂晓双²^,³, 叶奎材²^,³, 崔苑苑¹(), 乔玉琴²^,⁴()

1.上海大学材料科学与工程学院, 上海 200444
2.中国科学院上海硅酸盐研究所, 高性能陶瓷与超微结构国家重点实验室, 上海 200050
3.中国科学院大学材料科学与光电工程研究中心, 北京 100049
4.上海大学生命科学学院, 200444

收稿日期:2023-05-19 修回日期:2023-06-16 出版日期:2023-06-28 网络出版日期:2023-06-28
通讯作者: 乔玉琴, 副研究员. E-mail: qiaoyq@mail.sic.ac.cn;
崔苑苑, 副教授, E-mail: cui-yy@shu.edu.cn
作者简介:上官丽(1997-), 女, 硕士研究生. E-mail: 20724695@shu.edu.cn
基金资助:
国家自然科学基金(32171344);上海自然科学基金面上项目(20ZR1465100);上海市生物医药科技支撑专项(20S31903300)

Effects of Surface Wettability of Titanium Oxide Coatings on Osteoimmunomodulatory Properties

SHANGGUAN Li¹^,²(), NIE Xiaoshuang²^,³, YE Kuicai²^,³, CUI Yuanyuan¹(), QIAO Yuqin²^,⁴()

1. School of Materials Science and Engineering, Shanghai University, Shanghai 200444, China
2. State Key Laboratory of High Performance Ceramics and Superfine Microstructure, Shanghai Institute of Ceramics, Chinese Academy of Sciences, Shanghai 200050, China
3. Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing 100049, China
4. School of Life Sciences, Shanghai University, Shanghai 200444, China

Received:2023-05-19 Revised:2023-06-16 Published:2023-06-28 Online:2023-06-28
Contact: QIAO Yuqin, associate professor. E-mail: qiaoyq@mail.sic.ac.cn;
CUI Yuanyuan, associate professor. E-mail: cui-yy@shu.edu.cn
About author:SHANGGUAN Li (1997-), female, Master candidate. E-mail: 20724695@shu.edu.cn
Supported by:
National Natural Science Foundation of China(32171344);Shanghai Natural Science Foundation(20ZR1465100);Shanghai Science and Technology Pillar Program for Biomedicine(20S31903300)

摘要/Abstract

摘要：

钛及其合金以其优异的机械性能和生物相容性而被广泛应用作硬组织植入器械, 但其表面缺乏生物活性以及植入后的炎症反应易导致骨整合不佳。本研究利用不同气氛中的热处理工艺调控氧化钛涂层的润湿性, 并研究表面润湿性能对免疫反应和成骨性能的影响规律。研究结果表明, 与亲水(接触角~10º)的氧化钛涂层相比, 处于亲水/疏水临界状态的氧化钛涂层(接触角’90º), 在仅培养巨噬细胞时, 抑制了巨噬细胞向M1促炎方向极化; 在共培养小鼠骨髓间充质干细胞和巨噬细胞时, 促进了巨噬细胞向M2抗炎方向极化, 同时显著上调了骨髓间充质干细胞成骨相关标记物的基因表达, 显示出更好的免疫促成骨性能。

关键词: 表面润湿性, 氧化钛涂层, 退火, 免疫成骨

Abstract:

Titanium and its alloys have been widely used as hard tissue implants due to their excellent mechanical properties and biocompatibilities. However, the lack of biological activity on its surface and the inflammatory reaction after implantation can easily lead to unsatisfactory osseointegration. In this work, the wettability of titanium oxide coatings was modulated by annealing in different atmospheres, and the effects of surface wettability on polarization of macrophages and osteogenic differentiation of mBMSCs were studied. The results showed that, compared to the hydrophilic titanium oxide coating (~10º, PEO-A), the titanium oxide coating with contact angle about 90º (PEO-A-V) inhibited the polarization of macrophages towards M1 pro-inflammatory direction under the mono-culture condition. However, under the co-culture condition, the titanium oxide coating with contact angle about 90º promoted macrophage polarization towards M2 and significantly upregulated gene expressions of osteogenic markers related to mBMSCs, indicating better immunomodulatory effects on osteogenic differentiation of mBMSCs.

Key words: surface wettability, titanium oxide coating, annealing, osteoimmunomodulatory effect

中图分类号:

R318

上官丽, 聂晓双, 叶奎材, 崔苑苑, 乔玉琴. 氧化钛涂层润湿性对免疫成骨性能的影响规律[J]. 无机材料学报, 2023, 38(12): 1457-1565.

SHANGGUAN Li, NIE Xiaoshuang, YE Kuicai, CUI Yuanyuan, QIAO Yuqin. Effects of Surface Wettability of Titanium Oxide Coatings on Osteoimmunomodulatory Properties[J]. Journal of Inorganic Materials, 2023, 38(12): 1457-1565.

图/表 10

表1 RT-qPCR所用引物序列

Table 1 Primers for real-time quantitative polymerase chain reaction (RT-qPCR)

Gene	Primer sequence (F: forward; R: Reverse; 5ʹ-3ʹ)
Arg1	F: GCC AGG GAC TGA CTA CCT TAA R: AGT TCT GTC TGC TTT GCT GTG
IL4	F: TCA TCC TGC TCT TCT TTC TCG R: CTT CTC CTG TGA CCT CGT TCA
CD206	F: AGG GAA GAG AAG AAG ATC CAG R: TGG GAG AAG ATG AAG TCA AAC
TNF-α	F: TAG CCA GGA GGG AGA ACA GA R: CCA GTG AGT GAA AGG GAC AGA
IL6	F: ACC AAG ACC ATC CAA TTC ATC R: CTG ACC ACA GTG AGG AAT GTC
CD86	F: TCT CCA ACA GCC TCT CTC TTT R: ATC TTC ATT GAC TCC GTT TCC
OCN	F: ACC GCC TAC AAA CGC ATC TA R: AGA GGA CAG GGA GGA TCA AGT
OPN	F: CTT GAG CAT TCC AAA GAG AGC R: CTT GTG GCT GTG AAA CTT GTG
BMP2	F: TAA GTT CTG TCC CCA GTG ACG R: TTC GGT GCT GGA AAC TAC TGT
TGF-β1	F: AAC CAA GGA GAC GGA ATA CA R: CGT GGA GTT TGT TAT CTT TGC
Runx2	F: GCA GCA CGC TAT TAA ATC CAA R: GCC AAA CAG ACT CAT CCA TTC

图1 样品的(a)表面形貌、(b)元素组成、(c)物相组成、(d)电化学极化曲线和(e)接触角

Fig. 1 (a) Surface morphologies, (b) element compositions, (c) phase compositions, (d) electrochemical polarization curves, and (e) water contact angles of the samples Coloful figures are available on website

图2 在不同样品表面培养1和4 d的Raw264.7的增殖结果

Fig. 2 Proliferation of Raw264.7 incubated on different samples for 1 and 4 d Data are presented as the mean±SD, n = 3. ****: p < 0.0001 Coloful figures are available on website

图3 Raw264.7在不同样品表面培养4 h、1 d和4 d的细胞形态

Fig. 3 Morphologies of Raw264.7 cultured on different samples for 4 h, 1 d and 4 d

图4 在不同样品表面培养4 d的Raw264.7的炎症相关基因的表达水平

Fig. 4 Relative mRNA levels of inflammatory genes of Raw264.7 incubated on different samples for 4 d (a) Arg-1; (b) IL-4; (c) CD206; (d) TNF-α; (e) IL-6; (f) CD86 Data are presented as the mean±SD, n = 3. ****: p <0.0001; ***: p <0.001; **: 0.05< p <0.01 Coloful figures areavailable on website

图5 在不同样品表面培养不同时间的mBMSCs的细胞增殖结果

Fig. 5 Cell proliferation of mBMSCs incubated on different samples for different time Data are presented as the mean±SD, n = 3. ****: p <0.0001; **: 0.05< p <0.01; *: p<0.05 Coloful figures areavailable on website

图6 在不同样品表面培养4、12和24 h的mBMSCs细胞骨架染色

Fig. 6 Cytoskeleton staining of mBMSCs incubated on different samples for 4, 12 and 24 h

图7 在不同样品表面培养10 d的mBMSCs的成骨相关基因(a) OPN, (b) Runx2, (c) OCN和(d) BMP2的表达水平

Fig. 7 Relative mRNA levels of osteogenic genes of mBMSCs cultured on different samples for 10 d (a) OPN; (b) Runx2; (c) OCN; (d) BMP2 Data are presented as the mean±SD, n = 3. ****: p <0.0001; ***: p <0.001; **: 0.05< p <0.01; *: p <0.05

图8 在不同样品表面上与mBMSCs共培养3 d的Raw264.7炎症相关基因的表达水平

Fig. 8 Relative mRNA levels of inflammatory genes of Raw264.7 co-cultured with mBMSCs on different samples for 3 d (a) Arg-1; (b) IL-4; (c) CD206; (d) TNF-α; (e) IL-6; (f) CD86 Data are presented as mean±SD, n = 3. ****: p <0.0001; ***: p <0.001; **: 0.05< p <0.01; *: p <0.05

图9 在不同样品上和Raw264.7共培养3 d的mBMSCs的成骨相关基因表达水平

Fig. 9 Relative mRNA levels of osteogenic genes of mBMSCs co-cultured with Raw264.7 on different samples for 3 d (a) OPN; (b) TGF-β1; (c) OCN; (d) BMP2 Data are presented as the mean±SD, n = 3. ****: p <0.0001; ***: p <0.001; **: 0.05< p <0.01; *: p <0.05

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氧化钛涂层润湿性对免疫成骨性能的影响规律

Effects of Surface Wettability of Titanium Oxide Coatings on Osteoimmunomodulatory Properties

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