氧化锆基微量元素共掺杂羟基磷灰石增韧涂层研究

doi:10.15541/jim20190053

摘要/Abstract

摘要：

氧化锆陶瓷具有良好的力学性能和生物相容性, 是一种应用前景广阔的硬组织植入体材料。为促进植入体与骨组织形成稳定的骨结合, 本研究利用等离子喷涂制备了氧化锆增韧的锶、硅、氟微量掺杂羟基磷灰石(ZrO₂-DHA)涂层, 对涂层物相、形貌以及力学性能和体外生物学性能进行了研究。结果表明, 锶、硅、氟的共掺杂通过成骨分化的信号转导通路提高了羟基磷灰石涂层对成骨细胞的黏附和分化等生物学性能; 不同复合组分的ZrO₂-DHA涂层均不同程度地促进了小鼠成骨前体细胞的细胞活力和成骨分化相关基因的表达。在细胞培养的第7 d, DHA含量为70%的ZrO₂-DHA涂层(7DHA) Alp和Col-I的相对表达量分别是对照组的约2.8倍和2.3倍; ZrO₂-DHA涂层的力学性能随氧化锆组分的增加而增强, DHA涂层和7DHA涂层的硬度和结合强度分别为250.8、313HV和25.1、31.8 MPa; 7DHA涂层中的交织网络结构, 对残余热应力、压应力和拉伸力的承受能力较DHA涂层明显提升, 满足植入体应用需求。

关键词: 锶、硅、氟微量掺杂羟基磷灰石, 等离子喷涂, 氧化锆增韧, 力学性能, 成骨

Abstract:

Zirconia is a kind of prospective hard tissue implant materials, which possesses superior mechanical properties and excellent biocompatibilities. To promote the stable osseointegration between Zirconia implants and tissue, zirconia-toughened trace Sr-, Si- and F- co-doped hydroxyapatite (ZrO₂-DHA) was coated by plasma spraying on zirconia. The phase composition and structure of coatings were characterized, and the mechanical properties and in vitro biological properties were investigated. The results show that co-doping of trace Sr, Si and F enhances the biological properties of coatings on adhesion and differentiation of osteoblasts through the signal transduction pathway of osteogenic differentiation. All of ZrO₂-DHA coatings promote the cell viability and gene expression in osteogenic differentiation of MC3T3-E1. On the 7th day of cell culture, the relative expression levels of Alp and Col-I in the ZrO₂-DHA coating containing 70% DHA (7DHA) were about 2.8 times and 2.3 times higher than those in the ZrO₂ substrate group, respectively. Mechanical properties of ZrO₂-DHA coatings are improved with the increment of zirconia ratio. Hardness of DHA coating and 7DHA coating are 250.8 and 313.8HV respectively and their bond strength are 25.1 and 31.8 MPa, respectively. 7DHA coatings with network structure possess both excellent mechanical and biological performance for the implant coating application.

Key words: Sr/Si/F trace-doped hydroxyapatite, plasma spraying, zirconia reinforcing, mechanical property, osteogenesis

中图分类号:

TQ174

代钊,王铭,王双,李静,陈翔,汪大林,祝迎春. 氧化锆基微量元素共掺杂羟基磷灰石增韧涂层研究[J]. 无机材料学报, 2020, 35(2): 179-186.

DAI Zhao,WANG Ming,WANG Shuang,LI Jing,CHEN Xiang,WANG Da-Lin,ZHU Ying-Chun. Zirconia Reinforced Trace Element Co-doped Hydroxyapatite Coating[J]. Journal of Inorganic Materials, 2020, 35(2): 179-186.

图/表 10

表1 喷涂参数

Table 1 Plasma spraying parameters

Parameter	HA/DHA coating	ZrO₂-DHA coatings
Primary gas flow rate/(L?min^-1)	Ar/40-41	Ar/40-42
Auxiliary gas flow rate/(L?min^-1)	H₂/6-7	H₂/8-9
Carrier gas flow rate/(L?min^-1)	Ar/6-6.5	Ar/6-6.5
Powder feed rate/(g?min^-1)	18	20
Current/A	650	570
Gun transverse speed/(mm?s^-1)	500	500
Standoff distance/mm	100	100

表2 引物序列

Table 2 Sequence of primers

Gene	Sequence
Alp	F: 5-AACGTGGCCAAGAACATCATCA-3 R: 5-TGTCCATCTCCAGCCGTGTC-3
Col-I	F: 5-CCAGAAGAACTGGTACATCAGCAA-3 R: 5-CGCCATACTCGAACTGGAATC-3
Gapdh	F: 5-GGCATTGCTCTCAATGACAA-3 R: 5-TGTGAGGGAGATGCTCAGTG-3

图1 氧化锆基材(a)和DHA涂层表面(b)的SEM照片

Fig. 1 SEM images of ZrO2 substrate (a) and as-sprayed DHA coating (b)

图2 7DHA涂层的SEM照片(a)和元素分布扫描照片(Ca(b)和Zr(c)); 涂层平面(d~f)和截面(g~i)的SEM照片: DHA(d, g)、7DHA (e, h)、3DHA (f, i)

Fig. 2 SEM images (a) and elemental mapping of Ca (green) (b) and Zr (blue) (c) of 7DHA coating, and SEM images of surface (d-f) and cross sections (g-i) of DHA (d, g), 7DHA (e, h) and 3DHA (f, i)

图3 HA和DHA粉体的XRD图谱(a)及FT-IR谱图 (b), DHA涂层的XRD图谱(c)及7DHA涂层的放大XRD图谱(d)

Fig. 3 XRD patterns of HA and DHA powders with inset showing the enlarged parts (a), FT-IR spectra of HA and DHA powders (b), and XRD patterns of 7DHA, 5DHA and 3DHA coatings (c) with enlarged pattern of 7DHA coating (d)

图4 涂层显微硬度、结合强度(a)以及拉伸断面照片(b)

Fig. 4 Microhardness and bond strength of coatings (a), and the section image of stretching test (b)

图5 细胞活力实验OD值

Fig. 5 OD values of cell viability test (*p<0.05)

图6 细胞黏附实验的荧光照片(a~f)和细胞黏附数量(g)

Fig. 6 Fluorescence images (a-f) and cell numbers of cell adhesion test (g) (*p<0.05)

图7 MC3T3-E1细胞茜素红染色照片(a~f)和ALP活性比较(g)

Fig. 7 Optical images of MC3T3-E1 cell Alizarin red S staining (a-f) and comparison of ALP activity (g) (*p<0.05)

图8 成骨相关基因表达随时间变化图

Fig. 8 Osteogenesis-related genes expression with culture time (a) Alp expression; (b) Col-I expression (*p<0.05)

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