纳米形貌钛种植体对骨质疏松大鼠骨结合的影响

doi:10.13701/j.cnki.kqyxyj.2022.06.012

摘要/Abstract

摘要： 目的:探讨钛种植体表面改性后形成的纳米形貌对骨质疏松大鼠骨结合性能的影响。方法:钛片和钛种植体经碱热和水热反应处理后为纳米结构组,未经表面处理为对照组。采用扫描电镜、能量色散光谱仪、接触角测量仪、蛋白吸附试验分析样品表面形貌、元素组成、亲水性和蛋白吸附能力。将骨质疏松大鼠骨髓间充质干细胞接种至样品表面,通过对细胞肌动蛋白和细胞核染色,观察细胞粘附形态;通过CCK-8法检测细胞增殖活性;通过qRT-PCR法检测细胞ALP、BMP-2、COL-1、OPN和OCN的基因表达。此外,采用双侧卵巢切除术建立大鼠骨质疏松模型,将两组钛种植体植入骨质疏松大鼠股骨髁,通过micro-CT分析种植体在体内骨结合情况。结果:表面改性后钛表面形成了纳米尺度形貌,具有较好的亲水性(P<0.01)和蛋白质吸附能力(P<0.05)。纳米形貌的形成显著促进了细胞粘附,细胞在纳米形貌钛片上展现出更好的增殖活性(P<0.05)。纳米形貌也促进了骨髓间充质干细胞ALP、BMP-2、COL-1、OPN和OCN的基因表达(P<0.01)。表面改性的钛种植体在骨质疏松大鼠体内有更好的种植体-骨接触率(P<0.01)。结论:表面改性后钛表面形成了具有优异亲水性能和蛋白吸附性能的纳米形貌,显著促进了骨质疏松大鼠骨髓间充质干细胞的粘附、增殖和成骨分化,改善了种植体在骨质疏松环境下骨结合。

关键词: 骨结合, 骨质疏松, 表面改性, 纳米形貌, 碱热反应

Abstract: Objective: To investigate the effects of nano-topography of titanium implant formed by surface modification on osseointegration in osteoporotic rats. Methods: Titanium slices and titanium implants were treated with alkali-heat and hydrothermal treatment as the nanostructure group, and without surface treatment as the control group. Scanning electronic microscopy, energy dispersive spectroscopy, contact angle measuring instrument, and protein adsorption assay were used to analyze the surface morphology, element composition, hydrophilcity, and protein adsorption capacity of the samples. The bone marrow mesenchymal stem cells of osteoporotic rats were seeded on the sample surfaces. The actin and nucleus were stained to observe cell adhesion. The cell proliferation activity was assessed using cell counting kit-8 assay. The gene expression of ALP, BMP-2, COL-1, OPN, and OCN were examined by qRT-PCR. In addition, rat osteoporosis model was established by bilateral ovariectomy. Two groups of titanium implants were implanted into the femoral condyles of osteoporotic rats. The osseointegration of the implants in vivo was analyzed by micro-CT. Results: After surface modification, nano-scale morphology was formed on the surface of titanium, which had better hydrophilcity (P < 0.01) and protein adsorption ability (P<0.05). The formation of nano-topography effectively improved cell adhesion, and the cells exhibited better proliferation activity on the nano-morphologic titanium slices (P<0.05). The gene expression of ALP, BMP-2, COL-1, OPN, and OCN of bone marrow mesenchymal stem cells were also promoted by the nano-morphology (P<0.01). Surface modified implants had better bone-implant contact in vivo in osteoporotic rats (P<0.01). Conclusion: Alkali-heat and hydrothermal treatment formed the nano-topography with excellent hydrophilicity and protein adsorption ability, which significantly promoted the adhesion, proliferation, and osteogenic differentiation of bone marrow mesenchymal stem cells of osteoporotic rats, and improved the osseointegration of implants under osteoporosis.

Key words: osseointegration, osteoporosis, surface modification, nano-morphology, alkali-heat treatment

张成, 耿藤瑜, 王晶, 袁长永, 王鹏来. 纳米形貌钛种植体对骨质疏松大鼠骨结合的影响[J]. 口腔医学研究, 2022, 38(6): 545-552.

ZHANG Cheng, GENG Tengyu, WANG Jing, YUAN Changyong, WANG Penglai. Effects of Nano-morphologic Titanium Implant on Osteointegration in Osteoporostic Rats[J]. Journal of Oral Science Research, 2022, 38(6): 545-552.

参考文献

[1] Buser D, Sennerby L, De Bruyn H. Modern implant dentistry based on osseointegration: 50 years of progress, current trends and open questions [J]. Periodontol 2000, 2017, 73(1):7-21.
[2] Aghaloo T, Pi-Anfruns J, Moshaverinia A, et al. The effects of systemic diseases and medications on implant osseointegration: A systematic review [J]. Int J Oral Maxillofac Implants, 2019, 34:s35-s49.
[3] Noh JY, Yang Y, Jung H. Molecular mechanisms and emerging therapeutics for osteoporosis [J]. Int J Mol Sci, 2020, 21(20):7623.
[4] Merheb J, Temmerman A, Rasmusson L, et al. Influence of skeletal and local bone density on dental implant stability in patients with osteoporosis [J]. Clin Implant Dent Relat Res, 2016, 18(2):253-260.
[5] Beppu K, Kido H, Watazu A, et al. Peri-implant bone density in senile osteoporosis-changes from implant placement to osseointegration [J]. Clin Implant Dent Relat Res, 2013, 15(2):217-226.
[6] Souza J, Sordi MB, Kanazawa M, et al. Nano-scale modification of titanium implant surfaces to enhance osseointegration [J]. Acta Biomater, 2019, 94:112-131.
[7] Zhao Q, Yi L, Jiang L, et al. Surface functionalization of titanium with zinc/strontium-doped titanium dioxide microporous coating via microarc oxidation [J]. Nanomedicine, 2019, 16:149-161.
[8] Kunrath MF, Diz FM, Magini R, et al. Nanointeraction: The profound influence of nanostructured and nano-drug delivery biomedical implant surfaces on cell behavior [J]. Adv Colloid Interface Sci, 2020, 284:102265.
[9] Li K, Liu S, Hu T, et al. Optimized nanointerface engineering of micro/nanostructured titanium implants to enhance cell-nanotopography interactions and osseointegration [J]. ACS Biomater Sci Eng, 2020, 6(2):969-983.
[10] Cui J, Xia L, Lin K, et al. In situ construction of a nano-structured akermanite coating for promoting bone formation and osseointegration of Ti-6Al-4V implants in a rabbit osteoporosis model [J]. J Mater Chem B, 2021, 9(46):9505-9513.
[11] Wang H, Liu J, Wang C, et al. The synergistic effect of 3D-printed microscale roughness surface and nanoscale feature on enhancing osteogenic differentiation and rapid osseointegration [J]. J Mater Sci Technol, 2021, 63:18-26.
[12] Maimaiti B, Zhang N, Yan L, et al. Stable ZnO-doped hydroxyapatite nanocoating for anti-infection and osteogenic on titanium [J]. Colloids Surf B Biointerfaces, 2020, 186:110731.
[13] Li Y, He S, Hua Y, et al. Effect of osteoporosis on fixation of osseointegrated implants in rats [J]. J Biomed Mater Res B Appl Biomater, 2017, 105(8):2426-2432.
[14] de Medeiros F, Kudo G, Leme BG, et al. Dental implants in patients with osteoporosis: a systematic review with meta-analysis [J]. Int J Oral Maxillofac Surg, 2018, 47(4):480-491.
[15] Zhang C, Zhang T, Geng T, et al. Dental implants loaded with bioactive agents promote osseointegration in osteoporosis: A review [J]. Front Bioeng Biotechnol, 2021, 9:591796.
[16] Zeng Y, Yang Y, Chen L, et al. Optimized surface characteristics and enhanced in vivo osseointegration of alkali-treated titanium with nanonetwork structures [J]. Int J Mol Sci, 2019, 20(5):1127.
[17] Wang H, Zhang X, Wang H, et al. Enhancing the osteogenic differentiation and rapid osseointegration of 3D printed Ti6Al4V implants via nano-topographic modification [J]. J Biomed Nanotechnol, 2018, 14(4):707-715.
[18] Liu J, Pathak JL, Hu X, et al. Sustained release of zoledronic acid from mesoporous TiO₂-layered implant enhances implant osseointegration in osteoporotic condition [J]. J Biomed Nanotechnol, 2018, 14(11):1965-1978.
[19] Lin G, Zhou C, Lin M, et al. Strontium-incorporated titanium implant surface treated by hydrothermal reactions promotes early bone osseointegration in osteoporotic rabbits [J]. Clin Oral Implants Res, 2019, 30(8):777-790.
[20] Barik A, Chakravorty N. Targeted drug delivery from titanium implants: A review of challenges and approaches [J]. Adv Exp Med Biol, 2020, 1251:1-17.
[21] Eastell R, O'Neill TW, Hofbauer LC, et al. Postmenopausal osteoporosis [J]. Nat Rev Dis Primers, 2016, 2:16069.
[22] Huang T, Yu Z, Yu Q, et al. Inhibition of osteogenic and adipogenic potential in bone marrow-derived mesenchymal stem cells under osteoporosis [J]. Biochem Biophys Res Commun, 2020, 525(4):902-908.
[23] Zhang J, Zhou W, Wang H, et al. 3D-printed surface promoting osteogenic differentiation and angiogenetic factor expression of BMSCs on Ti6Al4V implants and early osseointegration in vivo [J]. J Mater Sci Technol, 2019, 35(2):336-343.