聚乙二醇化聚癸二酸甘油酯 /β-磷酸三钙涂层改性镁合金膜的体外生物学评价

doi:10.19439/j.sjos.2023.04.002

上海口腔医学 ›› 2023, Vol. 32 ›› Issue (4): 342-350.doi: 10.19439/j.sjos.2023.04.002

聚乙二醇化聚癸二酸甘油酯 /β-磷酸三钙涂层改性镁合金膜的体外生物学评价

张成龙^1,*, 张昌入^2,3,*, 司家文¹, 袁媛², 于洪波¹, 沈洪洲^1#, 沈国芳^1,4#

1.上海交通大学医学院附属第九人民医院口腔颅颌面科,上海交通大学口腔医学院,国家口腔医学中心,国家口腔疾病临床研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011;
2.华东理工大学医用生物材料教育部工程研究中心,上海 200237;
3.上海交通大学转化医学研究院,上海 200240;
4.上海健康医学院,上海 201318

收稿日期:2023-01-09 修回日期:2023-04-20 发布日期:2023-09-07
通讯作者: 沈国芳,E-mail：shengf@sumhs.edu.cn;沈洪洲,E-mail：shenhongzhou0389@163.com。^#共同通信作者
作者简介:张成龙（1997-）,男,在读硕士研究生,E-mail：zcl1123581321@163.com;张昌入（1994-）,男,硕士,E-mail：zcr_ecust@163.com。^*并列第一作者
基金资助:
国家自然科学基金（81970973）; 上海交通大学医学院多中心临床研究（DLY201808）

In vitro biological evaluation of PEGylated poly(glycerol sebacate)/β-TCP-coating modified magnesium alloy

ZHANG Cheng-long¹, ZHANG Chang-ru^2,3, SI Jia-wen¹, YUAN Yuan², YU Hong-bo¹, SHEN Hong-zhou¹, SHEN Guo-fang^1,4

1. Department of Oral and Craniomaxillofacial Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology. Shanghai 200011;
2. Engineering Research Center for Biomaterials of Ministry of Education, East China University of Science and Technology. Shanghai 200237;
3. Institute of Translational Medicine, Shanghai Jiao Tong University. Shanghai 200240;
4. Shanghai University of Medicine & Health Sciences.Shanghai 201318, China;

Received:2023-01-09 Revised:2023-04-20 Published:2023-09-07

摘要/Abstract

摘要： 目的通过在镁锌钆合金（MZG）膜表面形成乙二醇化聚癸二酸甘油酯/β-磷酸三钙（PEGS/β-TCP）涂层,制备PEGS/β-TCP改性镁合金（PEGS/β-TCP/MZG）膜,使用材料浸提液对PEGS/β-TCP/MZG复合膜的成骨诱导活性和免疫调节性能进行评价。方法采用溶胶凝胶法在MZG膜表面制备PEGS/β-TCP涂层,制备PEGS/β-TCP/MZG复合膜,并与PEGS/β-TCP和MZG膜进行对比,检测其形貌、成分和亲水性。检测材料浸提3天后的镁离子释放量和pH值。体外细胞实验观察成骨细胞MC3T3-E1细胞在浸提液中的增殖活性、碱性磷酸酶（alkaline phosphatase,ALP）和矿化结节形成情况,以及RAW264.7巨噬细胞在浸提液中的增殖活性、极化和细胞内相关因子的表达。采用GraphPad Prism 9.0软件包对数据进行统计学分析。结果制备出PEGS/β-TCP涂层与MZG膜紧密嵌合的PEGS/β-TCP/MZG复合膜,具有良好亲水性,可缓慢降解。细胞实验结果显示, PEGS/β-TCP/MZG复合膜浸提液对MC3T3-E1和巨噬细胞的增殖活性无明显影响。PEGS/β-TCP/MZG组显著增强MC3T3-E1的ALP表达和矿化结节形成,PEGS/β-TCP组和PEGS/β-TCP/MZG组在巨噬细胞极化模式上虽无明显差异,但PEGS/β-TCP/MZG膜在PEGS/β-TCP涂层的免疫调节基础上,可进一步降低炎症相关肿瘤坏死因子α（tumor necrosis factor-α,TNF-α）表达,提高成骨相关转化生长因子β（transforming growth factor-β,TGF-β）表达。结论 PEGS/β-TCP复合膜改性的MZG可为骨组织工程的发展提供新的材料选择。

关键词: 镁合金, PEGS/β-TCP, 骨再生, 免疫调节

Abstract: PURPOSE: To prepare PEGS/β-TCP modified magnesium alloy (PEGS/β-TCP/MZG) membranes by forming a glycolated poly(sebacate)/β-tricalcium phosphate (PEGS/β-TCP) coating on the surface of magnesium-zinc-gadolinium alloy (MZG) membranes, and to evaluate the osteogenic induction activity and immunomodulatory properties of PEGS/β-TCP/MZG using the material extract medium. METHODS: PEGS/β-TCP coating was prepared on the surface of MZG by solvent method, and the PEGS/β-TCP/MZG membrane was fabricated and compared with PEGS/β-TCP and MZG to examine the morphology, composition, and hydrophilicity. The amount of magnesium ions released and the pH value of the materials were tested after 3 days of immersion. The cell viability and osteogenic differentiation of MC3T3 cells induced by extract medium were investigated by CCK-8 assay, ALP and mineralized nodule staining. The cell viability and polarization of RAW cells induced by extract medium were then investigated. The expression of macrophage-secreted cytokines was examined by PCR analysis. GraphPad Prism 9.0 software package was used for statistical analysis. RESULTS: PEGS/β-TCP/MZG membranes with PEGS/β-TCP coating tightly embedded with MZG were successfully fabricated, and the material had good hydrophilicity. The results of degradation experiments indicated that the PEGS/β-TCP coating effectively slowed down the degradation rate of MZG, leading to a lower pH value and concentration of Mg²⁺ ion in the extract medium of PEGS/β-TCP/MZG group. The results of in vitro cell experiments showed that PEGS/β-TCP/MZG had no significant effect on the proliferation activity of both MC3T3-E1 and macrophages. PEGS/β-TCP/MZG significantly enhanced the expression of ALP and mineralized nodule staining in MC3T3-E1. Although there was no significant difference in macrophage polarization pattern between PEGS/β-TCP and PEGS/β-TCP/MZG groups, PEGS/β-TCP/MZG further reduced inflammation based on the immunomodulation of PEGS/β-TCP coating related TNF-α expression and increased osteogenesis related TGF-β expression. CONCLUSIONS: MZG membrane modified by PEGS/β-TCP may provide a new material option for the development of bone tissue engineering.

Key words: Magnesium alloy, PEGS/β-TCP, Bone regeneration, Immunoregulatory property

中图分类号:

R783.1

张成龙, 张昌入, 司家文, 袁媛, 于洪波, 沈洪洲, 沈国芳. 聚乙二醇化聚癸二酸甘油酯 /β-磷酸三钙涂层改性镁合金膜的体外生物学评价[J]. 上海口腔医学, 2023, 32(4): 342-350.

ZHANG Cheng-long, ZHANG Chang-ru, SI Jia-wen, YUAN Yuan, YU Hong-bo, SHEN Hong-zhou, SHEN Guo-fang. In vitro biological evaluation of PEGylated poly(glycerol sebacate)/β-TCP-coating modified magnesium alloy[J]. Shanghai Journal of Stomatology, 2023, 32(4): 342-350.

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聚乙二醇化聚癸二酸甘油酯 /β-磷酸三钙涂层改性镁合金膜的体外生物学评价

In vitro biological evaluation of PEGylated poly(glycerol sebacate)/β-TCP-coating modified magnesium alloy

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