中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (21): 3291-3297.doi: 10.12307/2022.634

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

基于三周期极小曲面β-磷酸三钙仿生骨支架设计和生物活性的检测

王金斯1，王胜法2，吴柱国1，何晓玲1，王馨钰2，罗小钰1，招  轶1，张静莹1   

  1. 1广东医科大学，广东医科大学附属东莞第一医院，广东省东莞市  523808；2大连理工大学国际信息与软件学院，辽宁省大连市  116620
  • 收稿日期:2021-06-28 接受日期:2021-08-12 出版日期:2022-07-28 发布日期:2022-01-27
  • 通讯作者: 张静莹，博士，副教授，广东医科大学，广东医科大学附属东莞第一医院，广东省东莞市 523808
  • 作者简介:王金斯，男，1997年生，黑龙江省佳木斯市人，汉族，广东医科大学在读硕士，主要从事骨组织工程方面的研究。
  • 基金资助:
    广东省基础与应用基础研究基金联合基金(2020B1515120001)，项目负责人：张静莹；广东省普通高校重点领域资助项目(2020ZDZX2013)，项目负责人：张静莹；广东医科大学学科建设项目(4SG21019G)，项目负责人：张静莹；广东医科大学学科建设项目(4SG21015G)，项目负责人：吴柱国

Design and biological activity of beta-tricalcium phosphate biomimetic bone scaffold based on triply periodic minimal surfaces

Wang Jinsi1, Wang Shengfa2, Wu Zhuguo1, He Xiaoling1, Wang Xinyu2, Luo Xiaoyu1, Zhao Yi1, Zhang Jingying1   

  1. 1Dongguan First Hospital Affiliated to Guangdong Medical University, Dongguan 523808, Guangdong Province, China; 2DUT-RU International School of Information and Software Engineering, Dalian University of Technology, Dalian 116620, Liaoning Province, China
  • Received:2021-06-28 Accepted:2021-08-12 Online:2022-07-28 Published:2022-01-27
  • Contact: Zhang Jingying, MD, Associate professor, Dongguan First Hospital Affiliated to Guangdong Medical University, Dongguan 523808, Guangdong Province, China
  • About author:Wang Jinsi, Master candidate, Dongguan First Hospital Affiliated to Guangdong Medical University, Dongguan 523808, Guangdong Province, China
  • Supported by:
    the Basic and Applied Basic Research of Guangdong Province, No. 2020B1515120001 (to ZJY); Key Fields Foundation of Colleges and Universities in Guangdong Province, No. 2020ZDZX2013 (to ZJY); Discipline Construction Project of Guangdong Medical University, No. 4SG21019G (to ZJY); Discipline Construction Project of Guangdong Medical University, No. 4SG21015G (to WZG)

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摘要：

文题释义：
β-磷酸三钙仿生骨支架：指采用基于数字激光加工的增材制造技术制备出的、具有三周期极小曲面仿生结构的β-磷酸三钙生物陶瓷骨支架，具有可定制、精度高的优点。
三周期极小曲面：指在欧氏空间沿X轴、Y轴、Z轴呈周期性变化的，平均曲率为零的曲面，因其结构与松质骨相仿而具有作为骨组织工程支架的应用潜力。

背景：增材制造技术可以精确、个性化地定制具有复杂多孔结构的骨支架，达到恢复临界骨缺损区域松质骨结构和功能的效果。
目的：通过材料学和细胞学表征，明确三周期极小曲面结构β-磷酸三钙生物陶瓷骨支架的机械性能及生物活性，揭示三周期极小曲面结构对成骨细胞的调控效果。
方法：通过Matlab R2020a软件设计330，420，510 μm 3种孔径的三周期极小曲面G曲面支架，Inspire 2018软件分析支架结构设计。以三周期极小曲面结构导出后的STL文件为蓝本，通过基于数字激光加工的增材制造技术制备3种β-磷酸三钙生物陶瓷支架，采用扫描电镜观测支架表面形貌，X射线衍射仪检测物相组成，万能材料试验机检测支架的力学强度。将MC3T3-E1细胞与3种支架共培养，检测细胞的增殖活性、黏附能力与碱性磷酸酶活性。
结果与结论：①Inspire 2018软件显示，三周期极小曲面呈现出光滑、连续、均一的贯通式多孔结构；②扫描电镜下可见，基于数字激光加工的增材制造技术成功实现了三周期极小曲面结构的精确成型；③X射线衍射结果证实，支架由纯β-磷酸三钙晶相组成；④3组支架的压缩强度和弹性模量均处于或接近松质骨力学强度范围，且支架的压缩强度与孔径大小呈反比；⑤CCK8实验显示，MC3T3-E1细胞在3种支架上生长良好，生物活性与孔径呈现剂量依赖关系，510 μm孔径促进细胞增殖效果最佳；⑥活细胞成像仪和激光共聚焦显微镜下可见，MC3T3-E1细胞能够在3种支架上实现早期黏附，且黏附量随孔径增大而增加；⑦碱性磷酸酶活性分析表明，420 μm孔径支架上细胞的碱性磷酸酶活性最高；⑧结果表明，三周期极小曲面结构β-磷酸三钙生物陶瓷支架表现出较好的机械性能与生物活性，其中420 μm孔径有利于促进细胞分化，510 μm孔径有利于细胞增殖，具有修复临界骨缺损的应用潜力。

https://orcid.org/0000-0002-4061-7051 (王金斯)

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: 三周期极小曲面, 生物陶瓷, β-磷酸三钙, 增材制造, 数字激光加工, 临界骨缺损, 孔径, MC3T3-E1细胞

Abstract: BACKGROUND: The bone scaffold with complex porous structure can be customized accurately and individually by additive manufacturing technology, so as to achieve the dual bionics of the structure and function of cancellous bone at critical-sized bone defects.
OBJECTIVE: To clear the mechanical properties and biological activity of β-tricalcium phosphate bioceramic bone scaffold with triply periodic minimal surface structure, and reveal the regulatory effect of triply periodic minimal surface structure on osteoblasts through material and cytological characterizations. 
METHODS: Three kinds of triply periodic minimal surface G surface structure β-tricalcium phosphate bioceramic bone scaffolds with different pore sizes of 330, 420, and 510 μm were designed by Matlab R2020a software, and the design drawings were analyzed by Inspire 2018 software. The STL file exported based on the triply periodic minimal surface structure was the blueprint. β-Tricalcium phosphate scaffolds were fabricated by additive manufacturing technology based on digital laser processing. The surface morphology was observed by scanning electron microscope. The phase composition was detected by X-ray diffractometer, and the mechanical strength was detected by universal material testing machine. MC3T3-E1 cells were co-cultured with scaffolds. Cell proliferation, cell adhesion, and alkaline phosphatase activity were detected.   
RESULTS AND CONCLUSION: (1) Inspire 2018 software showed that triply periodic minimal surface presented a smooth, continuous and uniform porous structure. (2) Scanning electron microscope confirmed that the additive manufacturing technology based on digital laser processing successfully realized the accurate molding of triply periodic minimal surface structure. (3) X-ray diffraction confirmed that the scaffold was composed of pure β-tricalcium phosphate crystal phase. (4) The compressive strength and elastic modulus of the three groups of scaffolds were all within or near the range of cancellous bone, and the compressive strength of the scaffolds was inversely proportional to the pore size. (5) CCK8 assay showed that MC3T3-E1 cells grew well on the scaffolds; the biological activity of the scaffolds was dose-dependent with the pore size; and the 510 μm pore diameter scaffold had the best effect on promoting proliferation. (6) Living cell imager and laser confocal microscope showed that MC3T3-E1 cells could adhere to the scaffold in the early stage, and the amount of adhesion increased with the increase of pore size. (7) Alkaline phosphatase activity analysis showed that the alkaline phosphatase activity of cells on 420 μm scaffold was the highest. (8) The results show that the mechanical properties and biological activity of the triply periodic minimal surface structure β-tricalcium phosphate bioceramic scaffold are excellent; the pore size of 420 μm is beneficial to cell differentiation and 510 μm is beneficial to cell proliferation, which has the potential to repair critical bone defects.

Key words: triply periodic minimal surfaces, bioceramic, β-tricalcium phosphate, additive manufacturing, digital laser processing, critical-sized bone defect, pore size, MC3T3-E1 cell

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