3D打印HA微球支架的制备与表征

doi:10.15541/jim20200499

无机材料学报 ›› 2021, Vol. 36 ›› Issue (6): 601-607.DOI: 10.15541/jim20200499

所属专题：【生物材料】骨骼与齿类组织修复；【虚拟专辑】增材制造及3D打印(2021-2022)

3D打印HA微球支架的制备与表征

吴重草¹^,²^,³(), 郇志广²^,³, 朱钰方²^,³, 吴成铁²^,³()

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

收稿日期:2020-08-28 修回日期:2020-10-03 出版日期:2021-06-20 网络出版日期:2020-10-10
通讯作者: 吴成铁, 研究员. E-mail: chengtiewu@mail.sic.ac.cn
作者简介:吴重草(1995-), 女, 硕士研究生. E-mail: wuzhongcao@student.sic.ac.cn
基金资助:
国家自然科学基金(51761135103)

3D Printing and Characterization of Microsphere Hydroxyapatite Scaffolds

WU Zhongcao¹^,²^,³(), HUAN Zhiguang²^,³, ZHU Yufang²^,³, WU Chengtie²^,³()

1. School of Materials Science and Engineering, Shanghai University, Shanghai 200444, China
2. State Key Lab 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

Received:2020-08-28 Revised:2020-10-03 Published:2021-06-20 Online:2020-10-10
Contact: WU Chengtie, professor. E-mail: chengtiewu@mail.sic.ac.cn
About author:WU Zhongcao(1995-), female, Master candidate. E-mail: wuzhongcao@student.sic.ac.cn
Supported by:
National Natural Science Foundation of China(51761135103)

摘要/Abstract

摘要：

生物材料表面微结构对于成骨具有重要的影响, 该研究以不同粒径(< 60 μm)的羟基磷灰石(HA)微球状粉体为原料, 通过3D打印技术制备了一系列(HA0、HA10、HA30、HA50)生物陶瓷支架。不同支架具有相似的理化性能, 由于微球粒径不同形成了不同的微结构, 对其生物学性能造成不同的影响。相比传统非微球颗粒打印的支架(HA0), HA微球构成的支架能够提供更多细胞粘附和生长位点, 24 h的粘附实验显示HA30支架能显著促进骨髓间充质干细胞的伪足伸长; 培养5 d的细胞增殖实验显示, 微球支架上的细胞数量与HA0支架出现显著性差异, 表面微球结构与细胞尺度相当的HA30支架具有最好的促增殖效果。因此, 3D打印技术在可控制备HA支架宏观结构的同时, 还可以通过控制生物陶瓷粉体的颗粒形貌, 调控3D打印支架的表面微结构, 从而优化其生物学效应, 在骨组织工程领域具有良好的应用前景。

关键词: HA微球, 3D打印, 支架, 骨组织工程

Abstract:

Surface microstructure of biomaterials plays an important role in osteogenesis. In this study, we prepared a series of hydroxyapatite (HA) scaffolds (HA0, HA10, HA30, HA50) by 3D printing microspheres with different particle sizes (<60 μm). These scaffolds have similar physical and chemical properties, but form different microstructures due to their different particle size of microspheres, which result in certain impact on their biological properties. Compared with the traditional scaffolds without micro-sphere structure (HA0), the microsphere-based hydroxyapatite scaffolds provide improved adhesion and growth sites for bone mesenchymal stem cells (BMSCs) after 24 h culture, among which HA30 scaffold significantly promotes the pseudopodia elongation of BMSCs. After 5 d of cell culture on the scaffolds, the micro-sphere based HA scaffolds significantly stimulate proliferation of the BMSCs in contrast to the HA0 group. HA30 scaffolds with microsphere at similar size to the cell size have the best promoting effect on cell proliferation. Therefore, 3D printing technology can not only control the macrostructure of HA scaffolds, but also construct the microstructure on the surface of HA scaffolds by controlling the particle sizes of bioceramic powders to optimize the biological effects, which shows great potential application in the field of bone tissue engineering.

Key words: hydroxyapatite microspheres, 3D printing, scaffolds, bone tissue engineering

中图分类号:

TQ174

吴重草, 郇志广, 朱钰方, 吴成铁. 3D打印HA微球支架的制备与表征[J]. 无机材料学报, 2021, 36(6): 601-607.

WU Zhongcao, HUAN Zhiguang, ZHU Yufang, WU Chengtie. 3D Printing and Characterization of Microsphere Hydroxyapatite Scaffolds[J]. Journal of Inorganic Materials, 2021, 36(6): 601-607.

图/表 6

图1 HA普通粉体与微球粉体(< 60 μm)的(a)光学显微镜照片和(b)扫描电镜照片((i) HA0, (ii) HA10, (iii) HA30, (iv) HA50)及其(c)逐级过筛后的HA微球粒径统计

Fig. 1 (a) Optical micrographs and (b) SEM images of HA powders with and without microsphere structure (< 60 μm) ((i) HA0, (ii) HA10, (iii) HA30, (N) HA50), and their (c) particle size statistics after successive sieving

图2 (a)两种形貌的HA粉体在1150 ℃烧结前后的XRD图谱和(b)烧结后的支架再粉碎后的SEM照片及其EDS图谱

Fig. 2 (a) XRD characterization of HA powder with two morphologies before and after sintered at 1150 ℃, and (b) SEM image with EDS pattern of scaffolds after sintering and grinding.

图3 (a)不同粒径3D打印HA微球支架及 (b) HA0、(c) HA10、(d) HA30和(e) HA50的微观结构

Fig. 3 (a) 3D printing HA microsphere scaffolds with different particle sizes and microstructure of (b) HA0, (c) HA10, (d) HA30, and (e) HA50 scaffolds

图4 HA支架的离子累积释放量

Fig. 4 Accumulative ion release of HA scaffolds

图5 rBMSCs在(a) HA0、(b) HA10、(c) HA30和(d) HA50支架上的生长状态(箭头所指为细胞伪足)

Fig. 5 Growth states of rBMSCs on (a) HA0, (b) HA10, (c) HA30 and (d) HA50 scaffolds with arrows pointing to the cell pseudopodia

图6 rBMSC细胞在各支架上增殖的结果

Fig. 6 Proliferation of rBMSCs on scaffolds (a) Quantitative statistics of cell proliferation, and (b) fluorescence photograghs of cell proliferation with DAPI staining

参考文献 20

[1]	WEINER S, WAGNER H D. The material bone: structure-mechanical dunction relations. Annual Review of Materials Science, 2003,28(1):271-298. DOI URL
[2]	WILSON C E, BRUIJN J D D, BLITTERSWIJK C A V, et al. Design and fabrication of standardized hydroxyapatite scaffolds with a defined macro-architecture by rapid prototyping for bone-tissue- engineering research. Journal of Biomedical Materials Research Part A, 2004,68a(1):123-132.
[3]	MALAFAYA P B, PEDRO A J, PETERBAUER A, et al. Chitosan particles agglomerated scaffolds for cartilage and osteochondral tissue engineering approaches with adipose tissue derived stem cells. Journal of Materials Science: Materials in Medicine, 2005,16(12):1077-1085. DOI URL
[4]	COX S C, THORNBY J A, GIBBONS G J, et al. 3D printing of porous hydroxyapatite scaffolds intended for use in bone tissue engineering applications. Materials Science & Engineering C Materials for Biological Applications, 2015,47:237-247.
[5]	LIU A, XUE G H, SUN M, et al. 3D Printing surgical implants at the clinic: a experimental study on anterior cruciate ligament reconstruction. Scientific Reports, 2016,6:21704 DOI URL
[6]	SHIMOMURA K, MORIGUCHI Y, MURAWSKI C D, et al. Osteochondral tissue engineering with biphasic scaffold: current strategies and techniques. Tissue Engineering Part B Reviews, 2014,20(5):468-476. DOI URL
[7]	WEI G, MA P X. Structure and properties of nano-hydroxyapatite/ polymer composite scaffolds for bone tissue engineering. Biomaterials, 2004,25(19):4749-4757. DOI URL
[8]	LEUKERS B, GÜLKAN H, IRSEN S H, et al. Hydroxyapatite scaffolds for bone tissue engineering made by 3D printing. Journal of Materials Science Materials in Medicine, 2005,16(12):1121-1124. DOI URL
[9]	WOODARD J R, HILLDORE A J, LAN S K, et al. The mechanical properties and osteoconductivity of hydroxyapatite bone scaffolds with multi-scale porosity. Biomaterials, 2007,28(1):45-54. DOI URL
[10]	LI S H, DE WIJN J R, LAYROLLE P, et al. Synthesis of macroporous hydroxyapatite scaffolds for bone tissue engineering. Journal of Biomedical Materials Research, 2010,61(1):109-120. DOI URL
[11]	SILVA M L A, CRAWFORD A, MUNDY J M, et al. Chitosan/ polyester-based scaffolds for cartilage tissue engineering: assessment of extracellular matrix formation. Acta Biomaterialia, 2010,6(3):1149-1157. DOI URL
[12]	FENG C, ZHANG W, DENG C, et al. 3D printing of lotus root-like biomimetic materials for cell delivery and tissue regeneration. Advanced Science, 2017,4(12):1700401. DOI URL
[13]	LUO Y, LODE A, WU C, et al. Alginate/nanohydroxyapatite scaffolds with designed core/shell structures fabricated by 3D plotting and in situ mineralization for bone tissue engineering. ACS Applied Materials & Interfaces, 2015,7(12):6541-6549.
[14]	DENG C J, YANG Q, SUN X L, et al. Bioactive scaffolds with Li and Si ions-synergistic effects for osteochondral defects regeneration. Applied Materials Today, 2018,10:203-216. DOI URL
[15]	DALBY M J, RIEHLE M O, YARWOOD S J, et al. Nucleus alignment and cell signaling in fibroblasts: response to a micro-grooved topography. Experimental Cell Research, 2003,284(2):274-282.
[16]	FU YA-KANG, ZHOU XUE, XIAO DONG-QIN, et al. Influence of micro-nano structure of haydroxyapatite particles on protein adsorption. Journal of Inorganic Materials, 2015,30(5):523-528. DOI URL
[17]	ZHAO C C, WANG X Y, GAO L, et al. The role of the micro-pattern and nano-topography of hydroxyapatite bioceramics on stimulating osteogenic differentiation of mesenchymal stem cells. Acta Biomaterialia, 2018,73:509 DOI URL
[18]	ZHAO C C, XIA L G, ZHAI D, et al. Designing ordered micropatterned hydroxyapatite bioceramics to promote the growth and osteogenic differentiation of bone marrow stromal cells. Journal of Materials Chemisty B, 2015,3(6):968-976.
[19]	ZHOU P Y, WU J H XIA Y, et al. Loading BMP-2 on nanostructured hydroxyapatite microspheres for rapid bone regeneration. International Journal of Nanomedicine, 2018,13:4083-4092. DOI URL
[20]	SHI Z L, HUANG X, CAI Y R, et al. Size effect of hydroxyapatite nanoparticles on proliferation and apoptosis of osteoblast-like cells. Acta Biomaterialia, 2009,5(1):338-345. DOI URL

3D打印HA微球支架的制备与表征

3D Printing and Characterization of Microsphere Hydroxyapatite Scaffolds

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