双相磷酸镁钙微球体外成血管和促成骨研究

doi:10.15541/jim20220662

无机材料学报 ›› 2023, Vol. 38 ›› Issue (7): 830-838.DOI: 10.15541/jim20220662 CSTR: 32189.14.10.15541/jim20220662

所属专题：【生物材料】骨骼与齿类组织修复(202506)

双相磷酸镁钙微球体外成血管和促成骨研究

吴未¹^,²(), BAKHET Shahd², ASANTE Naomi Addai², KAREEM Shefiu², KOMBO Omar Ramadhan³, 李宾斌², 戴红莲¹^,²()

1.武汉理工大学深圳研究院, 深圳 518000
2.武汉理工大学材料复合新技术国家重点实验室, 湖北省生物材料工程研究中心, 武汉 430070
3.姆贝亚科技大学医学科学与技术系, 姆贝亚, 坦桑尼亚

收稿日期:2022-11-05 修回日期:2022-12-18 出版日期:2023-03-20 网络出版日期:2023-03-20
通讯作者: 戴红莲, 教授. E-mail: daihonglian@whut.edu.cn
作者简介:吴未(1998-), 女, 硕士. E-mail: 2625276216@qq.com

In vitro Study of Biphasic Calcium Magnesium Phosphate Microspheres for Angiogenesis and Bone Formation

WU Wei¹^,²(), BAKHET Shahd², ASANTE Naomi Addai², KAREEM Shefiu², KOMBO Omar Ramadhan³, LI Binbin², DAI Honglian¹^,²()

1. Shenzhen Institute of Wuhan University of Technology, Shenzhen 518000, China
2. State Key Laboratory of Advanced Technology for Materials Synthesis and Processing, Biomedical Materials and Engineering Research Center of Hubei Province, Wuhan University of Technology, Wuhan 430070, China
3. Department of Medical Science and Technology, Mbeya University of Science and Technology, Mbeya, Tanzania

Received:2022-11-05 Revised:2022-12-18 Published:2023-03-20 Online:2023-03-20
Contact: DAI Honglian, professor. E-mail: daihonglian@whut.edu.cn
About author:WU Wei (1998-), male, Master. E-mail: 2625276216@qq.com
Supported by:
National Key Research and Development Program of China(2022YFB4601402);National Natural Science Foundation of China(32201109);Guangdong Basic and Applied Basic Research Foundation(2021A1515110557);Foshan Xianhu Laboratory of the Advanced Energy Science and Technology Guangdong Laboratory(XHT2020-008)

摘要/Abstract

摘要：

磷酸三钙(β-TCP)陶瓷替代材料由于其与骨矿物成分相近及良好的生物相容性和骨传导性, 近年来被广泛关注, 常以纳米颗粒、支架和微球等形式用于骨修复。本研究制备了五种不同的磷酸三钙/磷酸三镁(TMP) (TCP、25% TMP、50% TMP、75% TMP和TMP)复合微球并作了相应表征。随着复合微球中TMP含量增加, 微球释放的Mg²⁺和Ca²⁺的累积浓度增加, 且TMP可以调节复合微球的降解速率。以小鼠胚胎成骨细胞前体细胞(MC3T3-E1)和人脐静脉内皮细胞(HUVECs)为模型, 评价了该复合微球的生物相容性、成血管和成骨作用。结果表明, 与TCP、TMP和75% TMP相比, 25% TMP和50% TMP复合微球具有更好的细胞相容性, 对HUVECs有一定的促增殖作用。因此, 含25% TMP和50% TMP的复合微球对血管生成和成骨具有更积极的作用。

关键词: β-TCP, TMP, 骨缺损, 陶瓷, 微球

Abstract:

Beta tricalcium phosphate (β-TCP) ceramic substituted materials have attracted a large amount of attention in the last decades because of their chemical similarity with bone inorganic components, good biocompatibility, and osteoconductivity. Such materials can be used for bone replacement and bone formation in various forms, such as nanoparticles, scaffolds and microspheres. In this study, five different microsphere materials of tricalcium phosphate/trimagnesium phosphate (TMP) (TCP, 25% TMP, 50% TMP, 75% TMP, and TMP) composites were prepared and characterized. With the increase of TMP content in the composite microspheres, the cumulative concentration of Mg²⁺and Ca²⁺released from the microspheres increased, indicating that TMP can regulate the degradation rate of the composite microspheres. The osteoblast precursor cell line (MC3T3-E1 cells) and human umbilical vein endothelial cells (HUVECs) were used as models to evaluate the biocompatibility, angiogenesis and osteogenesis of the composite microspheres. The results showed that compared with TCP, TMP and 75% TMP group, 25% TMP and 50% TMP composite microspheres had better cell compatibility and had a certain proliferative effect on HUVECs. Therefore, composite microspheres of 25% TMP and 50% TMP have more significant positive effects on angiogenesis and osteogenesis.

Key words: β-TCP, TMP, bone defect, ceramics, microsphere

中图分类号:

TB321
R318

吴未, BAKHET Shahd, ASANTE Naomi Addai, KAREEM Shefiu, KOMBO Omar Ramadhan, 李宾斌, 戴红莲. 双相磷酸镁钙微球体外成血管和促成骨研究[J]. 无机材料学报, 2023, 38(7): 830-838.

WU Wei, BAKHET Shahd, ASANTE Naomi Addai, KAREEM Shefiu, KOMBO Omar Ramadhan, LI Binbin, DAI Honglian. In vitro Study of Biphasic Calcium Magnesium Phosphate Microspheres for Angiogenesis and Bone Formation[J]. Journal of Inorganic Materials, 2023, 38(7): 830-838.

图/表 11

Table 1 Primer sequences used in RT-qPCR

Gene	Primer sequence
VEGF	AGGAGTACCCCGACGAGATAGA CACATCTGCTGTGCTGTAGGAA
FGF	ACAGGAGCGACCAGCACATT TTGGTGTCTGCGAGCCGTAT
COL I	CACTGCAAGAACAGCGTAGC AAGTTCCGGTGTGACTCGTG
OPN	ACACTTTCACTCCAATCGTCCCTAC GGACTCCTTAGACTCACCGCTCTT

Fig. 1 XRD patterns of prepared TCP/TMP composite materials

Fig. 2 SEM images of different microsphere composites with insets showing their corresponding particle size distributions (a)TCP; (b) 25% TMP; (c) 50% TMP; (d) 75% TMP; (e) TMP

Fig. 3 TG/DTA/DTG curves of microspheres prepared by water in oil emulsion cross-linking method (a) TCP; (b) 25%TMP; (c) 50%TMP; (d) 75%TMP; (e) TMP

Fig. 4 Released Ca2+and Mg2+ concentrations from materials immersed in tris-HCl solution

Fig. 5 Cell viabilities of (a) MC3T3-E1 and (b) HUVECs assessed by CCK-8 assay (a) MC3T3-E1 and (b) HUVECs assayed on day 1, 3, 5 cultured with different microspheres concentration extracts *: p < 0.01; **: p < 0.005; ***: p < 0.0002; Colorful figures are available on website

Fig. 6 ALP activity and ARS staining of MC3T3-E1 cells cultured with microspheres extract compared with the control on the 7th and 14th day

Fig. 7 (a) VEGF and (b) FGF of HUVECs cultured with microspheres extracts on the 1st and 3rd day

Fig. 8 (a) COLI and (b) OPN genes expression of MC3T3-E1 which cultured with microspheres extract compared to the control on the 3rd and 7th day

Fig. S1 SEM images of different microsphere composites with Bars at 200 μm (up row) and 10 μm (down row)

Fig.S2 XRD patterns of TMP (a) and β-TCP (b)

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双相磷酸镁钙微球体外成血管和促成骨研究

In vitro Study of Biphasic Calcium Magnesium Phosphate Microspheres for Angiogenesis and Bone Formation

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