中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (26): 6710-6718.doi: 10.12307/2026.804

• 功能性生物材料Functional biomaterials • 上一篇    下一篇

淫羊藿苷缓释微球三维支架的体外促成骨与抗炎性能

赵张红1，金东升1，阮世强1，黄文良2，万  喻1，田仁元1，邓  江1   

  1. 1遵义医科大学第三附属医院(遵义市第一人民医院)骨科，贵州省遵义市  563000；2贵州省茅台医院骨科，贵州省仁怀市  564500
  • 接受日期:2025-12-11 出版日期:2026-09-18 发布日期:2026-03-10
  • 通讯作者: 邓江，教授，主任医师，遵义医科大学第三附属医院(遵义市第一人民医院)骨科，贵州省遵义市 563000
  • 作者简介:赵张红，男，1997年生，贵州省遵义市人，汉族，遵义医科大学在读硕士，医师，主要从事骨缺损基础研究。
  • 基金资助:
    国家自然科学基金项目(81660367)，项目负责人：邓江；贵州省科技计划项目(黔科合基础-ZK(2021)一般387)，项目负责人：黄文良；贵州省科技计划项目(黔科合成果-LC[2024]019)，项目负责人：黄文良

In vitro osteogenic and anti-inflammatory properties of icariin sustained-release microsphere three-dimensional scaffolds

Zhao Zhanghong1, Jin Dongsheng1, Ruan Shiqiang1, Huang Wenliang2, Wan Yu1, Tian Renyuan1, Deng Jiang1   

  1. 1Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University (First People’s Hospital of Zunyi City), Zunyi 563000, Guizhou Province, China; 2Department of Orthopedics, Moutai Hospital of Guizhou Province, Renhuai 564500, Guizhou Province, China
  • Accepted:2025-12-11 Online:2026-09-18 Published:2026-03-10
  • Contact: Deng Jiang, Professor, Chief physician, Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University (First People’s Hospital of Zunyi City), Zunyi 563000, Guizhou Province, China
  • About author:Zhao Zhanghong, Master candidate, Physician, Department of Orthopedics, Third Affiliated Hospital of Zunyi Medical University (First People’s Hospital of Zunyi City), Zunyi 563000, Guizhou Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81660367 (to DJ); Guizhou Provincial Science and Technology Project, No. ZK(2021)387 (to HWL); Guizhou Provincial Science and Technology Project, No. LC[2024]019 (to HWL)

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摘要：

文题释义：
淫羊藿苷：是从传统中草药淫羊藿(Epimedium)中提取的有效活性成分。淫羊藿苷与其代谢产物具有增加骨密度、降脂、抗肿瘤、抗炎、神经保护等多种药理作用。
缓释微球：是指药物溶解或分散在高分子材料基质中形成的粒径尺寸在5-250 μm之间的球状实体。

背景：淫羊藿苷具有促进骨形成和抑制骨吸收的双重活性，但临床应用存在生物利用度低、剂量控制难度高、不良反应风险大等问题。
目的：制备淫羊藿苷缓释微球三维支架，表征该材料的体外促成骨作用。
方法：分别制备丝素蛋白/壳聚糖/纳米羟基磷灰石三维支架(记为SF/CS/nHA支架)、淫羊藿苷缓释微球与负载淫羊藿苷缓释微球的丝素蛋白/壳聚糖/纳米羟基磷灰石三维支架(记为SF/CS/nHA-ICA支架)。表征淫羊藿苷缓释微球的载药率、包封率与体外药物缓释作用以及两种支架的孔径、孔隙率、吸水膨胀率与热水溶失率。将兔骨髓间充质干细胞(或人类风湿关节炎成纤维样滑膜细胞)分别接种于SF/CS/nHA支架与SF/CS/nHA-ICA支架上，以单独培养的细胞为对照，扫描电镜下观察两种细胞的黏附情况，CCK-8法、活/死染色、F-actin染色检测两种细胞的增殖与活性，RT-qPCR、Western blot检测骨髓间充质干细胞内Runx-2、骨钙素、Ⅰ型胶原的mRNA与蛋白表达。
结果与结论：①淫羊藿苷缓释微球的载药率、包封率分别为(29.38±0.04)%，(52.01±0.09)%，体外可持续释放淫羊藿苷达90 d以上；②扫描电镜下可见两组支架内部呈蜂窝状多孔结构，孔隙互连，两组支架的孔径、孔隙率、吸水膨胀率与总热水溶失率比较差异均无显著性意义(P > 0.05)；③扫描电镜下可见两种细胞紧密附着于支架表面和孔隙，其中SF/CS/nHA-ICA支架表面的细胞伪足伸展；CCK-8检测、活/死染色与F-actin染色显示，与对照组、SF/CS/nHA支架相比，SF/CS/nHA-ICA支架可提升兔骨髓间充质干细胞的增殖与活力，抑制人类风湿关节炎成纤维样滑膜细胞的增殖与活力；④RT-qPCR与Western blot检测显示，与对照组、SF/CS/nHA组比较，SF/CS/nHA-ICA组Runx-2、骨钙素、Ⅰ型胶原的mRNA与蛋白表达升高(P < 0.05)；⑤结果表明，淫羊藿苷缓释微球三维支架具有良好的细胞相容性、体外促成骨与抗炎作用。

https://orcid.org/0009-0003-1326-9316(赵张红)；https://orcid.org/0000-0003-0123-1559(邓江)
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: 淫羊藿苷, 缓释微球, 骨组织工程支架, 细胞相容性, 骨髓间充质干细胞, 成纤维样滑膜细胞, 生物材料

Abstract: BACKGROUND: Icariin has the dual activity of promoting bone formation and inhibiting bone resorption, but its clinical application is plagued by low bioavailability, difficulty in controlling dosage, and a high risk of adverse reactions.
OBJECTIVE: To prepare a three-dimensional scaffold containing icariin sustained-release microspheres and characterize their osteogenic activity in vitro.
METHODS: A silk fibroin/chitosan/nanohydroxyapatite three-dimensional scaffold (SF/CS/nHA scaffold), icariin sustained-release microspheres, and a silk fibroin/chitosan/nanohydroxyapatite three-dimensional scaffold loaded with icariin sustained-release microspheres (SF/CS/nHA-ICA scaffold) were prepared. The drug loading efficiency, encapsulation efficiency, and in vitro drug release of the icariin sustained-release microspheres were characterized. The pore size, porosity, water swelling rate, and hot water solubility of the two scaffolds were also characterized. Rabbit bone marrow mesenchymal stem cells (or human rheumatoid arthritis fibroblast-like synoviocytes) were seeded onto SF/CS/nHA scaffolds and SF/CS/nHA-ICA scaffolds, respectively. Cells cultured alone served as controls. Adhesion of the two cell types was observed under a scanning electron microscope. Cell proliferation and activity were assessed using CCK-8 assay, live/dead staining, and F-actin staining. The mRNA and protein expressions of Runx-2, osteocalcin, and type I collagen in bone marrow mesenchymal stem cells were determined using RT-qPCR and western blot assay.
RESULTS AND CONCLUSION: (1) The drug loading rate and encapsulation efficiency of icariin sustained-release microspheres were (29.38±0.04)% and (52.01±0.09)%, respectively, and icariin could be released sustainably in vitro for more than 90 days. (2) Scanning electron microscopy revealed that the interior of the two scaffolds showed a honeycomb-like porous structure with interconnected pores. There were no significant differences in pore size, porosity, water absorption expansion rate, and total hot water solubility between the two scaffolds (P > 0.05). (3) Scanning electron microscopy revealed that both cell types were tightly attached to the scaffold surface and pores, with extended pseudopodia on the surface of the SF/CS/nHA-ICA scaffold. CCK-8 assay, live/dead staining, and F-actin staining showed that the SF/CS/nHA-ICA scaffold enhanced the proliferation and viability of rabbit bone marrow mesenchymal stem cells and inhibited the proliferation and viability of human rheumatoid arthritis fibroblast-like synoviocytes compared with the control group and the SF/CS/nHA scaffold. (4) RT-qPCR and western blot assay revealed that the mRNA and protein expressions of Runx-2, osteocalcin, and type I collagen were increased in the SF/CS/nHA-ICA group compared with the control group and the SF/CS/nHA group (P < 0.05). (5) The results indicate that the icariin sustained-release microsphere three-dimensional scaffold has good cytocompatibility, in vitro osteogenesis, and anti-inflammatory effects.


Key words: icariin, sustained-release microspheres, bone tissue engineering scaffold, cytocompatibility, bone marrow mesenchymal stem cell, fibroblastic synoviocyte, biomaterial

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