中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (20): 5188-5200.doi: 10.12307/2026.690

• 水凝胶材料Hydrogel materials • 上一篇    下一篇

双离子时序释放多功能水凝胶修复感染性骨缺损

陈伟飞，梅远东，巨积辉   

  1. 扬州大学教学医院苏州瑞华骨科医院，江苏省苏州市  215000
  • 接受日期:2025-05-29 出版日期:2026-07-18 发布日期:2025-11-27
  • 通讯作者: 巨积辉，主任医师，副教授，扬州大学教学医院苏州瑞华骨科医院，江苏省苏州市 215000
  • 作者简介:陈伟飞，男，1985年生，江苏省常熟市人，汉族，硕士，副主任医师，主要从事骨感染再生修复及相关基础研究。
  • 基金资助:
    苏州市科技计划项目-医学应用基础研究(SKYD2023026)，项目负责人：巨积辉

Repair of infected bone defect with dual-ion time-sequenced release multifunctional hydrogels

Chen Weifei, Mei Yuandong, Ju Jihui   

  1. Suzhou Ruihua Orthopedic Hospital, Teaching Hospital of Yangzhou University, Suzhou 215000, Jiangsu Province, China
  • Accepted:2025-05-29 Online:2026-07-18 Published:2025-11-27
  • Contact: Ju Jihui, Chief physician, Associate professor, Suzhou Ruihua Orthopedic Hospital, Teaching Hospital of Yangzhou University, Suzhou 215000, Jiangsu Province, China
  • About author:Chen Weifei, MS, Associate chief physician, Suzhou Ruihua Orthopedic Hospital, Teaching Hospital of Yangzhou University, Suzhou 215000, Jiangsu Province, China
  • Supported by:
    Suzhou Science and Technology Plan - Basic Research in Medical Applications, No. SKYD2023026 (to JJH)

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摘要：

文题释义：
感染性骨缺损：是由严重创伤、术后感染、慢性骨髓炎或骨肿瘤切除术后导致的骨组织缺损，同时伴随细菌感染、生物膜形成及骨组织修复受损。感染性骨缺损的病理特征包括局部炎症反应增强、骨组织破坏加剧及骨再生受阻。
多功能水凝胶：是一种集抗菌、抗炎、血管生成、成骨诱导等两种或多种功能于一体的生物材料，因高含水性、良好的生物相容性和可控释放能力在骨修复与感染控制领域具有广泛应用。近年来，功能化水凝胶通过整合抗菌金属离子、生物活性分子或纳米材料能够在感染性骨缺损修复中发挥抗菌与促进骨再生的双重作用。

背景：开发既能在早期有效抗菌又能在后期促进骨再生的时序性功能生物材料，是解决感染性骨缺损治疗难题的关键。
目的：构建双离子时序释放多功能水凝胶，评估该水凝胶的抗菌与促骨再生功能。
方法：①采用表面活性剂模板法合成含锶介孔生物玻璃，通过金属-配体配位交联原理制备硫代透明质酸；将硫代透明质酸溶解于去离子水中，逐滴加入含锶介孔生物玻璃，混合均匀后缓慢滴加硝酸银溶液，搅拌15 min后得到双离子时序释放多功能水凝胶(记为SHA-Ag/SBG水凝胶)。表征SHA-Ag/SBG水凝胶的微观形貌、Sr2+及Ag+缓释性能与溶胀性能。②将金黄色葡萄球菌或大肠杆菌分别与SHA-Ag/SBG水凝胶、透明质酸水凝胶、硫代透明质酸水凝胶、载Ag+硫代透明质酸水凝胶共培养，以单独培养的细菌为对照，通过细菌涂板实验评估水凝胶的抗菌性能。③将大鼠骨髓间充干细胞分别与SHA-Ag/SBG水凝胶、透明质酸水凝胶、硫代透明质酸水凝胶、载Ag+硫代透明质酸水凝胶共培养，以单独培养的细胞为对照，成骨诱导后，通过活/死染色评估水凝胶的细胞相容性，通过碱性磷酸酶染色、茜素红染色、成骨相关基因检测评估水凝胶的促成骨性能。④在40只SD大鼠股骨髁内侧制备直径2 mm贯穿皮质骨的圆形骨缺损，随机分4组干预：正常对照组(n=10)不进行感染处理，模型组(n=10)骨缺损处注射金黄色葡萄球菌悬液模拟感染，对照组(n=10)骨缺损处注射金黄色葡萄球菌悬液后注射载Ag+硫代透明质酸水凝胶，实验组(n=10)骨缺损处注射金黄色葡萄球菌悬液后注射SHA-Ag/SBG水凝胶。术后4，12周取材，分别进行Micro-CT检测与苏木精-伊红、Masson、Giemsa染色。
结果与结论：①SHA-Ag/SBG水凝胶呈现良好的三维多孔结构，具有良好的溶胀性能；离子释放率实验显示，SHA-Ag/SBG水凝胶中Ag+早期释放较快，3 d后逐渐稳定；Sr2+释放较缓慢，7 d后才开始逐步释放，展示出双离子顺序释放抗菌促骨再生特性。②SHA-Ag/SBG水凝胶、载Ag+硫代透明质酸水凝胶可显著抑制金黄色葡萄球菌与大肠杆菌的生长，而透明质酸水凝胶、硫代透明质酸水凝胶无明显的抑菌作用。③活/死染色显示4组均无明显的细胞毒性作用。碱性磷酸酶、茜素红染色与成骨相关基因检测显示，SHA-Ag/SBG水凝胶可促进大鼠骨髓间充质干细胞的成骨分化。④Micro-CT检测显示，与其他3组比较，SHA-Ag/SBG水凝胶能够有效促进大鼠感染性骨缺损的骨再生；苏木精-伊红、Masson和Giemsa染色显示，SHA-Ag/SBG水凝胶具有良好的抗菌作用，能够显著促进骨组织的恢复。
https://orcid.org/0009-0004-7246-1161(陈伟飞)

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: 金属-配体配位交联, 透明质酸, 介孔生物玻璃, 感染性骨缺损, 抗菌, 骨再生, 生物材料

Abstract: BACKGROUND: The development of time-sequenced functional biomaterials that can effectively inhibit bacteria early and promote bone regeneration later is key to solving the treatment challenges of infected bone defects. 
OBJECTIVE: To construct a dual-ion time-sequenced release multifunctional hydrogel and evaluate its antibacterial and bone regeneration-promoting properties. 
METHODS: (1) Strontium-containing mesoporous bioglass was synthesized using a surfactant template method, and sulfohyaluronic acid was prepared via metal-ligand coordination crosslinking. Sulfohyaluronic acid was dissolved in deionized water, and the strontium-containing mesoporous bioglass was added dropwise. After mixing thoroughly, silver nitrate solution was slowly added dropwise and stirred for 15 minutes to obtain a multifunctional hydrogel with a timed release of dual ions (denoted as SHA-Ag/SBG hydrogel). The micromorphology, sustained release of Sr2+ and Ag+, and swelling properties of the SHA-Ag/SBG hydrogel were characterized. (2) Staphylococcus aureus or Escherichia coli were co-cultured with SHA-Ag/SBG hydrogels, hyaluronic acid hydrogels, sulfo-hyaluronic acid hydrogels, and Ag+-loaded sulfo-hyaluronic acid hydrogels, respectively. Bacterial plating experiments were used to evaluate the antibacterial properties of the hydrogels, using bacteria cultured alone as controls. (3) Rat bone marrow mesenchymal stem cells were co-cultured with SHA-Ag/SBG hydrogels, hyaluronic acid hydrogels, sulfo-hyaluronic acid hydrogels, and Ag+-loaded sulfo-hyaluronic acid hydrogels, respectively. Cells cultured alone served as controls. After osteogenic induction, the cytocompatibility of the hydrogels was evaluated by live/dead staining, and the osteogenic properties of the hydrogels were evaluated by alkaline phosphatase staining, Alizarin red staining, and expression of osteogenesis-related genes. (4) Circular bone defects with a diameter of 2 mm penetrating the cortical bone were created on the medial side of the femoral condyle in 40 SD rats. The rats were randomly divided into four intervention groups. The normal control group (n=10) received no infection treatment. The model group (n=10) received a Staphylococcus aureus suspension injected into the bone defect to simulate infection. The control group (n=10) received a Staphylococcus aureus suspension injected into the bone defect followed by an Ag+-loaded sulfo-hyaluronic acid hydrogel. The experimental group (n=10) received a Staphylococcus aureus suspension injected into the bone defect followed by an SHA-Ag/SBG hydrogel. Samples were obtained 4 and 12 weeks after surgery and subjected to micro-CT, hematoxylin-eosin, Masson, and Giemsa staining, respectively.
RESULTS AND CONCLUSION: (1) SHA-Ag/SBG hydrogels exhibited a well-defined three-dimensional porous structure and excellent swelling properties. Ion release experiments showed that Ag+ was released rapidly in the early stages of the SHA-Ag/SBG hydrogels and gradually stabilized after 3 days. Sr2+ was released more slowly, beginning gradually after 7 days, demonstrating the dual-ion sequential release and antibacterial and bone regeneration-promoting properties. (2) SHA-Ag/SBG hydrogels and Ag+-loaded sulfo-hyaluronic acid hydrogels significantly inhibited the growth of Staphylococcus aureus and Escherichia coli, whereas hyaluronic acid hydrogels and sulfo-hyaluronic acid hydrogels showed no significant antibacterial effect. (3) Live/dead staining revealed no significant cytotoxicity in the four groups. Alkaline phosphatase and Alizarin red staining, as well as osteogenesis-related gene expression assay demonstrated that SHA-Ag/SBG hydrogels promoted the osteogenic differentiation of rat bone marrow mesenchymal stem cells. (4) Micro-CT examination showed that the SHA-Ag/SBG hydrogel effectively promoted bone regeneration in infected bone defects of rats compared with the other three groups. Hematoxylin-eosin staining, Masson staining, and Giemsa staining demonstrated that the SHA-Ag/SBG hydrogel exhibited excellent antibacterial activity and significantly promoted bone tissue recovery.

Key words: metal-ligand coordination crosslinking, hyaluronic acid, mesoporous bioactive glass, infected bone defect, antibacterial, bone regeneration, biomaterial

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