中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (14): 3636-3642.doi: 10.12307/2025.586

• 复合支架材料 composite scaffold materials • 上一篇    下一篇

3D打印甲基丙烯酸酯化明胶/壳聚糖支架:抗菌、力学性能及细胞相容性评价

李  梁1,杨  翰2,索海瑞2,关  路1,王振林1   

  1. 1中国人民解放军联勤保障部队第九〇六医院,浙江省宁波市   315040;2杭州电子科技大学自动化学院,浙江省杭州市   310018
  • 收稿日期:2024-12-20 接受日期:2025-02-20 出版日期:2026-05-18 发布日期:2025-09-11
  • 通讯作者: 王振林,副主任医师,中国人民解放军联勤保障部队第九〇六医院,浙江省宁波市 315040
  • 作者简介:李梁,男,1982年生,浙江省宁波市人,汉族,临床医学博士,副主任医师,主要从事骨外科学、生物3D打印与组织工程方面的研究。
  • 基金资助:
    2021宁波市自然科学基金项目(2021J238),项目名称:3D打印梯度孔径支架复合滑膜间充质干细胞构建组织工程仿生软骨的实验研究,项目负责人:李梁

3D printed methacrylated gelatin/chitosan scaffolds: evaluation of antibacterial, mechanical properties and cytocompatibility

Li Liang1, Yang Han2, Suo Hairui2, Guan Lu1, Wang Zhenlin1    

  1. 1No. 906 Hospital of People’s Liberation Army, Ningbo 315040, Zhejiang Province, China; 2School of Automation, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China
  • Received:2024-12-20 Accepted:2025-02-20 Online:2026-05-18 Published:2025-09-11
  • Contact: Wang Zhenlin, Associate chief physician, No. 906 Hospital of People’s Liberation Army, Ningbo 315040, Zhejiang Province, China
  • About author:Li Liang, MD, Associate chief physician, No. 906 Hospital of People’s Liberation Army, Ningbo 315040, Zhejiang Province, China
  • Supported by:
    2021 Ningbo Natural Science Foundation Project, No. 2021J238 (to LL)

摘要:

文题释义:
组织工程支架:是用于支持细胞生长、增殖和组织再生的三维结构材料。在组织工程中,支架充当“细胞容器”,为细胞提供必要的机械支持和微环境,促进组织的形成和功能恢复。不同特性的支架可以应用于不同的组织工程方向,此次研究中的甲基丙烯酸酯化明胶/壳聚糖组织工程支架具有中等机械性能、良好的抗菌性能和细胞相容性,可应用于软骨组织工程和难愈创面的皮肤修复和抗感染组织工程。
可打印性能:在3D打印生物医学领域,生物墨水的可打印性能是指在打印过程中实现精确成型、稳定打印和功能性表现的能力。此次研究中3D打印针头处挤出物呈现液滴状态难以成型和针头堵塞均代表生物墨水的可打印性差,而喷头处出丝流畅且形成丝状结构,能打印出具有清晰孔隙和三维结构的支架,代表生物墨水的可打印性良好。

背景:3D打印是制造组织工程支架的重要手段,但如何提高支架的抗菌性是目前研究中容易忽略的重要环节。
目的:采用3D打印构建甲基丙烯酸酯化明胶/壳聚糖抗菌组织工程支架,评估支架的抗菌性能、力学性能和细胞相容性。
方法:将不同浓度的甲基丙烯酸酯化明胶(3%-15%)与壳聚糖(0%-5%)混合制备甲基丙烯酸酯化明胶/壳聚糖复合生物墨水,在常温下测试生物墨水的3D打印性,选择适宜的生物墨水进行后续实验。将10%甲基丙烯酸酯化明胶分别与不同浓度(0%,1%,3%,5%)壳聚糖混合作为生物墨水,3D打印甲基丙烯酸酯化明胶/壳聚糖支架,分别记为G10、G10C1、G10C3、G10C5,检测4种生物墨水与4种支架对大肠杆菌生长的作用,检测4种支架的抗压强度与压缩模量。将小鼠胚胎成纤维细胞NIH3T3分别接种于4种支架表面,培养3 d后进行活死染色,观察细胞活性。
结果与结论:①纯甲基丙烯酸酯化明胶的打印区间在7%-13%之间,壳聚糖的加入使甲基丙烯酸酯化明胶的打印区间向低浓度扩展,最低可至4%;高浓度甲基丙烯酸酯化明胶中加入壳聚糖会使其黏度增加而挤出困难,最终确定甲基丙烯酸酯化明胶浓度10%甲基丙烯酸酯化明胶分别与0%,1%,3%,5%壳聚糖混合作为生物墨水。②随着壳聚糖浓度的增加,生物墨水与3D打印支架的抗菌性均增强,3D打印支架的抗压强度与压缩模量均升高,3D打印支架上的NIH3T3细胞活性呈先增长后降低的趋势,其中G10C3支架上的细胞活性最好。③结果表明,通过调节壳聚糖的浓度、结合3D打印技术可以具备优异抗菌性能、力学性能及细胞相容性的甲基丙烯酸酯化明胶/壳聚糖组织工程支架。
https://orcid.org/0009-0004-4263-0585 (李梁) 

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: 3D打印, 组织工程支架, 甲基丙烯酸酯化明胶, 壳聚糖, 抗菌性, 工程化骨材料

Abstract: BACKGROUND: 3D printing is an important means to manufacture tissue engineering scaffolds, but how to improve the antibacterial property of scaffolds is an important link that is easily overlooked in current research.
OBJECTIVE: To construct methacrylated gelatin/chitosan antibacterial tissue engineering scaffolds by 3D printing and evaluate the antibacterial properties, mechanical properties, and cell compatibility of the scaffolds.
METHODS: Methacrylated gelatin (3%-15%) and chitosan (0%-5%) were mixed to prepare methacrylated gelatin/chitosan composite bio-inks. The 3D printability of the bio-inks was tested at room temperature, and the appropriate bio-inks were selected for subsequent experiments. 10% methacrylated gelatin was mixed with chitosan of different concentrations (0%, 1%, 3%, and 5%) as bio-inks, and methacrylated gelatin/chitosan scaffolds were 3D printed, which were recorded as G10, G10C1, G10C3, and G10C5, respectively. The effects of the four kinds of bio-inks and four kinds of scaffolds on the growth of Escherichia coli were detected, and the compressive strength and compression modulus of the four kinds of scaffolds were detected. Mouse embryonic fibroblasts NIH3T3 were inoculated on the surface of four scaffolds. The live-dead staining was performed after culturing for 3 days to observe cell viability.
RESULTS AND CONCLUSION: (1) The printing range of pure methacrylated gelatin was between 7% and 13%. The addition of chitosan expanded the printing range of methacrylated gelatin to a lower concentration, which could be as low as 4%. The addition of chitosan to high-concentration methacrylated gelatin increased its viscosity and made it difficult to extrude. Finally, methacrylated gelatin with a concentration of 10% was mixed with 0%, 1%, 3%, and 5% chitosan as bio-ink. (2) With the increase of chitosan concentration, the antibacterial properties of bio-ink and 3D printed scaffolds were enhanced; the compressive strength and compression modulus of 3D printed scaffolds were increased, and the viability of NIH3T3 cells on 3D printed scaffolds increased first and then decreased, among which the cell viability on G10C3 scaffold was the best. (3) The results show that by adjusting the concentration of chitosan and combining 3D printing technology, methacrylated gelatin/chitosan tissue engineering scaffolds with excellent antibacterial properties, mechanical properties and cell compatibility can be obtained.

Key words: 3D printing, tissue engineering scaffold, methacrylated gelatin, chitosan, antibacterial property, engineered bone material

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