中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (28): 4477-4482.doi: 10.12307/2022.301

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

京尼平交联3D打印胶原/壳聚糖支架的表征

张佳颖1，索海瑞1，2，徐铭恩1，2，王  玲1，2   

  1. 1杭州电子科技大学自动化学院，浙江省杭州市  310018；2浙江省医学信息与生物三维打印重点实验室，浙江省杭州市  310018
  • 收稿日期:2021-01-19 接受日期:2021-03-06 出版日期:2022-10-08 发布日期:2022-03-21
  • 通讯作者: 王玲，教授，杭州电子科技大学自动化学院，浙江省杭州市 310018；浙江省医学信息与生物三维打印重点实验室，浙江省杭州市 310018
  • 作者简介:张佳颖，女，1996年生，江西省丰城市人，汉族，杭州电子科技大学在读硕士，主要从事生物3D打印方面研究。
  • 基金资助:
    国家重点研发计划(2017YFC1103400)，项目负责人：徐铭恩；国家自然科学基金(61675059)，项目负责人：王玲

3D printed collagen/chitosan scaffolds crosslinked by genipin

Zhang Jiaying1, Suo Hairui1, 2, Xu Mingen1, 2, Wang Ling1, 2    

  1. 1School of Automation, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China; 2Key Laboratory of Medical Information and 3D Bioprinting of Zhejiang Province, Hangzhou 310018, Zhejiang Province, China
  • Received:2021-01-19 Accepted:2021-03-06 Online:2022-10-08 Published:2022-03-21
  • Contact: Wang Ling, Professor, School of Automation, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China; Key Laboratory of Medical Information and 3D Bioprinting of Zhejiang Province, Hangzhou 310018, Zhejiang Province, China
  • About author:Zhang Jiaying, Master candidate, School of Automation, Hangzhou Dianzi University, Hangzhou 310018, Zhejiang Province, China
  • Supported by:
    the National Key Research & Development Program of China, Grant No. 2017YFC1103400 (to XME); the National Natural Science Foundation of China, No. 61675059 (to WL)

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摘要：

文题释义：
京尼平：是一种天然的交联剂，在碱性环境下，京尼平受到OH-攻击开环形成单个醛基中间体，开环的京尼平单分子发生自聚合反应后形成稳定的长链结构，聚合物末端的醛基结合胶原和壳聚糖中的-NH2生成Schiff键，形成一种可靠的交联网状结构。
壳聚糖：是一种天然活性多糖，结构和糖胺聚糖相似，具备良好的生物相容性，在体内会被溶菌酶缓慢降解，形成能被人体吸收的单糖或寡糖，与胶原蛋白复合可改善其力学性能和降解性能，因此被广泛应用于组织工程支架研究。

背景：胶原/壳聚糖支架需交联才能达到相应力学性能，有研究表示调节交联剂浓度可以在一定范围内调控支架的理化性能。
目的：探究京尼平浓度对胶原/壳聚糖支架理化性能的影响，制备理化性能可调节的组织工程支架。
方法：将胶原和壳聚糖粉末分别溶于弱酸后混合均匀，作为打印墨水，利用生物3D打印机低温打印胶原支架与胶原/壳聚糖支架，经冻干、中和处理后分别以1，3，5 mmol/L的京尼平进行交联。检测各组支架的表观结构稳定性、抗拉能力、溶胀性能、降解性能与生物相容性。
结果与结论：①将支架在PBS中浸泡3 d后，对比未交联的冻干支架，交联后胶原支架表面维持规则的孔结构，但是支架出现明显变形；交联后胶原/壳聚糖支架表面结构规则，仅1 mmol/L京尼平交联的胶原/壳聚糖支架存在轻微变形。②随着京尼平浓度的增加，各组支架的力学性能增加，并且对应交联浓度下的胶原/壳聚糖支架力学性能好于胶原支架。③随着京尼平浓度的增加，胶原支架的溶胀率下降，胶原/壳聚糖支架的溶胀率无明显变化。④浸泡于胶原酶溶液中后，不同浓度京尼平交联的胶原支架在1 h内被完全降解，胶原/壳聚糖支架的降解速率随京尼平浓度的增加而降低，均呈现先快速后平缓的趋势。⑤将骨髓间充质干细胞接种于各组交联支架3 d后，1，3 mmol/L京尼平交联的胶原/壳聚糖支架(或胶原支架)上的细胞数量明显多于5 mmol/L京尼平交联的胶原/壳聚糖支架(P < 0.05)。⑥结果表明，京尼平可在一定范围调节胶原/壳聚糖支架理化性能，其中3 mmol/L京尼平交联的胶原/壳聚糖支架具有较好的力学性能、抗酶解能力与生物相容性。

https://orcid.org/0000-0001-6122-3724 (张佳颖) 

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: 交联, 胶原, 壳聚糖, 低温3D打印, 京尼平, 组织工程, 支架, 细胞活性

Abstract: BACKGROUND: Collagen/chitosan scaffolds need to be cross-linked to achieve corresponding mechanical properties. Studies have shown that the regulation of crosslinking agent concentration can control the physical and chemical properties of collagen/chitosan scaffolds. 
OBJECTIVE: To explore the effect of genipin concentration on physical and chemical properties of collagen/chitosan scaffolds, and manufacture tissue engineering scaffolds with adjustable physical and chemical properties. 
METHODS: Collagen and chitosan were separately dissolved in weak acid and then mixed as printing ink. The collagen/chitosa scaffolds were printed using biological 3D printer at low-temperature, then lyophilized and neutralized, and crosslinked with 1, 3 and 5 mmol/L genipin. The apparent structural stability, tensile strength, swelling performance, degradation performance and biocompatibility of scaffolds of each group were detected. 
RESULTS AND CONCLUSION: (1) After the scaffold was soaked in PBS for 3 days, compared with the uncrosslinked freeze-dried scaffold, the surface of the collagen scaffold maintained regular pore structure after crosslinking, but the scaffold showed obvious deformation. The surface structure of the collagen/chitosan scaffold was regular after crosslinking. The collagen/chitosan scaffold cross-linked with only 1 mmol/L genipin had slight deformation. (2) With the increased concentration of genipin, the mechanical properties of scaffolds of each group increased, and the mechanical properties of collagen/chitosan scaffolds at the corresponding crosslinking concentration were better than those of collagen scaffolds. (3) With the increased concentration of genipin, the swelling rate of the collagen scaffold decreased, and the swelling rate of the collagen/chitosan scaffold did not remarkably change. (4) After soaking in collagenase solution, the collagen scaffolds crosslinked with different concentrations of genipin were totally degraded within 1 hour. The degradation rate of collagen/chitosan scaffold decreased with the increased concentration of genipin, showing a trend of rapid first and then smooth. (5) After inoculating bone marrow mesenchymal stem cells on the crosslinked scaffolds for 3 days, the number of cells on the 1 and 3 mmol/L genipin crosslinked collagen/chitosan scaffold (or collagen scaffold) was significantly more than that on the 5 mmol/L genipin crosslinked collagen/chitosan scaffold (P < 0.05). (6) Results have suggested that genipin can adjust the physical and chemical properties of collagen/chitosan scaffolds within a certain extent. Among them, collagen/chitosan scaffold crosslinked with 3 mmol/L genipin has good mechanical properties, resistance to enzymatic hydrolysis and biocompatibility.

Key words: crosslinking, collagen, chitosan, low-temperature 3D printing, genipin, tissue engineering, scaffold, cell viability

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