中国组织工程研究

中国组织工程研究 ›› 2020, Vol. 24 ›› Issue (4): 524-531.doi: 10.3969/j.issn.2095-4344.1883

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

仿生可降解组织工程纤维环支架的制备与评估

张维昊1，2，徐宝山2，马信龙2，张  杨2，郭  悦2，杜立龙2，许海委2，张凯辉1，2，夏金健1，2，邵鹏飞1，2   

  1. 1天津医科大学研究生院，天津市  300070；2天津医院微创脊柱外科，天津市  300211
  • 收稿日期:2019-02-27 修回日期:2019-03-09 接受日期:2019-06-04 出版日期:2020-02-08 发布日期:2019-12-31
  • 通讯作者: 徐宝山，博士，主任医师，天津医院微创脊柱外科，天津市 300211
  • 作者简介:张维昊，男，1992年生，河北省唐山市人，汉族，2019年天津医科大学毕业，硕士，主要从事椎间盘修复方向研究。
  • 基金资助:
    国家自然科学基金面上项目(31670983)；国家自然科学基金青年科学基金项目(31500781)；天津市应用基础与前沿技术研究计划项目(15JCYBJC25300)

Fabrication and evaluation of biomimetic biodegradable tissue-engineered annulus fibrosus scaffold

Zhang Weihao1,2, Xu Baoshan2, Ma Xinlong2, Zhang Yang2, Guo Yue2, Du Lilong2, Xu Haiwei2, Zhang Kaihui1,2,  Xia Jinjian1,2, Shao Pengfei1,2#br#   

  1. 1Graduate School of Tianjin Medical University, Tianjin 300070, China; 2Department of Minimally Invasive Spine Surgery, Tianjin Hospital, Tianjin 300211, China
  • Received:2019-02-27 Revised:2019-03-09 Accepted:2019-06-04 Online:2020-02-08 Published:2019-12-31
  • Contact: Xu Baoshan, MD, Chief physician, Department of Minimally Invasive Spine Surgery, Tianjin Hospital, Tianjin 300211, China
  • About author:Zhang Weihao, Master, Graduate School of Tianjin Medical University, Tianjin 300070, China; Department of Minimally Invasive Spine Surgery, Tianjin Hospital, Tianjin 300211, China
  • Supported by:
    the National Natural Science Foundation of China (General Program), No. 31670983; the National Natural Science Foundation of China (Youth Program), No. 31500781; Applied Basic and Frontier Technology Research Project of Tianjin, No.15JCYBJC25300

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摘要：

文题释义：

纤维环：是保持椎间盘强度与脊柱稳定性层面的重要组成部分，纤维环的胶原纤维在椎间盘内呈同心圆排列，每层纤维环间呈60°夹角斜行排列，这类特殊的交错网状架构，令纤维环拥有较强的抗拉性能与压缩性，能够预防髓核往外突出，对保持椎间盘的稳定起着重要的作用。

背景：构建既具有仿生结构又具备合适可降解性和良好生物相容性的组织工程纤维环支架仍是难点。

目的：以聚己内酯和聚二恶烷酮为原材料制备仿生可降解支架并评估其作为组织工程纤维环支架的可行性。

方法：通过熔融纺丝技术制备5组不同比例支架(聚己内酯组、聚己内酯/聚二恶烷酮分别为70/30、50/50、30/70组、聚二恶烷酮组)。扫描电子显微镜观察其结构、纤维直径、孔径；测量支架的力学性能和接触角；通过体外模拟和皮下埋植观察支架体外、体内降解情况；检测降解组织周围炎症因子白细胞介素1β和肿瘤坏死因子α的表达情况。接种人脐带间充质干细胞培养7 d，通过CCK-8和死活细胞检测细胞增殖和存活情况。实验于2016-03-02经天津市天津医院医学伦理委员会批准。

结果与结论：①扫描电子显微镜观察显示各组支架纤维粗细均匀，纤维成60°角；②力学性能分析显示单纯聚二恶烷酮支架的拉伸和压缩模量最低，不符合纤维环力学要求；聚己内酯组的力学性能最佳，聚己内酯/聚二恶烷酮为70/30和50/50的支架力学性能适中；③亲水性检测结果表明聚二恶烷酮含量越多，支架亲水性越好；④支架降解情况分析显示，对于组织工程纤维环再生修复来说，单纯聚二恶烷酮和聚己内酯/聚二恶烷酮为30/70支架降解过快，聚己内酯组的降解过慢，聚己内酯/聚二恶烷酮为70/30和50/50的支架降解速率较合适；⑤降解组织周围炎症反应分析显示支架中聚己内酯比例越高炎症反应越严重；⑥CCK-8和死活细胞结果显示人脐带间充质干细胞在聚己内酯/聚二恶烷酮三组混合支架具有良好的增殖活性并且存活率高；⑦结果表明，采用熔融纺丝技术制备的聚己内酯/聚二恶烷酮为70/30和50/50两组支架能够模拟天然纤维环结构，同时具备合适可降解性、优越的力学性能和良好生物相容性，适合用于组织工程纤维环支架的构建。

ORCID: 0000-0001-9088-4222(张维昊)

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: 聚己内酯, 聚二恶烷酮, 纤维环, 仿生结构, 支架, 熔融纺丝, 微米纤维, 椎间盘

Abstract:

BACKGROUND: It is still difficult to construct tissue-engineered anulus fibrosus scaffolds which have bionic structure, suitable biodegradability and good biocompatibility.

OBJECTIVE: To fabricate bionic biodegradable scaffolds with polycaprolactone (PCL) and polydioxanone (PDS) and evaluate the feasibility as a tissue-engineered annulus fibrosus scaffold.

METHODS: Five groups of scaffolds at different PCL/PDS proportions were prepared by melt spinning technique: PCL, PCL/PDS70/30, PCL/PDS50/50, PCL/PDS30/70, and PDS groups. Scanning electron microscopy was used to observe the structure and measure the fiber diameter and pore size of these prepared scaffolds. The mechanical properties and contact angle of the scaffolds were measured. The in vitro and in vivo biodegradation of the scaffolds were observation by in vitro simulation and subcutaneous implantation. The expression of inflammatory factors interleukin-1β and tumor necrosis factor-α in the biodegraded tissues was detected. Human Wharton’s jelly mesenchymal stem cells were cultured for 7 days. Cell viability and proliferation was determined by live/dead cell staining. This study was approved by the Medical Ethics Committee of Tianjin Hospital, China on March 2, 2016.

RESULTS AND CONCLUSION: Scanning electron microscopy results showed that the thickness of the scaffold fibers was uniform and the angle between fibers was 60°. The mechanical properties analysis showed that the tensile and compressive modulus of the PDS group was the lowest, which did not meet the mechanical requirements of the anulus fibrosus; the tensile and compressive modulus in the PCL group was the highest, and those in the PCL/PDS70/30 and PCL/PDS50/50 group were moderate. Hydrophilicity test showed that higher PDS proportion led to better hydrophilicity. Biodegradation test showed that the biodegradation of pure PDS and PCL/PDS30/70 was too fast, that of PCL was too slow, and that of PCL/PDS70/30 and PCL/PDS50/50 was appropriate. Analysis of inflammatory response around the biodegraded tissue showed that higher proportion of PCL in the scaffold resulted in more severe inflammatory response. CCK-8 and live/dead cell staining showed that human Wharton’s jelly mesenchymal stem cells had good proliferative activity and high survival rate in the PCL/PDS70/30, PCL/PDS50/50, and PCL/PDS30/70 groups. These results suggest that scaffolds in the PCL/PDS70/30 and PCL/PDS50/50 groups can simulate the structure of natural annulus fibrosus, have appropriate biodegradability, excellent mechanical properties and good biocompatibility, which make it a suitable candidate for tissue-engineered annulus fibrosus scaffold.

Key words: polycaprolactone, polydioxanone, anulus fibrosus, bionic structure, scaffold, melt spinning, microfiber, intervertebral disc

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