天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (4): 337-340.doi: 10.11958/20180103

• 细胞与分子生物学 •    下一篇

聚己内酯/海藻酸钠/壳聚糖材料制备 组织工程椎间盘双相支架

李麒峰 1，2，徐宝山 2△，杨强 2，马信龙 2，张杨 2，郭悦 2，杨阳 2，张凯辉 1，2，夏金健 1，2，张维昊 1，2   

  1. 基金项目：国家自然科学基金资助项目（31670983，81272046）；天津市自然科学基金项目（15JCYBJC25300）；天津市卫生局攻关课题 （14KG121） 作者单位：1天津医科大学研究生院（邮编300070）；2天津市天津医院 作者简介：李麒峰（1989），男，硕士在读，主要从事脊柱外科、组织工程椎间盘研究 △通讯作者 E-mail: xubaoshan99@126.com
  • 收稿日期:2018-01-17 修回日期:2018-03-03 出版日期:2018-04-15 发布日期:2018-04-15
  • 通讯作者: 李麒峰 E-mail:774099942@qq.com
  • 基金资助:
    仿生斜交叠层结构的纳米PCL/SF纤维环支架复合人脐带间充质干细胞修复纤维 环缺损;动态力学刺激对纤维环-髓核组织工程化的影响;明胶海绵和丝素复合自体骨髓干细胞修复椎间盘手术缺损;明胶海绵和丝素复合自体骨髓干细胞修复椎间盘手术缺损

Tissue-engineered intervertebral disc biphasic scaffolds prepared by poly (ɛ-caprolactone)/sodium alginate/chitosan materials

LI Qi-feng1,2, XU Bao-shan2△, YANG Qiang2, MA Xin-long2, ZHANG Yang2, GUO Yue2, YANG Yang2, ZHANG Kai-hui1,2, XIA Jin-jian1,2, ZHANG Wei-hao1,2   

  1. 1 Graduate School of Tianjin Medical University, Tianjin 300070, China; 2 Tianjin Hospital △Corresponding Author E-mail: xubaoshan99@126.com
  • Received:2018-01-17 Revised:2018-03-03 Published:2018-04-15 Online:2018-04-15
  • Contact: Qi-Feng LI E-mail:774099942@qq.com
  • Supported by:
    Engineered nanofibrous bionic angle-ply laminate PCL/SF scaffold and hWJ-MSCs for the repair of annulus fibrosus defect

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摘要： 摘要：目的 以聚己内酯（PCL）、海藻酸钠、壳聚糖为材料，研制椎间盘双相支架，并评估其作为组织工程椎间盘 的可行性。方法 聚己内酯作为原料，采用熔融电纺法制备取向性多孔纤维环支架，将海藻酸钠/壳聚糖水凝胶注入 到中空的纤维环（AF）支架中央合成双相椎间盘支架。通过体式显微镜、扫描电镜观测双相支架的结构、孔径、孔隙 率；人脐带干细胞复合双相支架体外培养7 d，用死活细胞染色法评价生物相容性，CCK-8实验测定细胞增殖情况，力 学加载仪器测量双相支架的压缩弹性模量。结果 体式显微镜和扫描电镜可见纤维环相成菱形多孔结构，髓核相 （NP）呈不规则多孔结构；纤维环相和髓核相孔径分别为（225.6±3.9）μm、（205.5±5.2）μm，孔隙率分别为（74.17± 0.39）%、（85.52±0.48）%，支架扫描电镜可见细胞黏附在支架表面，周围有细胞外基质分泌；死活细胞染色显示无死 细胞；CCK-8检测结果显示人脐带干细胞具有良好的增殖活性，压缩弹性模量（173.24±44.93）kPa。结论 以聚己内 酯、海藻酸钠、壳聚糖为材料制备的椎间盘双相支架，具有良好的孔径、孔隙率和细胞相容性，支架间结合紧密，具有 三维网络结构，优良的力学特性，是构建组织工程椎间盘理想载体。

关键词: 组织工程, 椎间盘, 聚己内酯, 海藻酸钠, 壳聚糖

Abstract: Abstract: Objective To evaluate the feasibility of a tissue-engineered novel intervertebral disc biphasic scaffold, which used poly (ɛ-caprolactone) (PCL), sodium alginate and chitosan as materials. Methods PCL was used as material of melt spinning preparation of oriented porous annular fibrosus (AF) scaffolds, and the hydrogel was injected into hollow AF central composite biphasic scaffolds. The size and porosity of the biphasic scaffolds were determined by stereomicroscope and scanning electron microscopy (SEM). The human umbilical cord stem cells were inoculated to the corresponding sites of scaffolds and cultured in vitro for 7 days. CCK-8 assay was used to assess cell proliferation. The compression modulus of the biphasic stent was measured. Results Stereomicroscope and SEM showed that the AF formed diamond-shaped porous structure, nucleus pulposus (NP) phase formed irregularity porous structure. AF and NP phase pore sizes were (225.6±3.9) μm and (205.5±5.2) μm respectively, and porosity rates were (74.17±0.39)% and (85.52±0.48)% respectively. SEM showed that cell adhesion to the rack surface and surrounding cells were secreted by extracellular matrix. Live/dead cell staining showed that the cells showed good activity on the scaffold without dead cells. CCK-8 proliferation assay showed that human umbilical cord stem cells showed good proliferative activity. In addition, biphasic scaffold showed impressive mechanical properties (173.24 ± 44.93) kPa. Conclusion The biomimetic biphasic disc stents prepared from polycaprolactone and hydrogels have good pore size, porosity and cell compatibility. Those materials’scaffolds are closely integrated with each other, have a three-dimensional good network structure, and are excellent mechanical properties. So it is an ideal vehicle for constructing intervertebral disc of tissue engineering.

Key words: tissue engineering, intervertebral disk, poly(?-caprolactone), sodium alginate, chitosan