口腔疾病防治 ›› 2021, Vol. 29 ›› Issue (10): 669-676.DOI: 10.12016/j.issn.2096-1456.2021.10.004

• 基础研究 • 上一篇    下一篇

细胞膜片复合3D打印马鹿角粉/丝素蛋白/聚乙烯醇支架对羊下颌骨缺损的修复效果

张凯(),刘小元,李蕾,李君,韩祥祯,何惠宇()   

  1. 新疆医科大学第一附属医院(附属口腔医院)口腔修复科·新疆维吾尔自治区口腔医学研究所,新疆维吾尔自治区 乌鲁木齐(830054)
  • 收稿日期:2021-03-01 修回日期:2021-04-28 出版日期:2021-10-20 发布日期:2021-06-23
  • 通讯作者: 何惠宇
  • 作者简介:张凯,硕士研究生,Email: 442107538@qq.com
  • 基金资助:
    新疆维吾尔自治区科技支疆项目(2018E02060)

Effect of cell sheet combined with 3D printing an antler powder/silk fibroin/polyvinyl alcohol scaffold on the repair of mandibular defects in sheep

ZHANG Kai(),LIU Xiaoyuan,LI Lei,LI Jun,HAN Xiangzhen,HE Huiyu()   

  1. Xinjiang Uygur Autonomous Region Institute of Stomatology, Department of Prosthodontics, the First Affiliated Hospital(Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, China
  • Received:2021-03-01 Revised:2021-04-28 Online:2021-10-20 Published:2021-06-23
  • Contact: Huiyu HE
  • Supported by:
    Xinjiang Uygur Autonomous Region Science and Technology Support Project(2018E02060)

摘要:

目的 探讨细胞膜片复合马鹿角粉/丝素蛋白/聚乙烯醇支架修复下颌骨极限性骨缺损的效果。方法 通过3D打印制备马鹿角粉/丝素蛋白/聚乙烯醇支架及纳米级羟基磷灰石/丝素蛋白/聚乙烯醇支架,采用全骨髓培养法制备阿勒泰大尾羊髂骨骨髓血细胞膜片;按观察时间1、2、3个月分别建立阿勒泰大尾羊双侧下颌骨极限性缺损模型4只,实验组为细胞膜片包裹马鹿角粉/丝素蛋白/聚乙烯醇支架,对照组为细胞膜片包裹纳米级羟基磷灰石/丝素蛋白/聚乙烯醇支架,阴性对照组为细胞膜片包裹无支架的凝胶海绵;按动物双侧下颌骨缺损区自身对照的方法分别植入包裹细胞膜的支架或包裹细胞膜的凝胶海绵。在支架材料植入后第1、2、3个月末处死实验动物,拍摄锥形束CT(cone beam CT,CBCT)、取组织行石蜡切片HE染色观察,评价不同支架材料的骨再生效果与支架的降解。结果 扫描电镜结果显示:两组支架孔隙规则,连续性好,孔径及孔隙率比较均无差别(P > 0.05)。CBCT结果显示,术后3个月实验组对骨缺损的修复效果明显优于对照组,且支架降解速率与骨组织修复速率较为匹配,缺损中央骨密度高于对照组,与正常骨组织接近。实验组在各时间点的骨缺损区中央骨密度计量均高于对照组、阴性对照组,且差异具有统计学意义(P<0.05)。HE染色结果显示:实验组成骨细胞较为活跃,有较多新生毛细血管及骨小梁形成,同时支架材料的吸收多于对照组。 结论 细胞膜片复合马鹿角粉/丝素蛋白/聚乙烯醇支架能够促进临界骨缺损的修复,其降解性与骨组织愈合速率较为匹配,有望成为修复骨缺损的新方法。

关键词: 马鹿角粉, 丝素蛋白, 聚乙烯醇, 羟基磷灰石, 组织工程骨, 细胞膜片, 3D打印, 支架材料, 降解, 成骨, 骨缺损

Abstract:

Objective To explore the feasibility of antler powder/silk fibroin/polyvinyl alcohol scaffolds as tissue engineering bone scaffolds and the relationship between their degradation performance and the healing speed of bone defects. Methods Antler powder/silk fibroin/polyvinyl alcohol scaffolds and nano hydroxyapatite/silk fibroin/polyvinyl alcohol scaffolds were prepared by 3D printing. The whole bone marrow culture method was used to prepare blood cell sheets of Altay big tail sheep’s iliac bone marrow. With observation times of 1, 2 and 3 months, the mandibular defects of 4 sheep were established. The experimental group was coated with antler powder/silk fibroin/polyvinyl alcohol scaffolds. The control group was coated with nanohydroxyapatite/silk fibroin/polyvinyl alcohol scaffolds. The negative control group was coated with gel-free sponges. According to the self-control method of the bilateral mandible defect area, scaffolds wrapped with cell membranes or gel sponges wrapped with cell membranes were implanted. At the ends of the first, second and third months after implantation, the experimental animals were killed, cone beam CT was performed, and paraffin sections were taken for HE staining to evaluate the effect of different scaffold materials on bone regeneration and scaffold degradation. Results Scanning electron microscopy showed that both groups had regular pores and good continuity, and there was no difference in pore size and porosity between the two groups (P > 0.05). The results of CBCT imaging showed that in 3 months after operation, the experimental group had significantly better repair effects on bone defects than the control group, and the degradation rate matched the bone repair rate. The bone mineral density in the center of the defect was higher than that of the control group, which was close to that of normal bone tissue. The central bone mineral density of the experimental group at each time point was higher than those of the control group and the negative control group, and the difference was statistically significant (P < 0.05). HE staining results showed that the bone cells in the experimental group were more active, with more new capillaries and bone trabeculae formed, and the scaffold material absorbed more than the control group. Conclusion The antler powder/silk fibroin/polyvinyl alcohol scaffold can promote the repair of critical bone defects. Its degradability matches its bone tissue healing rate. It is expected to become a promising scaffold material for bone tissue engineering.

Key words: antler powder, Silk fibroin, Polyvinyl alcohol, hydroxyapatite, tissue engineered bone, cell sheet, 3D print, scaffolds, degradation, osteogenesis, bone defect

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