中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (14): 2197-2202.doi: 10.3969/j.issn.2095-4344.0769

• 药物控释材料 drug delivery materials • 上一篇    下一篇

脂质体介导温敏型海藻酸/磷酸三钙/胶原Ⅰ水凝胶的性能

谢宜刚1，2，李小丽3，陈昌盛3，廖振华3，刘伟强1，3   

  1. 1清华大学生物医学工程系，北京市 100084；2清华大学深圳研究生院生物医学工程系，广东省深圳市 518055；3深圳清华大学研究院生物医用材料及植入器械重点实验室，广东省深圳市 518057
  • 收稿日期:2017-12-14 出版日期:2018-05-18 发布日期:2018-05-18
  • 通讯作者: 刘伟强，博士，教授，清华大学，北京市 100084；深圳清华大学研究院，广东省深圳市 518055
  • 作者简介:谢宜刚，男，1991年生，汉族，重庆市人，清华大学在读硕士，主要从事可注射骨修复材料的研究。
  • 基金资助:

    国家自然科学基金(51403116)；广东省自然科学基金(2015A030313698)

Properties of a liposomes-mediated thermo-sensitive alginate/alpha-tricalcium phosphate/collagen I composite hydrogel

Xie Yi-gang1, 2, Li Xiao-li3, Chen Chang-sheng3, Liao Zhen-hua3, Liu Wei-qiang1, 3   

  1. 1Department of Biomedical Engineering, Tsinghua University, Beijing 100084, China; 2Department of Biomedical Engineering, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, Guangdong Province, China; 3Key Laboratory of Biomedical Materials and Implant Devices, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057, Guangdong Province, China
  • Received:2017-12-14 Online:2018-05-18 Published:2018-05-18
  • Contact: Liu Wei-qiang, M.D., Professor, Department of Biomedical Engineering, Tsinghua University, Beijing 100084, China; Key Laboratory of Biomedical Materials and Implant Devices, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057, Guangdong Province, China
  • About author:Xie Yi-gang, Master candidate, Department of Biomedical Engineering, Tsinghua University, Beijing 100084, China; Department of Biomedical Engineering, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 51403116; the Natural Science Foundation of Guangdong Province, No. 2015A030313698

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摘要：

文章快速阅读：

 

文题释义：
脂质体：系指将药物包封于类脂质双分子层内而形成的微型泡囊体。脂质体按照所包含类脂质双分子层的层数不同，分为单室脂质体和多室脂质体，小单室脂质体粒径0.02-0.08 μm；大单室脂质体为单层大泡囊，粒径在0.1-1 μm。多层双分子层的泡囊称为多室脂质体，粒径在1-5 μm之间。脂质体按照结构可分单室脂质体、多室脂质体、多囊脂质体，按照电荷可分为中性脂质体、负电荷脂质体、正电荷脂质体、按照性能可分为一般脂质体、特殊功效脂质体。 
海藻酸/α-磷酸三钙/胶原Ⅰ复合体系的温敏性：这种复合体系在35 ℃以下显液态，具有良好的流动性，可用于注射使用；在36 ℃以上，从液态向弹性凝胶转变。


背景：温敏水凝胶具有仿细胞外基质、良好生物相容性、微创可注射和原位成型等特点，在组织工程中广泛使用。
目的：制备脂质体介导的温敏型海藻酸/磷酸三钙/胶原Ⅰ(Alg/TCP/Col)水凝胶，并对其进行表征。
方法：通过交错融合制备包载不同浓度钙离子的脂质体Ca-IFVs，综合考察脂质体包封率和载药率，筛选最佳钙离子浓度的脂质体，用于以下实验。将Alg/TCP/Col前体溶液(海藻酸或海藻酸/磷酸三钙前体溶液)与脂质体Ca-IFVs以不同的配比(5，10，15，20)在37 ℃下混合，制备温敏型水凝胶，观察水凝胶的结构、流变性、体积溶胀比及机械性能。将MC3T3-E1细胞分别接种于Alg/TCP/Col水凝胶、海藻酸水凝胶、海藻酸/磷酸三钙水凝胶表面，接种1，3，7 d后，采用共聚焦显微镜观察细胞形态。
结果与结论：①冻干水凝胶中孔的尺寸在50-100 µm范围，α-磷酸三钙的颗粒均匀地附着在海藻酸/α-磷酸三钙水凝胶的表面上；与海藻酸/α-磷酸三钙水凝胶相比，Alg/TCP/Col水凝胶是具有胶原纤维的致密聚集体；②Alg/TCP/Col水凝胶具有在35-39 ℃之间适当的相转变温度Tm；③随着复合材料凝胶中Ca-IFVs体积的增加，凝胶体积溶胀比增加，当将α-磷酸三钙复合物加入到海藻酸/α-磷酸三钙水凝胶中时，溶胀比略有降低；与海藻酸/α-磷酸三钙水凝胶相比，Alg/TCP/Col水凝胶的溶胀比增加；④在前体溶液与脂质体配比为10时，Alg/TCP/Col水凝胶、海藻酸/α-磷酸三钙水凝胶的压缩模量高于海藻酸水凝胶(P < 0.05)；⑤在前体溶液与脂质体配比为10时，海藻酸水凝胶表面的MC3T3-E1细胞呈圆形；海藻酸/α-磷酸三钙水凝胶中的细胞形态趋于扩展，散布在水凝胶孔表面上；在Alg/TCP/Col水凝胶表面的细胞具有应力延长形态，细胞形态趋势相似，并向凝胶内部延伸，同时数量显著增加；⑥结果表明脂质体介导的Alg/TCP/Col水凝胶，具有良好的机械性能及细胞相容性。
ORCID: 0000-0003-4812-3475(谢宜刚)

关键词: 水凝胶, 组织修复, 温度敏感, 脂质体, 胶原, 国家自然科学基金, 生物材料

Abstract:

BACKGROUND: The thermo-sensitive composite hydrogels have gained increasing interest in bone regeneration domain due to their biomimetic extracellular matrix (ECM) structure, good biocompatibility, minimal invasive performance and in situ molding.

OBJECTIVE: To prepare a thermo-sensitive injectable alginate/α-tricalcium phosphate (α-TCP)/collagen I (Alg/TCP/Col) composite hydrogel and explore its characterization.
METHODS: Ca-carrying interdigitation-fusion vesicles (Ca-IFVs) were prepared. The liposomes carrying the optimal concentration of calcium ions were selected for the following experiments by investigating their encapsulation efficiency and drug loading rate. Alg/TCP/Col precursor solution (Alg or Alg/TCP precursor solution) was mixed with Ca-IFVs at 37 ℃ in different proportions (5, 10, 15, 20) to prepare thermosensitive hydrogels. The structure, rheology behavior, volume swelling ratio, and mechanical properties of the composite hydrogel were observed. MC3T3-E1 cells were co-cultured with Alg/TCP/Col, Alg, and Alg/TCP hydrogels, respectively. Then, morphology of the cells was observed by confocal microscopy at 1, 3, 7 days after co-culture.

RESULTS AND CONCLUSION: (1) The pore size of the freeze-dried hydrogel was 50-100 μm, and TCP particles uniformly adhered to the surface of the Alg/TCP hydrogel surface. The Alg/TCP/Col hydrogel was a dense aggregate with collagen fibers in contrast to the Alg/TCP hydrogel. (2) The Alg/TCP/Col hydrogel exhibited a suitable phase transition temperature (Tm) between 35-39 ℃. (3) The volume swelling ratio of the hydrogel was increased with the increase of Ca-IFVs size. When the α-TCP complex was added into the Alg/TCP hydrogel, the swelling ratio decreased slightly. Alg/TCP/Col hydrogel exhibited a higher swelling ratio than the Alg/TCP hydrogel. (4) When the mixture ratio of precursor solution to liposome was 10, the compressive modulus of Alg/TCP/Col hydrogel and Alg/TCP hydrogel was significantly higher than that of the Alg hydrogel (P < 0.05). (5) When the mixture ratio of precursor solution to liposome was 10, round MC3T3-E1 cells were observed on the Alg hydrogel; the cells on the surface of the Alg/TCP hydrogel were scattered and tended to extend; the cells on the surface of the Alg/TCP/Col hydrogel had a stress-extended morphology, and grew into the hydrogel, and meanwhile, the cell number increased significantly. To conclude, the liposome-mediated Alg/TCP/Col has good mechanical properties and cytocompatibility.

Key words: Hydrogel, Collagen, Liposomes, Calcium phosphates, Tissue Engineering

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