Manufacture and characterisation of EmDerm-novel hierarchically structured bio-active scaffolds for tissue regeneration
Manufacture and characterisation of EmDerm-novel hierarchically structured bio-active scaffolds for tissue regeneration
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DOI：
10.1007/s10856-018-6060-6
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发表时间：
2018-06-01
影响因子：
3.7
通讯作者：
Dye, Julian F.
中科院分区：
文献类型：
--
作者：
Lim, Xuxin;Potter, Matthew;Dye, Julian F.
使用乳液模板作为一种制造大孔蛋白质支架的方法面临着巨大的挑战。问题包括蛋白质在疏水界面上的吸附变性，乳状液的不稳定性，蛋白质凝胶后的油滴和表面活性物质的去除，以及兼容的交联法。我们研究了一种用表面活性剂混合物稳定的水包油大乳液，作为制备蛋白质基纳米结构生物智能支架(EmDerm)的模板，该支架具有可调的微尺度孔隙率，用于组织再生。采用热凝胶法、纤维蛋白法、酶促凝固法或胶原-纤维蛋白复合法制备Emderm支架原型。孔大小通过表面活性剂与油相的比例来控制。用EDC/NHS进行不同时间的交联化稳定支架。用扫描电子显微镜对支架的微结构和孔隙率进行了表征，用张力仪对支架的力学性能进行了表征。水解性和蛋白水解性降解谱通过质量随时间的减少来量化。以人真皮成纤维细胞、内皮细胞和骨髓间充质干细胞为材料，研究其细胞毒性和细胞增殖情况。Emderm支架具有纳米级的分级结构，平均孔径在40-100微米之间。弹性模量范围为1.1~2.9兆帕，极限抗拉强度为4~16兆帕。降解率与交联时间有关。与标准材料Integra(TM)和Matriderm(TM)相比，每种Emderm支架支持出色的细胞进入和增殖。乳液模板法是一种快速制备微孔纳米纤维蛋白支架的新方法。这些支架在真皮和其他软组织的再生方面具有很好的临床潜力，例如用于烧伤或慢性伤口治疗。
There are significant challenges for using emulsion templating as a method of manufacturing macro-porous protein scaffolds. Issues include protein denaturation by adsorption at hydrophobic interfaces, emulsion instability, oil droplet and surfactant removal after protein gelation, and compatible cross-linking methods. We investigated an oil-in-water macro-emulsion stabilised with a surfactant blend, as a template for manufacturing protein-based nano-structured bio-intelligent scaffolds (EmDerm) with tuneable micro-scale porosity for tissue regeneration. Prototype EmDerm scaffolds were made using either collagen, through thermal gelation, fibrin, through enzymatic coagulation or collagen-fibrin composite. Pore size was controlled via surfactant-to-oil phase ratio. Scaffolds were crosslink-stabilised with EDC/NHS for varying durations. Scaffold micro-architecture and porosity were characterised with SEM, and mechanical properties by tensiometry. Hydrolytic and proteolytic degradation profiles were quantified by mass decrease over time. Human dermal fibroblasts, endothelial cells and bone marrow derived mesenchymal stem cells were used to investigate cytotoxicity and cell proliferation within each scaffold. EmDerm scaffolds showed nano-scale based hierarchical structures, with mean pore diameters ranging from 40-100 microns. The Young's modulus range was 1.1-2.9 MPa, and ultimate tensile strength was 4-16 MPa. Degradation rate was related to cross-linking duration. Each EmDerm scaffold supported excellent cell ingress and proliferation compared to the reference materials Integra (TM) and Matriderm (TM). Emulsion templating is a novel rapid method of fabricating nano-structured fibrous protein scaffolds with micro-scale pore dimensions. These scaffolds hold promising clinical potential for regeneration of the dermis and other soft tissues, e.g., for burns or chronic wound therapies.