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Structural and functional considerations in the design of collagen-based electrospun scaffolds.

机译：在设计基于胶原蛋白的电纺支架时的结构和功能考虑。

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摘要

Electrospinning can be used to selectively process a variety of natural and synthetic polymers into highly porous scaffolds composed of nano-to-micron diameter fibers. This process shows great potential as a gateway to the development of physiologically relevant tissue engineering scaffolds. In this study we examine the structural and functional considerations regarding electrospun scaffolds for dermal template applications using novel quantification techniques. In order to characterize scaffold structure, a technique utilizing the fast Fourier transform was developed to systematically quantify fiber alignment and evaluate how different electrospinning parameters impact the structure and material properties of an electrospun scaffold. Gelatin was suspended at varying concentrations (80, 100, 130 and 150 mg/ml) and electrospun from 2,2,2 trifluoroethanol onto a rotating mandrel (200-7000 RPM). Scaffold anisotropy developed as a function of fiber diameter and mandrel speed and the induction of varying degrees of anisotropy imparted distinctive material properties to the electrospun scaffolds. Fiber alignment was the variable most closely associated with the regulation of peak stress, peak strain and modulus of elasticity. Next, we examined how the chemical and physical composition of the local microenvironment and the unmasking of possible RGD sensitive binding sites through collagen denaturation, independent of scaffold architecture and porosity, impacts cellular processes. We cultured human dermal fibroblasts on electrospun nylon coated with a variety of non-denatured and thermally denatured collagen-based proteins, as well as recovered electrospun collagen and gelatin (in an effort to examine if the electrospinning process degrades the collagen alpha chain). Differences in adhesion, proliferation and migration were exhibited between collagen-based proteins. Adhesion inhibition assays using a cyclic RGD peptide demonstrated no change in cell adhesion on non-denatured proteins and a significant drop in cell adhesion on thermally denatured proteins. Based on gel analysis and the results of our functional assays we conclude that collagen a chain structure is not directly altered by the electrospinning process. Overall, these results are critical to the understanding of how structure and architecture contribute to the overall properties of a scaffold, as well as how molecular variations can modulate scaffold functionality in a cellular environment.
机译：静电纺丝可用于选择性地将各种天然和合成聚合物加工成由纳米至微米直径的纤维组成的高度多孔的支架。该过程显示出巨大的潜力，可作为开发生理相关组织工程支架的途径。在这项研究中，我们使用新颖的定量技术研究了用于真皮模板应用的电纺支架的结构和功能方面的考虑。为了表征支架结构，开发了一种利用快速傅立叶变换的技术，以系统地量化纤维排列并评估不同的电纺参数如何影响电纺支架的结构和材料性能。将明胶以不同的浓度（80、100、130和150 mg / ml）悬浮，并从2,2,2三氟乙醇中静电纺丝到旋转的心轴上（200-7000 RPM）。支架各向异性随纤维直径和芯轴速度的变化而发展，并且不同程度的各向异性的诱导为电纺支架赋予了独特的材料特性。纤维排列是与峰值应力，峰值应变和弹性模量的调节最紧密相关的变量。接下来，我们研究了局部微环境的化学和物理组成以及通过胶原蛋白变性（与支架结构和孔隙率无关）对可能的RGD敏感结合位点的揭示如何影响细胞过程。我们在涂有多种非变性和热变性胶原蛋白的静电纺尼龙上培养了人类皮肤成纤维细胞，以及回收的静电纺胶原和明胶（以检查静电纺丝过程是否降解胶原α链）。胶原蛋白之间存在粘附，增殖和迁移的差异。使用环状RGD肽的粘附抑制试验表明，非变性蛋白的细胞粘附没有改变，而热变性蛋白的细胞粘附则明显下降。根据凝胶分析和我们的功能测定结果，我们得出结论，静电纺丝过程不会直接改变胶原蛋白的链结构。总体而言，这些结果对于理解结构和体系结构如何对支架的整体特性以及分子变异如何调节细胞环境中的支架功能至关重要。

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  • 作者

    Ayres, Chantal Emma.;

  • 作者单位

    Virginia Commonwealth University.;

  • 授予单位 Virginia Commonwealth University.;
  • 学科 Engineering Biomedical.
  • 学位 Ph.D.
  • 年度 2009
  • 页码 242 p.
  • 总页数 242
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物医学工程;
  • 关键词

  • 入库时间 2022-08-17 11:38:26

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