Acellular Tissue Matrix for Bioprosthetic Corneas

用于生物假体角膜的脱细胞组织基质

基本信息

项目摘要

DESCRIPTION (provided by applicant): Restricted availability of donor tissue limits access to corneal transplantation for millions of individuals worldwide. Efforts are under way to produce artificial keratoprostheses and bio-engineered corneas for transplantation, however, currently there is no ideal material to serve as stroma. Synthetic substrates do not integrate well into the ocular environment and artificial collagen gels lack adequate strength and transparency. The long term goals of the project are to develop prosthetic cornea assembled from living corneal cells. This initial phase tests the hypothesis that acellular non-human corneal stromal matrix will provide an effective scaffolding on which a corneal equivalent can be assembled invitro. The project will identify decellularization and extraction procedures for bovine stromal tissue that eliminates cellular debris, maximize tissue repopulation by keratocytes, and supports expression of keratocyte differentiated function. Three specific aims will test these properties of stromal matrix in increasingly complex biological environments: (1) Examine the influence of acellular stromal matrix prepared by various procedures in promotion of repopulation and differentiated function of keratocytes in vitro. Corneas will be decellularized by a combination of freezing, hypotonic lysis, enzyme digestion, and detergent extraction. Associated tissue antigens and growth factors will be removed by a variety of extraction procedures. These decellularized tissues will then be used as culture matrix for primary and passaged keratocytes. Expression of markers of keratocyte differentiated phenotype will be examined. (2) Assess the role of epithelial and endothelial cells in the determining the differentiated functions of keratocytes in acellular stromal matrix. Matrix populated with keratocytes will be assembled into a three dimensional corneal equivalent with primary epithelial and endothelial cells. Stromal deturgescence, epithelial differentiation and basal lamina formation will be examined in addition to keratocyte function. (3) Compare the re-cellularization, transparency, and tissue rejection of acellular stromal matrix preparations in interlamellar grafts in vivo in mice. Acellular matrix successful in previous procedures will be transplanted in a mouse interlamellar model. The transplants will be examined for tissue rejection, recellularization, and transparency. These three aims will establish the suitability of stromal matrix as a scaffold for prosthetic corneas and determine the most effective methods for its preparation.
描述(由申请人提供):供体组织的有限可用性限制了全球数百万人接受角膜移植。人们正在努力生产用于移植的人工角膜假体和生物工程角膜,然而,目前还没有理想的材料用作基质。合成基质不能很好地整合到眼部环境中,并且人工胶原蛋白凝胶缺乏足够的强度和透明度。该项目的长期目标是开发由活角膜细胞组装的人工角膜。这个初始阶段测试的假设,脱细胞的非人角膜基质基质将提供一个有效的支架上,角膜等同物可以体外组装。该项目将确定牛基质组织的脱细胞和提取程序,以消除细胞碎片,最大限度地增加角膜细胞的组织再增殖,并支持角膜细胞分化功能的表达。本研究的主要目的是在日益复杂的生物学环境中检测基质基质的这些特性:(1)检测不同方法制备的脱细胞基质对体外培养的角膜基质细胞增殖和分化的影响。将通过冷冻、低渗裂解、酶消化和去污剂提取的组合对角膜进行脱细胞。 相关的组织抗原和生长因子将通过各种提取程序去除。 然后将这些脱细胞组织用作原代和传代角膜细胞的培养基质。 将检查角膜细胞分化表型标志物的表达。 (2)评估上皮细胞和内皮细胞在决定细胞分化功能中的作用。 脱细胞基质基质中的角膜基质细胞。 填充有角膜细胞的基质将组装成具有原代上皮细胞和内皮细胞的三维角膜等同物。 除了角膜细胞功能外,还将检查基质消肿、上皮分化和基底层形成。(3)比较脱细胞基质基质制备物在小鼠体内板层间移植物中的再细胞化、透明性和组织排斥。 在之前的手术中成功的脱细胞基质将被移植到小鼠层间模型中。将检查移植物的组织排斥、再细胞化和透明度。 这三个目标将确定基质基质作为人工角膜支架的适用性,并确定最有效的制备方法。

项目成果

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JAMES L FUNDERBURGH其他文献

JAMES L FUNDERBURGH的其他文献

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{{ truncateString('JAMES L FUNDERBURGH', 18)}}的其他基金

Bioengineering Cornea with Autologous Stem Cells
自体干细胞生物工程角膜
  • 批准号:
    9113712
  • 财政年份:
    2016
  • 资助金额:
    $ 14.85万
  • 项目类别:
A Pluripotent Stem Cell Reagent for Corneal Regeneration
用于角膜再生的多能干细胞试剂
  • 批准号:
    8626779
  • 财政年份:
    2013
  • 资助金额:
    $ 14.85万
  • 项目类别:
A Pluripotent Stem Cell Reagent for Corneal Regeneration
用于角膜再生的多能干细胞试剂
  • 批准号:
    8776715
  • 财政年份:
    2013
  • 资助金额:
    $ 14.85万
  • 项目类别:
Stem Cells for Corneal Engineering
角膜工程干细胞
  • 批准号:
    7221869
  • 财政年份:
    2006
  • 资助金额:
    $ 14.85万
  • 项目类别:
Stem Cells for Corneal Engineering
角膜工程干细胞
  • 批准号:
    7101389
  • 财政年份:
    2006
  • 资助金额:
    $ 14.85万
  • 项目类别:
Stem Cells for Corneal Engineering
角膜工程干细胞
  • 批准号:
    9212811
  • 财政年份:
    2006
  • 资助金额:
    $ 14.85万
  • 项目类别:
Stem Cells for Corneal Engineering
角膜工程干细胞
  • 批准号:
    8655870
  • 财政年份:
    2006
  • 资助金额:
    $ 14.85万
  • 项目类别:
Stem Cells for Corneal Engineering
角膜工程干细胞
  • 批准号:
    8245697
  • 财政年份:
    2006
  • 资助金额:
    $ 14.85万
  • 项目类别:
Stem Cells for Corneal Engineering
角膜工程干细胞
  • 批准号:
    7613878
  • 财政年份:
    2006
  • 资助金额:
    $ 14.85万
  • 项目类别:
Stem Cells for Corneal Engineering
角膜工程干细胞
  • 批准号:
    7394345
  • 财政年份:
    2006
  • 资助金额:
    $ 14.85万
  • 项目类别:
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