A biodegradable, acellular xenogeneic scaffold for regeneration of the vocal fold lamina propria
A biodegradable, acellular xenogeneic scaffold for regeneration of the vocal fold lamina propria
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DOI：
10.1089/ten.2006.0169
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发表时间：
2007-03-01
影响因子：
--
通讯作者：
Tirunagari, Neeraj
中科院分区：
文献类型：
--
作者：
Xu, Chet C.;Chan, Roger W.;Tirunagari, Neeraj
开发了一种制备用于组织工程的无细胞异种细胞外基质支架的新方法。牛声带固有层标本经高浓度氯化钠、核酸消化、乙醇脱水处理，脱细胞并去除免疫原性外来表位。将原代培养的人声带成纤维细胞接种到无细胞支架上并培养 21 天。通过光学显微镜、荧光显微镜和扫描电子显微镜检查脱细胞和再细胞化支架。成纤维细胞的胶原合成和释放通过 Sircol 测定进行定量，而透明质酸、核心蛋白聚糖和纤连蛋白的合成和释放通过酶联免疫吸附测定进行评估。支架的粘弹性剪切特性通过单剪切流变仪在高达 250 Hz 的频率下进行量化。初步结果表明，设计了一种可生物降解的、无细胞的细胞外基质支架，具有完整的基底膜和基质蛋白的三维结构。声带成纤维细胞容易附着并渗透支架，具有高活力和活性蛋白质合成，证明了生物相容性。无细胞支架和成纤维细胞再生支架的弹性剪切模量和动态粘度与人声带覆盖物相当。这些发现支持支架作为异种移植物用于声带重建和再生的潜力。
A novel method for preparing an acellular xenogeneic extracellular matrix scaffold for tissue engineering was developed. Bovine vocal fold lamina propria specimens were treated with high-concentration sodium chloride, nucleic acid digestion, and ethanol dehydration for decellularization and removal of immunogenic foreign epitopes. Human vocal fold fibroblasts from primary culture were seeded onto the acellular scaffolds and cultured for 21 days. The decellularized and the recellularized scaffolds were examined by light microscopy, fluorescent microscopy, and scanning electron microscopy. Collagen synthesis and release by fibroblasts were quantified by the Sircol assay, whereas the synthesis and release of hyaluronic acid, decorin, and fibronectin were assessed by enzyme-linked immunosorbent assays. Viscoelastic shear properties of the scaffolds were quantified by a simple-shear rheometer at frequencies of up to 250 Hz. Preliminary results showed that a biodegradable, acellular extracellular matrix scaffold with an intact basement membrane and 3-dimensional structure of the matrix proteins was engineered. Vocal fold fibroblasts readily attached to and infiltrated the scaffold with high viability and active protein synthesis, demonstrating the biocompatibility. The elastic shear modulus and dynamic viscosity of the acellular scaffold and the fibroblast-repopulated scaffold were comparable to those of the human vocal fold cover. These findings support the potential of the scaffold as a xenograft for vocal fold reconstruction and regeneration.