A reproducible, high throughput method for fabricating fibrin gels.
A reproducible, high throughput method for fabricating fibrin gels.
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DOI：
10.1186/1756-0500-5-423
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发表时间：
2012-08-08
影响因子：
1.8
通讯作者：
Leach JK
中科院分区：
文献类型：
--
作者：
Murphy KC;Leach JK
纤维蛋白凝胶是一种很有前途的组织工程生物材料。然而，目前的制备方法是时间密集型的，存在固有的变化。迫切需要开发新的和一致的方法来生产用于检测的纤维蛋白水凝胶。我们开发了一种高通量的方法，使用聚二甲基硅氧烷(PDMS)制造的模具来创建纤维蛋白凝胶。纤维蛋白凝胶是通过将纤维蛋白原和凝血酶的溶液添加到PDMS薄片上的圆柱形缺陷中而制备的。通过移除床单来收集未受干扰的凝胶，并对纤维蛋白凝胶进行表征。通过测量包裹的人骨髓间充质干细胞(MSCs)的压缩硬度和成骨反应，将所得凝胶的特性与公开发表的数据进行比较。凝胶的压缩模数与我们之前报道的制备方法几乎相同。碱性磷酸酶活性是MSCs成骨分化的早期标志，其趋势也与先前的数据一致。这些发现证明了一种简化的纤维蛋白凝胶生产方法，这种方法大大减少了制造纤维蛋白凝胶所需的时间，同时还减少了凝胶批次之间的变异性。这种制造技术为以经济有效的方式产生大量凝胶提供了有价值的工具。
Fibrin gels are a promising biomaterial for tissue engineering. However, current fabrication methods are time intensive with inherent variation. There is a pressing need to develop new and consistent approaches for producing fibrin-based hydrogels for examination. We developed a high throughput method for creating fibrin gels using molds fabricated from polydimethylsiloxane (PDMS). Fibrin gels were produced by adding solutions of fibrinogen and thrombin to cylindrical defects in a PDMS sheet. Undisturbed gels were collected by removing the sheet, and fibrin gels were characterized. The characteristics of resulting gels were compared to published data by measuring compressive stiffness and osteogenic response of entrapped human mesenchymal stem cells (MSCs). Gels exhibited compressive moduli nearly identical to our previously reported fabrication method. Trends in alkaline phosphatase activity, an early marker of osteogenic differentiation in MSCs, were also consistent with previous data. These findings demonstrate a streamlined approach to fibrin gel production that drastically reduces the time required to make fibrin gels, while also reducing variability between gel batches. This fabrication technique provides a valuable tool for generating large numbers of gels in a cost-effective manner.