Avidin-Conjugated Nanofibrillar Cellulose Hydrogel Functionalized with Biotinylated Fibronectin and Vitronectin Promotes 3D Culture of Fibroblasts,Biomacromolecules

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Avidin-Conjugated Nanofibrillar Cellulose Hydrogel Functionalized with Biotinylated Fibronectin and Vitronectin Promotes 3D Culture of Fibroblasts
Biomacromolecules ( IF 5.5 ) Pub Date : 2021-09-20 , DOI: 10.1021/acs.biomac.1c00579
Jenni Leppiniemi ₁ , Zeeshan Mutahir _{1,

2} , Alexander Dulebo ₃ , Piia Mikkonen ₄ , Markus Nuopponen ₄ , Paula Turkki _{1,

5} , Vesa P Hytönen _{1,

5}

Affiliation

The future success of physiologically relevant three-dimensional (3D) cell/tissue models is dependent on the development of functional biomaterials, which can provide a well-defined 3D environment instructing cellular behavior. To establish a platform to produce tailored hydrogels, we conjugated avidin (Avd) to anionic nanofibrillar cellulose (aNFC) and demonstrated the use of the resulting Avd-NFC hydrogel for 3D cell culture, where Avd-NFC allows easy functionalization via biotinylated molecules. Avidin was successfully conjugated to nanocellulose and remained functional, as demonstrated by electrophoresis and titration with fluorescent biotin. Rheological analysis indicated that Avd-NFC retained shear-thinning and gel-forming properties. Topological characterization using AFM revealed the preserved fiber structure and confirmed the binding of biotinylated vitronectin (B-VN) on the fiber surface. The 3D cell culture experiments with mouse embryonic fibroblasts demonstrated the performance of Avd-NFC hydrogels functionalized with biotinylated fibronectin (B-FN) and B-VN. Cells cultured in Avd-NFC hydrogels functionalized with B-FN or B-VN formed matured integrin-mediated adhesions, indicated by phosphorylated focal adhesion kinase. We observed significantly higher cell proliferation rates when biotinylated proteins were bound to the Avd-NFC hydrogel compared to cells cultured in Avd-NFC alone, indicating the importance of the presence of adhesive sites for fibroblasts. The versatile Avd-NFC allows the easy functionalization of hydrogels with virtually any biotinylated molecule and may become widely utilized in 3D cell/tissue culture applications.

中文翻译：

生物素化纤连蛋白和玻连蛋白功能化的抗生物素蛋白偶联纳米原纤维纤维素水凝胶促进成纤维细胞的 3D 培养

与生理相关的三维 (3D) 细胞/组织模型的未来成功取决于功能生物材料的开发，它可以提供一个明确定义的 3D 环境来指导细胞行为。为了建立一个生产定制水凝胶的平台，我们将抗生物素蛋白 (Avd) 与阴离子纳米原纤维纤维素 (aNFC) 结合，并展示了所得 Avd-NFC 水凝胶用于 3D 细胞培养的用途，其中 Avd-NFC 允许通过以下方式轻松功能化生物素化分子。亲和素成功地与纳米纤维素结合并保持功能，如电泳和荧光生物素滴定所示。流变分析表明，Avd-NFC 保留了剪切稀化和凝胶形成特性。使用 AFM 的拓扑表征揭示了保留的纤维结构，并证实了生物素化玻连蛋白 (B-VN) 在纤维表面的结合。小鼠胚胎成纤维细胞的 3D 细胞培养实验证明了用生物素化纤连蛋白 (B-FN) 和 B-VN 功能化的 Avd-NFC 水凝胶的性能。在用 B-FN 或 B-VN 功能化的 Avd-NFC 水凝胶中培养的细胞形成成熟的整合素介导的粘附，由磷酸化粘着斑激酶指示。我们观察到，与单独在 Avd-NFC 中培养的细胞相比，当生物素化蛋白与 Avd-NFC 水凝胶结合时，细胞增殖率明显更高，这表明存在粘附位点对成纤维细胞的重要性。多功能 Avd-NFC 允许使用几乎任何生物素化分子轻松功能化水凝胶，并可能广泛用于 3D 细胞/组织培养应用。

更新日期：2021-10-12

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