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用于药物释放和三维细胞培养的可调谐重组蛛丝蛋白水凝胶

Tuneable Recombinant Spider Silk Protein Hydrogels for Drug Release and 3D Cell Culture.

作者信息

Arndt Tina, Chatterjee Urmimala, Shilkova Olga, Francis Juanita, Lundkvist Johan, Johansson Daniel, Schmuck Benjamin, Greco Gabriele, Nordberg Åsa Ekblad, Li Yan, Wahlberg Lars U, Langton Maud, Johansson Jan, Götherström Cecilia, Rising Anna

机构信息

Department of Biosciences and Nutrition Karolinska Institutet Neo Huddinge 14152 Sweden.

Sinfonia Biotherapeutics Hälsovägen 7 Huddinge 14157 Sweden.

出版信息

Adv Funct Mater. 2024 Aug 28;34(35):2303622. doi: 10.1002/adfm.202303622. Epub 2023 May 26.

Abstract

Hydrogels are useful drug release systems and tissue engineering scaffolds. However, synthetic hydrogels often require harsh gelation conditions and can contain toxic by-products while naturally derived hydrogels can transmit pathogens and in general have poor mechanical properties. Thus, there is a need for a hydrogel that forms under ambient conditions, is non-toxic, xeno-free, and has good mechanical properties. A recombinant spider silk protein-derived hydrogel that rapidly forms at 37 °C is recently developed. The temperature and gelation times are well-suited for an injectable in situ polymerising hydrogel, as well as a 3D cell culture scaffold. Here, it is shown that the diffusion rate and the mechanical properties can be tuned by changing the protein concentration and that human fetal mesenchymal stem cells encapsulated in the hydrogels show high survival and viability. Furthermore, mixtures of recombinant spider silk proteins and green fluorescent protein (GFP) form gels from which functional GFP is gradually released, indicating that bioactive molecules are easily included in the gels, maintain activity and can diffuse through the gel. Interestingly, encapsulated ARPE-19 cells are viable and continuously produce the growth factor progranulin, which is detected in the cell culture medium over the study period of 31 days.

摘要

水凝胶是有用的药物释放系统和组织工程支架。然而,合成水凝胶通常需要苛刻的凝胶化条件,并且可能含有有毒副产物,而天然衍生的水凝胶可能传播病原体且通常机械性能较差。因此,需要一种在环境条件下形成、无毒、无动物源且具有良好机械性能的水凝胶。最近开发了一种由重组蜘蛛丝蛋白衍生的水凝胶,它在37°C时能快速形成。该温度和凝胶化时间非常适合用于可注射的原位聚合水凝胶以及3D细胞培养支架。在此表明,通过改变蛋白质浓度可以调节扩散速率和机械性能,并且封装在水凝胶中的人胎儿间充质干细胞具有高存活率和活力。此外,重组蜘蛛丝蛋白和绿色荧光蛋白(GFP)的混合物形成凝胶,功能性GFP从该凝胶中逐渐释放,这表明生物活性分子很容易包含在凝胶中,保持活性并能扩散通过凝胶。有趣的是,封装的ARPE - 19细胞具有活力并持续产生生长因子前颗粒蛋白,在31天的研究期间可在细胞培养基中检测到该蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dadc/11440629/15ae51bfa7d7/ADFM-34-0-g002.jpg

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