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用于肾小球组织工程的一种独特弹性中链长度聚羟基脂肪酸酯的生物合成、表征及生物相容性

Biosynthesis, characterisation and biocompatibility of a unique and elastomeric medium chain-length polyhydroxyalkanoates for kidney glomerular tissue engineering.

作者信息

Syed Mohamed Syed Mohammad Daniel, Tuffin Jack, Watson Judy, Mele Andrea, Fricker Annabelle, Gregory David A, Elghazy Elbaraa, Saleem Moin A, Satchell Simon C, Welsh Gavin I, Roy Ipsita

机构信息

School of Chemical, Materials & Biological Engineering, Faculty of Engineering, University of Sheffield, Sheffield, S37HQ, UK.

PHAsT Limited, Sheffield, S1 4DP, UK.

出版信息

Mater Today Bio. 2025 May 31;33:101932. doi: 10.1016/j.mtbio.2025.101932. eCollection 2025 Aug.

Abstract

Polyhydroxyalkanoates (PHAs) are bacteria-derived polymers that are being actively explored for their potential in biomedical engineering applications. These polymers are not only highly biocompatible in nature but also sustainable, produced using renewable substrates, and hence considered future biomaterials. In addition to normal fermentation, PHAs can also be produced through a synthetic biology approach. This study explores a medium chain-length PHA (mcl-PHA) produced by CH50 by batch fermentation, fed with glucose as the sole carbon source. The polymer was extensively characterised, and it exhibited an elastomeric property of a typical mcl-PHA with 215 ± 52 % elongation at break. The mcl-PHA also had a low melting point, , of around 55 °C, making it processable with various fabrication methods. The extracted mcl-PHA was prepared as a solvent-cast film and tested as a potential cell culture substrate for human glomerular cells, the conditionally immortalised human podocytes (CiHP) and conditionally immortalised human glomerular endothelial cells (CiGEnC). Initial resazurin assay under proliferative conditions showed promising cell metabolic activities of the cells cultured on the mcl-PHA film, comparable with those cultured on tissue culture plastic (TCP). Despite the decreased expression of collagen IV under proliferative conditions, the differentiated co-cultured cells on mcl-PHA had comparable values with cells those grown on TCP. These promising results verified the biocompatibility of the mcl-PHA produced by CH50 and established its potential as a bio-based sustainable alternative in biomedical applications including glomerular tissue engineering.

摘要

聚羟基脂肪酸酯(PHA)是细菌衍生的聚合物,目前正被积极探索其在生物医学工程应用中的潜力。这些聚合物不仅本质上具有高度生物相容性,而且可持续,使用可再生底物生产,因此被视为未来的生物材料。除了常规发酵外,PHA还可以通过合成生物学方法生产。本研究探索了CH50通过分批发酵生产的中链长度PHA(mcl-PHA),以葡萄糖作为唯一碳源进行补料。对该聚合物进行了广泛表征,其表现出典型mcl-PHA的弹性体特性,断裂伸长率为215±52%。该mcl-PHA的熔点也较低,约为55℃,这使得它可以用各种制造方法进行加工。提取的mcl-PHA被制备成溶剂浇铸膜,并作为人肾小球细胞、条件永生化人足细胞(CiHP)和条件永生化人肾小球内皮细胞(CiGEnC)的潜在细胞培养底物进行测试。增殖条件下的初始刃天青测定显示,在mcl-PHA膜上培养的细胞具有良好的细胞代谢活性,与在组织培养塑料(TCP)上培养的细胞相当。尽管在增殖条件下IV型胶原的表达有所下降,但在mcl-PHA上分化的共培养细胞与在TCP上生长的细胞具有相当的值。这些有前景的结果证实了CH50生产的mcl-PHA的生物相容性,并确立了其作为生物基可持续替代品在包括肾小球组织工程在内的生物医学应用中的潜力。

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