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可生物降解的聚乳酸-乙醇酸共聚物支架作为基因修饰成纤维细胞的载体

Biodegradable poly (lactic acid-co-glycolic acid) scaffolds as carriers for genetically-modified fibroblasts.

作者信息

Perisic Tatjana, Zhang Ziyang, Foehr Peter, Hopfner Ursula, Klutz Kathrin, Burgkart Rainer H, Slobodianski Alexei, Goeldner Moritz, Machens Hans-Günther, Schilling Arndt F

机构信息

Experimental Plastic Surgery, Department for Plastic Surgery and Hand Surgery, Klinikum rechts der Isar, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.

Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.

出版信息

PLoS One. 2017 Apr 5;12(4):e0174860. doi: 10.1371/journal.pone.0174860. eCollection 2017.

Abstract

Recent advances in gene delivery into cells allow improved therapeutic effects in gene therapy trials. To increase the bioavailability of applied cells, it is of great interest that transfected cells remain at the application site and systemic spread is minimized. In this study, we tested clinically used biodegradable poly(lactic acid-co-glycolic acid) (PLGA) scaffolds (Vicryl & Ethisorb) as transient carriers for genetically modified cells. To this aim, we used human fibroblasts and examined attachment and proliferation of untransfected cells on the scaffolds in vitro, as well as the mechanical properties of the scaffolds at four time points (1, 3, 6 and 9 days) of cultivation. Furthermore, the adherence of cells transfected with green fluorescent protein (GFP) and vascular endothelial growth factor (VEGF165) and also VEGF165 protein secretion were investigated. Our results show that human fibroblasts adhere on both types of PLGA scaffolds. However, proliferation and transgene expression capacity were higher on Ethisorb scaffolds most probably due to a different architecture of the scaffold. Additionally, cultivation of the cells on the scaffolds did not alter their biomechanical properties. The results of this investigation could be potentially exploited in therapeutic regiments with areal delivery of transiently transfected cells and may open the way for a variety of applications of cell-based gene therapy, tissue engineering and regenerative medicine.

摘要

细胞基因递送方面的最新进展使得基因治疗试验的治疗效果得到改善。为了提高应用细胞的生物利用度,使转染细胞保留在应用部位并将全身扩散降至最低具有重要意义。在本研究中,我们测试了临床使用的可生物降解聚乳酸 - 乙醇酸共聚物(PLGA)支架(薇乔和依索博)作为基因修饰细胞的瞬时载体。为此,我们使用人成纤维细胞,在体外检测未转染细胞在支架上的附着和增殖情况,以及在培养的四个时间点(1、3、6和9天)支架的力学性能。此外,还研究了转染绿色荧光蛋白(GFP)和血管内皮生长因子(VEGF165)的细胞的黏附情况以及VEGF165蛋白的分泌情况。我们的结果表明,人成纤维细胞能附着在两种类型的PLGA支架上。然而,依索博支架上的细胞增殖和转基因表达能力更高,这很可能是由于支架结构不同所致。此外,在支架上培养细胞并未改变其生物力学性能。本研究结果可能会在局部递送瞬时转染细胞的治疗方案中得到应用,并可能为基于细胞的基因治疗、组织工程和再生医学的各种应用开辟道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6760/5381796/127cc429139d/pone.0174860.g001.jpg

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