Pozzobon Laura Gonçalves, Sperling Laura Elena, Teixeira Cristian E, Malysz Tais, Pranke Patricia
Hematology and Stem Cell Laboratory, Faculty of Pharmacy, Av. Ipiranga 2752, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; Stem Cell Laboratory, Fundamental Health Science Institute, Rua Sarmento Leite, 500, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Hematology and Stem Cell Laboratory, Faculty of Pharmacy, Av. Ipiranga 2752, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; Stem Cell Laboratory, Fundamental Health Science Institute, Rua Sarmento Leite, 500, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Chem Biol Interact. 2021 Oct 1;348:109621. doi: 10.1016/j.cbi.2021.109621. Epub 2021 Aug 25.
A promising alternative to conventional nerve grafting is the use of artificial grafts made from biodegradable and biocompatible materials and support cells. The aim of this study has been to produce a biodegradable nerve conduit and investigate the cytocompatibility with stem cells and its regeneration promoting properties in a rat animal model. A poly (lactic-co-glycolic acid) (PLGA) conduit of aligned nanofibers was produced by the electrospinning method, functionalized with gelatin and seeded either with mouse embryonic stem cells (mESCs) or with human mesenchymal stem cells (SHED). The cell proliferation and viability were analyzed in vitro. The conduits were implanted in a rat model of sciatic nerve lesion by transection. The functional recovery was monitored for 8 weeks using the Sciatic Functional Index (SFI) and histological analyses were used to assess the nerve regeneration. Scaffolds of aligned PLGA fibers with an average diameter of 0.90 ± 0.36 μm and an alignment coefficient of 0.817 ± 0.07 were produced. The treatment with gelatin increased the fiber diameter to 1.05 ± 0.32 μm, reduced the alignment coefficient to 0.655 ± 0.045 and made the scaffold very hydrophilic. The cell viability and Live/dead assay showed that the stem cells remained viable and proliferated after 7 days in culture. Confocal images of phalloidin/DAPI staining showed that the cells adhered and proliferated widely, in fully adaptation with the biomaterial. The SFI values of the group that received the conduit were similar to the values of the control lesioned group. In conclusion, conduits composed of PLGA-gelatin nanofibers were produced and promoted a very good interaction with the stem cells. Although in vitro studies have shown this biomaterial to be a promising biomaterial for the regeneration of nerve tissue, in vivo studies of this graft have not shown significant improvements in nerve regeneration.
一种有前景的替代传统神经移植的方法是使用由可生物降解和生物相容性材料以及支持细胞制成的人工移植物。本研究的目的是制备一种可生物降解的神经导管，并在大鼠动物模型中研究其与干细胞的细胞相容性及其促进再生的特性。通过静电纺丝法制备了具有排列纳米纤维的聚（乳酸-乙醇酸）（PLGA）导管，用明胶进行功能化处理，并接种小鼠胚胎干细胞（mESCs）或人间充质干细胞（SHED）。在体外分析细胞增殖和活力。将导管植入坐骨神经横断损伤的大鼠模型中。使用坐骨神经功能指数（SFI）监测8周的功能恢复情况，并通过组织学分析评估神经再生。制备了平均直径为0.90±0.36μm、排列系数为0.817±0.07的排列PLGA纤维支架。用明胶处理使纤维直径增加到1.05±0.32μm，排列系数降低到0.655±0.045，并使支架具有很强的亲水性。细胞活力和活/死检测表明，干细胞在培养7天后仍保持活力并增殖。鬼笔环肽/ DAPI染色的共聚焦图像显示，细胞广泛粘附并增殖，与生物材料完全适应。接受导管组的SFI值与损伤对照组的值相似。总之，制备了由PLGA-明胶纳米纤维组成的导管，其与干细胞促进了非常良好的相互作用。尽管体外研究表明这种生物材料是神经组织再生的有前景的生物材料，但这种移植物的体内研究尚未显示出神经再生有显著改善。
