Vonbrunn Eva, Mueller Marc, Pichlsberger Melanie, Sundl Monika, Helmer Alexander, Wallner Stefanie Angela, Rinner Beate, Tuca Alexandru-Cristian, Kamolz Lars-Peter, Brislinger Dagmar, Glasmacher Birgit, Lang-Olip Ingrid
Division of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.
Institute of Multiphase Processes, Leibniz University Hanover, Hannover, Germany.
Front Bioeng Biotechnol. 2020 Dec 16;8:604123. doi: 10.3389/fbioe.2020.604123. eCollection 2020.
Mesenchymal stem/stromal cells (MSCs) exert beneficial effects during wound healing, and cell-seeded scaffolds are a promising method of application. Here, we compared the suitability of a clinically used collagen/elastin scaffold (Matriderm) with an electrospun Poly(ε-caprolactone)/poly(l-lactide) (PCL/PLA) scaffold as carriers for human amnion-derived MSCs (hAMSCs). We created an epidermal-like PCL/PLA scaffold and evaluated its microstructural, mechanical, and functional properties. Sequential spinning of different PCL/PLA concentrations resulted in a wide-meshed layer designed for cell-seeding and a dense-meshed layer for apical protection. The Matriderm and PCL/PLA scaffolds then were seeded with hAMSCs, with or without Matrigel coating. The quantity and quality of the adherent cells were evaluated . The results showed that hAMSCs adhered to and infiltrated both scaffold types but on day 3, more cells were observed on PCL/PLA than on Matriderm. Apoptosis and proliferation rates were similar for all carriers except the coated Matriderm, where apoptotic cells were significantly enhanced. On day 8, the number of cells decreased on all carrier types except the coated Matriderm, which had consistently low cell numbers. Uncoated Matriderm had the highest percentage of proliferative cells and lowest apoptosis rate of all carrier types. Each carrier also was topically applied to skin wound sites in a mouse model and analyzed over 14 days via optical imaging and histological methods, which showed detectable hAMSCs on all carrier types on day 8. On day 14, all wounds exhibited newly formed epidermis, and all carriers were well-integrated into the underlying dermis and showing signs of degradation. However, only wounds treated with uncoated PCL/PLA maintained a round appearance with minimal contraction. Overall, the results support a 3-day culture of scaffolds with hAMSCs before wound application. The PCL/PLA scaffold showed higher cell adherence than Matriderm, and the effect of the Matrigel coating was negligible, as all carrier types maintained sufficient numbers of transplanted cells in the wound area. The anti-contractive effects of the PCL/PLA scaffold offer potential new therapeutic approaches to wound care.
间充质干/基质细胞(MSCs)在伤口愈合过程中发挥有益作用,而接种细胞的支架是一种很有前景的应用方法。在此,我们比较了临床使用的胶原/弹性蛋白支架(Matriderm)与电纺聚(ε-己内酯)/聚(L-丙交酯)(PCL/PLA)支架作为人羊膜来源间充质干细胞(hAMSCs)载体的适用性。我们制备了一种表皮样PCL/PLA支架,并评估了其微观结构、力学和功能特性。不同PCL/PLA浓度的顺序纺丝产生了一个用于细胞接种的宽网层和一个用于顶端保护的密网层。然后将Matriderm和PCL/PLA支架接种hAMSCs,有或没有基质胶包被。评估贴壁细胞的数量和质量。结果表明,hAMSCs可附着并浸润两种支架类型,但在第3天,在PCL/PLA上观察到的细胞比在Matriderm上更多。除包被的Matriderm外,所有载体的凋亡率和增殖率相似,包被的Matriderm上凋亡细胞显著增加。在第8天,除包被的Matriderm外,所有载体类型上的细胞数量均减少,包被的Matriderm细胞数量一直较低。未包被的Matriderm在所有载体类型中增殖细胞百分比最高,凋亡率最低。每种载体还局部应用于小鼠模型的皮肤伤口部位,并在14天内通过光学成像和组织学方法进行分析,结果显示在第8天所有载体类型上均可检测到hAMSCs。在第14天,所有伤口均出现新形成的表皮,所有载体均很好地整合到下层真皮中并显示出降解迹象。然而,只有用未包被的PCL/PLA处理的伤口保持圆形外观,收缩最小。总体而言,结果支持在伤口应用前将支架与hAMSCs进行3天的培养。PCL/PLA支架比Matriderm表现出更高的细胞粘附性,并且基质胶包被的效果可忽略不计,因为所有载体类型在伤口区域均保持足够数量的移植细胞。PCL/PLA支架的抗收缩作用为伤口护理提供了潜在的新治疗方法。