Noreikaitė Aurelija, Antanavičiūtė Ieva, Mikalayeva Valeryia, Darinskas Adas, Tamulevičius Tomas, Adomavičiūtė Erika, Šimatonis Linas, Akramienė Dalia, Stankevičius Edgaras
Institute of Physiology and Pharmacology, Medical Academy, Lithuanian University of Health Sciences, Kaunas, Lithuania.
Institute of Cardiology, Medical Academy, Lithuanian University of Health Sciences, Kaunas, Lithuania.
Medicina (Kaunas). 2017;53(3):203-210. doi: 10.1016/j.medici.2017.07.001. Epub 2017 Jul 13.
The aim of this study was to test polymeric materials (collagen, fibrin, polyimide film, and polylactic acid) for single- and multi-layer scaffold formation.
In our study, we used rabbit bone marrow stem cells (rBMSCs) and human mesenchymal stem cells (hMSCs) with materials of a different origin for the formation of an artificial scaffold, such as a collagen scaffold, fibrin scaffold produced from clotted rabbit plasma, electrospun poly(lactic acid) (PLA) mats, polyimide film (PI), and the combination of the latter two. Cell imaging was performed 3-14 days after cell cultivation in the scaffolds. Time-lapse imaging was used to determine hMSC mobility on the PI film.
Cell incorporation in collagen and clotted fibrin scaffolds was evaluated after 2-week cultivation in vitro. Histological analysis showed that cells penetrated only external layers of the collagen scaffold, while the fibrin clot was populated with rBMSCs through the entire scaffold thickness. As well, cell behavior on the laser micro-structured PI film was analyzed. The mobility of hMSCs on the smooth PI film and the micro-machined surface was 20±2μm/h and 18±4μm/h, respectively. After 3-day cultivation, hMSCs were capable of spreading through the whole 100±10μm-thick layer of the electrospun PLA scaffold and demonstrated that the multilayer scaffold composed of PI and PLA materials ensured a suitable environment for cell growth.
The obtained results suggest that electrospinning technology and femtosecond laser micro-structuring could be employed for the development of multi-layer scaffolds. Different biopolymers, such as PLA, fibrin, and collagen, could be used as appropriate environments for cell inhabitation and as an inner layer of the multi-layer scaffold. PI could be suitable as a barrier blocking cell migration from the scaffold. However, additional studies are needed to determine optimal parameters of inner and outer scaffold layers.
本研究旨在测试用于单层和多层支架形成的聚合材料（胶原蛋白、纤维蛋白、聚酰亚胺薄膜和聚乳酸）。
在我们的研究中，我们将兔骨髓干细胞（rBMSCs）和人间充质干细胞（hMSCs）与不同来源的材料用于构建人工支架，如胶原蛋白支架、由兔血浆凝块制成的纤维蛋白支架、电纺聚乳酸（PLA）垫、聚酰亚胺薄膜（PI）以及后两者的组合。在细胞接种到支架中3 - 14天后进行细胞成像。使用延时成像来测定hMSCs在PI薄膜上的迁移能力。
在体外培养2周后评估细胞在胶原蛋白和凝块纤维蛋白支架中的植入情况。组织学分析表明，细胞仅穿透胶原蛋白支架的外层，而纤维蛋白凝块在整个支架厚度内都有rBMSCs分布。同样，分析了细胞在激光微结构化PI薄膜上的行为。hMSCs在光滑PI薄膜和微加工表面上的迁移速度分别为20±2μm/h和18±4μm/h。培养3天后，hMSCs能够在整个100±10μm厚的电纺PLA支架层中铺展，并且由PI和PLA材料组成的多层支架为细胞生长提供了适宜的环境。
所得结果表明，电纺技术和飞秒激光微结构化可用于开发多层支架。不同的生物聚合物，如PLA、纤维蛋白和胶原蛋白，可作为细胞栖息的适宜环境以及多层支架的内层。PI可作为阻止细胞从支架迁移的屏障。然而，需要进一步研究以确定支架内层和外层的最佳参数。
