1Department of Biology, Laboratory of Animal Physiology, Aristotle University of Thessaloniki, 54124 Thessaloníki, Greece.
Biohellenika Biotechnology Company, 65 Leoforos Georgikis Scholis, 57001 Thessaloníki, Greece.
Tissue Eng Regen Med. 2019 Mar 18;16(2):161-175. doi: 10.1007/s13770-019-00185-z. eCollection 2019 Apr.
Μonocyte-derived multipotential cells (MOMCs) include progenitors capable of differentiation into multiple cell lineages and thus represent an ideal autologous transplantable cell source for regenerative medicine. In this study, we cultured MOMCs, generated from mononuclear cells of peripheral blood, on the surface of nanocomposite thin films.
For this purpose, nanocomposite Poly(e-caprolactone) (PCL)-based thin films containing either 2.5 wt% silica nanotubes (SiOntbs) or strontium hydroxyapatite nanorods (SrHAnrds), were prepared using the spin-coating method. The induced differentiation capacity of MOMCs, towards bone and endothelium, was estimated using flow cytometry, real-time polymerase chain reaction, scanning electron microscopy and fluorescence microscopy after cells' genetic modification using the Sleeping Beauty Transposon System aiming their observation onto the scaffolds. Moreover, Wharton's Jelly Mesenchymal Stromal Cells were cultivated as a control cell line, while Human Umbilical Vein Endothelial Cells were used to strengthen and accelerate the differentiation procedure in semi-permeable culture systems. Finally, the cytotoxicity of the studied materials was checked with MTT assay.
The highest differentiation capacity of MOMCs was observed on PCL/SiOntbs 2.5 wt% nanocomposite film, as they progressively lost their native markers and gained endothelial lineage, in both protein and transcriptional level. In addition, the presence of SrHAnrds in the PCL matrix triggered processes related to osteoblast bone formation.
To conclude, the differentiation of MOMCs was selectively guided by incorporating SiOntbs or SrHAnrds into a polymeric matrix, for the first time.
单核细胞衍生的多潜能细胞（MOMCs）包括能够分化为多种细胞谱系的祖细胞，因此代表了再生医学中理想的自体可移植细胞来源。在这项研究中，我们在纳米复合薄膜的表面培养了由外周血单核细胞产生的 MOMCs。
为此，我们使用旋涂法制备了含有 2.5wt%二氧化硅纳米管（SiOntbs）或锶羟基磷灰石纳米棒（SrHAnrds）的聚（己内酯）（PCL）基纳米复合薄膜。通过流式细胞术、实时聚合酶链反应、扫描电子显微镜和荧光显微镜，在使用 Sleeping Beauty 转座子系统对细胞进行遗传修饰后，估计 MOMCs 向骨和内皮的诱导分化能力，目的是观察它们在支架上的分化。此外，我们培养了 Wharton 果冻间充质基质细胞作为对照细胞系，同时使用人脐静脉内皮细胞在半渗透培养系统中加强和加速分化过程。最后，通过 MTT 测定检查研究材料的细胞毒性。
在 PCL/SiOntbs 2.5wt%纳米复合薄膜上观察到 MOMCs 的最高分化能力，因为它们逐渐失去其天然标志物并获得内皮谱系，在蛋白质和转录水平上均如此。此外，PCL 基质中 SrHAnrds 的存在引发了与成骨细胞骨形成相关的过程。
总之，首次通过将 SiOntbs 或 SrHAnrds 掺入聚合物基质中，选择性地指导 MOMCs 的分化。
