Forni Monica, Bernardini Chiara, Zamparini Fausto, Zannoni Augusta, Salaroli Roberta, Ventrella Domenico, Parchi Greta, Degli Esposti Micaela, Polimeni Antonella, Fabbri Paola, Fava Fabio, Prati Carlo, Gandolfi Maria Giovanna
Department of Veterinary Medical Sciences, University of Bologna, Ozzano Emilia, 40064 Bologna, Italy.
Laboratory of Biomaterials, Green Materials and Oral Pathology, School of Dentistry, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy.
Nanomaterials (Basel). 2020 Jan 29;10(2):243. doi: 10.3390/nano10020243.
Vascularization is a crucial factor when approaching any engineered tissue. Vascular wall-mesenchymal stem cells are an excellent in vitro model to study vascular remodeling due to their strong angiogenic attitude. This study aimed to demonstrate the angiogenic potential of experimental highly porous scaffolds based on polylactic acid (PLA) or poly-e-caprolactone (PCL) doped with calcium silicates (CaSi) and dicalcium phosphate dihydrate (DCPD), namely PLA-10CaSi-10DCPD and PCL-10CaSi-10DCPD, designed for the regeneration of bone defects. Vascular wall-mesenchymal stem cells (VW-MSCs) derived from pig thoracic aorta were seeded on the scaffolds and the expression of angiogenic markers, i.e. CD90 (mesenchymal stem/stromal cell surface marker), pericyte genes α-SMA (alpha smooth muscle actin), PDGFR-β (platelet-derived growth factor receptor-β), and NG2 (neuron-glial antigen 2) was evaluated. Pure PLA and pure PCL scaffolds and cell culture plastic were used as controls (3D in vitro model vs. 2D in vitro model). The results clearly demonstrated that the vascular wall mesenchymal cells colonized the scaffolds and were metabolically active. Cells, grown in these 3D systems, showed the typical gene expression profile they have in control 2D culture, although with some main quantitative differences. DNA staining and immunofluorescence assay for alpha-tubulin confirmed a cellular presence on both scaffolds. However, VW-MSCs cultured on PLA-10CaSi-10DCPD showed an individual cells growth, whilst on PCL-10CaSi-10DCPD scaffolds VW-MSCs grew in spherical clusters. In conclusion, vascular wall mesenchymal stem cells demonstrated the ability to colonize PLA and PCL scaffolds doped with CaSi-DCPD for new vessels formation and a potential for tissue regeneration.
血管化是构建任何工程组织时的一个关键因素。血管壁间充质干细胞因其强大的血管生成能力，是研究血管重塑的优秀体外模型。本研究旨在证明基于聚乳酸（PLA）或聚ε-己内酯（PCL）并掺杂硅酸钙（CaSi）和二水磷酸二钙（DCPD）的实验性高孔隙率支架（即PLA-10CaSi-10DCPD和PCL-10CaSi-10DCPD）的血管生成潜力，这些支架是为骨缺损再生而设计的。将源自猪胸主动脉的血管壁间充质干细胞（VW-MSCs）接种到支架上，并评估血管生成标志物的表达，即CD90（间充质干/基质细胞表面标志物）、周细胞基因α-SMA（α-平滑肌肌动蛋白）、PDGFR-β（血小板衍生生长因子受体-β）和NG2（神经胶质抗原2）。使用纯PLA和纯PCL支架以及细胞培养塑料作为对照（3D体外模型与2D体外模型）。结果清楚地表明，血管壁间充质细胞在支架上定植并具有代谢活性。在这些3D系统中生长的细胞显示出它们在对照2D培养中具有的典型基因表达谱，尽管存在一些主要的定量差异。DNA染色和α-微管蛋白的免疫荧光测定证实了两种支架上均有细胞存在。然而，在PLA-10CaSi-10DCPD上培养的VW-MSCs呈单个细胞生长，而在PCL-10CaSi-10DCPD支架上，VW-MSCs呈球形簇状生长。总之，血管壁间充质干细胞证明了能够在掺杂CaSi-DCPD的PLA和PCL支架上定植以形成新血管，并具有组织再生的潜力。
