Re Federica, Sartore Luciana, Pasini Chiara, Ferroni Matteo, Borsani Elisa, Pandini Stefano, Bianchetti Andrea, Almici Camillo, Giugno Lorena, Bresciani Roberto, Mutti Silvia, Trenta Federica, Bernardi Simona, Farina Mirko, Russo Domenico
Unit of Blood Diseases and Cell Therapies, Department of Clinical and Experimental Sciences, University of Brescia, "ASST-Spedali Civili" Hospital of Brescia, 25123 Brescia, Italy.
Centro di Ricerca Emato-Oncologica AIL (CREA), ASST Spedali Civili, 25123 Brescia, Italy.
J Funct Biomater. 2024 Jul 31;15(8):217. doi: 10.3390/jfb15080217.
Human mesenchymal stromal cells (hMSCs), whether used alone or together with three-dimensional scaffolds, are the best-studied postnatal stem cells in regenerative medicine. In this study, innovative composite scaffolds consisting of a core-shell architecture were seeded with bone-marrow-derived hMSCs (BM-hMSCs) and tested for their biocompatibility and remarkable capacity to promote and support bone regeneration and mineralization. The scaffolds were prepared by grafting three different amounts of gelatin-chitosan (CH) hydrogel into a 3D-printed polylactic acid (PLA) core (PLA-CH), and the mechanical and degradation properties were analyzed. The BM-hMSCs were cultured in the scaffolds with the presence of growth medium (GM) or osteogenic medium (OM) with differentiation stimuli in combination with fetal bovine serum (FBS) or human platelet lysate (hPL). The primary objective was to determine the viability, proliferation, morphology, and spreading capacity of BM-hMSCs within the scaffolds, thereby confirming their biocompatibility. Secondly, the BM-hMSCs were shown to differentiate into osteoblasts and to facilitate scaffold mineralization. This was evinced by a positive Von Kossa result, the modulation of differentiation markers (osteocalcin and osteopontin), an expression of a marker of extracellular matrix remodeling (bone morphogenetic protein-2), and collagen I. The results of the energy-dispersive X-ray analysis (EDS) clearly demonstrate the presence of calcium and phosphorus in the samples that were incubated in OM, in the presence of FBS and hPL, but not in GM. The chemical distribution maps of calcium and phosphorus indicate that these elements are co-localized in the same areas of the sections, demonstrating the formation of hydroxyapatite. In conclusion, our findings show that the combination of BM-hMSCs and PLA-CH, regardless of the amount of hydrogel content, in the presence of differentiation stimuli, can provide a construct with enhanced osteogenicity for clinically relevant bone regeneration.
人间充质基质细胞（hMSCs），无论是单独使用还是与三维支架一起使用，都是再生医学中研究最充分的出生后干细胞。在本研究中，将由骨髓来源的hMSCs（BM-hMSCs）接种到具有核壳结构的创新复合支架上，并测试其生物相容性以及促进和支持骨再生与矿化的显著能力。通过将三种不同量的明胶-壳聚糖（CH）水凝胶接枝到3D打印的聚乳酸（PLA）核（PLA-CH）中来制备支架，并分析其力学和降解性能。将BM-hMSCs在含有生长培养基（GM）或成骨培养基（OM）以及分化刺激物的支架中培养，并添加胎牛血清（FBS）或人血小板裂解物（hPL）。主要目的是确定BM-hMSCs在支架内的活力、增殖、形态和铺展能力，从而确认其生物相容性。其次，BM-hMSCs被证明可分化为成骨细胞并促进支架矿化。这通过阳性的冯·科萨结果、分化标志物（骨钙素和骨桥蛋白）的调节、细胞外基质重塑标志物（骨形态发生蛋白-2）和I型胶原蛋白的表达得以证明。能量色散X射线分析（EDS）结果清楚地表明，在含有FBS和hPL的OM中孵育的样品中存在钙和磷，但在GM中不存在。钙和磷的化学分布图表明这些元素共定位在切片的相同区域，证明了羟基磷灰石的形成。总之，我们的研究结果表明，在分化刺激存在的情况下，BM-hMSCs与PLA-CH的组合，无论水凝胶含量多少，都可为临床相关的骨再生提供具有增强成骨能力的构建体。
