Gardel Leandro S, Correia-Gomes Carla, Serra Luís A, Gomes Manuela E, Reis Rui L
3B's Research Groups: Biomaterials, Biodegradables and Biomimetics, Department of Polymer Engineering, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Taipas, Guimarães, Portugal; ICVS/3B's PT Government Associated Lab, AvePark, 4806-909, Braga, Portugal; Department of Clinic Veterinary, ICBAS-University of Porto, Porto, Portugal.
J Biomed Mater Res B Appl Biomater. 2013 Nov;101(8):1377-86. doi: 10.1002/jbm.b.32955. Epub 2013 May 17.
This works reports the development and preliminary assessment of a new bioreactor for culturing large-sized three-dimensional constructs in bone tissue engineering. The bidirectional continuous perfusion bioreactor (BCPB) promotes mechanical stimulation of cells through the creation of shear forces induced by flow perfusion. The main innovation consists in the possibility of culturing scaffolds of large dimensions that can be suitable for the regeneration of critical sized defects. The functionality of BCPB was preliminarily evaluated by culturing starch-polycaprolactone scaffolds/goat bone marrow stromal cells for 14 and 21 days. Cylindrical blocks were stacked (42 mm thick). Static culture was used as controls. The samples were collected for DNA, alkaline phosphatase (ALP), scanning electron microscopy (SEM), and histological analysis. The results showed higher ALP levels in the bioreactor cultures than those obtained under static conditions. The number of cells in constructs cultured in the bioreactor showed lower values compared to static cultures, suggesting that static conditions tend to privilege the metabolic path way for cellular proliferation while dynamic conditions tend to privilege the metabolic path for osteogenic differentiation. SEM observations show that, the migration and cell distribution was observed in the bioreactor. These results demonstrate the feasibility and the benefit of culturing constructs in BCPB.
本研究报告了一种用于骨组织工程中培养大型三维构建体的新型生物反应器的开发及初步评估。双向连续灌注生物反应器(BCPB)通过流动灌注产生的剪切力促进细胞的机械刺激。主要创新之处在于能够培养适用于临界尺寸缺损修复的大尺寸支架。通过将淀粉 - 聚己内酯支架/山羊骨髓基质细胞培养14天和21天,对BCPB的功能进行了初步评估。将圆柱形块堆叠(42毫米厚)。以静态培养作为对照。收集样品进行DNA、碱性磷酸酶(ALP)、扫描电子显微镜(SEM)和组织学分析。结果显示,生物反应器培养物中的ALP水平高于静态条件下获得的水平。与静态培养相比,在生物反应器中培养的构建体中的细胞数量较低,这表明静态条件倾向于有利于细胞增殖的代谢途径,而动态条件倾向于有利于成骨分化的代谢途径。SEM观察表明,在生物反应器中观察到了细胞迁移和分布。这些结果证明了在BCPB中培养构建体的可行性和益处。