Radi Sonya, EzEldeen Mostafa, Végvári Ákos, Coates Dawn, Jacobs Reinhilde, Bostanci Nagihan, Bao Kai
OMFS-IMPATH Research Group, Faculty of Medicine, Department of Imaging and Pathology, KU Leuven and Oral and Maxillofacial Surgery, University Hospitals Leuven, Leuven, Belgium.
Department of Oral Health Sciences, KU Leuven and Paediatric Dentistry and Special Dental Care, University Hospitals Leuven, Leuven, Belgium.
Sci Rep. 2025 Apr 9;15(1):12120. doi: 10.1038/s41598-025-96632-0.
Bone disorders represent a significant global burden. Currently, animal models are used to develop and screen novel treatments. However, interspecies variations and ethical concerns highlight the need for a more complex 3D bone model. In this study, we developed a simplified in vitro bone-like model using a U-CUP perfusion-based bioreactor system, designed to provide continuous nutrient flow and mechanostimulation through 3D cultures. An immortalized human fetal osteoblastic cell line was seeded on collagen scaffolds and cultured for 21 days in both a perfusion bioreactor system and in static cultures. PrestoBlue™ assay, scanning electron microscopy, and proteomics allowed monitoring of metabolic activity and compared morphological and proteome differences between both conditions. Results indicated an altered cellular morphology in the bioreactor compared to the static cultures and identified a total of 3494 proteins. Of these, 105 proteins exhibited significant upregulation in the static culture, while 86 proteins displayed significant downregulation. Enrichment analyses of these proteins revealed ten significant pathways including epithelial-mesenchymal transition, TNF-alpha signaling via NF-kB, and KRAS pathway. The current data indicated of osteogenic differentiation enhancement within the bioreactor on day 21 compared to static cultures. In conclusion, the U-CUP perfusion bioreactor is beneficial for facilitating osteogenic differentiation in 3D cultures.
骨骼疾病是一项重大的全球负担。目前，动物模型被用于开发和筛选新的治疗方法。然而，种间差异和伦理问题凸显了对更复杂的三维骨骼模型的需求。在本研究中，我们使用基于U-CUP灌注的生物反应器系统开发了一种简化的体外类骨模型，该系统旨在通过三维培养提供持续的营养物质流动和机械刺激。将永生化的人胎儿成骨细胞系接种在胶原蛋白支架上，并在灌注生物反应器系统和静态培养中培养21天。使用普洛麦格公司的碧云天试剂、扫描电子显微镜和蛋白质组学来监测代谢活性，并比较两种条件下的形态和蛋白质组差异。结果表明，与静态培养相比，生物反应器中的细胞形态发生了改变，并鉴定出总共3494种蛋白质。其中，105种蛋白质在静态培养中显著上调，而86种蛋白质显著下调。对这些蛋白质的富集分析揭示了十条重要途径，包括上皮-间质转化、通过核因子-κB的肿瘤坏死因子-α信号传导和KRAS途径。目前的数据表明，与静态培养相比，生物反应器在第21天促进了成骨分化。总之，U-CUP灌注生物反应器有利于促进三维培养中的成骨分化。
