Zenobi Eleonora, Gramigna Giulia, Scatena Elisa, Panizza Luca, Achille Carlotta, Pecci Raffaella, Convertino Annalisa, Del Gaudio Costantino, Lisi Antonella, Ledda Mario
E. Amaldi Foundation, Via del Politecnico snc, 00133 Rome, Italy.
Hypatia Research Consortium, Viale I Maggio, 156, Grottaferrata, 00046 Rome, Italy.
J Funct Biomater. 2025 Jul 24;16(8):271. doi: 10.3390/jfb16080271.
Three-dimensional cell culture systems are relevant in vitro models for studying cellular behavior. In this regard, this present study investigates the interaction between human osteoblast-like cells and 3D-printed scaffolds mimicking physiological and osteoporotic bone structures under simulated microgravity conditions. The objective is to assess the effects of scaffold architecture and dynamic culture conditions on cell adhesion, proliferation, and metabolic activity, with implications for osteoporosis research. Polylactic acid scaffolds with physiological (P) and osteoporotic-like (O) trabecular architectures were 3D-printed by means of fused deposition modeling technology. Morphometric characterization was performed using micro-computed tomography. Human osteoblast-like SAOS-2 and U2OS cells were cultured on the scaffolds under static and dynamic simulated microgravity conditions using a rotary cell culture system (RCCS). Scaffold biocompatibility, cell viability, adhesion, and metabolic activity were evaluated through Bromodeoxyuridine incorporation assays, a water-soluble tetrazolium salt assay, and an enzyme-linked immunosorbent assay of tumor necrosis factor-α secretion. Both scaffold models supported osteoblast-like cell adhesion and growth, with an approximately threefold increase in colonization observed on the high-porosity O scaffolds under dynamic conditions. The dynamic environment facilitated increased surface interaction, amplifying the effects of scaffold architecture on cell behavior. Overall, sustained cell growth and metabolic activity, together with the absence of detectable inflammatory responses, confirmed the biocompatibility of the system. Scaffold microstructure and dynamic culture conditions significantly influence osteoblast-like cell behavior. The combination of 3D-printed scaffolds and a RCCS bioreactor provides a promising platform for studying bone remodeling in osteoporosis and microgravity-induced bone loss. These findings may contribute to the development of advanced in vitro models for biomedical research and potential countermeasures for bone degeneration.
三维细胞培养系统是用于研究细胞行为的相关体外模型。在这方面，本研究调查了人成骨样细胞与模拟生理和骨质疏松骨结构的3D打印支架在模拟微重力条件下的相互作用。目的是评估支架结构和动态培养条件对细胞黏附、增殖和代谢活性的影响，这对骨质疏松症研究具有重要意义。采用熔融沉积建模技术3D打印具有生理（P）和骨质疏松样（O）小梁结构的聚乳酸支架。使用微型计算机断层扫描进行形态计量学表征。使用旋转细胞培养系统（RCCS）在静态和动态模拟微重力条件下将人成骨样SAOS-2和U2OS细胞培养在支架上。通过溴脱氧尿苷掺入试验、水溶性四唑盐试验和肿瘤坏死因子-α分泌的酶联免疫吸附试验评估支架生物相容性、细胞活力、黏附及代谢活性。两种支架模型均支持成骨样细胞黏附和生长，在动态条件下，高孔隙率O支架上的定植增加了约三倍。动态环境促进了表面相互作用的增加，放大了支架结构对细胞行为的影响。总体而言，细胞持续生长和代谢活性，以及未检测到炎症反应，证实了该系统的生物相容性。支架微观结构和动态培养条件显著影响成骨样细胞行为。3D打印支架和RCCS生物反应器的组合为研究骨质疏松症中的骨重塑和微重力诱导的骨质流失提供了一个有前景的平台。这些发现可能有助于开发用于生物医学研究的先进体外模型以及针对骨质退化的潜在对策。
