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Advances in Materials Science and Engineering
Volume 2015, Article ID 268930, 8 pages
Research Article

A 3D Osteoblast In Vitro Model for the Evaluation of Biomedical Materials

1Post-Graduation Program on Dentistry, Fluminense Federal University, 22245-100 Niterói, RJ, Brazil
2Clinical Research Unit, Antônio Pedro Hospital, Fluminense Federal University, 22245-100 Niterói, RJ, Brazil
3Post-Graduation Program on Science and Biotechnology, Fluminense Federal University, 22245-100 Niterói, RJ, Brazil
4Department of Molecular and Cell Biology, Institute of Biology, Fluminense Federal University, 22245-100 Niterói, RJ, Brazil

Received 29 September 2015; Revised 2 December 2015; Accepted 6 December 2015

Academic Editor: Massimiliano Barletta

Copyright © 2015 Luciana Restle et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Biomedical materials for bone therapy are usually assessed for their biocompatibility and safety employing animal models or in vitro monolayer cell culture assays. However, alternative in vitro models may offer controlled conditions closer to physiological responses and reduce animal testing. In this work, we developed a 3D spheroidal cell culture with potential to evaluate simultaneously material-cell and cell-cell interactions. Different cell densities of murine MC3T3-E1 preosteoblasts or human primary osteoblasts (HOb) were used to determine the ideal procedure of spheroidal cultures and their adequacy to material testing. Cells were seeded on 96-well plates coated with agar and incubated in agitation from 1 to 7 days. Aggregate morphology was qualitatively evaluated considering the shape, size, repeatability, handling, and stability of spheroids. Higher cell densities induced more stable spheroids, and handling was considered appropriate starting from 2 × 104 cells. Confocal microscopy and Scanning Electron Microscopy indicate that most cells within the aggregate core are viable. Exposure to positive controls has shown a dose dependent cell death as measured by XTT assay. Aggregates were stable and presented good viability when employed on standardized testing of metallic and polymer-based biomaterials. Therefore, osteoblast spheroids may provide a promising tool for material screening and biocompatibility testing.