88556 20230921135432.0 doi 10.3389/fbioe.2020.00336 sideral 117355 ART-2020-117355 eng Nasello, Gabriele Universidad de Zaragoza (orcid)0000-0002-0255-6200 Primary Human Osteoblasts Cultured in a 3D Microenvironment Create a Unique Representative Model of Their Differentiation Into Osteocytes 2020 Access copy available to the general public Unrestricted {Microengineered systems provide an in vitro strategy to explore the variability of individual patient response to tissue engineering products, since they prefer the use of primary cell sources representing the phenotype variability. Traditional in vitro systems already showed that primary human osteoblasts embedded in a 3D fibrous collagen matrix differentiate into osteocytes under specific conditions. Here, we hypothesized that translating this environment to the organ-on-a-chip scale creates a minimal functional unit to recapitulate osteoblast maturation toward osteocytes and matrix mineralization. Primary human osteoblasts were seeded in a type I collagen hydrogel, to establish the role of lower (2.5 × 10⁵ cells/ml) and higher (1 × 10⁶ cells/ml) cell density on their differentiation into osteocytes. A custom semi-automatic image analysis software was used to extract quantitative data on cellular morphology from brightfield images. The results are showing that cells cultured at a high density increase dendrite length over time, stop proliferating, exhibit dendritic morphology, upregulate alkaline phosphatase (ALP) activity, and express the osteocyte marker dental matrix protein 1 (DMP1). On the contrary, cells cultured at lower density proliferate over time, do not upregulate ALP and express the osteoblast marker bone sialoprotein 2 (BSP2) at all timepoints. Our work reveals that microengineered systems create unique conditions to capture the major aspects of osteoblast differentiation into osteocytes with a limited number of cells. We propose that the microengineered approach is a functional strategy to create a patient-specific bone tissue model and investigate the individual osteogenic potential of the patient bone cells. info:eu-repo/grantAgreement/EC/H2020/722535/EU/Predictive models and simulations in bone regeneration: a multiscale patient-specific approach/CuraBone This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 722535-CuraBone info:eu-repo/grantAgreement/ES/MINECO/DPI2017-84780-C2-1-R info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 5.89 2020 MULTIDISCIPLINARY SCIENCES 12 / 72 = 0.167 2020 Q1 T1 1.081 2020 Bioengineering 2020 Q1 Histology 2020 Q1 Biotechnology 2020 Q1 Biomedical Engineering 2020 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Alamán-Díez, Pilar Universidad de Zaragoza (orcid)0000-0003-1958-4432 Schiavi, Jessica Pérez, María Ángeles Universidad de Zaragoza (orcid)0000-0002-2901-4188 McNamara, Laoise García-Aznar, José Manuel Universidad de Zaragoza (orcid)0000-0002-9864-7683 5004 605 Universidad de Zaragoza Dpto. Ingeniería Mecánica Área Mec.Med.Cont. y Teor.Est. 8 (2020), 336 [14 pp] Front. Bioeng. Biotechnol. Frontiers in Bioengineering and Biotechnology 2296-4185 1359619 http://zaguan.unizar.es/record/88556/files/texto_completo.pdf Versión publicada 35422 http://zaguan.unizar.es/record/88556/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:88556 articulos driver 2023-09-21-13:30:24 ARTICLE