Bastos Ana Raquel, Raquel Maia F, Miguel Oliveira J, Reis Rui Luís, Correlo Vitor Manuel
3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimarães, Portugal; ICVS/3B's - PT Government Associated Laboratory, Portugal.
3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimarães, Portugal; ICVS/3B's - PT Government Associated Laboratory, Portugal.
Mater Sci Eng C Mater Biol Appl. 2021 Oct;129:112413. doi: 10.1016/j.msec.2021.112413. Epub 2021 Sep 3.
The scientific community has been doing significant efforts towards engineering new 3D bone models in recent years. Osteocytes are mechanosensitive cells that play significant roles in the maintenance of bone homeostasis. Currently, as far as we know, there are no 3D models that faithfully recapitulate a bone microenvironment capable of promoting the differentiation of osteoblasts towards osteocytes. Besides, in the existing models, the use of human cells does not prevail over the animal cell lines. For so, we propose a 3D model that may have important implications for ongoing efforts towards a better understanding of bone physiology and disease. The main aim of the current work was the promotion of an effective differentiation of osteoblasts into osteocytes by mean of using a 3D model composed of primary human osteoblasts (hOBs) cultured on Gellan Gum-Hydroxyapatite (GG-HAp) matrix under a long-term osteogenic culture. The results revealed that GG-HAp matrix stimulated a fast cell migration/entrapment, attachment, spreading, and mineralization. Moreover, the transition process from osteoblasts to osteocytes was confirmed by the expression of the osteogenic-related (ALP, Runx2, COL I, OC, OPN and OSX) and osteocyte-related (hPDPN) marker throughout the culture time. Overall, the developed 3D model holds a great promise for the treatment of various bone diseases, namely on diagnostic applications and for bone regeneration purposes.
近年来，科学界一直在为构建新型3D骨模型做出重大努力。骨细胞是机械敏感细胞，在维持骨稳态中发挥着重要作用。目前，据我们所知，尚无能够忠实地重现促进成骨细胞向骨细胞分化的骨微环境的3D模型。此外，在现有模型中，人类细胞的使用并不比动物细胞系占优势。因此，我们提出了一种3D模型，这可能对当前为更好地理解骨生理学和疾病所做的努力具有重要意义。当前工作的主要目的是通过使用一种3D模型促进成骨细胞有效地分化为骨细胞，该模型由在结冷胶-羟基磷灰石（GG-HAp）基质上培养的原代人成骨细胞（hOBs）组成，并处于长期成骨培养条件下。结果表明，GG-HAp基质刺激了细胞的快速迁移/滞留、附着、铺展和矿化。此外，在整个培养过程中，通过成骨相关（碱性磷酸酶、Runx2、I型胶原、骨钙素、骨桥蛋白和OSX）和骨细胞相关（人Podoplanin）标志物的表达，证实了从成骨细胞到骨细胞的转变过程。总体而言，所开发的3D模型在各种骨疾病的治疗方面，即在诊断应用和骨再生目的方面，具有巨大的潜力。
