In vitro bioprinted 3D model enhancing osteoblast-to-osteocyte differentiation - Archive ouverte HAL
Article Dans Une Revue Biofabrication Année : 2024

In vitro bioprinted 3D model enhancing osteoblast-to-osteocyte differentiation

Pragnere Sarah
  • Fonction : Auteur
Essayan Lucie
  • Fonction : Auteur
El-Kholti Naima
  • Fonction : Auteur
Petiot Emma
  • Fonction : Auteur
Paillet-Mattei Cyril
  • Fonction : Auteur

Résumé

In vitro bone models are pivotal for understanding tissue behavior and cellular responses, particularly in unravelling certain pathologies' mechanisms and assessing the impact of new therapeutic interventions. A desirable in vitro bone model should incorporate primary human cells within a 3D environment that mimics the mechanical properties characteristics of osteoid and faithfully replicate all stages of osteogenic differentiation from osteoblasts to osteocytes. However, to date, no bio-printed model using primary osteoblasts has demonstrated the expression of osteocytic protein markers. This study aimed to develop bio-printed in vitro model that accurately captures the differentiation process of human primary osteoblasts into osteocytes. Given the considerable impact of hydrogel stiffness and relaxation behavior on osteoblast activity, we employed three distinct cross-linking solutions to fabricate hydrogels. These hydrogels were designed to exhibit either similar elastic behavior with different elastic moduli, or similar elastic moduli with varying relaxation behavior. These hydrogels, composed of gelatin (5% w/v), alginate (1%w/v) and fibrinogen (2%w/v), were designed to be compatible with micro-extrusion bioprinting and proliferative. The modulation of their biomechanical properties, including stiffness and viscoelastic behavior, was achieved by applying various concentrations of cross-linkers targeting both gelatin covalent bonding (transglutaminase) and alginate chains' ionic cross-linking (calcium). Among the conditions tested, the hydrogel with a low elastic modulus of 8 kPa and a viscoelastic behavior over time exhibited promising outcomes regarding osteoblast-to-osteocyte differentiation. The cessation of cell proliferation coincided with a significant increase in alkaline phosphatase (ALP) activity, the development of dendrites, and the expression of the osteocyte marker PHEX. Within this hydrogel, cells actively influenced their environment, as evidenced by hydrogel contraction and the secretion of collagen I. This bio-printed model, demonstrating primary human osteoblasts expressing an osteocyte-specific protein, marks a significant achievement. We envision its substantial utility in advancing research on bone pathologies, including osteoporosis and bone tumors.
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Dates et versions

hal-04762570 , version 1 (31-10-2024)

Identifiants

Citer

Emma Petiot, Pragnere Sarah, Essayan Lucie, El-Kholti Naima, Petiot Emma, et al.. In vitro bioprinted 3D model enhancing osteoblast-to-osteocyte differentiation. Biofabrication, 2024, ⟨https://doi.org/10.1088/1758-5090/ad8ca6⟩. ⟨hal-04762570⟩
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