Biologically Relevant In Vitro 3D-Model to Study Bone Regeneration Potential of Human Adipose Stem Cells

• Published in: Biomolecules (Basel, Switzerland) Vol. 12; no. 2; p. 169
• Main Authors: van Santen, Victor J B, Bastidas Coral, Angela P, Hogervorst, Jolanda M A, Klein-Nulend, Jenneke, Bakker, Astrid D
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 21.01.2022; MDPI
• Subjects: adipose stem cells; Adipose Tissue; biphasic calcium phosphate; Body fat; Bone growth; Bone Regeneration; Bones; Calcium phosphates; Cbfa-1 protein; Cell culture; Cell Differentiation; Cell proliferation; Cells, Cultured; Cytokines; Experiments; Gene expression; Humans; Hypoxia; Inflammation; Interleukin 1; Interleukin 4; Interleukin 6; Mesenchymal stem cells; mesenchymal stromal cells; Mesenchyme; Metabolism; Osteogenesis; Proteins; Regeneration; Stem Cells; Stromal cells; Tissue engineering; Tumor necrosis factor-α; United States > US; Netherlands; adipose stem cells; hypoxia; proliferation; osteogenic differentiation; mesenchymal stromal cells; bone regeneration; cytokines; biphasic calcium phosphate
• ISSN: 2218-273X; 2218-273X
• DOI: 10.3390/biom12020169

Standard cell cultures may not predict the proliferation and differentiation potential of human mesenchymal stromal cells (MSCs) after seeding on a scaffold and implanting this construct in a bone defect. We aimed to develop a more biologically relevant in vitro 3D-model for preclinical studies on the bone regeneration potential of MSCs. Human adipose tissue-derived mesenchymal stromal cells (hASCs; five donors) were seeded on biphasic calcium phosphate (BCP) granules and cultured under hypoxia (1% O ) for 14 days with pro-inflammatory TNFα, IL4, IL6, and IL17F (10 mg/mL each) added during the first three days, simulating the early stages of repair (bone construct model). Alternatively, hASCs were cultured on plastic, under 20% O and without cytokines for 14 days (standard cell culture). After two days, the bone construct model decreased total DNA (3.9-fold), (9.8-fold), and expression (19.6-fold) and metabolic activity (4.6-fold), but increased expression (38.6-fold) in hASCs compared to standard cultures. After seven days, the bone construct model decreased expression (64-fold) and metabolic activity (2.3-fold), but increased (54.5-fold) and expression (5.7-fold) in hASCs compared to standard cultures. The effect of the bone construct model on hASC proliferation and metabolic activity could be largely mimicked by culturing on BCP alone (20% O , no cytokines). The effect of the bone construct model on expression could be mimicked by culturing hASCs under hypoxia alone (plastic, no cytokines). In conclusion, we developed a new, biologically relevant in vitro 3D-model to study the bone regeneration potential of MSCs. Our model is likely more suitable for the screening of novel factors to enhance bone regeneration than standard cell cultures.