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- W1879392941 abstract "Osteocytes are currently regarded as being pivotal in maintaining bone homeostasis.They differentiate from osteoblasts, are embedded in mineralised matrix and difficultto isolate. Current models for in vitro osteocyte studies are limited. Others havesuggested that osteoblasts in 3-dimensional (3D) cultures differentiate to osteocytes.This study aimed to develop 3D cultures enabling differentiation of osteoblasts toosteocytes, which could be used for studies of osteocyte differentiation andresponses to mechanical loading. Furthermore, the effects of external compounds onosteoblast differentiation in 3D were assessed.Mouse (MC-3T3, IDG-SW3) and human primary osteoblasts (hOBs) weremaintained in type I collagen gels in either non-osteogenic or osteogenic media, and+/- compounds such as insulin-like growth factor-1 (IGF-1). Furthermore,mechanical loading (5 mins, 10 Hz, 2.5 N) was applied to 3D cultures and responsescharacterised.Cells were viable in collagen gels for 25 days, and expressed mRNA for matureosteocyte markers e.g. sclerostin in osteogenic medium. Furthermore, IGF-1 upregulatedmRNA expression of osteocyte markers and other molecules (e.g. receptoractivator of the nuclear factor kappa-β ligand - RANKL - 43-fold) in MC-3T3 cells,indicating modulation of cell differentiation and function. Osteocyte markers wereexpressed earlier in IDG-SW3 cells in 3D compared to published marker expressionprofiles in 2D monolayer cultures. Following mechanical loading, known mechanosensitivemarkers were modulated in IDG-SW3 cells in 3D, for example, RANKLand vascular endothelial growth factor (VEGF) up-regulated and sclerostin downregulatedpost-loading.This 3D model enables differentiation of osteoblasts to osteocytes in an environmentakin to osteocytes in vivo. External compounds accelerated cell differentiation, andthis was also accelerated in 3D compared to monolayer. Furthermore, the 3D modelenabled osteocyte mechanical loading. This model can be used with human cells,will further our understanding of osteocyte differentiation, and inform on osteocytefunction including their responses to mechanical loading." @default.
- W1879392941 created "2016-06-24" @default.
- W1879392941 creator A5051198270 @default.
- W1879392941 date "2015-01-01" @default.
- W1879392941 modified "2023-09-27" @default.
- W1879392941 title "Differentiation of osteoblasts to osteocytes in 3D type I collagen gels - a novel tool to study osteocyte responses to mechanical loading" @default.
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