The effects of prostaglandin E2 on gene expression of IDG-SW3-derived osteocytes in 2D and 3D culture.
Biochemical and biophysical research communications(2022)
摘要
Prostaglandin E2 (PGE2) is a key signaling molecule produced by osteocytes in response to mechanical loading, but its effect on osteocytes is less understood. This work examined the effect of PGE2 on IDG-SW3-derived osteocytes in standard 2D culture (collagen-coated tissue culture polystyrene) and in a 3D degradable poly(ethylene glycol) hydrogel. IDG-SW3 cells were differentiated for 35 days into osteocytes in 2D and 3D cultures. 3D culture led to a more mature osteocyte phenotype with 100-fold higher Sost expression. IDG-SW3-derived osteocytes were treated with PGE2 and assessed for expression of genes involved in PGE2, anabolic, and catabolic signaling. In 2D, PGE2 had a rapid (1 h) and sustained (24 h) effect on many PGE2 signaling genes, a rapid stimulatory effect on Il6, and a sustained inhibitory effect on Tnfrsf11b and Bglap. Comparing culture environment without PGE2, osteocytes had higher expression of all four EP receptors and Sost but lower expression of Tnfrsf11b, Bglap, and Gja1 in 3D. Osteocytes were more responsive to PGE2 in 3D. With increasing PGE2, 3D led to increased Gja1 and decreased Sost expressions and a higher Tnfrsf11b/Tnfsf11 ratio, indicating an anabolic response. Further analysis in 3D revealed that EP4, the receptor implicated in PGE2 signaling in bone, was not responsible for the PGE2-induced gene expression changes in osteocytes. In summary, osteocytes are highly responsive to PGE2 when cultured in an in vitro 3D hydrogel model suggesting that autocrine and paracrine PGE2 signaling in osteocytes may play a role in bone homeostasis.
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