Effect of simulated microgravity on osteocytes responding to fluid shear stress

Abstract

Osteocytes, as most abundant cells and major mechanical sensor in bone, play an important role in the mechanism of microgravity-induced bone loss. The response of osteocytes to fluid flow stress under simulated microgravity was investigated in this study. MLO-Y4, an osteocyte-like cell line, was cultured under simulated microgravity condition for 5 days. Then cells were sheared at 15dyn/cm2 in flow chamber. After 15min shear, nitric oxide (NO) was examined by Griess Reagent and prostaglandin E2 (PGE2) by ELISA. After 6h shear, alkaline phosphatase (ALP) was examined by PNPP, osteocalcin (OC) and procollagen type I N propeptide (PINP) by ELISA. Cells were divided into four groups: CON (1G with no shear), CON-S (1G with shear), SM (simulated microgravity with no shear) and SM-S (simulated microgravity with shear). The results showed that (1) NO, ALP activity, OC and PINP increased significantly while PGE2 showed no change in SM compared with CON. (2) NO, PGE2, ALP activity and PINP increased significantly while OC decreased significantly in CON-S compared with CON. (3) NO in SM-S had no significant difference compared to SM, PGE2 and OC increased while ALP activity and PINP decreased significantly in SM-S compared with SM. (4) The increasing amplitude of PGE2 and OC, the decreasing amplitude of ALP activity in SM-S to SM was lower than that in CON-S to CON. In addition, some changes of F-actin cytoskeleton were observed by confocal microscopy. All results indicated that the response induced by fluid shear in osteocytes could be inhibited by simulated microgravity, namely the mechanosensibility of osteocytes decreased under simulated microgravity. This may partly contribute to the mechanism of microgravity-induced osteoporosis and will be helpful to find out effective description.