The effects of Pannexin1 in the osteogenic differentiation of the bone marrow mesenchymal stem cells with mechanical stimulation / 中华骨科杂志

ABSTRACT

Objective To explore the effects of Pannexin1 (Px1) channel protein on osteogenic differentiation of mesen?chymal stem cells (MSCs) under mechanical stress stimulation. Methods Bone marrow was extracted from Sprague Dawley (SD) rats (3 weeks, weighing 100-120 g) and cultured following 1st generation cellular technology. The safe dose of CBX (an inhibitor of Px1 channel protein) on MSCs was determined by CCK8 method and 100 μmol/L was used. The MSCs were cultured by the whole marrow culture method. When the cell was passaged to 3-4 generation, high purity of the MSCs were harvested. MSCs were divided into three groupscontrol group, stress?stimuli group (4 000μstrain) and stress stimuli+CBX group. The duration of stress was 15 min and CBX pre?treatment time point was 3 h, 6 h, 12 h, and 24 h, respectively. The alkaline phosphatase (ALP) activity, type I collagen expression, intracellular calcium ion (Ca2+) concentration, and Px1 channel protein, p?p38MAPK and p?ERK ex?pression were determined. Results ALP activities were highest in the stress group, and it was reduced by pretreatment of CBX. Similarly, stress increased the expression of type I collagen, concentration of Ca2+, and expressions of Px1 channel protein and p?p38MAPK was reduced by CBX. p?ERK was down?regulated by stress but not affected by CBX treatment. Conclusion Mechani?cal stress could promote osteogenic differentiation of MSCs and this promotion was inhibited by pretreatment of CBX, which might result in regulation of p?p38MAPK.