Integrinβ1 (INT-β1) blocking disables the morphology and function of osteocytic cells cultured on normal matrices. (A) SEM and confocal images illustrating distinct morphological changes induced by integrinβ1 blocking in osteocytic cells cultured on the normal matrices (NM). (Scale bars: 50 μm). (B) Shape factor determined on the osteocytic cells cultured on normal matrices upon integrinβ1 blocking. (C) Cell adhesion determined on the osteocytic cells grown on the normal matrices upon integrinβ1 blocking (D to G) Gene expression profiles of osteocyte markers as determined by qRT-PCR in the presence of integrinβ1 blocking antibodies on normal matrix. The results are expressed as relative gene expression after normalisation using the 18srRNA and GAPDH housekeeping gene. (H) Osteocytic cells grown on the normal matrices were treated with integrinβ1 blocking antibody and incubated for 24 hours, and then detection of apoptosis was performed by flow cytometry. (I) Zymographic analysis shows the expression of MMP2 in osteocytic cell conditioned media collected after 24 hours of incubation after culturing on normal matrices in the presence or absence of integrinβ1 blocking antibody. All images and graphs are repremsentative of experiments performed on osteocytic cells cultured on matrices derived from three different normal patients. Results are shown as mean ± SD, * P ≤0.05 by Student’s t-test.
Prasadam et al. Arthritis Research & Therapy 2013 15:R150 doi:10.1186/ar4333