IADR Abstract Archives
Mast Cells Increase Gene Expression Of Osteoclastogenic Markers In SHR
Objectives: Spontaneously hypertensive rats (SHR) present genetic alterations in bone metabolism and in adaptive immunity. Mast cells (MCs) and their secreted products play a fundamental role in inflammatory response, tissue repair and defense of the organism and may also be involved in bone metabolism which play a potentially pathogenic role in osteoporosis bone loss. The aim of this study was to evaluate the influence of mast cells in the gene expression of bone resorption markers in osteosclasts cells from bone marrow of SHR in vitro.
Methods: Cells extracted from bone marrow of femur of 5 week-old male SHR were incubated in α-MEM medium with soluble macrophage colony-stimulating factor (M-CSF 100ng/mL) for 3 days. Then, these cells were plated in 24-well culture plates at density of 1x104 cells/well where they received osteclastogenic medium (OCM, M-CSF 30ng/mL; RANK-L 100ng/mL) for 7 days. The RBL-2H3 MC lines were added in inserts at density of 2x104 cells/insert at the 4th day of osteoclast culture, forming co-culture. Were evaluated the gene expression of osteoclastogenic markers at day 7 by qRT-PCR.
Results: The addition of MCs in the osteoclasts culture decreased gene expression of transcription factor evaluated as NFATc1 (p<0.05) and osteoclast-associated receptor (OSCAR). On the other hand, the co-culture increased the expression of tartrate-resistant acid phosphatase (TRAP), cathepsin-K (CtsK), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and suppressor of cytokine signaling 1 (SOCS-1) (p<0.001).
Conclusions: Indirect mast cell co-culture decreased gene expression of transcription factors involved in the osteoclast differentiation and increased the gene expression profile of important osteoclastogenic markers involved in the matrix degradation and bone resorption process.
Methods: Cells extracted from bone marrow of femur of 5 week-old male SHR were incubated in α-MEM medium with soluble macrophage colony-stimulating factor (M-CSF 100ng/mL) for 3 days. Then, these cells were plated in 24-well culture plates at density of 1x104 cells/well where they received osteclastogenic medium (OCM, M-CSF 30ng/mL; RANK-L 100ng/mL) for 7 days. The RBL-2H3 MC lines were added in inserts at density of 2x104 cells/insert at the 4th day of osteoclast culture, forming co-culture. Were evaluated the gene expression of osteoclastogenic markers at day 7 by qRT-PCR.
Results: The addition of MCs in the osteoclasts culture decreased gene expression of transcription factor evaluated as NFATc1 (p<0.05) and osteoclast-associated receptor (OSCAR). On the other hand, the co-culture increased the expression of tartrate-resistant acid phosphatase (TRAP), cathepsin-K (CtsK), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and suppressor of cytokine signaling 1 (SOCS-1) (p<0.001).
Conclusions: Indirect mast cell co-culture decreased gene expression of transcription factors involved in the osteoclast differentiation and increased the gene expression profile of important osteoclastogenic markers involved in the matrix degradation and bone resorption process.