IADR Abstract Archives
The Administration of 2-PCPA to Prevent Bone Damage by Smoking
Objectives: Trans-2-Phenylcyclopropylamin (2-PCPA) is FDA-approved antidepressant. Recently, it was found to accelerate the osteogenic differentiation of human adipocytes derived stem as histone lysine-specific demethylase 1 (LSD1) inhibitor. Our study aimed to explore whether 2-PCPA administration could prevent inflammatory bone damage caused by smoking.
Methods: The effect of smoking on osteogenic differentiation of osteoblastic cells was investigated with murine pre-osteoblasts (Mc3T3-E1) and human bone marrow stroma cells (hBMCs). These cells were pretreated with cigarette smoking extract (CSE), then treated with different dosage of 2-PCPA for 24 hours before their osteogenic induction. The cell cultures are fixed for ALP and mineralization staining after osteogenic induction at 7 and 14 days. Meanwhile, their protein and total RNA were extracted at the different time points, and the expression of osteogenic markers and proinflammatory cytokines were examined by western blotting and Real-time PCR. All the experiments were repeated three times and student t-test was applied for the statistical analysis.
Results: Our experiment showed that administration of 2-PCPA on osteogenic cells increased their ALP staining and mineralization in a dosage-dependent pattern. Both the protein and mRNA expression of osteogenic genes such as Runx-2, ALP, BSP were also significantly increased. CSE treatment decreased the osteogenic gene expressions, while elevated mRNA expression of proinflammatory cytokines such as IL-1, IL-6, and TNF-α. Interestingly, CSE treatment suppress the endogenous protein level of H3k4me2, an important histone methyltransferase regulating osteogenic differentiation. After administration of 2-PCPA in CSE-treated osteoblastic cells, the suppressed osteogenic genes expression were partially restored. Coincidently, H3K4me2 protein level suppressed by CSE was partially restored by 2-PCPA as well.
Conclusions: Our findings suggest that 2-PCPA protected the osteogenic differentiation of osteoblastic cells against smoking through its LSD1 inhibition in vitro. Future work will be largely directed at investigating the optimum dosage of 2-PCPA applied in smoking animal models in vivo.
Methods: The effect of smoking on osteogenic differentiation of osteoblastic cells was investigated with murine pre-osteoblasts (Mc3T3-E1) and human bone marrow stroma cells (hBMCs). These cells were pretreated with cigarette smoking extract (CSE), then treated with different dosage of 2-PCPA for 24 hours before their osteogenic induction. The cell cultures are fixed for ALP and mineralization staining after osteogenic induction at 7 and 14 days. Meanwhile, their protein and total RNA were extracted at the different time points, and the expression of osteogenic markers and proinflammatory cytokines were examined by western blotting and Real-time PCR. All the experiments were repeated three times and student t-test was applied for the statistical analysis.
Results: Our experiment showed that administration of 2-PCPA on osteogenic cells increased their ALP staining and mineralization in a dosage-dependent pattern. Both the protein and mRNA expression of osteogenic genes such as Runx-2, ALP, BSP were also significantly increased. CSE treatment decreased the osteogenic gene expressions, while elevated mRNA expression of proinflammatory cytokines such as IL-1, IL-6, and TNF-α. Interestingly, CSE treatment suppress the endogenous protein level of H3k4me2, an important histone methyltransferase regulating osteogenic differentiation. After administration of 2-PCPA in CSE-treated osteoblastic cells, the suppressed osteogenic genes expression were partially restored. Coincidently, H3K4me2 protein level suppressed by CSE was partially restored by 2-PCPA as well.
Conclusions: Our findings suggest that 2-PCPA protected the osteogenic differentiation of osteoblastic cells against smoking through its LSD1 inhibition in vitro. Future work will be largely directed at investigating the optimum dosage of 2-PCPA applied in smoking animal models in vivo.