Abstract
Background: This study investigates the effect of tannic acid (TA) combined with pamidronate (PAM) on a human osteoblast cell line. Methods: EC50 for TA, PAM, and different combination ratios of TA and PAM (25:75, 50:50, 75:25) were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The combination index value was utilized to analyze the degree of drug interaction, while trypan blue assay was applied to analyze the cells proliferation effect. The mineralization and detection of bone BSP and Osx genes were determined via histochemical staining and PCR test, respectively. Results: The EC50 of osteoblasts treated with TA and a 75:25 ratio of TA and PAM were more potent with lower EC50 at 0.56 µg/mL and 0.48 µg/mL, respectively. The combination of TA and PAM (75:25) was shown to have synergistic interaction. On Day 7, both TA and PAM groups showed significantly increased proliferation compared with control and combination groups. On Day 7, both the TA and combination-treated groups demonstrated a higher production of calcium deposits than the control and PAM-treated groups. Moreover, on Day 7, the combination-treated group showed a significantly higher expression of BSP and Osx genes than both the TA and PAM groups. Conclusion: Combination treatment of TA and PAM at 75:25 ameliorated the highest enhancement of osteoblast proliferation and mineralization as well as caused a high expression of BSP and Osx genes.
Highlights
Bone is a dynamic tissue that constantly undergoes two processes throughout life, namely, modeling and remodeling, to grow or change shape
MTT assay was done to investigate the effect of tannic acid (TA) and different combinations of TA and PAM toward the hFOB 1.19 cell line compared to PAM as positive control
The MTT assay illustrates that the TA and combinations of TA with PAM stimulated better cellular viability compared with PAM alone as the control drug
Summary
Bone is a dynamic tissue that constantly undergoes two processes throughout life, namely, modeling and remodeling, to grow or change shape. An active osteoblast secretes the earliest bone formation marker, which is osterix (Osx). This will reduce the number of osteoblasts by converting it to osteocytes [5]. The mineralization and detection of bone BSP and Osx genes were determined via histochemical staining and PCR test, respectively. On Day 7, both TA and PAM groups showed significantly increased proliferation compared with control and combination groups. On Day 7, the combination-treated group showed a significantly higher expression of BSP and Osx genes than both the TA and PAM groups. Conclusion: Combination treatment of TA and PAM at 75:25 ameliorated the highest enhancement of osteoblast proliferation and mineralization as well as caused a high expression of BSP and Osx genes
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