METHODS: Prior to analyzing the ability of this novel combined herbal therapy to promote aspects of bone regeneration, its cytotoxicity was determined using MC3T3-E1 cells (pre-osteoblast model). Cell proliferation was evaluated using phase-contrast microscopy and cell differentiation was estimated using alkaline phosphatase activity. The effect of the combined herbal therapy (CUR + FLL) was also assessed in terms of mineralization in the extracellular matrix (ECM) of cultured cells. Further, to explore the molecular mechanisms of bone formation, time-dependent expression of bone-regulating protein biomarkers was also evaluated.
RESULTS: Combined herbal therapy (CUR + FLL) significantly upregulated the viability, proliferation and differentiation of MC3T3-E1 cells compared to the monotherapy of CUR or FLL. The magnitude of ECM mineralization (calcium deposition) was also higher in MC3T3-E1 cells treated with combined therapy. The time-dependent expression of bone-forming protein biomarkers revealed that the tendency of expression of these bone-regulating proteins was remarkably higher in cells treated with combined therapy.
CONCLUSION: The co-administration of CUR and FLL had superior promotion of elements of bone regeneration in cultured cells, thus could be a promising alternative herbal therapy for the management of bone erosive disorders such as osteoporosis.
Materials and Methods: Extracted human teeth were biomechanically prepared, vertically sectioned, placed in the tissue culture wells exposing the root canal surface to E. faecalis to form a biofilm. At the end of the third week, all groups were treated for 15 min with the test solutions and the control. The results were analyzed both quantitatively and qualitatively.
Results: Statistical analysis was performed by using one-way analysis of variance and compared by the Mann-Whitney test using the Statistical Package for the Social Sciences (SPSS) software, version 20.0. The qualitative assay with the 3-week biofilm on the canal portion showed complete inhibition of bacterial growth for NaOCl, whereas samples treated with herbal solutions showed significant reduction of bacterial growth compared to control group, which showed 139.9 × 109 CFU/mL among the experimental herbal solutions groups. P. amarus has shown maximum bacterial count followed by C. longa and T. indica.
Conclusion: NaOCl 5% showed maximum antibacterial activity against 3-week biofilm on tooth substrate. T. indica, P. amarus, and C. longa showed statistically significant antibacterial activity against 3-week biofilm. The use of herbal alternatives might prove to be advantageous considering the several undesirable characteristics of NaOCl.