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Enhanced osteogenic differentiation potential of Arnica montana and Bellis perennis in C3H10T1/2 multipotent mesenchymal stem cells

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Abstract

Background

Arnica montana and Bellis perennis are two medicinal plants that are thought to accelerate bone repair in homoeopathic literature. Mesenchymal stem cells (MSCs) are multipotent stem cells with the ability to differentiate and regenerate bone or osteogenesis. Hence, we aimed to determine the role of Arnica montana and Bellis perennis on the osteogenic differentiation of the C3H10T1/2 stem cell line.

Methods and Results

The cell proliferation of Arnica montana and Bellis perennis was evaluated by MTT assay. Osteogenic differentiation of C3H10T1/2 was induced by the addition of β-glycerophosphate, ascorbic acid and dexamethasone in the differentiation medium over 3 weeks. Cells were treated with Arnica montana and Bellis perennis individually as well as in combination. The osteogenic differentiation potential of Arnica montana and Bellis perennis to differentiate C3H10T1/2 into osteoblasts was measured by alkaline phosphatase activity, alizarin red staining and the expression of Osteocalcin using immunostaining and qRT-PCR.

Arnica montana and Bellis perennis could enhance C3H10T1/2 cell proliferation at 1600 µg. Further, the compound showed the ability to augment osteogenesis as confirmed by increased expression of alkaline phosphatase and enhanced calcium accumulation as seen by the Alizarin Red staining and quantification. Enhanced osteogenesis was further supported by the increased expression of osteocalcin in the treated cells with individual and combined doses of Arnica montana and Bellis perennis. Therefore, the findings provide additional support for the positive impact of Arnica montana and Bellis perennis on bone formation.

Conclusions

Our findings suggest that homoeopathic compounds Arnica montana and Bellis perennis can augment osteogenesis individually as well as in combination.

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Data availability

No Data is associated in the manuscript.

Abbreviations

ALP:

Alkaline phosphatase

ARS:

Alizarin red staining

DAPI:

4′,6-Diamidino-2-phenylindole

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

Dimethyl sulfoxide

FBS:

Foetal bovine serum

MSCs:

Mesenchymal stem cells

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

OCN:

Osteocalcin

ODM:

Osteogenic differentiation medium

PBS:

Phosphate buffered saline

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Acknowledgements

The authors would like to thank the Yenepoya Research Centre, Yenepoya Deemed to be University Mangalore, for its infrastructure and core facility support for conducting this research. We also thank Mrs. Yashaswini Devi G V, PhD scholar, Yenepoya Research Centre, Yenepoya (Deemed to be University) for the timely help provided during the staining experiments.

Funding

This research was funded by Yenepoya University seed grant YU/Seed Grant/078-2019. awarded to the Principal Investigator (Sudheer Shenoy P).

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Author notes

  1. Bhat K. Deepika and Nagendra H. Apoorva have equally contributing first authors.

Authors and Affiliations

  1. Stem Cells and Regenerative Medicine Centre, Yenepoya Research Centre, Yenepoya Deemed to Be University, University Road, Deralakatte, Mangalore, Karnataka, 575018, India

    Bhat K. Deepika, Nagendra H. Apoorva, Pinto R. Joel, Bipasha B. & Shenoy P. Sudheer

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  1. Bhat K. Deepika
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Contributions

DBK, AHN and JRP were responsible for the study design, development of experiments, data collection, statistical analysis, preparing of figures and initial manuscript writing. BB and SS guided the design and implementation of the whole research, analysed data, prepared manuscript-quality figures, wrote and edited the manuscript. All authors reviewed the results and approved the final version of the manuscript.

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Correspondence to Shenoy P. Sudheer.

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Deepika, B.K., Apoorva, N.H., Joel, P.R. et al. Enhanced osteogenic differentiation potential of Arnica montana and Bellis perennis in C3H10T1/2 multipotent mesenchymal stem cells. Mol Biol Rep 51, 596 (2024). https://doi.org/10.1007/s11033-024-09509-2

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