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Photobiomodulatory effect delivered by low-level laser on dental pulp stem cell differentiation for osteogenic lineage

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Abstract

Background

Photobiomodulation (PBM) therapy has attracted major interest in the field of tissue engineering as it can enhance stem cell differentiation. It has been shown that PBM therapy can stimulate differentiation of cells in culture by exerting biomodulatory effect. Recent evidences show that PBM therapy can positively modulate dental pulp stem cell (DPSC) properties. Combination of PBM therapy with growth factors and biomaterials can possibly accelerate osteogenic differentiation of dental pulp stem cells.

Aims

To evaluate the biomodulatory effect of low-level laser dose on dental pulp stem cells in the presence of hydroxyapatite-based scaffold particle for osteogenic differentiation.

Materials and methods

DPSCs were harvested from human premolar teeth and expanded using mesenchymal stem cell medium. Characterization of DPSCs was done using fluorescence-activated cell sorting with CD105, CD44, CD34, and CD45 markers. Cultured DPSCs along with the N-acetylcysteine-labeled hydroxyapatite (NAC-HA) particles and osteogenic differentiation media were exposed to gallium-aluminum-arsenide (Ga-Al-As) diode laser at 810 nm. Cells were divided into 3 groups: L1 (single exposure), L2 (double exposure), and control (no exposure). Osteodifferentiation after PBM therapy was assessed using Alizarin red S staining, Alkaline phosphatase activity (ALP), and by osteopontin expression. Differences between groups at each time point were analyzed using the Mann–Whitney U test. A level of significance of 5% was adopted (p < 0.05).

Results

DPSCs grown on NAC-HA polymers show increased cell adhesion and proliferation. Double irradiated groups were consistent with increased calcium (71%) and alkaline phosphatase activity (75%) when compared with single-irradiated groups. mRNA expression of osteopontin was relatively increased in a significant (p < 0.001) manner in L2 when compared with L1 group. Alizarin red S and ALP positive staining confirmed the presence of calcium deposition in the test samples. The osteopontin expression of L2 (216.681) as compared with L1 (123.276) group prove the efficacy of double exposures over a single dose of PBM therapy.

Conclusion

The result envisages the enhanced osteogenic potential of PBM therapy on the differentiation of DPSCs in NAC-HA scaffolds. Double exposure of PBM therapy expresses better biomodulatory effect on DPSCs as compared with the single dose.

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Acknowledgments

The authors acknowledge PMS College of Dental Science and Research for facilitating the infrastructure. The authors would like to profoundly thank Dr. KiranNayak for providing the laser device.

Funding

Partly funded by Department of Biotechnology, Government of India, New Delhi.

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Authors and Affiliations

  1. Department of Oral Pathology and Microbiology, PMS College of Dental Science and Research, Thiruvananthapuram, Kerala, India

    Trivandrum T. Sivakumar, Anna P. Joseph & A. Reshmi

  2. Center of Laser Surgery and Laser Therapy, Universita Degli Studi di Genova, Genova, Italy

    Alex M. Muruppel

  3. Biogenix Research Center, Center for Molecular Biology and Applied Science, Thiruvananthapuram, Kerala, India

    Rajesh Ramachandran

  4. Division of Tissue Engineering & Regeneration Technologies., BMT Wing, Sree Chitra Tirunal Institute for Medical Sciences & Technology, Poojapura, Thiruvananthapuram, Kerala, India

    Prabha D. Nair

  5. Department of Oral Pathology, RMDC, Annamalai University, Chidambaram, Tamil Nadu, India

    Sunil P. Mohan

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  1. Trivandrum T. Sivakumar
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  2. Alex M. Muruppel
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  3. Anna P. Joseph
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Correspondence to Trivandrum T. Sivakumar.

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The authors declare that they have no conflict of interest.

Ethical approval

Ethical clearance was obtained from the institutional ethical committee with IEC number: PMS/IEC/2012/25.

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Informed consent was obtained from all individual participants included in the study.

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Sivakumar, T.T., Muruppel, A.M., Joseph, A.P. et al. Photobiomodulatory effect delivered by low-level laser on dental pulp stem cell differentiation for osteogenic lineage. Laser Dent Sci 3, 175–181 (2019). https://doi.org/10.1007/s41547-019-00066-7

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