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The effects of melatonin on the viability and osteogenic/odontogenic differentiation of human stem cells from the apical papilla

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Abstract

Background

An experimental study was conducted to examine whether melatonin influences osteogenic/odontogenic differentiation of human stem cells derived from the apical papilla (hSCAPs).

Materials and methods

In order to isolate hSCAPs, the undeveloped root of a third molar of a human tooth was used. Melatonin was administered to the experimental groups in an osteogenic medium. No treatment was administered to the control group. The methyl thiazolyl tetrazolium (MTT) assay was performed on days 1, 2, and 3 to assess cell viability (n = 8). A determination of odontogenic/osteogenic differentiation was accomplished using alkaline phosphatase (ALP) activity alizarin red staining (ARS) (n = 6), and the expression of osteogenic genes by real-time polymerase chain reaction (RT-PCR) (n = 3) on days 1, 2, and 7. Evaluation of the data was conducted using SPSS version 18. All experiments were conducted at least three times. The Mann Whitney U test, the ANOVA analysis, Tukey’s test, and t-test was implemented to analyze the data (α = 0.05).

Results

After 24 h, 48 h, and 72 h, No significant difference was observed between the control group and the melatonin treatment group in terms of viability of hSCAPs. (from 1 up to 10 µg/ml) (P > 0.05). The assessment of ARS and ALP activity showed that melatonin treatment enhanced osteogenic differentiation of hSCAPs (P < 0.001). Melatonin treatment caused hSCAPs to show an increase of genes related to osteogenic/odontogenic differentiation. These genes included ALP, dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP-1), and bone sialoprotein (BSP) (P < 0.001).

Conclusions

Melatonin treatment enhanced osteogenic/odontogenic differentiation of hSCAPs with a dose dependent effect on cell viability.

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Data Availability

The corresponding author, Elham Khoshbin, possesses the entire documentation of the study participant and other data, which can be accessed through a reasonable request.

Abbreviations

hSCAPs:

Human Stem cells from the apical papilla

MTT:

Methyl thiazolyl tetrazolium

ISSCR:

International Society for Stem Cells Research

qrt-PCR:

Quantitative reverse-transcription polymerase chain reaction

ARS:

Alizarin red staining

LLL:

Low level laser

LLLI:

Low level laser irradiation

LLLT:

Low level laser therapy

ALP:

Alkaline phosphatase

DSPP:

Dentin sialophosphoprotein

DMP1:

Dentin matrix protein 1

BSP:

Bone sialoprotein

DPSCs:

Dental pulp stem cells

hPDL:

Human periodontal ligament

hPDLSCs:

Human periodontal ligament stem cells

LED:

Light-emitting diode

PBS:

Phosphate-buffered saline

SPSS:

Statistical Package of the Social Sciences

SEM:

Standard error of the mean

Fig:

Figure

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Acknowledgements

The Vice-Chancellor of Research and Technology of the Hamadan University of Medical Sciences provided funding for this study (140111119663).

Funding

The Vice-Chancellor of Research and Technology of the Hamadan University of Medical Sciences provided funding for this study (140111119663).

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Author notes

  1. Elham Khoshbin

    Present address: Hamadan Dental School, Shahid Fahmideh Street, PO Box 6517838677, Hamadan, Iran

Authors and Affiliations

  1. Department of Endodontics, Dental Research Center, Hamadan University of Medical Sciences, Hamadan, Iran

    Hamed Karkehabadi

  2. Department of Endodontics, Dental School, Hamadan University of Medical Sciences, Hamadan, Iran

    Roshanak Abbasi & Elham Khoshbin

  3. Department of Medical Molecular & Genetics, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

    Rezvan Najafi

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Contributions

Hamed Karkehabadi: Conceptualization; Study selection; Data collection; Quality assessment; Methodology; Project administration; Resources; Supervision; Validation; Visualization; Writing (review/ editing). Roshanak Abbasi: Conceptualization; Study selection; Data collection; Quality assessment; Methodology; Project administration; Resources; Supervision; Validation; Visualization; Writing (original draft). Rezvan Najafi: Conceptualization; Study selection; Data collection; Quality assessment; Methodology; Project administration; Resources; Supervision; Validation; Visualization: Elham Khoshbin: Conceptualization; Study selection; Data collection; Quality assessment; Methodology; Project administration; Resources; Supervision; Validation; Visualization; Writing (review/ editing). The final manuscript was read and approved by all authors.

Corresponding author

Correspondence to Elham Khoshbin.

Ethics declarations

Ethics approval and consent to participate

The research was carried out in accordance with the ISSCR Guidelines for Stem Cell Research and Clinical Translation, and received approval from the ethics committee of Hamadan University of Medical Sciences (IR.UMSHA.REC.1401.605). The study’s procedure was conducted following ethical standards established in the 1964 Helsinki Declaration and its subsequent amendments.

Informed consent

The patient who was enrolled in the trial received a thorough explanation of the procedures involved. The participant signed an informed permission form for the express aim of using the extracted teeth for study prior to the extraction at the Department of Oral and Maxillofacial Surgery, Hamadan University of Medical Sciences.

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Not applicable.

Competing interests

The authors have nothing significant to disclose in terms of financial or non-financial interests. The material discussed in this article is not subject to any financial or proprietary interests held by the authors.

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Karkehabadi, H., Abbasi, R., Najafi, R. et al. The effects of melatonin on the viability and osteogenic/odontogenic differentiation of human stem cells from the apical papilla. Mol Biol Rep 50, 8959–8969 (2023). https://doi.org/10.1007/s11033-023-08747-0

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