Fluence-Dependent Effects of 940-nm Photobiomodulation on the Proliferation and Osteoblastic Differentiation of Human Periodontal Ligament Stem Cells.
6 affiliations
- 1Clinic for Orthodontics, School of Dental Medicine, University of Belgrade, 11000 Belgrade, Serbia.
- 2Institute for the Application of Nuclear Energy, University of Belgrade, 11000 Belgrade, Serbia.
- 3Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, 11000 Belgrade, Serbia.
- 4Clinic for Stomatology, Medical Faculty of the Military Medical Academy, University of Defense, 11154 Belgrade, Serbia.
- 5Serbian Academy of Sciences and Arts, Knez Mihailova 35, 11000 Belgrade, Serbia.
- 6Faculty of Medicine Foča, University of East Sarajevo, 73300 Foča, Bosnia and Herzegovina.
Abstract
Background/Objectives: Photobiomodulation (PBM) has been proposed to regulate periodontal tissue remodeling, but its effects on osteoblastogenesis in periodontal ligament stem cells (PDLSCs) remain highly dependent on laser parameters. This study investigated the fluence-dependent effects of 940 nm diode laser irradiation on the proliferation and osteoblastic differentiation of human PDLSCs. Methods: Six primary human PDLSC cultures were established and characterized using flow cytometry and multilineage differentiation assays. Cell proliferation was assessed after a single irradiation at fluences of 1, 2, 4, and 10 J/cm². Osteoblastogenesis was induced using repeated irradiation regimens initiated at 2, 4, or 10 J/cm² under suboptimal or complete osteogenic conditions and evaluated by Alizarin Red staining on day 21. The expression of osteogenic genes (RUNX2, BMP2, SPP1, TGFB1, COL1A1, ALPL, and BGLAP) was evaluated by qRT-PCR at different time points. Culture supernatants were analyzed for soluble osteogenic mediators. Results: The established PDLSC cultures had an MSC-like phenotype, robust osteogenic potential, moderate chondrogenic potential, and weak or no adipogenic potential. Proliferation of PDLSCs was considerably increased by PBM at 2 J/cm² on day 7, whereas fluences of 4 and 10 J/cm² inhibited proliferation. Osteoblastic mineralization was increased with the 2 J/cm² irradiation regimen but was markedly reduced with the 10 J/cm² regimen. Gene expression analysis confirmed this fluence-dependent response under optimal culture conditions. The 2 J/cm² irradiation regimen significantly upregulated each analyzed osteogenic gene at one or more time points, whereas the 10 J/cm² regimen downregulated RUNX2, BMP2, SPP1, COL1A1, and BGLAP. PBM at 2 J/cm² also increased production of soluble BMP-2, OPG, PDGF-BB, ALP, and Leptin in a time-dependent manner. Conclusions: PBM exerts a fluence-dependent effect on human PDLSCs. A single irradiation at 2 J/cm² increased proliferation, whereas the irradiation regimen initiated at 2 J/cm² promoted osteoblastogenesis. In contrast, higher-fluence conditions, particularly the regimen initiated at 10 J/cm², were inhibitory.
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BibTeX
@article{markovic2026fluencedependent,
title = {{Fluence-Dependent Effects of 940-nm Photobiomodulation on the Proliferation and Osteoblastic Differentiation of Human Periodontal Ligament Stem Cells.}},
author = {Jovan Marković and Milan Marković and Jelena Đokić and Sergej Tomić and Luka Pavlović and Mile Eraković and Zorana Stamenković and Miodrag Čolić},
journal = {Dentistry journal},
year = {2026},
month = oct,
doi = {10.3390/dj14100676},
url = {https://europepmc.org/article/MED/42930442},
note = {CC BY},
}
RIS
TY - JOUR
TI - Fluence-Dependent Effects of 940-nm Photobiomodulation on the Proliferation and Osteoblastic Differentiation of Human Periodontal Ligament Stem Cells.
AU - Jovan Marković
AU - Milan Marković
AU - Jelena Đokić
AU - Sergej Tomić
AU - Luka Pavlović
AU - Mile Eraković
AU - Zorana Stamenković
AU - Miodrag Čolić
PY - 2026
DA - 2026/10/16
JO - Dentistry journal
DO - 10.3390/dj14100676
UR - https://europepmc.org/article/MED/42930442
AB - Background/Objectives: Photobiomodulation (PBM) has been proposed to regulate periodontal tissue remodeling, but its effects on osteoblastogenesis in periodontal ligament stem cells (PDLSCs) remain highly dependent on laser parameters. This study investigated the fluence-dependent effects of 940 nm diode laser irradiation on the proliferation and osteoblastic differentiation of human PDLSCs. Methods: Six primary human PDLSC cultures were established and characterized using flow cytometry and multilineage differentiation assays. Cell proliferation was assessed after a single irradiation at fluences of 1, 2, 4, and 10 J/cm². Osteoblastogenesis was induced using repeated irradiation regimens initiated at 2, 4, or 10 J/cm² under suboptimal or complete osteogenic conditions and evaluated by Alizarin Red staining on day 21. The expression of osteogenic genes (RUNX2, BMP2, SPP1, TGFB1, COL1A1, ALPL, and BGLAP) was evaluated by qRT-PCR at different time points. Culture supernatants were analyzed for soluble osteogenic mediators. Results: The established PDLSC cultures had an MSC-like phenotype, robust osteogenic potential, moderate chondrogenic potential, and weak or no adipogenic potential. Proliferation of PDLSCs was considerably increased by PBM at 2 J/cm² on day 7, whereas fluences of 4 and 10 J/cm² inhibited proliferation. Osteoblastic mineralization was increased with the 2 J/cm² irradiation regimen but was markedly reduced with the 10 J/cm² regimen. Gene expression analysis confirmed this fluence-dependent response under optimal culture conditions. The 2 J/cm² irradiation regimen significantly upregulated each analyzed osteogenic gene at one or more time points, whereas the 10 J/cm² regimen downregulated RUNX2, BMP2, SPP1, COL1A1, and BGLAP. PBM at 2 J/cm² also increased production of soluble BMP-2, OPG, PDGF-BB, ALP, and Leptin in a time-dependent manner. Conclusions: PBM exerts a fluence-dependent effect on human PDLSCs. A single irradiation at 2 J/cm² increased proliferation, whereas the irradiation regimen initiated at 2 J/cm² promoted osteoblastogenesis. In contrast, higher-fluence conditions, particularly the regimen initiated at 10 J/cm², were inhibitory.
SN - 2304-6767
ER -