# Infrared low-level laser therapy enhances proliferation and viability in murine osteoblasts in vitro

**Authors:** Brenda Lizbeth Arroyo Reyes, Luis G. Vázquez-de-Lara-Cisneros, Fabian Galindo Ramírez, Ruben Ramos García, P. Zaca Morán

PMC · DOI: 10.1007/s10103-026-04802-x · 2026-01-22

## TL;DR

Infrared low-level laser therapy boosts the growth and survival of bone cells in lab tests, offering a potential method for bone regeneration.

## Contribution

This study identifies an optimal laser therapy protocol for enhancing osteoblast proliferation without toxicity.

## Key findings

- Three 10 J/cm² infrared LLLT sessions increased osteoblast proliferation significantly.
- Reactive oxygen species levels peaked after the third session but did not cause cytotoxicity.
- LLLT reduced apoptosis in early stages, showing a temporary protective effect on cells.

## Abstract

Infrared low-level laser therapy (LLLT) has shown great promise in promoting cell proliferation and viability, making it a valuable tool in regenerative medicine. This study investigated how the interval between sessions shapes the response to 970 nm LLLT in murine osteoblast cultures by delivering three 10 J/cm² sessions separated by 24–48 h and measuring proliferation, reactive oxygen species (ROS), cytotoxicity, and apoptosis, with the goal of informing protocol design for bone regeneration.

Two osteoblast cultures were used, one control and the other LLL-treated group. The latter consisted of three irradiation sessions (10 J/cm2 each) applied at 24, 48, and 96 h.

The experimental results showed a significant increase in cell proliferation after two and three sessions (p < 0.05), while ROS levels progressively accumulated, peaking after the third session (p < 0.001). Cell viability remained above 90% in both groups during the first 48 h; however, a slight but significant reduction was observed in the LLLT group at 96 h. Apoptosis levels were lower in LLLT-treated cells during early phases (24–48 h), suggesting a transient cytoprotective effect that diminished after the third session. These findings indicate that infrared LLLT promotes cell proliferation without inducing cytotoxicity or programmed cell death.

The results demonstrate that applying three infrared LLLT sessions of 10 J/cm² applied at 24, 48, and 96 h promotes osteoblastic proliferation and viability without inducing cytotoxicity or apoptosis. The proposed protocol, defined by energy dose and irradiation timing, provides a safe and effective strategy for bone tissue engineering.

## Full-text entities

- **Genes:** Cox4i1 (cytochrome c oxidase subunit 4I1) [NCBI Gene 12857] {aka COX, COX IV-1, COXIV, Cox4, Cox4a, IV-1}, Nfkb1 (nuclear factor of kappa light polypeptide gene enhancer in B cells 1, p105) [NCBI Gene 18033] {aka NF-KB1, NF-kappaB, NF-kappaB1, p105, p50, p50/p105}
- **Diseases:** LLLT (MESH:D009800), pain (MESH:D010146), inflammation (MESH:D007249), bone defects (MESH:D001847), Cytotoxicity (MESH:D064420), knee osteoarthritis (MESH:D020370)
- **Chemicals:** amphotericin B (MESH:D000666), CO2 (MESH:D002245), streptomycin (MESH:D013307), propidium iodide (MESH:D011419), penicillin (MESH:D010406), ROS (MESH:D017382), ATP (MESH:D000255), DMEM (-), EthD-1 (MESH:C018533), ascorbic acid (MESH:D001205), phenol red (MESH:D010637), calcium (MESH:D002118), Calcein AM (MESH:C085925), PI (MESH:D010716), glucose (MESH:D005947)
- **Species:** Homo sapiens (human, species) [taxon 9606], Mus musculus (house mouse, species) [taxon 10090]
- **Cell lines:** MG-63 — Homo sapiens (Human), Osteosarcoma, Cancer cell line (CVCL_0426), MC3T3-E1 — Mus musculus (Mouse), Spontaneously immortalized cell line (CVCL_0409)

## Figures

6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12823643/full.md

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Source: https://tomesphere.com/paper/PMC12823643