The effect of 648 nm diode laser irradiation on second messengers in senescent human keratinocytes

D. Hawkins Evans; H. Abrahamse

doi:10.1117/12.809009

23 February 2009 The effect of 648 nm diode laser irradiation on second messengers in senescent human keratinocytes

D. Hawkins Evans, H. Abrahamse

Proceedings Volume 7165, Mechanisms for Low-Light Therapy IV; 716509 (2009) https://doi.org/10.1117/12.809009
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Background/purpose: Stress induced premature senescence (SIPS) is defined as the long-term effect of subcytotoxic stress on proliferative cell types. Cells in SIPS display differences at the level of protein expression which affect energy metabolism, defense systems, redox potential, cell morphology and transduction pathways. This study aimed to determine the effect of laser irradiation on second messengers in senescent cells and to establish if that effect can be directly linked to changes in cellular function such as cell viability or proliferation. Materials and Methods: Human keratinocyte cell cultures were modified to induce premature senescence using repeated sub-lethal stresses of 200 uM H₂O₂ or 5% OH every day for four days with two days recovery. SIPS was confirmed by senescence-associated β-galactosidase staining. Control conditions included normal, repeated stress of 500 uM H₂O₂ to induce apoptosis and 200 uM PBN as an anti-oxidant or free radical scavenger. Cells were irradiated with 1.5 J/cm² on day 1 and 4 using a 648 nm diode laser (3.3 mW/cm²) and cellular responses were measured 1 h post irradiation. The affect on second messengers was assessed by measuring cAMP, cGMP, nitric oxide and intracellular calcium (Ca²⁺) while functional changes were assessed using cell morphology, ATP cell viability, LDH membrane integrity and WST-1 cell proliferation. Results: Results indicate an increase in NO and a decrease in cGMP and Ca²⁺ in 200 uM H2O2 irradiated cells while PBN irradiated cells showed a decrease in cAMP and an increase in ATP viability and cell proliferation. Conclusion: Laser irradiation influences cell signaling which ultimately changes the biological function of senescent cells. If laser therapy can stimulate the biological function of senescent cells it may be beneficial to conditions such as immune senescence, skin ageing, muscle atrophy, premature ageing of arteries in patients with advanced heart disease, neurodegenerative disorders and chronic renal failure.

Citation Download Citation

D. Hawkins Evans and H. Abrahamse "The effect of 648 nm diode laser irradiation on second messengers in senescent human keratinocytes", Proc. SPIE 7165, Mechanisms for Low-Light Therapy IV, 716509 (23 February 2009); https://doi.org/10.1117/12.809009

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