Low-energy helium neon laser irradiation does not alter human keratinocyte differentiation.

There are reports that low-energy HeNe irradiation can enhance wound healing in vivo. We have previously demonstrated that HeNe irradiation increases the motility of human epidermally derived keratinocytes in vitro. Here we investigate whether HeNe irradiation alters normal keratinocyte differentiation, which is essential for the formation of a normal, functioning epidermis. Subconfluent keratinocyte cultures were irradiated three times within 24 h with either 0, 0.8, 3, or 7.2 J/cm2. After cultures reached post-confluence, parameters of growth and differentiation, such as cell number, cornified envelope (CE) formation, and transglutaminase activity were measured. No significant differences were found between the control (0 J) and irradiated cultures in these assays. We also examined the pattern of newly synthesized keratins in cultures irradiated with 7.2 J/cm2 three times within a 24-h period. Both control and irradiated cultures exhibited similar keratin patterns. These results provide evidence that HeNe irradiations of up to 7.2 J/cm2 have no direct deleterious effect on normal keratinocyte differentiation needed for the formation of a functional epidermis. Hence, it is anticipated that the clinical use of the HeNe laser irradiance that enhances keratinocyte migration in vitro (0.8 J/cm2) to promote wound healing in vivo will not alter the ultimate integrity or differentiated function of the epidermis that migrates to cover the wounded area.

Low-energy helium-neon laser irradiation increases the motility of cultured human keratinocytes.

Helium-neon (HeNe) laser irradiation is known to stimulate wound healing. We investigated whether the biostimulatory effects of HeNe irradiation result from enhancement of keratinocyte proliferation or motility. HeNe effects on keratinocyte motility were evaluated by irradiating a "wounded" culture with 0.8 J/cm2 3 times over a 20-h period. At 20 h post-irradiation, videocinemicroscopy and sequential quantitative measurements of the leading edge were taken over a 6-h period. There was a significant difference in migration of the leading edge in irradiated "wounds" compared to non-irradiated "wounded" controls (12.0 microns/h vs 4.0 microns/h, p less than 0.0001). To determine if the increase in migration observed in irradiated cultures resulted from a proliferative effect of HeNe irradiation, subconfluent human keratinocyte cultures were irradiated with single or multiple doses of different fluences of HeNe irradiation (0.4 to 7.2 J/cm2) and evaluated 72 h post-irradiation. Irradiated and non-irradiated keratinocyte cultures grown on a microporous membrane surface were co-cultured with irradiated and non-irradiated fibroblasts to determine if HeNe irradiation induced a paracrine effect on keratinocyte proliferation. No significant increase in keratinocyte proliferation was demonstrated in any of these treatments. The biostimulatory effects of HeNe irradiation may now be extended to include enhancement of keratinocyte motility in vitro; this may contribute to the efficacy of HeNe irradiation in wound healing.