During the last years, several attempts have been accomplished to improve the wound healing. Device application aimed at enhancing skin ability to reconstruct its damaged sites through a proper dermal regenerative process. In particular, Q-switched
Nd-YAG laser (Medlite C6
laser, Conbio, USA) applied with a fluence of 8 J/cm2, a pulse width of 5 ns, and a spot size of 4 mm exerts a photo-mechanical action that improve skin repair. Besides,
hyaluronan hybrid cooperative complexes (HCC) widely exploited in dermoesthetic applications proved specific actions on keratinocytes and fibroblasts monolayer repair. We evaluated this specific
laser treatment in vitro on a wound healing model based on human keratinocytes (HaCaT) alone and in combination with HCC. In addition, we evaluated key
biomarkers of dermal repair. Scratched HaCaT monolayers were treated with
laser and successively with HA-based formulations (HHA and HCC). For each treatment and the control samples, at least 3 different wells were analyzed.
Wound closure was quantified, measuring five view filed for each well at increasing incubation time, exploiting time lapse videomicroscopy and image analysis, permitting to compare the different healing rate of treatments respect to control. By real-time PCR and western blotting, we evaluated
biomarkers of
wound regeneration, such as
integrins,
aquaporin three (AQP3), and proinflammatory
cytokines. The ANOVA test was used to assess statistical significance of the results obtained.
Laser-treated cells achieved
wound closure in about 37 h, faster than the control, while when coupled to HCC, the complete reparation was obtained in 24 h.
Integrin αV was upregulated by treatments, with in particular about four-fold increase respect to the control when HCC + laser was used. In addition,
integrin β3 was upregulated by all treatments especially with the combination of
laser and HCC proved more efficient than others (~ 14-folds). A slighter but significant increase of AQP3 gene expression of 61% was found for
laser treatment while the latter combined with HCC determined an upregulation of 72%. By coupling
laser treatment and HCC, further healing improvement and consistent
biomarker modulation was observed. Our results may support clinical implementation of new dermatology protocols conjugating
laser treatments with topical or injective HA formulations as a valid tool in treatments to repair
scars or other skin defects.