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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\u00a0J\/cm<jats:sup>2<\/jats:sup>, a pulse width of 5\u00a0ns, and a spot size of 4\u00a0mm 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\u00a0h, faster than the control, while when coupled to HCC, the complete reparation was obtained in 24\u00a0h. Integrin \u03b1V was upregulated by treatments, with in particular about four-fold increase respect to the control when HCC\u2009+\u2009laser was used. In addition, integrin \u03b23 was upregulated by all treatments especially with the combination of laser and HCC proved more efficient than others (~\u200914-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.<\/jats:p>","DOI":"10.1007\/s10103-020-03145-5","type":"journal-article","created":{"date-parts":[[2020,9,26]],"date-time":"2020-09-26T10:02:44Z","timestamp":1601114564000},"page":"1047-1057","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Q-switched Nd-YAG laser alone and in combination with innovative hyaluronic acid gels improve keratinocytes wound healing in vitro"],"prefix":"10.1007","volume":"36","author":[{"given":"Anna","family":"de Filippis","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonella","family":"D\u2019Agostino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna Virginia Adriana","family":"Pirozzi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Antonietta","family":"Tufano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1066-843X","authenticated-orcid":false,"given":"Chiara","family":"Schiraldi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adone","family":"Baroni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,9,26]]},"reference":[{"key":"3145_CR1","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1056\/NEJM199909023411006","volume":"341","author":"AJ Singer","year":"1999","unstructured":"Singer AJ, Clark RA (1999) Cutaneous wound healing. 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In fact, here we used a commercially available cell line of human keratinocyte.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Compliance with ethical standards"}},{"value":"The treatments are among approved medical device for human use in Europe (CE marked), and therefore applicable to in vitro test on human cells.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Compliance with ethical standards"}},{"value":"The authors declare that they have no conflict of interest.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"This experimental work was accomplished using immortalized human keratinocytes obtained by Istituto Zooprofilattico Bologna, Italy. The research did not involve any human and animal species, it is part of the main research activity of the Department of Experimental Medicine, and thus does not need any approval from the University Ethical committee.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}},{"value":"Not required. In fact, this research is not inherent to any clinical study; there were no patients involved and therefore the request of informed consent does not apply.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Informed consent"}}]}}