title: In vitro analysis of the effects on wound healing of high- and low-molecular weight chains of hyaluronan and their hybrid H-HA/L-HA complexes.
abstract: BACKGROUND
  Recent studies have reported the roles of Hyaluronic acid (HA) chains of diverse length in wound repair, especially considering the simultaneous occurrence in vivo of both high- (H-HA) and low-molecular weight (L-HA) hyaluronan at an injury site. It has been shown that HA fragments (5 ≤ MW ≤ 20 kDa) usually trigger an inflammatory response that, on one hand, is the first signal in the activation of a repair mechanism but on the other, when it's overexpressed, it may promote unwanted side effects. The present experimental research has aimed to investigate H-HA, L-HA and of a newly developed complex of the two (H-HA/L-HA) for stability (e.g. hyaluronidases digestion), for their ability to promote wound healing of human keratinocytes in vitro and for their effect on cellular biomarker expression trends.
  
  
  RESULTS
  Time-lapse video microscopy studies proved that the diverse HA was capable of restoring the monolayer integrity of HaCat. The H-HA/L-HA complex (0.1 and 1%w/v) proved faster in regeneration also in co-culture scratch test where wound closure was achieved in half the time of H-HA stimulated cells and 2.5-fold faster than the control. Gene expression was evaluated for transformation growth factor beta 1 (TGF-β1) proving that L-HA alone increased its expression at 4 h followed by restoration of similar trends for all the stimuli. Depending on the diverse stimulation (H-HA, L-HA or the complex), metalloproteinases (MMP-2, -9, -13) were also modulated differently. Furthermore, type I collagen expression and production were evaluated. Compared to the others, persistence of a significant higher expression level at 24 h for the H-HA/L-HA complex was found.
  
  
  CONCLUSIONS
  The outcomes of this research showed that, both at high and low concentrations, hybrid complexes proved to perform better than HA alone thus suggesting their potential as medical devices in aesthetic and regenerative medicine.
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. antonella.dagostino@unina2.it.
authors.forename: Antonella
authors.initials: A
authors.lastname: D'Agostino
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. antonietta.stellavato@unina2.it.
authors.forename: Antonietta
authors.initials: A
authors.lastname: Stellavato
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. teresa.busico@alice.it.|ALFATESTLAB s.r.l, Via Pelizza da Volpedo,, 59 20092, Cinisello Balsamo, MI, Italy. teresa.busico@alice.it.
authors.forename: Teresa
authors.initials: T
authors.lastname: Busico
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. agata.papa@alfatestlab.it.|ALFATESTLAB s.r.l, Via Pelizza da Volpedo,, 59 20092, Cinisello Balsamo, MI, Italy. agata.papa@alfatestlab.it.
authors.forename: Agata
authors.initials: A
authors.lastname: Papa
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. virginia.tirino@unina2.it.
authors.forename: Virginia
authors.initials: V
authors.lastname: Tirino
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. gianpaolo.papaccio@unina2.it.
authors.forename: Gianpaolo
authors.initials: G
authors.lastname: Papaccio
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. annalisa.lagatta@unina2.it.
authors.forename: Annalisa
authors.initials: A
authors.lastname: La Gatta
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. mario.derosa@unina2.it.
authors.forename: Mario
authors.initials: M
authors.lastname: De Rosa
authors.affiliation: Department of Experimental Medicine, Section of Biotechnology, Medical Histology and Molecular Biology "A. Cascino", Second University of Naples, Via De Crecchio 7, 80138, Naples, Italy. chiara.schiraldi@unina2.it.
authors.forename: Chiara
authors.initials: C
authors.lastname: Schiraldi
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D018920:Coculture Techniques; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000235:genetics / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D018715:Microscopy, Video; D008970:Molecular Weight; D013291:Streptococcus / Q000378:metabolism; D059008:Time-Lapse Imaging; D059467:Transcriptome / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism; D014783:Viscosity; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D024042:Collagen Type I; D053773:Transforming Growth Factor beta1; D006820:Hyaluronic Acid; D053509:Matrix Metalloproteinase 13; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9