An artificial wound in a confluent monolayer of human keratinocyte HaCaT cells or mouse embryofibroblast Swiss NIH 3T3 cells was used to analyze the effects of the nitric oxide (NO) chemical donor, S-nitroso-N-acetylpenicillamine (SNAP). SNAP exposure promoted an enhanced rate of wound closure and accelerated motility of both keratinocytes andfibroblasts compared to control cells. The wounded monolayer cultures of HaCaT and NIH 3T3 cells, treated with or without SNAP, were monitored under a phase contrast microscope. Structural and ultrastructural modifications were analyzed by scanning electron microscopy (SEM). The images were captured by a digital camera at different time points (0–28 h) and the wound area was analyzed through software included in Matlab. As early as 15 min, SNAP induced significant cytoskeletal remodeling, as shown by immunostaining (phalloidin-labelling), which in turn was associated with increased filopodium number and length rise. NO donor treatment also induced overexpression of Ki-67 protein, a typical marker of cell proliferation, as shown by immunostaining. Both SNAP-induced migration and proliferation were antagonized by the NO-sensitive GC inhibitor 1H-[1,2,4]oxadiazolo[-4,3-a]quinoxalin-1-one (ODQ), which suggests activation of the NO/cGMP signalling cascade in the observed SNAP-induced effects in the early stages of the healing process. Moreover, we provide evidence that PPAR-bantagonist (GSK0660) may interfere with NO-mediated wound healing process.

Nitric Oxide Chemical Donor Affects the Early Phases of In Vitro Wound Healing Process.

LA TORRE, CRISTINA
;
CINQUE, BENEDETTA;LOMBARDI, FRANCESCA;MICONI, GIANFRANCA;PALUMBO, PAOLA;EVTOSKI, ZORAN;PLACIDI, GIUSEPPE;FANINI, DONATELLA;CIMINI, Anna Maria;BENEDETTI, ELISABETTA;GIULIANI, Maurizio;CIFONE, MARIA GRAZIA
2016-01-01

Abstract

An artificial wound in a confluent monolayer of human keratinocyte HaCaT cells or mouse embryofibroblast Swiss NIH 3T3 cells was used to analyze the effects of the nitric oxide (NO) chemical donor, S-nitroso-N-acetylpenicillamine (SNAP). SNAP exposure promoted an enhanced rate of wound closure and accelerated motility of both keratinocytes andfibroblasts compared to control cells. The wounded monolayer cultures of HaCaT and NIH 3T3 cells, treated with or without SNAP, were monitored under a phase contrast microscope. Structural and ultrastructural modifications were analyzed by scanning electron microscopy (SEM). The images were captured by a digital camera at different time points (0–28 h) and the wound area was analyzed through software included in Matlab. As early as 15 min, SNAP induced significant cytoskeletal remodeling, as shown by immunostaining (phalloidin-labelling), which in turn was associated with increased filopodium number and length rise. NO donor treatment also induced overexpression of Ki-67 protein, a typical marker of cell proliferation, as shown by immunostaining. Both SNAP-induced migration and proliferation were antagonized by the NO-sensitive GC inhibitor 1H-[1,2,4]oxadiazolo[-4,3-a]quinoxalin-1-one (ODQ), which suggests activation of the NO/cGMP signalling cascade in the observed SNAP-induced effects in the early stages of the healing process. Moreover, we provide evidence that PPAR-bantagonist (GSK0660) may interfere with NO-mediated wound healing process.
File in questo prodotto:
File Dimensione Formato  
Torre_et_al-2016-Journal_of_Cellular_Physiology.pdf

solo utenti autorizzati

Tipologia: Documento in Pre-print
Licenza: Creative commons
Dimensione 1.6 MB
Formato Adobe PDF
1.6 MB Adobe PDF   Visualizza/Apri   Richiedi una copia

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11697/101166
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 13
  • ???jsp.display-item.citation.isi??? 13
social impact