Amina Khadjavi1, Chiara Magnetto2, Alice Panariti3, Monica Argenziano4, Giulia Rossana Gulino5, Ilaria Rivolta3, Roberta Cavalli4, Giuliana Giribaldi5, Caterina Guiot1, Mauro Prato6. 1. Dipartimento di Neuroscienze, Università di Torino, Torino, Italy. 2. Istituto Nazionale di Ricerca Metrologica (INRIM), Torino, Italy. 3. Dipartimento di Scienze della Salute, Università di Milano Bicocca, Monza, Italy. 4. Dipartimento di Scienza e Tecnologia del Farmaco, Università di Torino, Torino, Italy. 5. Dipartimento di Oncologia, Università di Torino, Torino, Italy. 6. Dipartimento di Neuroscienze, Università di Torino, Torino, Italy; Dipartimento di Scienze della Sanità Pubblica e Pediatriche, Università di Torino, Torino, Italy. Electronic address: mauro.prato@unito.it.
Abstract
BACKGROUND: In chronic wounds, efficient epithelial tissue repair is hampered by hypoxia, and balances between the molecules involved in matrix turn-over such as matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are seriously impaired. Intriguingly, new oxygenating nanocarriers such as 2H,3H-decafluoropentane-based oxygen-loaded nanodroplets (OLNs) might effectively target chronic wounds. OBJECTIVE: To investigate hypoxia and chitosan-shelled OLN effects on MMP/TIMP production by human keratinocytes. METHODS: HaCaT cells were treated for 24h with 10% v/v OLNs both in normoxia or hypoxia. Cytotoxicity and cell viability were measured through biochemical assays; cellular uptake by confocal microscopy; and MMP and TIMP production by enzyme-linked immunosorbent assay or gelatin zymography. RESULTS: Normoxic HaCaT cells constitutively released MMP-2, MMP-9, TIMP-1 and TIMP-2. Hypoxia strongly impaired MMP/TIMP balances by reducing MMP-2, MMP-9, and TIMP-2, without affecting TIMP-1 release. After cellular uptake by keratinocytes, nontoxic OLNs abrogated all hypoxia effects on MMP/TIMP secretion, restoring physiological balances. OLN abilities were specifically dependent on time-sustained oxygen diffusion from OLN core. CONCLUSION: Chitosan-shelled OLNs effectively counteract hypoxia-dependent dysregulation of MMP/TIMP balances in human keratinocytes. Therefore, topical administration of exogenous oxygen, properly encapsulated in nanodroplet formulations, might be a promising adjuvant approach to promote healing processes in hypoxic wounds.
BACKGROUND: In chronic wounds, efficient epithelial tissue repair is hampered by hypoxia, and balances between the molecules involved in matrix turn-over such as matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are seriously impaired. Intriguingly, new oxygenating nanocarriers such as 2H,3H-decafluoropentane-based oxygen-loaded nanodroplets (OLNs) might effectively target chronic wounds. OBJECTIVE: To investigate hypoxia and chitosan-shelled OLN effects on MMP/TIMP production by human keratinocytes. METHODS: HaCaT cells were treated for 24h with 10% v/v OLNs both in normoxia or hypoxia. Cytotoxicity and cell viability were measured through biochemical assays; cellular uptake by confocal microscopy; and MMP and TIMP production by enzyme-linked immunosorbent assay or gelatin zymography. RESULTS: Normoxic HaCaT cells constitutively released MMP-2, MMP-9, TIMP-1 and TIMP-2. Hypoxia strongly impaired MMP/TIMP balances by reducing MMP-2, MMP-9, and TIMP-2, without affecting TIMP-1 release. After cellular uptake by keratinocytes, nontoxic OLNs abrogated all hypoxia effects on MMP/TIMP secretion, restoring physiological balances. OLN abilities were specifically dependent on time-sustained oxygen diffusion from OLN core. CONCLUSION: Chitosan-shelled OLNs effectively counteract hypoxia-dependent dysregulation of MMP/TIMP balances in human keratinocytes. Therefore, topical administration of exogenous oxygen, properly encapsulated in nanodroplet formulations, might be a promising adjuvant approach to promote healing processes in hypoxic wounds.
Authors: Lohrasb R Sayadi; Derek A Banyard; Mary E Ziegler; Zaidal Obagi; Jordyne Prussak; Michael J Klopfer; Gregory Rd Evans; Alan D Widgerow Journal: Int Wound J Date: 2018-01-05 Impact factor: 3.315
Authors: Lohrasb R Sayadi; Rebecca Rowland; Alexandra Naides; Luke Tomlinson; Adrien Ponticorvo; Anthony J Durkin; Alan D Widgerow Journal: Ann Plast Surg Date: 2021-10-01 Impact factor: 1.763