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In vitro comparison of human plasma-based and self-assembled tissue-engineered skin substitutes:two different manufacturing processes for the treatment of deep and difficult to heal injuries

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Burns & Trauma 2023年第1期11卷 800-813页

作者： Álvaro Sierra-Sánchez Brice Magne Etienne Savard Christian Martel Karel Ferland Martin A.Barbier Anabelle Demers Danielle Larouche Salvador Arias-Santiago lucie germain LOEX Tissue Engineering Laboratory and Department of Surgery Faculty of MedicineUniversitéLaval140118e rueQuébec(Québec)G1J 1Z4Canada CHU de Québec-UniversitéLaval Research Center Division of Regenerative Medicine140118e rueQuébec(Québec)G1J 1Z4Canada Unidad de Producción Celular e Ingeniería Tisular(UPCIT) Virgen de las Nieves University Hospitalibs.GRANADAAndalusian Network for the design and translation of Advanced TherapiesAv.de las Fuerzas ArmadasNº24ªPlanta Ed.de Gobierno18014GranadaSpain Department of Dermatology Virgen de las Nieves University HospitalAv.MadridNº11-1518012GranadaSpain Department of Dermatology Faculty of MedicineUniversity of GranadaAv.de la InvestigaciónNº1118016GranadaSpain

Background:The aim of this in vitro study was to compare side-by-side two models of human bilayered tissue-engineered skin substitutes(hbTESSs)designed for the treatment of severely burned *** are the scaffold-free self-assembled skin substitute(SASS)and the human plasma-based skin substitute(HPSS).Methods:Fibroblasts and keratinocytes from three humans were extracted from skin biopsies(N=3)and cells from the same donor were used to produce both hbTESS *** SASS manu-facture,keratinocytes were seeded over three self-assembled dermal sheets comprising fibroblasts and the extracellular matrix they produced(n=12),while for HPSS production,keratinocytes were cultured over hydrogels composed of fibroblasts embedded in either plasma as unique biomaterial(Fibrin),plasma combined with hyaluronic acid(Fibrin-HA)or plasma combined with collagen(Fibrin-Col)(n/biomaterial=9).The production time was 46-55 days for SASSs and 32-39 days for *** were characterized by histology,mechanical testing,PrestoBlue™-assay,immunofluorescence(Ki67,Keratin(K)10,K15,K19,Loricrin,type IV collagen)and Western blot(type I and IV collagens).Results:The SASSs were more resistant to tensile forces(p-value<0.01)but less elastic(p-value<0.001)compared to HPSSs.A higher number of proliferative Ki67+cells were found in SASSs although their metabolic activity was *** epidermal differentiation,no significant difference was observed in the expression of K10,K15,K19 and ***,the production of type I and type IV collagens and the adhesive strength of the dermal-epidermal junction was higher in ***:This study demonstrates,for the first time,that both hbTESS models present similar in vitro biological ***,mechanical properties differ and future in vivo experiments will aim to compare their wound healing potential.

关键词： Biomaterial Burn Fibrin Human plasma Self-assembly Skin substitute Tissue engineering

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Donkey pericardium as a select sourcing to manufacture percutaneous heart valves:Decellularization has not yet demonstrated any clear cut advantage to glutaraldehyde treatment

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Medicine in Novel Technology and Devices 2019年第4期4卷 38-46页

作者： Aisa Rassoli Yiming Li Xianhao Bao Fabien Kawecki Xinzhe Zhao Daniel Chappard Gaetan Le-Bel Jiaxuan Feng Benedikt Weber Nasser Fatouraee Ze Zhang Zaiping Jing lucie germain Lu Wang Robert Guidoin Department of Surgery Faculty of MedicineUniversite Laval and Regenerative Medicine AxisCHU de Quebec Research CenterQuebecCanada Department of Biomedical Engineering Amirkabir University of TechnologyTehranIran Departnent of Vascular Surgery Changhai Hospital of Naval Military Medical UniversityShanghaiChina Key Laboratory of Textile Sciences and Technology Ministry of EducationColle of TextilesDonghua UniversityShanghaiChina GEROM Institut de Biologie en SanteCHU D'AngersAngersFrance Department of Dermatology Medical University of ViennaViennaAustria

Background:The donkey pericardium is considered a good candidate to manufacture percutaneous heart valves based upon its thinness,low cellularity and undulating collagen bundles and *** represents an avenue worth exploring,should its superiority to glutaraldehyde-treated pericardium be *** and methods:Donkey pericardium was divided into two groups:regular glutaraldehyde fixation and mild *** treated pericardia were observed using scanning electron microscopy,histology and transmission electron *** tests were performed along the axial and perpendicular directions,with the data fitted into both the Gasser–Ogden–Holzapfel(GOH)material model and the Fung’s anisotropic ***:The microstructures of the pericardia processed by the two protocols were similar,showing collagen bundles and laminates free of *** decellularization eliminated most of the cells,however leaving the structure somehow *** collagen filaments in bundles were slightly *** anisotropy rates of the non-decellularized specimens were almost identical to the decellularized *** decellularized pericardium appeared ***:The decellularization proved to be ***,it makes the tissue stiffer,which may lead to higher shear concentration during cardiac cycles and reduce its wavy ***,it appears premature to select decellularized donkey pericardium to manufacture heart valves.

关键词： Donkey pericardium Decellularization Valve leaflet Transcatheter aortic valve implantation Wavy microstructure

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*** of a new role for the WNK1 kinase in corneal wound healing

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Annals of Eye Science 2019年第1期 196-196页

作者： Pascale Desjardins Camille Couture lucie germain Sylvain Guérin Département d'ophtalmologie UniversitéLavalCUO-recherche/LOEXCentre de recherche du CHU de Québec-UniversitéLavalQuébecQCCanada Département de chirurgie UniversitéLavalCentre de recherche en organogénèse expérimentale de l’UniversitéLaval/LOEXCentre de recherche du CHU de Québec-UniversitéLavalQuébecQCCanada

Background:Damage to the corneal epithelium triggers important changes in the composition of the extracellular matrix(ECM)to which the basal human corneal epithelial cells(hCECs)*** changes are perceived by integrins,a family of trans-membrane receptors that activate different intracellular signalling pathways,ultimately leading to re-epithelialization of the injured *** goal is to study the impact of the pharmacological inhibition/activation of specific signal transduction mediators on corneal wound healing using both monolayers of hCECs and tissue-engineered human corneas(hTECs)as in vitro ***:hTECs were produced by the self-assembly approach and wounded with a 8-mm biopsy *** RNA and proteins were isolated from the wounded and unwounded hTECs to conduct gene profiling analyses and protein kinase *** wounded tissues were then incubated either with the lysine deficient protein kinase 1(WNK1)inhibitor WNK463,the WNK1 indirect agonist AM1241,or with the vehicle alone(DMSO;negative control)and wound healing was monitored for 6 *** impact of WNK1 inhibition/activation on hCECs monolayers was determined using scratch wound ***:Gene profiling analyses and protein kinases arrays revealed that expression and activity of several mediators from the integrin-dependent signalling pathways were altered in response to the ECM changes taking place during corneal wound *** of the WNK1 kinase turned out to be the most striking activation event occurring during wound *** the pharmacological inhibition of WNK1 by WNK463 significantly reduced the rate of corneal wound closure in our hTECs and hCECs monolayers compared to their respective negative controls,we believe that the pharmacological activation of WNK1 could turn out to be an interesting avenue to accelerate corneal wound ***:These results will contribute to a better understanding of the cellular and molecular mechanisms

关键词： Cornea wound healing lysine deficient protein kinase 1(WNK1)

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*** of WNK1 inhibition on corneal wound healing using a model of human tissue-engineered cornea

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Annals of Eye Science 2018年第1期 495-495页

作者： Pascale Desjardins Camille Couture Karine Zaniolo lucie germain Sylvain Guérin Département d'ophtalmologie CUO-recherche/LOEXCentre de recherche du CHU de QuébecUniversitéLavalQuébecQCCanada Département de chirurgie Centre de recherche en organogénèse expérimentale de l’UniversitéLaval/LOEXCentre de recherche du CHU de QuébecUniversitéLavalQuébecQCCanada

Background:Because of its superficial anatomical localization,the cornea is particularly vulnerable to abrasive forces and various traumas,which can lead to significant visual *** injury of the corneal epithelium,there are important changes that occur in the composition of the underlying extracellular matrix(ECM).Those changes are perceived by the integrins that recognize the ECM components as their ligand and activate different intracellular signalling pathways,ultimately leading to reepithelialisation and reorganization of the injured epithelium,both of which are necessary in order to restore the visual properties of the *** goal of this study was to analyse the impact of the pharmacological inhibition of specific signal transduction mediators of integrin-dependant signalling pathways on corneal wound healing using both monolayers of hCECs and tissue-engineered human corneas(hTECs)as in vitro ***:hTECs were produced by the self-assembly approach and wounded with a 8-mm diameter biopsy *** RNA and proteins were isolated from the wounded and unwounded hTECs to conduct gene profiling analyses and protein kinase *** wounded tissues were then incubated with the WNK1 inhibitor WNK463 and wound healing was monitored over a period of 6 *** corneas were incubated with the vehicle alone(DMSO).The impact of WNK1 inhibition on hCECs monolayers was determined using a scratch wound ***:Gene profiling analyses and protein kinases arrays revealed important alterations in the expression and activity of several mediators from the integrin-dependent signalling pathways in response to the ECM changes taking place during corneal wound *** these,WNK1 is considerably activated through phosphorylation during corneal wound *** pharmacological inhibition of WNK1 by WNK463 significantly reduced the dynamic of corneal wound closure in our hTECs and hCECs monolayers compared to their respective negative control

关键词： Cornea tissue engineering wound healing signaling pathways integrins

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*** of cyclic-AMP-response element binding protein and its impact on corneal wound healing in vitro and in vivo

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Annals of Eye Science 2019年第1期 195-195页

作者： Camille Couture Pascale Desjardins Karine Zaniolo Richard Bazin lucie germain Sylvain Guérin Département d'ophtalmologie UniversitéLavalCUO-recherche/LOEXCentre de recherche du CHU de Québec-UniversitéLavalQuébecQCCanada Département de chirurgie UniversitéLavalCentre de recherche en organogénèse expérimentale de l’UniversitéLaval/LOEXCentre de recherche du CHU de Québec-UniversitéLavalQuébecQCCanada

Background:The cornea composes the outer surface of the eye and its transparency is required to allow light transmission to the ***,because of its position,the cornea is subjected to chemical and mechanical injuries that may lead to *** studies conducted using the human tissue-engineered cornea(hTEC)as a model provided evidence that the cyclic-AMP-response element binding protein(CREB)pathway is repressed during closure of corneal *** on these results,we hypothesized that closure of corneal wounds can be enhanced by preventing activation of CREB with the pharmacological inhibitor *** goals were to proceed to the pharmacological inhibition of CREB(I)in vitro using the hTECs as a model,and then(II)in vivo using the rabbit as a ***:The self-assembly approach was used to create hTECs,that were then wounded with an 8-mm diameter biopsy punch to create an epithelial *** tissues were then incubated with 10μM of C646(n=8).DMSO was used alone as a negative control(n=4).Closure of the wounds was monitored over a period of 5 ***,the cornea of New Zealand white rabbits was debrided with an ethanol 70%solution to create an epithelial defect of 8-mm *** concentrations of C646(1,10,100μM et 1 mM)were applied as eye drops 3 times a day for up to 7 *** wounded corneas(n=4 per concentration)were stained with fluorescein and photographed every ***:In vitro pharmacological inhibition of CREB with C646 considerably accelerated wound closure of all treated hTECs(4 days)compared to the control group(7 days).Moreover,the in vivo C646 treatment also accelerated wound healing of the corneas compared to the control *** most effective concentration of C646 tested was the lowest(1μM),as it considerably enhanced the wound healing ***:This study demonstrates that wound healing both in vitro and in vivo can be enhanced by preventing activation of CREB using a pharmacological inhibition appr

关键词： Cyclic-AMP-response element binding protein(CREB) protein kinase B(AKT) healing corneal wound tissue-engineering

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*** tissues bank for vision health research

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Annals of Eye Science 2018年第1期 499-499页

作者： Kim Santerre Mathieu Thériault Sylvain Chemtob lucie germain Claude Giasson Sylvain Guérin Solange Landreville Patrick JRochette Christian Rochette Mike Sapieha Élodie Boisselier Vincent Pernet Stéphanie Proulx Centre de Recherche du CHU de Québec-UniversitéLaval Axe Médecine RégénératriceQuébecQCCanada Centre de Recherche en Organogénèse Expérimentale de l'UniversitéLaval/LOEX QuébecQCCanada Département d'Ophtalmologie et d'Oto-Rhino-Laryngologie-Chirurgie Cervico-Faciale Facultéde MédecineUniversitéLavalQuébecQCCanada Département d’ophtalmologie Universitéde MontréalQCCanada Maisonneuve-Rosemont Hospital Research Center MontrealQCCanada Département de chirurgie Facultéde MédecineUniversitéLavalQuébecQCCanada École d’optométrie Universitéde MontréalQuébecQCCanada

Background:This infrastructure delivers biological material necessary for several research projects to Vision Health Research Network investigators(VHRN).Methods:Héma-Québec is the organism in charge obtaining consent and retrieving donor eyes for patient treatment or for *** Quebec City,donor eyes are sent to the eye bank of the“Centre Universitaire d’Ophtalmologie”(CUO)of Saint-Sacrement *** at the eye bank evaluate the quality of the *** unfit for graft are transferred to the infrastructure where the coordinator encodes samples prior to their ***:Between 2013 and 2017,27 fundamental investigators,clinical investigators and collaborators supported by 60 students,trainees and laboratory assistants used this infrastructure to move forward their *** 2013,results from those projects generated 21 scientific publications and 232 *** infrastructure helped VHRN investigators obtain near 4 million dollars in grants from many organisms(CIHR,NSERC,Foundations,etc.).These grants allowed recruitment and formation of highly qualified *** year(April 2016 to March 2017),189 corneas and 23 eyes transited through the ***:This infrastructure is available for all investigators that are members of the *** original projects have been elaborated thanks to the human ocular tissues provided by this *** projects will advance our knowledge in vision health.A better understanding of eye functions will lead to new treatments for eye diseases.

关键词： Human ocular tissues for research fresh tissues

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