EffectsofnegativepressuresonepithelialtightjunctionsandmigrationinwoundhealingChih-ChinHsu,1,2,3Wen-ChungTsai,2,3,4CarlPai-ChuChen,4Yun-MeiLu,3andJong-ShyanWang31DepartmentofPhysicalMedicineandRehabilitation,ChangGungMemorialHospitalatKeelung,Keelung;2SchoolofTraditionalChineseMedicine,ChangGungUniversity,Taoyuan;3GraduateInstituteofRehabilitationScience,CollegeofMedicine,ChangGungUniversity,Taoyuan,and4DepartmentofPhysicalMedicineandRehabilitation,ChangGungMemorialHospitalatLinkou,Taoyuan,TaiwanSubmitted12November2009;acceptedinfinalform30April2010HsuCC,TsaiWC,ChenCP,LuYM,WangJS.Effectsofnegativepressuresonepithelialtightjunctionsandmigrationinwoundhealing.AmJPhysiolCellPhysiol299:C528–C534,2010.FirstpublishedMay5,2010;doi:10.1152/ajpcell.00504.2009.—Neg-ative-pressurewoundtherapyhasrecentlygainedpopularityinchronicwoundcare.Thisstudyattemptedtoexploreeffectsofdifferentnegativepressuresonepithelialmigrationinthewound-healingprocess.Theelectriccell-substrateimpedancesensing(ECIS)techniquewasusedtocreatea5�10�4cm2woundintheMadin-Darbycaninekidney(MDCK)andhumankeratinocyte(HaCaT)cells.Thewoundedcellswereculturedinanegativepressureincubatoratambientpressure(AP)andnegativepressuresof75mmHg(NP75),125mmHg(NP125),and175mmHg(NP175).Theeffectivetime(ET),completewoundhealingtime(Tmax),healingrate(Rheal),celldiam-eter,andwoundareaovertimeatdifferentpressureswereevaluated.Traditionalwound-healingassayswerepreparedforfluorescentstain-ingofcellsviability,celljunctionproteins,includingZO-1andE-cadherin,andactins.AmountofcelljunctionproteinsatAPandNP125wasalsoquantified.InMDCKcells,theET(1.25�0.27h),Tmax(1.76�0.32h),andRheal(2.94�0.62�10�4cm2/h)atNP125weresignificantly(P�0.01)differentfromthoseatthreeotherpressureconditions.InHaCaTcells,theTmax(7.34�0.29h)andRheal(6.82�0.26�10�5cm2/h)atNP125weresignificantly(P�0.01)differentfromthoseatNP75.Prominentcellmigrationfeatureswereidentifiedincellsatthespecificnegativepressure.Cellmigra-tionactivitiesatdifferentpressuresc...
