Received:6November2019Revised:3December2019Accepted:5December2019
DOI:10.1002/jbio.201960177
Polarizedlighttherapy:Shiningalightonthemechanism underlyingitsimmunomodulatoryeffects
JackFeehan1,2* |NicholasTripodi3,4 |SarahFraser3 | KathleenMikkelsen3 |AprilThewlis1 |DimitriosKiatos1,3 |MajaHusaric3,4 | VassoApostolopoulos3*
1OsteopathyGroup,CollegeofHealthand Biomedicine,VictoriaUniversity,Victoria, Australia
2AustralianInstituteforMusculoskeletal Science(AIMSS),UniversityofMelbourne andWesternHealth,StAlbans,Victoria, Australia
3InstituteforHealthandSport,Victoria University,Victoria,Australia
4FirstYearCollege,VictoriaUniversity, Victoria,Australia
*Correspondence
JackFeehan,OsteopathyGroup,College ofHealthandBiomedicine,Victoria University,Victoria,Australia.
Email:jfeehan@student.unimelb.edu.au
VassoApostolopoulos,InstituteforHealth andSport,VictoriaUniversity,Victoria, Australia.
Email:vasso.apostolopoulos@vu.edu.au
Abstract
Thisstudyinvestigatestheimmunomodulatoryeffectsofpolychromatic polarizedlighttherapy(PLT)on humanmonocytecells.Whilethereis someevidencedemonstratingaclinicaleffectinthetreatmentofcertain conditions,thereislittleresearchinto itsmechanismofaction.Herein,U937 monocytecellswereculturedand exposedtoPLT.Thecellswerethen analyzedforchangeinexpressionof genesandcellsurfacemarkersrelatingto inflammation.Itwasnotedthat6hours ofPLTreducedtheexpressionoftheCD14,MHCIandCD11breceptors,and increasedtheexpressionofCD86.ItwasalsoshownthatPLTcauseddownregulationofthegenesIL1B,CCL2, NLRP3andNOD1,andupregulationof NFKBIAandTLR9.ThesefindingsimplythatPLThasthecapacityforimmunomodulationinhumanimmunecells,possiblyexertingananti-inflammatoryeffect.
KEYWORDS
inflammation,phototherapy,polarizedlight,polarizedlighttherapy
1 | INTRODUCTION
Inflammationisaprocessheavilyimplicatedinpathological statesofallkinds,fromallergyandneoplasia,toinfection. Despiteitscentralroleinthepathophysiologyofmany commonconditions,currentmethodsofmanaginginflammationarefraughtwithproblems,particularlyinachronic setting.Treatmenttypicallyinvolvespharmacologicalinterventions,manyofwhichexertarangeofunwantedeffects
inadditiontotheirtherapeuticaction[1].Thecontinuing evolutionanddevelopmentofnoveltherapeuticapproaches tothemanagementofinflammationisessentialinorderto advancepatientcare.Evaluationofnonpharmacological anti-inflammatorytreatmentsisakey,butrelativelyunderrepresentedpartofthisprocess.Asmallbutgrowingbody ofevidenceindicatesthatphototherapiessuchaspolarized lighttherapy(PLT)areapromisingavenueofexploration inthisarea[2,3].
ThisisanopenaccessarticleunderthetermsoftheCreativeCommonsAttribution-NonCommercialLicense,whichpermitsuse,distributionandreproductioninany medium,providedtheoriginalworkisproperlycitedandisnotusedforcommercialpurposes.
©2019TheAuthors. JournalofBiophotonics publishedbyWILEY-VCHVerlagGmbH&Co.KGaA,Weinheim
J.Biophotonics. 2019;e201960177.
www.biophotonics-journal.org 1of9 https://doi.org/10.1002/jbio.201960177
Thetherapeuticbenefitsoflighttherapyhavebeen reportedsincethelate1960swithapplicationsrangingfrom neonataljaundicetopsoriasisandvitiligo[3,4].Thetheoreticalbasisofphototherapyinvolvestheuseoflightto inducephysiologicalchangewithinatargettissue[5],with subsequenttherapeuticeffects.Thereareseveraltypesof phototherapiesdifferentiatedbythespecificphysicalqualitiesofthelightused,withthemostcommonbeinglowlevellasertherapy(LLLT)andultraviolet(UV)therapies. Inrecentyears,however,ithasbeenproposedthatabroad lightspectrumcoveringalllightcolors,andpolarization arecrucialelementsinlighttherapy[6].Polarizedlight (PL)isformedbythefilteringoflightwavessothatthey arealignedandvibratedinasingleplane(Figure1).Once polarized,lighthastheabilitytopenetratefurtherintotissuesthanitsunpolarizedcounterpart[7].
Broad,visiblespectrumPLTdiffersfromotherformsof phototherapyasitusesamuchwiderrangeofwavelengths thanothermodalitiessuchas,LLLTorUV.Consequently, thedevicesusedinPLTaregenerallylessexpensiveand relativelyeasytouse.PLThasbeensuggestedbyadvocates anddevicemanufacturersforuseinseveralcontextssuch aspain,woundhealing,skinconditionsandinflammatory arthritis.Preliminaryclinicalevidenceindicatesefficacyin themanagementofskinulcers[8–10],burns[11–13],musculoskeletalinjuries[14–18]aswellassurgicalandnonhealingwounds[6,19–22].Despitethis,therearelittle scientificdataregardingthephysiologicalmechanisms underlyingthesechangesinvitroorinvivo[2].Ithas suggestedthattheinteractionsoflightwiththekeymitochondrialenzymecytochromeoxidaseCareresponsible forthebeneficialchangesobservedinLLLT[23–25];however,littleinformationhasbeendocumentedexploringthe mechanismsofPLT.
Withagingpopulations,inmostdevelopedeconomies, andtheassociatedincreaseindemandsonhealthbudgets,noninvasive,nonpharmacologicalavenuesofdisease treatmentwillberequiredtocountertheinevitableburdenofinflammatorydisease.PLThasthepotentialtobe aninexpensive,technicallysimpleandsafephototherapeuticintervention.Thisstudyaimedtodetermine whetherPLinducescellularchangestothehumanmonocytecellline,toprovideinsightintothepossiblemechanismsofactionunderpinningitsclinicaluse.Itwas hypothesizedthatadecreaseintheinflammatoryprofile ofimmunecellsmaybepartofthebiologicalactionof PLT.Specifically,thisinvolvedtheexposureofacultured U937monocyte/macrophagecelllinetoPL,andsubsequentassessmentofcellsurfacemarkersandgeneexpressionrelatingtoinflammation.Weshowedthatexposure toPLdecreasestheexpressionofproinflammatorycell surfacemarkersandcauseschangesingeneexpression consistentwithadecreasedinflammatoryprofile.
2 | MATERIALSANDMETHODS
AllproceduresinthisstudywereperformedinPC2laboratoriesattheWerribeecampusofVictoriaUniversity, andtheWesternCentreforHealthResearchandEducation(WesternCHRE)atSunshinehospitalunderstandardlaboratoryconditions.Allcellcultureworkwas doneunderasepticconditionsinaclassIIbiosafetycabinet.Trypanbluestainingwasusedbeforeandafterilluminationtoensurecellviabilitythroughoutthe experiments.Noethicswasrequiredforthisresearch.
2.1 | Cellcultureandcelldifferentiation
TheU937monocytecelllinewasculturedinRoswell ParkMemorialInstitutemedia,supplementedwith10% fetalbovineserum(InterpathServicesPty.Ltd.),1%penicillin/streptomycin(Sigma-Aldrich)and0.1%glutamine (Sigma-Aldrich),andincubatedat37 C,5%CO2 asper standardcultureprotocol.Thecellswerepassagedto50% to90%confluencyandmediareplenishedevery48hours untilanadequatenumberofcellswereobtained.Astock ofexperimentalcells(cellbank)wasthencreatedand storedinliquidnitrogen.Onevialfromthecellbankwas thawedeachtimetheexperimentwasrepeated,toensure allexperimentalcellswereatthesamepassagethroughouttheexperiments.U937cellsarepromonocyticcells andaredifferentiatedintomonocyte/macrophagecells viatheadditionofvitaminD3 tothemediatoafinalconcentrationof100nм for72hours[26].
2.2 | Polarizedlightexposure
ThetherapeuticPLsourceemployedwasusingaBioptron MedAllPAG-960lamp(Bioptron,Switzerland)witha wavelengthrangefrom400to3400nm.Thelightsource hasanaveragepowerdensityof40mW/cm2 evenlydistributedacrossthewavelengthspectrum.Theentireapparatus,includinglampandtripod,washousedcompletely withinanincubator(Figure2).Theincubatorandmedia temperaturesweremonitoredtoensuretherewasnooverheatingofthecultureduetothelightexposure.Thisisto ensurethatobservedeffectsstemmedfromcellularinteractionswithlight,ratherthanathermiceffect.Thelight fromthelampwasprojectedperpendicularlyontoa5cm cellcultureplateatadistanceof10cm,accordingtomanufacturer'sinstructions,withfulllightcoverageofthe sample.Thelidoftheculturedishcontainingtheexperimentalcellswasremoved,toavoidinterferenceofthe lightpassingthrough.Controlcellswerekeptinthesame incubator,separatedbyasolid,stainlesssteelpartitionto
FIGURE1 A,Summary figureofdifferentphysical propertiesofphototherapeutic light.B,Summaryoftheprocessof lightpolarization
ensurenospill-overlightinteractedwiththecontrolsample.Thelidofthecontrolcellplatewasalsoremovedto ensureinteractionswiththeatmospherewithintheincubatordidnotaccountforthechangesseen.Ininitial experiments,cellswereexposedfor5minutes,15minutes or6hours,followedbyincubationfor24hoursbefore beingpreparedforflowcytometricanalysisofcellsurface markerexpression.Thecellsusedforanalysisofgene expressionwereexposedtoPLfor6hours,immediately snapfrozen,andtheirRNAisolatedpriortotheir
preparationforPCRgenearrayanalysis.Allanalysiswas performedintriplicateastechnicalreplicates.
2.3 | Flowcytometry
Thecellswerelabeledforflowcytometryaccordingto manufacturer'sinstructions,andstandardprotocols.All antibodiesweretitratedpriortotheexperimenttoensure optimalworkingconcentrations.Briefly,cellswereplated
ina96-wellU-bottomplateandtreatedwithFcRblockingreagentfor10minutesatroomtemperature.The cellswerethenincubatedwiththeirspecificconjugated antibody(CD86-Alexa-Fluor488,MajorHistocompatibilityComplexClass[MHC]I,II-BV510,CD206-PE-Cy7, CB11b-PEorCD14-BV421)for45minutesinthedarkat 4 Catapredeterminedconcentration.Otherantibodies againstCD40,CD83andCD209werenegativeonthis celllineandweresubsequentlynotused.Cellswere transportedafterlabelinginthedarkonicepriortoanalysis.AnalysiswasperformedwithaBDFACSCantoII flowcytometer,withthreelasers(violet-405nm,blue –488nmandred – 633nm).Isotypecontrolswereusedto controlforbackgroundfluorescence.AllconjugatedantibodiesweresourcedfromBDBiosciences(SanJose,California)toensurecompatibilitywiththeflowcytometer used.AnalysisofdatawasdonewithBDFACSDivasoftware(Version3.0).Themonocytepopulationwasgated onforwardandsidescatterplotsandanalyzedforfluorescenceintensityagainstunilluminatedcontrolcells. Resultsarereportedasflowcytometricdotplotswithgatingstrategies,overlayhistogramsofcontrolandtreated cells,andmedianfluorescenceintensity,asrecommendedbytheInternationalSocietyfortheAdvancementofCytometrydatastandardstaskforce[27].
2.4 | Genearrays
AllreagentsandconsumablesusedinRNAextraction, cDNAsynthesisandgeneexpressionanalysiswereacquired fromQIAGEN(Hilden,Germany).Thearrayusedwasthe RT2 ProfilerPCRarray “HumanInnateandAdaptive ImmuneResponses” whichanalyses84genesrelatedto immuneandinflammatoryresponses,includingkeycytokines,chemokinesandimmuneactivesurfacemembrane receptors.RNAqualityandconcentrationwereassessedvia Agilent2100Bioanalyzer,Qubitfluorimeter(Invitrogen)and spectrophotometer(DeNovix).RNAextractionwas
performedusingtheRNeasyMinikitaccordingtothemanufacturer'sinstructions.Briefly,cellswerelysedinRLT buffercontaining β-mercaptoethanol(10 μL/mL)and centrifugedthroughaQIAshreddercolumn.Lysateswere passedthroughRNeasyspincolumns,treatedon-column withDNase,followingwhichtheRNAwascollectedin RNasefreewaterforpreparationofcDNA.cDNAwaspreparedinathermalcyclerusingtheQiagenRT2 FirstStrand kit.Briefly,RNA(500ng)wasincubatedat42 Cfor 2minutesinthepresenceofGEbuffer(containingDNase) toeliminateanyremaininggenomicDNA,beforebeing immediatelytransferredtoice.cDNAwasreversetranscribedfor15minutesat42 C,andat92 Cfor3minutesto terminatethereactions.cDNAwasthenaddedtotheRT2 SYBRgreenqPCRmastermixanddispensedtothe96-well genearrayplateaspermanufacturer'sinstructions.Thegene arraywasruninBioradCFX96real-timePCRcyclerwithan initialdenaturationstepof10minutesat95 C,followedby 40cyclesof15secondsat95 Cand1minuteat60 C.Once theRT-PCRwascomplete,meltcurveanalysiswasperformedtoensurethepresenceofasinglePCRproductfor eachgene.GenearraydatawereanalyzedusingCFXMaestro(BioRad).Anunpaired t testwasusedtocompare groups;a P value ≤.05anda2-foldchangeinexpression weresetasthresholdsforsignificance.Geneswereexcluded fromfurtheranalysisifthecrossingpoint(Cq)valuewas greaterthan34and/orauniquemeltingtemperaturewas notobserved.
3 | RESULTS
3.1 | Polarizedlightdecreasesthe expressionofcellsurfacemarkersrelated toinflammation
Gatingusedforanalysiswasperformedagainstappropriateisotypecontrolstoaccountforbackgroundfluorescence.ExamplegatingstrategyisshowninFigure3.
Nochangetocellsurfacemarkerexpressionwasseen after5or30minutesofPLexposure.Thecellsurface markerexpressionchangeafter6hoursexposuretoPL isshown(Figure4).SixhoursofexposuretoPLcaused ameandecreaseinthemedianfluorescenceintensityof 23%forCD11b,39%forCD14,27%inMHCIand35% inMHCII,thoughMHCIIexpressionwaslowatbaseline(Figure4).Conversely,therewasameanincrease inthemedianfluorescenceof20%inCD86.Therewere noconsistentchangesseeninCD206expression.
3.2 | Polarizedlightdecreasesgenes relatedtoinflammation
Allexperimentalandcontrolsamplespassedtheinbuilt qualitycontrolmeasuresinthegenearray.Normalizationwasperformedagainstthetwomoststablehousekeepinggenes GAPDHandRPLP0.Cutoffpointswere setat2-foldupordownregulation,and P valuescalculatedwithsignificancelevelsetat P <.05.Theresultsare summarizedinFigures5and6.Figure5showsgenes withsignificantchangeinregulation,andmorethan 2-foldregulation,andthetotalchangesforallexpressed
FIGURE3 Examplegatingstrategy.Left-handpanel,doubletdiscriminationstrategy;middlepanel,monocytesgatedusingsizeand density;right-handpanel,fluorescenceintensityofthegivenantibodywithquadrantsforvisualinspection.FSC-A,forwardscatterarea; FFC-H,forwardscatterheight;SSC-A:Sidescatterarea
FIGURE4 Changesincellsurfacemarkerexpressionasassessedbyflowcytometryfollowing6hoursexposuretopolarizedlight therapy.LivecellsweregatedonFSCvsSSCprofileandisotypecontrolantibodieswereusedasbackgroundcontrol.Shownarevalues abovethebackgroundisotypecontrols.Experimentswereperformedintriplicate;representativesamplesaredisplayedindotplotsand histograms.MFI,medianfluorescenceintensity
FIGURE5 Geneexpressionchangesfollowing6hoursofPLT.TopLeft,foldregulationandsignificanceofgenes;topright,scatterplot offoldregulation;bottom,clusteredheatmap.Experimentswererepeatedthreetimesandmeanofthreerepeatsareshown
FIGURE6 Bargraphsofrelativenormalizedfoldregulationofsignificantgenescomparedtoreferencepointof1.0. *P <.05; **P <.01; ***P <0.001
genesarepresentedasascatterplotandheatmap. Figure6showsthespecificchangeinexpressionofthe sixgenesthatreachedgreaterthan2-foldup-ordownregulation,andstatisticalsignificance.SixhoursofPLT causedupregulationofNFKBIAandTLR9,anddownregulationofIL1B,CCL2,NLRP3andNOD1.NFKBIA isakeyinflammatoryinhibitorofcytokineproduction, andTLR9codesfortheproductionofthetoll-likereceptor9,akeymembranereceptorinvolvedinpathogen recognitionandimmuneactivation.IL1BandCCL2are importantcytokines,whileNLRP3andNOD1aremembranereceptorsinvolvedininflammatorysignaling.All othergenesassessedinthearraydidnotreachthesignificanceandfoldregulationcutoffs.
4 | DISCUSSION
ThisstudydemonstratesthatPLTcanexertameasurable effectonthehumanimmunesystem,givingmuch neededmechanisticevidenceforitsclinicaleffects.While phototherapiessuchaslow-levellaserhavebeenpreviouslyshowntohaveasuppressiveeffectoninflammation andimmuneresponse[2],verylimitedevidenceexists investigatingwhetherPLThasthesamecapacity.The resultsofthisstudydemonstratechangesingeneexpressionandcellsurfacemarkerexpressionindifferentiated U937cellsfollowing6hoursexposureofPL.ThissuggeststhatPLThasthecapacitytocausemodulationof thebehaviorofactivatedmonocytesinvitro.Thiscould aidinexplainingthedemonstratedeffectsofPLTin improvingwoundhealinganddecreasingpainand inflammationinvivo.
Thecellsurfacemarkersstudiedhereareknownto beinvolvedinarangeofprocessesvitaltotheimmune andinflammatoryresponse,includingrecognitionofself, Tcellactivation,recognitionandphagocytosisofpathogensandimmunecellinfiltrationandaccumulation.Itis difficulttopredicthowtheobservedchangesdemonstratedinthisresearchtranslateintotheinfinitelymore complexcellularenvironmentsurroundinganinflammatoryorhealingprocess,howeverpossibleinvivoeffects maybeinferred.TheMHCIandMHCIIreceptorsare involvedinantigenpresentation[28]andCD11breceptor ininflammatorycellaccumulation[29]andtheCD14 receptorisassociatedwiththedetectionandphagocytosis ofbacteriallipopolysaccharides,andotherproteinaceous debrisbymacrophages[30].Theirsuppressionmaysignifyadecreaseintheinflammatoryactivityofthesecells andhavetheneteffectofdampeningtheintensityand hasteningtheresolutionofinflammationinvivo.Similar changesinimmunecellmarkerexpressionhavebeen showninothercelltypesfollowingexposuretoLLLT,
suggestingthepossibilityofcommonmechanismsforthe actionsofphototherapies[31,32].CD86hasbeenshown toplayanimportantroleintheamplificationandmaintenanceofaninflammatorystateinvivo[33,34].This couldresultinfasterclearanceofdebrisandpathogens andahasteningoftheresolutionoftheinflammatory response,leadingtoenhancedhealingtimes,correlating withtheobservedclinicaleffectsofPLT.
Whilethereductionincellsurfacemarkerssuggestsa functionalimmunomodulatoryeffect,changesingene expressionprovideamechanisticinsightintothecellular responsetoPLT.ThedownregulationofIL1B,CCL2, NLRP3andNOD1suggestsadampeningoftheimmune andinflammatoryresponse.Thesegenesandtheirassociatedproductsareimplicatedinlymphocytedifferentiationandproliferation,inflammasomeactivation[35], systemicinflammation[36],immunecellmigrationand accumulation[37],antigenrecognition[38,39]and phagocyteactivity[39].Othershavedescribeddecreases inIL1BinmurinewoundmodelsandaorticsmoothmusclecellsfollowingexposuretoLLLT[40,41]andLED therapy[42].Additionally,LLLThasbeenshownto increaseordecreasetheexpressionofCCL2atdifferent intensitiesintheTHP-1monocytecellline,suggestinga dose-dependenteffect[43].Thefindingsinthisstudyprovideatheoreticalmechanismfortheirinvivofindings. Decreasingtheexpressionofthesegenesinactivated monocyte/macrophagecellsislikelytocauseafunctional reductionininflammation.Theobservedincreasein NFKBIAfitswithinthepictureofimmunesuppression, asthisgenehasbeenshowntomodulateanddecrease theeffectsofmalignant[44],inflammatory[45]and auto-immunedisease[46].TheoutcomeofthePLmediatedincreaseinTLR9expressionmightplayislessclear. Itisinvolvedintherecognitionandbindingofbacterial andviralDNAandhasbeenassociatedwithautoimmunediseaseandinflammation[47],buthowan upregulationoftheseprocessesmightexertanantiinflammatoryorincreasedhealingstateisunclear.
Whilethisresearchprovidesproofofconceptfora physiologicalmechanismbywhichPLTmayexertitsclinicaleffects,manyquestionsremain.Firstly,whilstmonocyte/macrophagecellsareimportantcontributorsto inflammationandhealingprocesses,thereareahostof othercellsinvolvedinthenaturalsetting.Furtherstudies toevaluatewhetherPLTexertsachangeonotherimportantimmune,connectivetissuesandstemcellsarenecessarytogiveamoreintegratedpictureoftheeffectsofPLT ontissuehealing.Additionally,whilecelllinesareaconvenientandusefulresearchtools,theirbehaviormaynot perfectlyreflectthatoftheirinvivocounterparts.Consequently,evaluationofhowPLTmayaffectthediverse rangeofnativecellularinteractionsinvolvedinhealing
andinflammationshouldbeexaminedthroughanimal modelresearch.Theeffectoflightpenetrancemustalso beevaluatedtoidentifywhethertheeffectsshowninthis studyareequivalenttothosethatmightoccurinaclinical settingwheretheincidentlightisobstructedbyoverlying tissues.WhilePLhasbeenshowntopenetratetoadepth ofupto5centimeters[7],itiscurrentlyunclearifitsabilitytoexertaphysiologicalchangeisaffectedbydistance fromthelightsourceorthelighttransmittingproperties ofthetissuebeingtreated.Finally,thedoseresponseof PLTmustbeevaluatedfurther.Thisstudyidentified 6hoursofPLTasbeingabletogenerateacellular response,butfortranslationintoclinicaluse,minimum effectivetimeframesmustbeidentified,andtheeffectsof repeateddosagesmustalsobeexploredfurther.
5 | CONCLUSION
Noninvasive,nonpharmacologicalinterventionstotreat inflammationandtoassistintheprocessoftissuehealing havethepotentialtorevolutionizethetreatmentofa widerangeofconditions.Thisstudyprovidespreliminary evidencesuggestingthatPLTcouldbeacandidatetofill thisgapintherapeuticapproaches.
Thisstudyprovideskeymechanisticevidenceforthe capacityofPLTtoimpartaphysiologicalchange.The changesingeneandcellsurfacemarkerexpression observedafter6hoursofPLTweresuggestiveofimmune andinflammatorysuppression,providingpotentialfor clinicaluseinahostofillnessesandinjuries.ThesefindingssupportthereportedresultsofPLTseeninclinical practice,whereitisusedtoassistinthehealingof woundsandinjuries.Futureresearchonothercelltypes shouldbeperformedtoestablishaclearmechanism,and doseresponseexperimentsdonetoidentifyoptimaltreatmentpractices.Further,studiescomparingtheeffectof theindividualwavelengthsthatmakeupPLTcould allowformoretargetedtherapeuticapproaches.
CONFLICTOFINTEREST
Theauthorshavenoconflictofinteresttodeclare.
AUTHORCONTRIBUTIONS
J.F.,N.T.andK.M.performedtheexperimentsunderthe supervisionofV.A.,S.F.andM.H.;J.F.,D.K.and V.A.wrotethearticleandJ.F.,V.A.,S.F.,M.H.,D.K.and A.T.editedandreviewedthearticle.
ACKNOWLEDGMENTS
TheauthorswouldliketothanktheImmunologyand TranslationalGroupwithintheInstituteforHealthand Sport,VictoriaUniversity,forsupportanddiscussions.
V.A.wouldliketothanktheVictoriaUniversityCollege ofHealthandBiomedicinestart-upfundsforfinancial support.J.F.wassupportedbytheAustralianGovernmentResearchTrainingProgramandMKbytheVictoria UniversityVice-ChancellorsPostgraduateScholarship.
ORCID
JackFeehan https://orcid.org/0000-0002-9627-1299 VassoApostolopoulos https://orcid.org/0000-0001-67882771
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Howtocitethisarticle: FeehanJ,TripodiN, FraserS,etal.Polarizedlighttherapy:Shininga lightonthemechanismunderlyingits immunomodulatoryeffects. J.Biophotonics .2019; e201960177. https://doi.org/10.1002/jbio.201960177