3/21/2016 Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoralcavitycancer:[PAUTHORS],IndianJournalofDentalResearch(IJDR)

ORIGINALRESEARCH
Year:2008|Volume:19|Issue:3|Page:213218

Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoral
cavitycancer
SChitra1,CSShyamalaDevi2,
1DepartmentofBiotechnology,SriRamachandraCollegeofBiomedicalScience,TechnologyandResearch,SriRamachandraUniversityandResearchInstitute,Porur,Chennai600116,India
2DepartmentofBiochemistryandMolecularBiology,UniversityofMadras,GuindyCampus,Chennai600025,TamilNadu,India

CorrespondenceAddress:
SChitra
DepartmentofBiotechnology,SriRamachandraCollegeofBiomedicalScience,TechnologyandResearch,SriRamachandraUniversityandResearchInstitute,Porur,Chennai600116
India

Abstract
Objective:The objective of the present study was to evaluate early and late effects of radiation and atocopherol on the secretion rate of saliva and on selected saliva salivary
parametersinoralcavitycancerpatients.Patients&Methods:Eightyninehistologicallyconfirmedoralcavitycancerpatients(OCC)wereenrolledinthestudy.Restingwholesaliva
wascollectedbefore,duringandattheendoftheradiationtherapy(RT)andsimultaneoussupplementationwithtocopheroltotheradiationtreatedpatients(RT+AT).Results:
Salivaryflowrate,pH,amylaseactivity,totalprotein,sodiumandpotassiumwereanalyzed.IncreasedpH,potassiumanddecreasedflowrate,amylaseactivity,proteincontentand
sodiumwereobservedin6weeksofradiationtreatedpatientswhencomparedtoOCCpatients.Asignificantimprovementofthoseparameterswasobservedontocopherol
supplementationinRT+ATpatients.Conclusion:Supplementationwithtocopherolimprovesthesalivaryflowratethereby,maintainssalivaryparameters.

Howtocitethisarticle:
ChitraS,ShyamalaDeviCS.Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoralcavitycancer.IndianJDentRes
200819:213218

HowtocitethisURL:
ChitraS,ShyamalaDeviCS.Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoralcavitycancer.IndianJDentRes
[serialonline]2008[cited2016Mar20]19:213218
Availablefrom:http://www.ijdr.in/text.asp?2008/19/3/213/42953

http://www.ijdr.in/printarticle.asp?issn=09709290year=2008volume=19issue=3spage=213epage=218aulast=Chitra 1/5
3/21/2016 Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoralcavitycancer:[PAUTHORS],IndianJournalofDentalResearch(IJDR)

FullText
Salivahasanumberofbeneficialfunctionsintheoralcavitylikelubrication,protectionofmucosalintegrityandantimicrobialactivity.[1]Whileundernormalphysiologicalconditions,
theprotectivefunctionsofsalivaremainintact,theyseemtobedisturbedinoralcavitycancer(OCC).[2]Alteredsalivaryglandfunctioncanprovokeunpleasantoralsymptomsbut
equallyinsidiousistheabsenceofsymptomswhichmayresultinthecancergoingundiagnosed.Radiationeffectsonsalivaryglandsareofparticularinterestinclinicalradiotherapy
ofheadandnecktumors,wherethereducedflowrateandthealteredcompositionofsalivaresultindistress,oftenirreversiblecomplicationssuchasoraldryness,hamperedoral
functioning,nocturnaloraldiscomfort,burningmouth,impededsocialactivitiesandasusceptibilitytooralinfectionsanddentalcaries.[3]Significantsalivaryflowreductiondevelops
aftertheirradiationofsalivaryglandsinthetreatmentofheadandneckcancer.Theaveragesalivaryflowratedecreasesby57%afteroneweekofradiation,by67%aftersixweeks
ofradiationandby95%afterthreeyearsoftreatment.[4]Generally,thedamageisirreversibleinpatientswhoreceivedoses6000cGy.[5]

Theparotidglandsaremoreradiosensitivethanthesubmandibularorsublingualglands.[6]Radiationeffectsonparotidglandtissuearemainlyresponsibleforxerostomiaand
associatedsideeffects.Disruptionofmucosalintegrityasadirecteffectofradiationtherapy,demonstratesenhancedsensitivitytophysical,chemicalandmicrobialinsultsinthe
mouth. Pain and discomfort associated with hyposalivation and mucositis, may affect a patient's nutritional intake, oral function and quality of life. [7] OCC patients undergoing
radiationtherapynotonlyshowdrasticallydecreasedquantitiesofsaliva,butalsoshowqualitativechangessuchasalteredviscosity,pH,immunoglobulinandelectrolytelevelsof
saliva.[8]

RadiationtherapyinflictstremendousdamagetohealthycellssurroundingtumorcellsinOCCpatients.However,therearespecificnutrients,flavonoidsandherbsthathavebeen
showntoprotectthebodyagainstcancerradiationtherapy.AntioxidanttherapyisbeneficialtoavoidthedamagecausedbyradiotherapyinOCCpatients.VitaminEcanprotect
againstmutagenicand/orcarcinogeniceffectsofionizingradiationinbothanimalsandcellculture.[9]RadioprotectionbyvitaminEagainstsalivarydysfunction[10]andpulmonary
fibrosis[11]werestudiedinirradiatedrats.Inthepresentstudy,auniversallyacceptedantioxidantandfreeradicalscavenger,tocopherolwasusedasasupplementaryagentto
reducethetoxicsideeffectsofradiationinOCCpatients.

Thehypothesisofthepresentstudywasthatsalivaflowrateandconcentrationsofprotein,aamylaseandelectrolytesmaybedisturbedinsalivaryglandsbythephysiologicaleffects
causedbyradiationtherapyofOCC.Therefore,onthebasisofpreviousdata,ouraimwastoevaluatetheeffectsofatocopherol(AT)onrestingwholesalivasecretionrates,pHand
concentrationsofsalivaryamylase,proteinsandelectrolytesinpatientsbefore,duringandafterRTandRT+AT.

MaterialsandMethods

HistologicallyconfirmedpatientspresentingattheGovernmentArignarAnnaMemorialCancerResearchInstituteandHospitalinKancheepuram,TamilNadu,India,wereincludedin
thiscancerclinicaltrial.OCCpatients(agemeanSD=5015years)54malesand35females,hadcancerinvarioussitessuchasthecheek(n=28),alveolus(n=17),tongue(n
=13),floorofthemouth(n=11),lip(n=8),palate(n=7)andretromolartrigone(n=5).Thepatientshadvarioushabitsofchewingtobacco(n=48),smokingtobacco(n=29),and
alcoholconsumption(n=12).TumorstagingwasdoneaccordingtotheTumorNodularMetastasis(TNM)classificationoftheUICC(UnionInternationalContreLeCancer)into
stages:I(n=4),II(n=16),III(n=29)andIVA(n=40)[Table1].Allpeoplegaveinformedconsentpriortotheirinclusioninthestudyandamedicalpractitionermonitoredthewhole
experiment.StudieswereperformedinaccordancewiththeethicalstandardsoftheInstitution.

All89OCCpatientswereconsideredtobeingroup1beforeradiation.Thisgroupwasrandomlydividedintotwogroups,i.e.,2(aandb)and3(aandb).Group2aconsistedofOCC
patientstreatedwiththreeweeksofradiation(n=52)(nonATgroup)andgroup2bconsistedofOCCpatientstreatedwithsixweeksofradiation.(n=52).Group3aconsistedofOCC
patientstreatedwiththreeweeksofRTandsimultaneouslysupplementedwithatocopherol(RT+AT)(n=37)(RT+ATgroup)forthreeweeks.Group3bconsistedofOCCpatients
treatedwithsixweeksofRTandsimultaneouslysupplementedwithatocopherol(RT+AT)(n=37)(RT+ATgroup)forsixweeksduringtheentireperiodofradiotherapy[Flowchart
1].RTwasgivenwiththeaidofaTelecobaltbeamusinganteriorandlateralwedgepairorlateralparallelportals(Gammatron60COTheraton78060COPhoenix60CO)ata
dosageof6000cGyinfivefractionsperweekforaperiodofsixweeks.Tocopherol(BioEcapsules)wasgivenatadosageof400IUperdayforsixweeksfromthefirstdayofRTto

http://www.ijdr.in/printarticle.asp?issn=09709290year=2008volume=19issue=3spage=213epage=218aulast=Chitra 2/5
3/21/2016 Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoralcavitycancer:[PAUTHORS],IndianJournalofDentalResearch(IJDR)

theendofthetreatmentinRT+ATpatients.

Collectionofsalivasamples

Patientswerenotallowedtosmoke/chewtobaccoordrinkalcoholduringtheentireperiodofradiotherapy.Collectionofsalivatookplaceattheendofthird(2a)andsixth(2b)weeks
ofradiationtreatment.Similarlyingroup3,samplingwasdoneattheendofthird(3a)andsixth(3b)weeksofradiationtreatment,simultaneouslysupplementedwithTocopherol.
The participants had to refrain from eating and drinking water for a minimum of 90 minutes before saliva collection. The whole resting saliva was collected into widemouthed
graduatedtubesovera30minperiodaccordingtothemethodofNavazeshetal.[12]

Biochemicalassays

Thesalivaflowrate(inmilliliters/minute)ofrestingwholesalivawasmeasuredimmediatelyaftersalivacollection.SalivarypHwasdeterminedbyusingapHmeterandtheflowrate
wasmeasuredaccordingtothemethodofEpsteinetal.[13]Thesampleswerecentrifugedat1200gfor10minat4C.Aliquotsofcentrifugedsalivaneededfortheanalysiswere
transferredtoeppendorfvialsandstoredat20Cuntilanalysis.Allthesampleswereanalyzedwithin15daysofcollection.Salivaryaamylasewasassayedaccordingtoaslight
modificationofthemethodofBernfeld[14]andtotalproteinconcentrationwasmeasuredbyusingtheLowrymethod.[15]Sodiumandpotassiumconcentrationswereanalyzedusing
anatomicabsorptionspectrophotometer(PerkinandElmer2380).

Statisticalanalysis

StatisticalanalysisofthedatawasperformedusingtheMannWhitney'U'testandP0.05wasconsideredasstatisticallysignificant.[16]

Results

Salivaryflowrate,pH,aamylaseactivityandtotalprotein,sodiumandpotassiumlevelswereanalyzed.Decreasesintheactivityofamylase,flowrateandproteinlevels(PPPPP
PPPPPP[17],[18]Radiationinducedsalivaryglanddysfunctionisassociatedwithdecreasedsalivaryflow.[19]HyposalivationisprogressiveandpersistentinRTpatients.Salivary
flowmeasurablydecreaseswithinaweekafterstartingradiationtreatmentanddiminishesprogressivelywithcontinuedtreatment.[20]Irreversibledecreaseintheflowratemaybe
relatedtoalossoftissueintegrityandalterationsintheintralobularnerveendings,ratherthantoareducedpotentialoftheglandulartissue.[21]Unstimulatedwholesalivaryflowrate
[22]Xerostomiawasobservedaftersixweeksofradiationtreatmentinthepresentstudy.

Changesintheplasmamembranesofacinarcellsmayalsobethecauseofdisturbedsignaltransductionafterirradiation,particularlyasthepolyunsaturatedfattyacylchainsof
membrane phospholipids may be readily damaged by irradiation. [23] High levels of potassium and low levels of sodium in the saliva of OCC and RT patients could be due to
radiotherapyrelateddemineralization.Osteoradionecrosisisaseriouscomplicationforpatientswhoreceiveradiationforheadandnecktumors.[17]

ThehighactivityofamylaseandtotalproteincontentinOCCpatientshowever,canbeexplainedbythelowervolumeofsecretedsalivaratherthanbytherateofproteinsynthesisin
thesalivaryglands.Thisapproachhasbeenusedinanumberofstudies[19],[20],[21]toanalyzetheeffectofasystemicfactoronthesynthesisofsalivaryproteinsandsubsequent
salivarycomposition.

Amylaseissynthesizedprimarilyintheacinarcellsandlessconsistentlyintheproximalcellsoftheintercalatedductshence,gingivalcervicalfluid,arouteofentranceforsome
salivaryproteins,doesnotcontributetoamylaselevels.[24]Therefore,salivaryamylaseisagoodindicatorofthefunctionofsalivaryglands,andhence,ofthegeneralhealthofthe
salivaryglands.IncreasedactivityofamylaseobservedinOCCpatientsmightbeduetoanincreaseinthenumberofacinarcellsintheparotidgland.Highersecretionofamylase
hasbeenreportedinpathologicalconditionsand/orinflammation.[25]Decreasedaamylaseactivitywasobservedaftersixweeksofradiationtreatmentprobablyduetoareduction
inthenumberofacinarcells,incompletetissueregenerationandlatestromaleffectssuchasdelayedvasculardamageduetoradiation.Theparotidglandismoreradiosensitive[26]
http://www.ijdr.in/printarticle.asp?issn=09709290year=2008volume=19issue=3spage=213epage=218aulast=Chitra 3/5
3/21/2016 Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoralcavitycancer:[PAUTHORS],IndianJournalofDentalResearch(IJDR)

andtheamountofseroussecretionisdirectlyproportionaltotheamylasecontent.[27]Thechangesobservedinbothsalivaryconcentrationandrateofsecretionreflectradiation
induceddisturbancesintheacinarproteinsynthesis.amylaseisalsostronglyaffectedbychangesintheflowrateandtotalproteinconcentration[28]asobservedinthisstudy.

ReversalofchangesinpH,flowrate,aamylaseaswellasinprotein,sodiumandpotassiumlevelswereobservedontocopherolsupplementation.Theenhancementinthesalivary
secretionratereflectsanimprovementintissuerepairandarecoveryoftheinjuredcellsorarepopulationoftheglandulartissue.Antioxidantvitaminshavebeenknowntocause
extensivemorphologicalchangesintumorigenicacinarcellsandcanalsocausesignificantgrowthinhibitioninvitro.[29]VitaminEcanprotectagainstthecarcinogeniceffectsof
ionizingradiationinducedinanimalsandcellculture.[9]Radioprotectionagainstsalivarydysfunction[10]byvitaminEwasstudiedinirradiatedrats.Ourobservationwasconsistent
withothers'findings.

Conclusion

Toconcludethepresentstudy,hyposalivation(xerostomia)resultswithinsixweeksoftreatmentinRTpatients.AreducedsalivaryflowrateintheoralcavityofRTpatients,may
promotethegrowthofmicrobialfloraandcausesdifficultyinspeech,swallowingandultimately,affectsthepatient'squalityoflife.atocopherolincreasesthesalivaryflowrateand
andmaintainstheoralenvironmentinOCCpatientstreatedwithradiotherapy.

Acknowledgments

ThisworkwassupportedbytheIndianCouncilforMedicalResearch,NewDelhiandBioEcapsuleswereprovidedbytheAmericanRemedies,Chennaiandalsothehelprendered
byDr.Koteeswaran,DentalSurgeonwithoutwhichwasnotpossibleisgratefullyacknowledged.

References
1 ZellesT,PurashothamKR,MacauleySP,OxfordGE,HumphreysBeherMG.Salivaandgrowthfactors,thefountainofyouthresidesinusall.JDentRes199574:182632.
2 Dutta SK, Orestes M, Vengulekur S, Kwo P. Ethanol and human saliva: Effect of chronic alcoholism on flow rate, composition, and epidermal growth factor. Am J
Gastroenterol199287:3504.
3 FoxPC,VanderVanPF,SoniesBC,WeiffenbaumJM,BaumBJ.Xerostomia:Evaluationofasymptomwithincreasingsignificance.JAmDentAssoc1985110:51925.
4 DoseAM.Thesymptomexperienceofmucositis,stomatitisandxerostomia.SeminOncolNurs199511:24855.
5 BergerAM,KilroyTJ.Oralcomplicationsofcancertherapy.In:BergerA,PortenoyRK,WeissmanDE,editors.Principlesandpracticeofsupportiveoncology,Philadelphia:
LippincottRaven1998.p.223.
6 SrephensLC,SchulthesisIE,SmallSM,AngKK.PetersLJ.Responseofparotidorganculturetoradiation.RadiatRes1989120:14053.
7 DumbrigueHB,SandowPL,NguyenKT,HumphreysBeherMG.Salivaryepidermalgrowthfactorlevelsdecreaseinpatientsreceivingradiationtherapytotheheadand
neck.OralSurgOralMedOralPatholOralRadiolEndod200089:7106.
8 BergerAM,KilroyTJ.Oralcomplications.In:DeVitaBergerA,PortenoyRK,WeissmanDE,editors.Principlesandpracticeofsupportiveoncology,Philadelphia:Lippincott
Raven1998.p.223.
9 DirierA,AkmansuM,BoraH,GurerM.TheeffectofvitaminEonacuteskinradiationcausedbyradiotherapy.ClinExpDermatol200732:5713.
10 FlaviaMariaDM,AnjosPontualML,AlmeidaSM,BoscoloFN,TabchouryCP,NovaesPD.EvaluationofradioprotectiveeffectofvitaminEinsalivarydysfunctioninirradiated
rats.ArchOralBiol200651:96101.
11 BeseNS,MunzurogluF,UsluB,ArbakS,YesiladaliG,SutN,Altug,T,OberA.*VitaminEprotectstheratsagainstthedevelopmentofradiationinducedpulmonaryfibrosis.
ClinOncol200719:260264.

http://www.ijdr.in/printarticle.asp?issn=09709290year=2008volume=19issue=3spage=213epage=218aulast=Chitra 4/5
3/21/2016 Effectsofradiationandtocopherolonsalivaflowrate,amylaseactivity,totalproteinandelectrolytelevelsinoralcavitycancer:[PAUTHORS],IndianJournalofDentalResearch(IJDR)

12 NavazeshM,ChristensenC,BrightmanV.Clinicalcriteriaforthediagnosisofsalivaryglandhypofunction.JDentRes199271:13639.
13 EpsteinJB,EmertonS,GugliettaA,LeN.Assessmentofepidermalgrowthfactorinoralsecretionsofpatientsreceivingradiationtherapyforcancer.OralOncol199733:359
63.
14 BernfeldP.Amylasealphaandbeta.MethodsEnzymol19551:14958.
15 LowryOH.RoseboroughNJ,FairAL,RandallRJ.ProteinmeasurementwiththeFolinphenolreagent.JBiolChem1951193:26575.
16 MannHB,WhitneyDR.Onatestofwhetheroneoftworandomvariablesisstochasticallylargerthantheother.MathStat194718:5060.
17 ValdezIH,AtkinsonJC,ShipJA,FoxPC.Majorsalivaryglanddysfunctioninpatientswithradiationinducedxerostomia:Flowratesandsialochemistry.IntJRadiatOncol
BiolPhys199325:417.
18 ChometteG,AuriolM,VaillantJM,BertrandJC,ChenalC.Effectsofirradiationonthesubmandibularglandoftherat.VirchowsArchPatholAnat1981391:2919.
19 KonningsAW.Effectsofheatandirradiationonmammaliancells.RadiatPhysChem198730:33949.
20 BrandzaegP,FjellangerI,GjeruldsenST.HumansecretoryimmunoglobulinsIgA:Salivarysecretionsfromindividualswithnormalorlowlevelsofserumimmunoglobulins.
ScandJHaematolSuppl197012:383.
21 MandelID,BarrCE,TurgeonL.LongitudinalstudyofparotidsalivainHIV1infection.JOralPatholMed199221:20913.
22 van der Reijden WA, van der Kwaak JS, Veerman EC, Nieuw Amerongen AV. Analysis of the concentration and output of whole salivary constituents in patients with
Sjogren'ssyndrome.EurJOralSci1996104:33540.
23 Lenander LM, Puhakka T, Makela MJ, Vilja P, Ruuskanen O, Tenovuo J. Effects of the common cold and intranasal fluticasone propionate treatment on mucosal host
defenseassessedbyhumansaliva.OralSurgOralMedOralPatholOralRadiolEndod199987:6959.
24 LenanderLM,IhalinR,LahteenojaH.Changesinwholesalivainpatientswithceliacdisease.ArchOralBiol200045:34754.
25 SeifertG,DonathK.Morphologyofsalivaryglanddiseases.ArchOtolaryngol1976213:1118.
26 ShannonLL,TrodahlJN,StarckeEN.Radiosensitivityofthehumanparotidgland.ProcSocExpBiolMed1978:157:503.
27 VisinkA,GravenmadeEJ,LigeonEE,KonningAW.Afunctionalandchemicalstudyofradiationeffectsonratparotidandsubmandibularsublingualglands.RadiatRes
1990124:25965.
28 RantonenPJ,MeurmanHJ.Correlationsbetweentotalprotein,lysozyme,immunoglobulin,amylaseandalbumininstimulatedwholesalivaduringdaytime.ActaOdontol
Scand200058:1605.
29 PrasadKN,KumarR.Effectofindividualandmultipleantioxidantvitaminsongrowthandmorphologyofhumannontumorigenicandtumorigenicparotidacinarcellsin
culture.NutrCancer199626:119.

Sunday,March20,2016
SiteMap|Home|ContactUs|Feedback|CopyrightandDisclaimer

http://www.ijdr.in/printarticle.asp?issn=09709290year=2008volume=19issue=3spage=213epage=218aulast=Chitra 5/5