IntJPharmSciTech(2009)

Vol-3, Issue-2, July-Dec, 2009

ISSN: 0975-0525
RESEARCHARTICLE
InvivoevaluationandthestabilitystudyoftheformulationDenticapintendedforlocalapplicationto
relievedentalpain
GopaRoy,SomaGhosh,BiswadipSinhaandBiswajitMukherjee*
DepartmentofPharmaceuticalTechnology,JadavpurUniversity,Kolkata-700032,WestBengal,India
Received 10/11/09; accepted 22/12/09 ; published 28/12/09.
ABSTRACT
Lidocaineiswidelyusedintoothache,teething,torelievepainorirritationcausedbydenturesorotherdentalappliances,
includingbraces.Mostoftheavailabledentalformulationsareliquidandlastonlyfewhoursuponapplication.Toovercome
theabovementionedproblems,asoftpolymericmold,Denticap,containingLidocainehydrochlorideasmodeldrugwas
developedforsustaineddrugreleasetoprovidebetterreliefindentalpain.Twodifferentformulationswereprepared.One
witheudragitL100-55,carbopol971P,gumkarayapowderandethylcellulosewithLidocanehydrochlorideandanother
withcornzein,carbopol934P,gumkarayapowderandpoloxamer407withLidocanehydrochloride.Theywerenamed
asDenticapGandDenticapS,respectively.Theirphysico-chemicalpropertieshavebeenreportedearlier.Thestability
studyoftheDenticapswasperformedandthedatashowedthatthedrugwasstableintheformulationsfollowingthe
conditionsasperICHguideline.Short-termtoxicityforDenticapswasperformedonnormalhealthySwissalbinomice.In
thisstudythefindingssuggestthatthepolymersusedintheformulationofDenticapswerenontoxicandwelltoleratedfor
the30daystudyperiod.In vivostudyoftheDenticapswasperformedintenhealthyhumanvolunteers(bothmaleand
female).HeretheonsetanddurationoflocalanestheticeffectofLidocainehydrochloridereleasedfromtheDenticaps
placedonthetoothsurfaceofthevolunteerswereobserved.Theresultsweresatisfactoryandsupportedbyreported
findings.
KEYWORDS:Denticap,dentalpain,stabilitystudy,short-termtoxicitystudy,In vivostudy
Addressforcorrespondence:
DepartmentofPharmaceuticalTech-
nology,JadavpurUniversity,Kolkata-
700032,WestBengal,India
Tel:+91-33-24146677,
Fax:+91-33-24146393.
E-mail:biswajit55@yahoo.com
Vol-3, Issue-2, July-Dec, 2009 Page-18
INTRODUCTION:
Toothacheusuallyreferstopaininand/oraroundtheteeth
orjaws.Inmostinstances,toothachesarecausedbytoothor
gumproblems,suchasadentalcavity,acrackedtooth,an
exposedtoothroot,gumdisease,diseaseofthejawjoint
(temporal-mandibularjoint),orspasmsofthemusclesused
forchewing.Theseverityofatoothachecanrangefromchronic
andmildtosharpandexcruciating.Thepainmaybe
aggravatedbychewingorbycoldorheat(EdwardJ,
2000).
Lidocainehasbeenwidelyusedasanestheticagentfor
dentalscaling,rootplanning,painsensitivityandearly
woundhealingfollowingnonsurgicalperiodontaltherapy
(Stoltenberget al,2007;Kasajet al,2007).However
Lidocaineformulationsfordentalusearegenerallyliquid
andlastonlyfewhoursuponapplication.Sustained-release
deliverysystemscanbeusefultotreatdentalandperiodontal
diseasescomparedtotheconventionaldosageforms.
Fasterlocalactionascomparedtoslowonsetofactionby
oralrouteandavoidanceofhepaticfirst-passeffectare
importantadvantagesofthisformulation.Researchto
formulate sustained drug release devices for dental
applicationandtreatmentisarelativelynewareainthis
field(Ghoshet al,2009).
Vol-3, Issue-2, July-Dec, 2009 Page-19
IntJPharmSciTech(2009)
AsoftpolymericmoldcontainingLidocainehydrochloride
wasmadetoreleasethedruginasustainedmanner.Ithadan
appropriate consistency to adhere to the tooth and was
developedtoprovidethepatientcompliance.Wenamedit
Denticap(Mukherjeeet al,2009).TwotypesofDenticaps
wereprepared,onewitheudragitL100-55,carbopol971
P,gumkarayapowder,ethylcelluloseandanotherwithcorn
zein,carbopol934P,gumkarayapowder,poloxamer407
containingLidocainehydrochloride.Differentphysico-
chemicalcharacterizationstudiessuchastoothadhesive
strengthtest,waterabsorptioncapacity,swellingindex,weight
loss, surface pH, content uniformity related to those
formulationsandin vitrodrugreleasestudieshavealready
beenreportedbyus(Mukherjeeet al,2009;Ghoshet al,
2009).
HerestabilityforthesixmonthsasperICHguidelines,short-
termtoxicityandin vivostudieshavebeencarriedout.
DevelopmentofFormulationandtheirphysico-chemical
studies
MoldableDenticapswereformulatedwithcombinationof
polymerssuchaseudragitL100-55,carbopol971P,gum
karayapowder,ethylcelluloseandanotherwithcornzein,
carbopol934P,gumkaraya,poloxamer407,ethylcellulose
(TableI).Drugsandpolymersweremixedtogetherinethanol
(95%),usinghomogenizertoformapaste.Themixturewas
pouredinacap-shapedethanol-proofplasticmold(internal
diameter1cm,height1cm)andwasthensubjectedto
evaporationofsolventat37Cfor2h.Finally,theformulations
werecoatedfromallthesidesexceptthesidefordrugrelease
on the affected tooth (Figure1). Prior to coating, the
formulationswereremovedfromtheplasticmolds.The
coatingwasdoneusing5%ethanolicsolutionofethylcellulose
followingthemethodavailable(OhandLuner,1999)keeping
drugreleaseside-covered.
Thephysico-chemicalcharacteristicssuchasmucoadhesion
test,swellingindex,weightlossstudy,scanningelectron
microscopy,in vitrodrugreleaseprofilehavebeenstudied
andthefindingshavealreadybeenreported(Mukherjeeet
al,2009;Ghoshet al,2009).
Stabilitystudy
Denticap G and Denticap S were stored at elevated
temperaturesandrelativehumidity(RH)(302C/60%RH,
452C/75%RH)(DarwinChambersCompany,St.Louis,
USA)overaperiodof6monthsasperICHguideline(ICH,
2003).
Denticapsstoredat2-8Cwereusedascontrol.Thesurface
pH,drugcontent,toothadhesionstrength,in vitrorelease
profilesofthesampleskeptfor6monthsforstabilityanalysis
werecomparedwiththoseofthecontrolformulations(Garg
et al,2007).
Short-termtoxicitystudy
In vivoevaluationstudiesforDenticapswereperformedon
normalhealthySwissalbinomiceof6-8weeks,weighing
20to25geach.TheapprovaloftheInstitutionalAnimal
EthicsCommitteewasobtainedbeforestartingthestudy.
Thestudywasconductedinaccordancewithstandard
institutionalguidelines.Eighteenhealthymice(male)were
acclimatizedtolaboratoryconditionsfor7dayspriorto
initiationofdosing.Theywererandomlyassignedtothree
cages.Micewereassignedinthreegroups.Oneforcontrol
andanothertwoformixtureofpolymersofDenticapG
(eudragitL100-55,carbopol971P,gumkarayapowder
andethylcellulosein2:1:1:1ratiobyweight)andmixtureof
polymersofDenticapS(cornzein,carbopol934P,gum
karayaandpoloxamer407in8:8:4:1ratiobyweight).The
miceweredeprivedoffoodfor16hbeforeand3hafter
administrationofthetestsubstance.Asemisolidmassoftest
substancewasfedtothemad libitum.Observationsof
pharmacotoxicsignsweremadeat10,30,60and120
minutesandat4and6hoursafterdosingduringthefirstday
andoncedailythereafterfor30days.Thetimeofonset,
intensityanddurationofthesesymptoms,ifany,were
recorded.Allanimalswereobservedtwicedailyformortality
duringthe30dayperiodofstudy(BidheandGhosh,2004).
Attheendofthe30dayperiodtheanimalswerefasted
overnight.Thefollowingmorning,eachanimalweresacrificed
andbloodsampleswerecollected.Theplasmaactivitiesof
serumglutamicpyruvictransaminase(SGPT)andserum
glutamicoxaloacetictransaminase(SGOT)wereassayed
byusingLiquidgoldreagentsforAspartateaminotransferase
(AST)andalaninetransaminase(ALT)estimation(Span
DiagnosticsLtd,Surat,India).Thereagentswereprepared
accordingtotheguidelinesofthemanufacturer.Calculation
ASTactivity(IU/L)=A/minuteKineticfactor
Where,
A/minute=Changeinabsorbanceperminute
Kineticfactor(K)=1768
Kineticfactorisevaluatedbythefollowingformula
1 TV 1
K=10
6
M SV P
ALTactivity(IU/L)=A/minuteKineticfactor
Where,
A/minute=Changeinabsorbanceperminute
Kineticfactor(K)=1768
SurfacepH,ToothadhesivestrengthanddrugcontentofDenticapGandDenticapSStoredatDifferentTemperature
andHumidityConditionsfor6months
Table-2:
Vol-3, Issue-2, July-Dec, 2009 Page-20
IntJPharmSciTech(2009)
Kineticfactorisevaluatedbythe
followingformula
1 TV 1
K=10
6
M SV P
Where,
M=Molarextinctioncoefficientof
NADHandisequalto6.2210
3
lit/
mol/cmat340nm
TV= Sample volume + Working
Reagentvolume
SV=Samplevolume
P=Opticalpathlength
10
6
=Constant
In vivostudyoftheDenticaps
ThisstudywasconductedforbothDenticapGandDenticap
S.TheapprovaloftheInstitutionalEthicsCommitteewas
obtainedbeforestartingthestudy.TheDenticapGand
DenticapScontainingLidocainehydrochlorideshowed
goodandreproduciblein vitroreleaseprofileinsimulated
salivaat372Coveratimeperiodof24h.Toevaluate
thein vivoreleasestudy,10adulthumanvolunteers(both
maleandfemale)hadbeengiventheDenticapsinbuccal
cavityovertoothsurface(Figure2)andaskedtonotice
thelocalanestheticeffectofLidocainehydrochlorideover
atimeperiodof8-12h.Nobloodorhumansampleswas
collectedforthepurpose.Ourintentionwastoverifythe
sustainedlocaleffectofthedrugfromtheDenticaps.
Table:1:DrugpolymercompositionofDenticap
formulations
(DenticapG:eudragitL100-55,carbopol971P,gum
karaya powder, ethyl cellulose and Lidocaine
hydrochlorideintheratio1:1:0.5:0.5:0.3byweight,
DenticapS:containscornzein,carbopol934P,gum
karaya powder, poloxamer 407 and Lidocaine
hydrochloride8:8:4:1:2.1byweight,totalweightofthe
Denticapwas500mg)
Statistics:
Datawereassessedbyone-wayANOVAfollowedby
TukeyHSDtestusingVassarStatssoftware(USA).P<
0.05hasbeenconsideredasstatisticalsignificance.



Formulation

Ingredients (ratio by weight) Lidocaine
hydrochloride:
Polymer(by
weight)
Denticap G Eudragit L 100-55, Carbopol 971 P,
Gum karaya, Ethyl cellulose
(1 :1 :0.5 :0.5)
1:10
Denticap S Corn zein, Carbopol 934 P, Gum
karaya, and Poloxamer 407 (8:8:4:1;
ethyl cellulose was used for coating
only)
1:10
Table-3:In-vivotestingdata(onsetanddurationofactionofLidocainehydrochloridereleasedfromtheDenticapGand
DenticapSappliedonthetoothsurfaceofthevolunteers)
Fig-1:PictureofDenticapGandDenticapS(cylindri-
calinshape,diameter-1cm,height-1cm,volume-0.79
cm3DenticapG:eudragitL100-55,carbopol971P,
gumkarayapowder,ethylcelluloseandLidocainehy-
drochlorideintheratio1:1:0.5:0.5:0.3byweight,
DenticapS:containscornzein,carbopol934P,gum
karayapowder,poloxamer407andLidocainehydro-
chloride8:8:4:1:2.1byweight)
Fig-2:PictureofDenticapGandDenticapSapplied
onthetoothsurface(DenticapSatthelowerJawand
DenticapGattheupperjaw)
Vol-3, Issue-2, July-Dec, 2009 Page-21
IntJPharmSciTech(2009)
Results:
Table2representsthevariousdataofstabilitysamplesstoredat
differenttemperatureandhumidityconditionsfor6monthsas
perICHguideline(ICH,2003).
Figure3and4representtheFTIRdataofthestabilitystudiesof
DenticapGandDenticapSkeptat2-8C,302C/60%RH
and452C/75%RHfor6months.Figure5showsthedrug
releasestudyoftheDenticapGandDenticapS,respectively,
storedat2-8C,302C/60%RHand452C/75%RHfor
6months.Thedatasuggestthatthedrugwasstableinthe
formulationsunderthementionedstorageconditions.
Itwasobservedthattheanimalsfedwiththepolymermixture
werehealthy.Nounusualchangesinbehaviorandnosignsof
intoxication were observed during the 30 day period. No
differenceswerefoundingrowthbetweenthecontrolgroupand
theanimalsfedwithpolymermixtureofeudragitL100-55,
carbopol971P,ethylcelluloseandgumkarayaat2:1:1:1ratio
and corn zein, carbopol 934 P, gum karaya powder and
poloxamer407at8:8:4:1ratio.Thefoodconsumptionofmice
ofcontrolandexperimentalgroupswassimilar.Theplasma
activitiesofserumglutamicpyruvictransaminase(SGPT)and
serumglutamicoxaloacetictransaminase(SGOT)werenot
significantlydifferent(P>0.05)forbothcontrolandtestgroups
(Figure6).
Toevaluatethein vivoreleasestudy,10adulthumanvolunteers
(bothmaleandfemale)weredividedintwogroupsrandomly
andtheyhadbeengivenDenticapGinbuccalcavityovertooth
surface(onegroupinupperjawandotherinlowerjaw).They
wereaskedtonoticethelocalanestheticeffectofLidocaine
hydrochlorideoveratimeperiodof8-12h.Sameexperiment
wasrepeatedafterfewdaysforDenticapS.Ithadbeenobserved
thatincaseofDenticapGtheonsetofactionofLidocaine
hydrochlorideinupperjawandlowerjawwere3minutesand2
minutes,respectively.WhereasincaseofDenticapStheywere
3minutesand1minute,respectively.Durationofactionof
LidocainehydrochlorideinDenticapGwas120minutesand
105minutesforapplicationonthetoothsurfaceatupperjaw
andlowerjaw,respectively.WhereasDurationofactionof
LidocainehydrochlorideinDenticapSwas125and90minutes
forapplicationonthetoothsurfaceatupperjawandlowerjaw,
respectively.
Figure3:FTIRdataofstabilitystudyofDenticapGstoredat
differenttemperatureandhumidityconditionsfor6months.GA:
DenticapGwithoutdrugat2-8C,GB:DenticapGwithdrug
at2-8C,GC:DenticapGwithoutdrugat302C/60%RH,
GD:DenticapGwithdrugat302C/60%RH,GE:Denticap
Gwithoutdrugat452C/75%RH,GF:DenticapGwithdrug
at452C/75%RH
Figure4:FTIRdataofstabilitystudyofDenticapS
storedatdifferenttemperatureandhumidityconditions
for6months.(SA:DenticapSwithoutdrugat2-8C,
SB:DenticapSwithdrugat2-8C,SC:DenticapS
withoutdrugat302C/60%RH,SD:DenticapSwith
drugat302C/60%RH,SE:DenticapSwithoutdrug
at 452C/75% RH, GF: Denticap S with drug at
452C/75%RH
Figure-5:CumulativereleaseprofileofLidocainehydrochloride
fromDenticapGandDenticapSrespectivelyafterkeepingin
differenttemperatureandhumidconditionsfor6months(data
showmeanSD,n=3)
Figure6:Bardiagramofplasmaenzymeactivities
(SGOT:serumglutamicoxaloacetictransaminase,
SGPT:serumglutamicpyruvictransaminase)
Vol-3, Issue-2, July-Dec, 2009 Page-22
IntJPharmSciTech(2009)
Discussion
ResultsofthestabilitystudyoftheDenticapGand
DenticapSfor6monthsshowedthatthedrugcontent
andsurfacepHwerealmostsameasthoseofthecontrol
formulations, whereas the tooth adhesive strength
decreased.Thestabilitystudywascarriedoutfollowing
Vol-3, Issue-2, July-Dec, 2009 Page-23
IntJPharmSciTech(2009)
ICHguidelineforstabilitytestingofnewdrugsubstancesand
productsinacontainer-closuresystem.Sincethestudywas
conductedinacontainer-closuresystemtheproductcould
havelostapercentageofmoistureinthecontainer-closure
systemduringitsstoragefor6monthsat2-8Ctemperature.
Thispossiblyresultedinadecreasetoothadhesivestrength
valueascomparedtothefreshformulations(i.e.before
storage).FTIRspectraofallthesamplesundergonethestability
studyhadsimilaritywiththoseofthecontrolonesandthere
washardlyanyvariationamongthesamplesstoredatdifferent
temperatureandhumidityconditions.Thereleasepatternsfor
Lidocainehydrochloridealsofollowedthesamepatternas
thecontrolDenticaps.Initiallyfasterdiffusionofthedrug
moleculesfromthesurfaceoftheDenticapsshowedfirst-order
drugrelease.Butthedrugreleasepatternsvariedin5hfor
Lidocaine hydrochloride. It was noted that the release
graduallychangedfromtimedependenttotimeindependent
zero-ordertypeofreleaseastheinitialone(Mukherjeeet al,
2009;Ghoshet al,2009).Thereleasestudyofstability
samples can be ranked as 2-8C/6month>30C/
6month>45C/6monthforboththeformulations.Thiscould
beduetothehardeningofthematrixwithtime(Cetinet al,
2005). Therefore it is obvious from the results that the
Denticapswerestablechemicallyatthoseelevatedtemperature
andhumidityconditions.
Thepurposeoftoxicitystudywastolookatthetoxicityprofile
of the polymers used.A 30 day study is considered a
subchronicstudy,whichiswellacceptedforelicitinganytoxicity
onlong-termfeeding(BidheandGhosh,2004).Themean
foodconsumptionoftheanimalsinthecontrolandtestgroups
wassimilar.Thisfindingindicatesthatthefoodintakewasnot
affectedbytheintakeofthepolymers(eudragitL100-55,
carbopol971P,ethylcelluloseandgumkarayaandcornzein,
carbopol934P,poloxamer407,ethylcelluloseandgum
karaya).Therewerenodifferencesbetweenthecontroland
thetestwithrespecttonormalbehavior.Theplasmaactivities
ofserumglutamicpyruvictransaminase(SGPT)andserum
glutamicoxaloacetictransaminase(SGOT)werealsoassayed.
SerumSGOTandSGPTlevelsareelevatedinViralandother
formsofLiverdiseaseassociatedwithHepaticnecrosis(Keng-
Lianget al,2004).SGPTismoreLiverspecificenzyme,while
SGOTactivityincreasesrelativelymoreduringmyocardial
infraction.SGOTcatalysesthetransaminationofL-aspartate
and-ketoglutaratetoformL-glutamateandoxaloacetate.In
subsequentreaction,malatedehydrogenase(MDH)reduces
oxaloacetate to malate with simultaneous oxidation of
nicotinamideadeninedinucleotide[reduced](NADH)to
nicotinamide adenine dinucleotide (NAD). The rate of
oxidationofNADHismeasuredkineticallybymonitoringthe
decreaseinabsorbanceat340nmandisdirectlyproportional
toASTactivityinthesample.Lactatedehydrogenase(LD)
isaddedtoenzymesystemtopreventendogenouspyruvate
interference, which is normally present in the serum.
(Schumannet al,2002).
Alanine amino transferase (SGPT) catalyses the
transaminationofL-alanineand-ketoglutaratetoform
pyruvateandL-glutamate.Insubsequentreaction,lactate-
dehydrogenase (LD) reduces pyruvate to lactate with
simultaneousoxidationofnicotinamideadeninedinucleotide
[reduced](NADH)tonicotinamideadeninedinucleotide
(NAD). The rate of oxidation of NADH is measured
kineticallybymonitoringthedecreaseinabsorbanceat340
nm.LDrapidlyandcompletelyreducesendogenoussample
pyruvateduringtheinitialincubationperiod,sothatitdoes
notinterferewiththeassay(Eelkoet al,1998).
Plasmasamplesfromcontrolandtestmicewereassayed
forSGOTandSGPTactivity(Figure6).Averageplasma
SGOTactivityinuntreatedmicewas28.23IU/Landthe
valuewas18.54IU/LincaseofSGPTactivitywithan
SGOT/SGPTratioof1.19.IncaseofDenticapGaverage
plasmaSGOTactivityintreatedmicewas24.25IU/Land
21.42IU/LSGPTactivitywithanSGOT/SGPTratioof
1.13.InDenticapSinplasmafromuntreatedmicemean
SGOTactivitywas33.80IU/LandmeanSGPTactivity
was25.32IU/LforanSGOT/SGPTratioof1.33.No
statisticallysignificantdifferences(P<0.05)inSGOTand
SGPTactivitiesbetweentestandcontrolanimalswere
observed.ThecontroldataofSGOTandSGPTstudiesof
boththeDenticapsarealsosupportedbythereported
findings(Al-Bekairiet al,2004;Sharmaet al,2003).
Thus,ourfindingsindicatethatthepolymersusedinthe
formulationofDenticapsarenontoxicandwell-tolerated
forthe30daystudyperiodandthereforemayhavepotential
forsafeuseindentalformulations.
Inthe in vivostudy,theonsetanddurationoflocalanesthetic
effect of Lidocaine hydrochloride released from the
Denticapsplacedonthetoothsurfaceofthevolunteerswere
observed.DenticapGandDenticapSshowedalmostsame
onsetanddurationofaction.Theonsetofanestheticeffect
was1-5minutesanddurationwas105-125minutes(study
period). The data were satisfactory and supported by
reportedfindings(Simonet al,1997).
HenceapplicationsofDenticapGandSarenovelapproach
forlocaldeliveryofLidocainehydrochlorideforaprolonged
periodwhenappliedonanaffectedtoothtorelievedental
pain.
Acknowledgements
ThisworkwassupportedbygrantsfromIndianCouncilof
MedicalResearch(Grant
No.45/55/2004-PHA/BMS).
Vol-3, Issue-2, July-Dec, 2009 Page-24
IntJPharmSciTech(2009)
References
Al-BekairiAM,OsmanAM,HafeezMA,Al-GharablyNM,
Al-ShabanahOAandAl-HarbiMM.Effectofdesferrioxamine
onthehepatotoxicityofadriamycininnormalmice.DrugDev
Res.(2004)29:56-62.
BidheRMandGhoshS.Acuteandsubchronic(28-Day)oral
toxicitystudyinratsfedwithnovelsurfactants.AAPSPharm
SciTech.(2004)6:14.
CetinEO,BuduneliN,AtlihanEandKirilmazL.In vitro
studies of a degradable device for controlled-release of
meloxicam.JClinPeriodontol.(2005)32:773-77.
EdwardJ.Managingincompletetoothfractures.JAmDent
Assoc.(2000)131:1168-74.
EelkoG,MarcelT,JohannesP,JackTandVerripsT.Pyruvate
decarboxylasecatalyzesdecarboxylationofbranched-chain
2-oxoacidsbutisnotessentialforfuselalcoholproductionby
Saccharomyces cerevisiae.ApplEnvironMicrobiol.(1998)
64:13037.
GargM,DuttaTandJainNK.Stabilitystudyofstavudine-
loaded O-palmitoyl-anchored carbohydrate-coated
lyposomes.AAPSPharmSciTech.(2007)8:38.
GhoshS,RoyGandMukherjeeB.Dentalmold:anovel
formulationtotreatcommondentaldisorders.AAPSPharm
SciTech.(2009)10:692-702.
InternationalConferenceonHarmonisationofTechnical
RequirementsforRegistrationofPharmaceuticalsforHuman
Use.Stabilitytestingofnewdrugsubstancesandproducts
Q1A(R
2
).CurrentStep4versiondated6February2003.
KasajA,HeibAandWillershausenB.Effectivenessofatopical
salve(Dynexan)onpainsensitivityandearlywoundhealing
followingnonsurgicalperiodontaltherapy.EurJMedRes.
(2007)12:19699.
Keng-liang W, Sheng-nan L, Chi-sin C, King-wah C,
ChunghuangK,Seng-keeC,Jien-weiL,Meng-chihL,Hock-
liew E, Shun-sheng C, Chuan-mo L and Chao-long C.
Sequentialchangesofserumaminotransferaselevelsinpatients
withsevereacuterespiratorysyndrome.AmJTropMedHyg.
(2004)71:125-28.
MukherjeeB,RoyGandGhoshS.DevelopmentofDenticap,
amatrixbasedsustainedreleaseformulationfortreatmentof
toothache,dentalinfectionandothergumproblem.Curr
DrugDeliv.(2009)6:199-207.
OhEandLunerPE.Surfacefreeenergyofethylcellulose
filmsandtheinfluenceofplasticizers.IntJPharm.(1999)
188:20319.
SchumannG,BonoraR,CeriottiF,FrardG,FerreroCA,
Franck PF et al. International federation of clinical
chemistryandlaboratorymedicine.IFCCprimaryreference
procedures for the measurement of catalytic activity
concentrationsofenzymesat37degreesC.International
FederationofClinicalchemistryandlaboratorymedicine.
Part 5. Reference procedure for the measurement of
catalyticconcentrationofaspartateaminotransferase.
(2002)40:725-33.
SharmaS,RamyaTNC,SuroliaAandSuroliaN.Triclosan
asasystemicantibacterialagentinamousemodelofacute
bacterialchallenge.AntimicrobAgentsChemother.(2003)
47:3859-66.
SimonMAM,GielenMJM,AlberinkN,VreeTBand
EgmondJVan.Intravenousregionalanesthesiawith0.5%
articaine,0.5%lidocaine,or0.5%prilocaine:adouble-
blindrandomizedclinicalstudy.RegAnesPainMed.(1997)
22:29-34.
StoltenbergJL,OsbornJB,CarlsonJF,HodgesJSand
MichalowiczBS.Apreliminarystudyofintra-pockettopical
versusinjectedanaestheticforscalingandrootplaning.J