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A Review on Night Enhancement Eyedrops Using Chlorin e6 Licina, G; Tibbetts, J This work is licensed under the Creative Commons AttributionShareAlike 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/bysa/4.0/. Disclaimer The authors of this paper are writing this review for research and informative purposes only. Increased light amplification may have adverse effects on the cellular structure of the eye if improperly used and the some of the materials used in this mixture should not be used on humans or animals. Introduction Chlorin e6 (Ce6) has been used for many years as a therapy agent in cancer treatment 1,2 . However, in recent years other uses for ce6 have been found, the most notable in this case being its application into the conjuctival sac of the eye as a means of treating night blindness and improving the dim light vision of those with visual disturbances 3 . This preliminary study attempts to test the ability of a mixture containing Ce6 to improve the dim light vision of healthy adults. In 2012 a patent was filed based in some part on the work of Washington et al 4 . The patent claims that a mixture can be made which, when applied to the eye, will absorb to the retina and act to increase vision in low light. The mixture put forth in the patent is a simple combination of Ce6 and insulin in saline. It is mentioned in the same, that dimethlysulfoxide (DMSO) can be used in place of the insulin. We propose a combination of the two could lead to the most noted effects. For testing purposes, the mixture from the patent (Ce6, Saline, Insulin) was used with the addition of DMSO for increased permeability. Material Background Ce6 is a tetrapyrolle and a chlorophyll analog. As mentioned, it has historically been used as a photosensitizer in laser assisted cancer remediation. The light amplification properties of the Ce6 are used to use the energy from a low power light source to destroy cancerous cells with literal laser precision. The reaction creates oxygen species which induce apoptosis in tumor cells. This lead to the concerns about the mixture, as it would be possible that bright or even ambient daylight’s amplified effect in the eye may harm the cells, potentially causing permanent damage. The function of the insulin is not expressly mentioned in the patent or the journals papers. It has been shown that insulin downregulates the ABCG2 mediated transport pathway 5 . With ABCG2 downregulated, greater absorption is shown for photosensitizers like Ce6 6 . In the case of this solution, the insulin is used to allow absorption of the Ce6 into the chamber of the eye.

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  • AReviewonNightEnhancementEyedropsUsingChlorine6Licina,GTibbetts,J

    ThisworkislicensedundertheCreativeCommonsAttributionShareAlike4.0InternationalLicense.

    Toviewacopyofthislicense,visithttp://creativecommons.org/licenses/bysa/4.0/.

    DisclaimerTheauthorsofthispaperarewritingthisreviewforresearchandinformativepurposesonly.Increasedlightamplificationmayhaveadverseeffectsonthecellularstructureoftheeyeifimproperlyusedandthesomeofthematerialsusedinthismixtureshouldnotbeusedonhumansoranimals.IntroductionChlorine6(Ce6)hasbeenusedformanyyearsasatherapyagentincancertreatment1,2.However,inrecentyearsotherusesforce6havebeenfound,themostnotableinthiscasebeingitsapplicationintotheconjuctivalsacoftheeyeasameansoftreatingnightblindnessandimprovingthedimlightvisionofthosewithvisualdisturbances3.ThispreliminarystudyattemptstotesttheabilityofamixturecontainingCe6toimprovethedimlightvisionofhealthyadults.In2012apatentwasfiledbasedinsomepartontheworkofWashingtonetal4.Thepatentclaimsthatamixturecanbemadewhich,whenappliedtotheeye,willabsorbtotheretinaandacttoincreasevisioninlowlight.ThemixtureputforthinthepatentisasimplecombinationofCe6andinsulininsaline.Itismentionedinthesame,thatdimethlysulfoxide(DMSO)canbeusedinplaceoftheinsulin.Weproposeacombinationofthetwocouldleadtothemostnotedeffects.Fortestingpurposes,themixturefromthepatent(Ce6,Saline,Insulin)wasusedwiththeadditionofDMSOforincreasedpermeability.MaterialBackgroundCe6isatetrapyrolleandachlorophyllanalog.Asmentioned,ithashistoricallybeenusedasaphotosensitizerinlaserassistedcancerremediation.ThelightamplificationpropertiesoftheCe6areusedtousetheenergyfromalowpowerlightsourcetodestroycancerouscellswithliterallaserprecision.Thereactioncreatesoxygenspecieswhichinduceapoptosisintumorcells.Thisleadtotheconcernsaboutthemixture,asitwouldbepossiblethatbrightorevenambientdaylightsamplifiedeffectintheeyemayharmthecells,potentiallycausingpermanentdamage.Thefunctionoftheinsulinisnotexpresslymentionedinthepatentorthejournalspapers.IthasbeenshownthatinsulindownregulatestheABCG2mediatedtransportpathway5.WithABCG2downregulated,greaterabsorptionisshownforphotosensitizerslikeCe66.Inthecaseofthissolution,theinsulinisusedtoallowabsorptionoftheCe6intothechamberoftheeye.

  • DMSOisusedincellpreservationandinmedicationapplication.It'sprimaryability,inthisscope,istocauseincreasedpermeabilityofthecellularmembrane,allowingforfreepassageforanychemicalsthatcomeintocontactwiththedosedarea.Whileanecdotalreportsofhealingabilitiesortastinglemonjuicethroughonesskinaboundontheinternet,thehighriskofcellulartoxicityfromoutsidecontaminantsbeingabsorbedthroughtheskinmakethischemicalsomethingthatshouldonlybehandledwithcaution.ProcedureTheCe6(FrontierScientific,CAS:19660776),wasfoundtobeafineblackpowderwhichclungtoallsurfaces.Tomakemanipulatingthechemicaleasier,alargebatchofthetotalsolutionwasmadeandthenaliqoutedintoseparatecontainersforstorage.200mgofCe6wasmixedwith400units(4ml)ofinsulin(70/30Lantus).Tothiswasadded5.38mlofsterilesalinesolution(0.9%sodiumchloride).Themixturewassonicatedbriefly(30seconds)toallowforproperdispersalofthepowderintosaturatedsolutionandthen625lofDMSO(Amresco)wasadded.Thesolutionwassealedwithparafilmandsonicatedfor150seconds.Theresultingliquidwasthinandblackincolor.Solutionwaskeptinglassaliqoutswrappedinfoilat20C.Fortheapplication,thesubjectrestedsupineandhiseyeswereflushedwithsalinetoremoveanymicrodebrisorcontaminantsthatmightbepresent.Eyeswerepinnedopenwithasmallspeculumtoremovethepotentialforblinking,whichmayforceexcessliquidoutbeforeithadachancetoabsorb.Ce6solutionwasaddedtotheconjunctivalsacviamicropippetteat3dosesof50lintoeacheye.Aftereachapplication,pressurewasappliedtothecanthustostopliquidfrommovingfromtheeyetothenasalregion.Eachdosewasallowedtoabsorbbetweenreloadingthepippette,withtheblackcolordisappearingafteronlyafewseconds.Afterapplicationwascomplete,thespeculumwasremovedandblackscleralenseswereplacedintoeacheyetoreducethepotentiallightenteringtheeye.Blacksunglasseswerethenwornduringallbuttesting,toensureincreasedlowlightconditionsandreducethepotentialforbrightlightexposure.TestingTheCe6solutionhasbeenshowntoworkinaslittleasonehour,withtheeffectslastingformanyhoursafterwards3.After2hoursofadjustment,thesubjectand4controlsweretakentoadarkenedareaandsubjectedtotesting.Threeformsofsubjectivetestingwereperformed.Theseconsistedofsymbolrecognitionbydistance,symbolrecognitiononvaryingbackgroundcolorsatastaticdistance,andtheabilitytoidentifymovingsubjectsinavariedbackgroundatvarieddistances.Symbolrecognitionconsistedofplacingacollectionofobjectswithmarkingsonthem(numbers,letters,shapes).Subjectswerethenaskedtoidentifythemarkings,eachviewingtheobjectsfromthesamelocationatadistanceof10meters.Themarkingswerenotmadepriortothemomentoftesting.Forsubjectrecognition,individualswentmovedinasmallgroveoftrees.Theywereallowedtochosetheirownlocationindependently.Distancesrangedfrom25to50metersfromobservationpointandtreesandbrushwereusedforblending.Locationswerechosen

  • withoutbeingobservedbythetestsubjects.TheCe6subjectandcontrolswerehandedalaserpointerandaskedtoidentifythelocationofthepeopleinthegrove.AftertestingtheCe6subjectreplacedthesunglasseswhichwerenotremoveduntilsleep.Eyesightinthemorningseemedtohavereturnedtonormalandasof20days,therehavebeennonoticeableeffects.TheCe6subjectconsistentlyrecognizedsymbolsthatdidnotseemtobevisibletothecontrols.TheCe6subjectidentifiedthedistantfigures100%ofthetime,withthecontrolsshowinga33%identificationrate.DiscussionItisnotedthatmoretestingwillneedtobedoneonthisparticularproject.Currenttestingdonewassubjectiveinnature.AGanzfeldstimulatorandelectroretinigraphwillbeusedtomeasuretheactualamountofelectricalstimulationincreasefromtheeye,givingahardquantifiablenumbertothedegreeofamplification.Itisalsopossibletotestwhichrangesofvisionarebeingamplifiedaswell.However,giventhecurrentresultsandthepreviousbodyofworkonthetechnique,itseemsfairtosaythatthistechniqueissuccessfulinitclaimsforlowlightamplificationinthehumaneye.Thesefindingsaresubjectiveexperiences.Subjectexperiencednoadverseeffectsfollowingadministration.Preliminarytestingseemstoindicatethisincreaseindimlightvisiontobeoccurring.Furthertestingisneedtoconfirmandmeasurethedegreeofimprovementinhealthsubjects.Beingabletoaccesstheinformationputforthinjournalsandpatentsisextremelyimportantforfuturescientiststobeabletoworkwithandbuildfromtheknowledgethatwehavecurrently.Moreover,itisextremelyimportantforclearmethodstobeavailableforanyresearcherwhodesirestoreviewascientificpaper.Thelastyear(2014)hasshownmorescientificjournalarticlesrescindedthananyyearpreviously.CitizenscientistsandDIYbiologistsareundernopressuretoreachorholdapositionoftenureandoftendonothavetheneedtoproduceformonetaryreasons.Itispossiblethatthiswillallowforlessbiasinpublishingandamoreopenreleaseofworkduetothelackofexternalmotivators.Bymakinginformationaccessible,onecanpreemptscoopingandinsteadfocusoncollaboration.Duringthisresearch,wefeelwewerefortunatetobeoperatingfromjustsuchaposition.Thedisadvantagehoweverwasadecreasedavailabilityofaccesstomanyofthetoolsthatwouldallowustoverifyourfindingsquicklyandeasily.Ce6administeredasdescribedinthispaperinthedosagesdescribedhavesofarbeenwithoutanyadverseeffectsandshowgreatpotentialtoenhancethevisionofhealthyadultsindimlightsituations.Furtherstudiesshouldbeperformedinordertomeasuretheeffectsofthisce6solutionobjectively.

  • References1BachorR,ScholzM,SheaC,HasanT.Mechanismofphotosensitizationbymicrosphereboundchlorine6inhumanbladdercarcinomacells.CancerResearch,1991,51:44104414.2AkhlyninaT,RosenkranzA,JansD,SobolevA.InsulinmediatedIntracellularTargetingEnhancesthePhotodynamicActivityofChlorine6.CancerResearch,1995,55:101410193Shantha,T,inventor,assignee.MethodstoEnhanceNightVisionandTreatmentofNightBlindness.21June2012.Patent20120157377.4WashingtonI,ZhouJ,JockushS,TurroN,NakanishiK,SparrowJ.ChlorophyllDerivativesasaVisualPigmentforSuperVisionintheRed.Photochem.Photobiol.,2007,6:775779,DOI:10.1039/b618104j5LiuX,JingX,JinS,LiY,LiuL,YuY,LiuX,XieL.Insulinsuppressestheexpressionandfunctionofbreastcancerresistanceproteininprimaryculturesofratbrainmicrovesselendothelialcells.PharmacologicalReports,2011,63:4874936RobeyR,SteadmanK,PolgarO,BatesS.ABCG2MediatedTransportofPhotosensitizers:PotentialImpactonPhotodynamicTherapy,CancerBiology&Therapy,2005,4:2,195202,DOI:10.4161/cbt.4.2.1440


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