Stem Cells - Virginia Veterinary Medical Association … and PRP...Inside a Pathologic Joint •...
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1 Contributor: Dr.med.vet. J.Troillet; St. Petersburg 24.02.12 Victoria Maxwell, DVM, MBA 2018 Potomac Regional Veterinary Conference Hyatt Regency Inner Harbor Baltimore, Maryland Regenerative Medicine IRAP and PRP in the Equine Athlete Equine Regenerative Medicine • provide a scaffold for repair • provide cells for the healing process • provide growth factors to stimulate the healing process • Some regenerative therapies currently used in horses: – Stem Cells – Platelet-Rich Plasma (PRP) – Autologous Conditioned Serum (irap) Stem Cells • Cells that have the ability to divide and develop into many different cell types in the body during early life and growth. • May also help repair the body by dividing to replenish cells that are damaged by disease, injury, or normal wear. – When a stem cell divides, each new cell has the potential to either: • remain a stem cell • become another type of cell with a more specialized function, such as a nerve cell, a skin cell, or a red blood cell. Stem Cells • 3 types of stem cells identified in human and veterinary research: – Adult stem cells • Two of the most common sources of equine adult stem cells are bone marrow and adipose (fat)-derived mesesenchymal stem cells (MCS) – Human embryonic stem cells (hESC) • Come from embryos that develop from eggs that have been fertilized in vitro (in a test tube or another artificial environment). – Induced pluripotent stem cells (iPS) • Generated by reprogramming adult cells that have already differentiated into a specific cell, such as a skin cell. • FDA has not approved any stem cell-based products for use, except cord blood-derived hematopoietic progenitor cells (blood forming stem cells) for certain human indications including certain blood cancers and some inherited metabolic and immune system disorders . http://www.vet-stem.com/science.php Veterinary Medical Devices • Further, a device "does not achieve any of its principal intended purposes through chemical action within or on the body of man or other animals, and is not dependent upon being metabolized for the achievement of any of its principal intended purposes." Examples of devices include such things as needles, syringes, surgical instruments, prosthetic devices, X-ray equipment, certain diagnostic test kits, and dental appliances. • Pre-market Approval is Not Required: The FDA does not require submission of a 510(k) or formal pre-market approval for devices used in veterinary
Transcript of Stem Cells - Virginia Veterinary Medical Association … and PRP...Inside a Pathologic Joint •...
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Contributor:Dr.med.vet.J.Troillet;St.Petersburg24.02.12
VictoriaMaxwell,DVM,MBA2018PotomacRegionalVeterinaryConference
HyattRegencyInnerHarborBaltimore,Maryland
RegenerativeMedicineIRAPandPRPintheEquineAthlete
EquineRegenerativeMedicine• provideascaffoldforrepair• providecellsforthehealingprocess• providegrowthfactorstostimulatethehealingprocess• Someregenerativetherapiescurrentlyusedinhorses:
– StemCells– Platelet-RichPlasma(PRP)– AutologousConditionedSerum(irap)
StemCells• Cellsthathavetheabilitytodivideanddevelopinto
manydifferentcelltypesinthebodyduringearlylifeandgrowth.
• Mayalsohelprepairthebodybydividingtoreplenishcellsthataredamagedbydisease,injury,ornormalwear.– Whenastemcelldivides,eachnewcellhasthepotentialtoeither:
• remainastemcell• becomeanothertypeofcellwithamorespecializedfunction,suchasanervecell,askincell,oraredbloodcell.
StemCells• 3typesofstemcellsidentifiedinhumanandveterinaryresearch:
– Adultstemcells• Twoofthemostcommonsourcesofequineadultstemcellsarebonemarrowandadipose(fat)-derivedmesesenchymalstemcells(MCS)
– Humanembryonicstemcells(hESC)• Comefromembryosthatdevelopfromeggsthathavebeenfertilizedinvitro(inatesttubeoranotherartificialenvironment).
– Inducedpluripotentstemcells(iPS)• Generatedbyreprogrammingadultcellsthathavealreadydifferentiatedintoaspecificcell,suchasaskincell.
• FDAhasnotapprovedanystemcell-basedproductsforuse,exceptcordblood-derivedhematopoieticprogenitorcells(bloodformingstemcells)forcertainhumanindicationsincludingcertainbloodcancersandsomeinheritedmetabolicandimmunesystemdisorders.
http://www.vet-stem.com/science.php
VeterinaryMedicalDevices
• Further,adevice"doesnotachieveanyofitsprincipalintendedpurposesthroughchemicalactionwithinoronthebodyofmanorotheranimals,andisnotdependentuponbeingmetabolizedfortheachievementofanyofitsprincipalintendedpurposes."Examplesofdevicesincludesuchthingsasneedles,syringes,surgicalinstruments,prostheticdevices,X-rayequipment,certaindiagnostictestkits,anddentalappliances.
• Pre-marketApprovalisNotRequired:TheFDAdoesnotrequiresubmissionofa510(k)orformalpre-marketapprovalfordevicesusedinveterinary
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Orthokinevetirap60® Orthokinevetirap10®
MedicalDeviceforPreparationofAutologousConditionedSerum
irapTherapy
• Whatisirap?– Interleukin-1ReceptorAntagonistProtein(irap)– iraptherapyutilizesthepatient’sownbloodandnaturalhealingmechanisms
– ThepatentedirapsyringeproducesAutologousConditionedSerum(ACS),whichcontainsbeneficialanti-inflammatoryandregenerativecytokines.
Cytokine=immunoregulatoryproteins(asinterleukinorinterferon)secretedbycellsespeciallyoftheimmunesystem
AutologousConditionedSerum
Autologous:100%body’sowncomponentsConditioned:AchievedthroughphysicalprocessesSerum:Cell-freesupernatantofcoagulatedwholeblood
• Howdoesirapwork?– theirapsyringecontainsmedicalgradeglassbeads
– stimulatethewhitebloodcellstoproducetheanti-inflammatoryandregenerativecytokines
– wholebloodisincubatedandthencentrifugedtocollectserum
– theserum,(ACS),isthenre-injectedintothepatient.
irapTherapy
GlassBeads
MedicalGrade“A”GlassBeadsTheyareNOTcoatedwithanything
• Theconditioningofthebloodisachievedthroughexposureofthewholebloodtoaspecificsurfaceofborosilicateglassspheres
• Certaincellsaccumulatearoundthesesurfacesandbegintoproducecytokinesthatarebeneficialforregenerativeprocess
AutologousProteinProductioninWholeBloodModel:MonocyteAdherencetosurfaces
activemolecules
binding
e.g.glassspheres
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Monocyte
Macrophage
Lymphocyte
Neutrophil granulocyte
Immunologic cell
IL-1Ra Leukocytes(WBCs)
?
Dr.med.vet.J.Troillet;St.Petersburg24.02.12
HowdoesACSwork?
BiomechanicalFactorsActivationofcytokinenetworkcascade
Imbalanceinmatrixmetabolism
ACScanhelptoinhbitthedestructionofcartilageandbreakthisviciouscycle!
Pro-inflammatoryanddestructivecytokines
Destructionofhyalinecartilageandit‘smatrix
InflammatoryMediatorsOneResponsibleCytokine
• Pro-Inflammatorycytokine• Stimulationofcollagenaseandmetallo-proteinase• Destructionofhyalinecartilageanditsmatrix
Interleukin-1Receptorantagonist
• Anti-inflammatoryandchondroprotective
Interleukin-1
• TypeIandIIIL-1receptors
IL- 1
• Pro-inflammatory • Syngergy with TNF-α • Promotes destruction of
chondrocytes abd cartilage matrix
• IL-RA • Competitive inhibition of
IL-1 receptors • Anti-inflammatory and
chondroprotective
ACSModeofAction
IL-1TNFα
IL-1RAIL-10
JointhomeostasisAnabolismandCatabolismareinbalance
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DiseaseModeofAction
IL-1TNFα
IL-1RAIL-10
JointDiseaseAnabolismandCatabolismareinoutofbalance
DZ
ChronicDegeneration
• Suppressionandsynthesis– Proteoglycans– TypeIICollagenFibers
• IncreasedDeNovo– MMP’sandAggrecanase– IL-1onchondrocytes
Dr.med.vet.J.Troillet;St.Petersburg24.02.12
InsideaPathologicJoint
• Irapisusedtotreatavarietyofinflammatoryconditionsinhorses:– Jointlameness– Tendonandligamentinjuries– Muscleinjuries
irapTherapy
irapTherapy• ControlledstudywithOAmodelinhorsesatCSU
showedpositiveeffects• IL-1Raisgood• IL-1isbad
• Results*:↑ IL-1Ra(140fold)↑ IL-4(2.1fold) Anti-inflammatory↑ IL-10(2.2fold)NochangeIL-1beta NochangeTNFalpha Pro-inflammatory*FrisbeeD,KawcakC,WerpyN,ParkR,McIlwraithW,Clinical,biochemicalandhistologicaleffectsof
intra-articularadministrationofautologousconditionedseruminhorseswithexperimentallyinducedosteoarthritis.AmerJourVetResearch,Vol.68,No.3,2007.
irapTherapy-Equine• Hasirapbeentested?
– Thepatentediraptherapyhasbeenusedinthousandsofhorsesaroundtheworld.*
– ControlledstudyatColoradoStateUniversity*
• resultsindicatedthattherewassignificantclinicalandhistologicalimprovementinosteoarthritis-affectedjointsofhorsesfollowingtreatmentwithACS,comparedwithplacebotreatment.
• HorsestreatedwithACShadsignificantclinicalimprovementinlameness,unliketheplacebogroup.
*PerOrthogenVeterinary,Duesseldorf,Germany**FrisbeeD,KawcakC,WerpyN,ParkR,McIlwraithW,Clinical,biochemicalandhistologicaleffectsofintra-articularadministrationofautologousconditionedseruminhorseswithexperimentallyinducedosteoarthritis.AmerJourVetResearch,Vol.68,No.3,2007.
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irapTherapy-Equine
• 16horseswithexperimentallyinducedOA– 8horsestreatedw/6mlof
ACSon14,21,28&35– 8horsestreatedw/saline
• Clinicalexaminations• Synovialfluidanalysis• Grossexamination• Histology• Biochemical&biomarker
Clinical,biochemical,andhistologiceffectsofintra-articularadministrationofAutologousConditionedSeruminhorseswithexperimentallyinducedosteoarthritis
(Frisbieetal.2007)
Clinical,biochemical,andhistologiceffectsofintra-articularadministrationofAutologousConditionedSeruminhorseswithexperimentallyinducedosteoarthritis
(Frisbieetal.2007)
irapTherapy-Equine
• Noadversereaction• Improvedlameness• Decreasedsynovialmembranehyperplasia• Lessgrosscartilagefibrillation• Lesssynovialmembranehemorrhage
PlateletRichPlasma
OsteokinePRPWhatisPRP?• PRPisapreparationofautologousconcentratedplatelets(thrombocytes)• PRPproducedbyOsteokinecontainstherapeuticallysignificantconcentrationsof
platelets
Howdoesitwork?• Wholebloodistakenfromthehorseandplacedintothered-cappedOsteokine
bag• AfterprocessinginaspecialcentrifugeapelletofPRPisgeneratedwhichcanbe
re-suspendedtotreatthehorsefromwhichitwastaken
WhatcanPRPbeusedfor?• PRPhasbeenshowntoacceleratehealingininjuriesinvolvingfibroustissues
(tendons,ligaments,skin)andhasalsobeenusedinbonehealing.
Thrombocytes/Platelets• Plateletsdomorethancoagulation• α-granulescontainGFindispensablefornormalwoundhealing
• 1stphase:uponinjurytissuethromboplastininducesplateletcoagulationandreducesbleeding
• 2ndphase:plasmafibrinogenbreakstofibrin,furthercoagulationstopsbleeding.AggregationofplateletsstartsoffGFsecretion.GFinducecellproliferation,stemcelldifferentiationandmigration.Macrophagesmigratetothesiteandtakeovertheplaceofplateletsassourceofcytokines.
Platelets
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Osteokine• Concentratedplateletsarearichsourceof(autologous)growth
factors.
• Ifplateletsarefreshandfullyfunctional->optimalsequentialgrowthfactoroutput
• TotaloperatingtimetoretrievePRPisabout40min
• Averageplateletconcentrationof1,300,000µL±250,000/µlfrom50mLinafinalvolumeof5-10mL
• Concentrationfactorofplateletsroutinely3-6fold
Average platelet count in native blood: 275,000 / µl
Average platelet count in 8 mL of PRP yielded by Osteokine:
1,300,000 / µl
Characteristics of the Osteokine® PRP system
FactorspresentinOsteokine®PRP• PDGF• IGF• FGF• TGF• HGF• VEGF
• EGF• Pluscytokines• suchasIL-1Ra,IL-6
BenefitsofOsteokine®PRP• Acceleratedwoundhealinginmultipletissuesincludingskin,tendon,ligament
• Highercollagencontentinhealinginjuriesresultinginlowerscarformation
• Reducedperioperativetraumaticpain• Lowerinfectionrates• Injectimmediatelyintocorelesions/superficiallesion
• Improvedtissueformation(e.g.morecollagen)• improvedbonematuration
TwooptionstoLamenessOsteokine(PRP)• Useinsofttissue(fibrous)
injuries– Tendons– Ligaments– Skin,muscle&bone
• Easilyproducedin-clinicwithin30-40minutes
• Treatinjuredhorse1time• HighPRPpotency
irap• Useintra-articular(IA)totreat
osteo-arthritis• Processedin-clinicforeaseofuse• Usedinthousandsofhorses
aroundtheworldsince2004• ProveneffectivebyCSU• 3initialtreatmentsthen
maintenance(horsedictatesneed)
• Highpotencyandyield• Proven&trusted!
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Conclusion
• irapIAworkswithgreatsuccessagainstlamenesscausedbymildtomoderateOA
• OsteokineisafantasticPRPsystemtotreatsofttissueinjuries
• Onlyiraphasbeentestedincontrolledstudiesandinrealworldsituations.
Discussion
