ProductDescription
AdvancedBiomatrixoffersPhotoHA®,amethacrylatedhyaluronicacid(HA)forphotocrosslinkablehydrogels.Thesehydrogelsprovidenative-like3DHAgelswiththeuniqueattributestobepreparedatvariousconcentrationsandphotocrosslinkedtoprovidevariousgelstiffness.Thekitcomeswith100mgoflyophilizedmethacrylatedhyaluronicacidand100mgofLAPphotoinitiator(405nmbluelightphotocrosslinking).
| Item | CatalogNumber | PackageSize | StorageTemperature |
| MethacrylatedHyaluronicAcid | #5212 | 100mg | -20°C |
| LAPPhotoinitiator | #5269 | 100mg | 2-10°C |
Hyaluronicacidisthemostabundantglycosaminoglycaninthebodybeinganimportantcomponentofseveraltissuesthroughoutthebody.Whileitisabundantinextracellularmatrices,hyaluronicacidalsocontributestotissuehydrodynamics,movementandproliferationofcells,andparticipatesinanumberofcellsurfacereceptorinteractions.
Forthemajorityofcelltypes,itisrecommendedtoaddadditionalECMproteinstothehyaluronicacidhydrogels.Theproteinsprovideimportantcellbindingsites.
Storage:
Theproductshipsonfrozengelpacks.Uponreceipt,storethePhotoHA®at-20°C.StoretheLAPat2to10°C.Theproductandcomponentsarestableforaminimumof1yearatreceiptinpowderform.
Oncesolubilized,thePhotoHA®canbestoredat2-10°Cfor1month.Thephotoinitiatorcanbestoredfornomorethan2weeksoncesolubilized.
| Parameter,Testing,andMethod | MethacrylatedHyaluronicAcid#5212 |
| SterilizationMethod | Filtration |
| Sterility-USPmodified | Nogrowth |
| Form | LyophilizedPowder |
| PackageSize | 100mg |
| StorageTemperature | -20°C |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| ShelfLifeAfterReconstitution | 1Month |
| DegreeofMethacrylation | 45-65% |
| MolecularWeight | 100-150kDa |
NMRAnalysis | Characteristic |
HydrogelYoung"sModulusE(Pa) | Characteristic |
SwellingCharacteristicsofPhotoHA®:
50μLhydrogelsfabricatedin4.7mmdiametermoldswereimagedbeforeandafterincubationinphosphatebufferedsalineat25°Cfor24hours.ThediameterofhydrogelswerequantifiedusingImageJsoftware.Statisticalcomparisonsbetweengroups(n=3)wereperformedviaone-wayANOVAwithposthoctestingandsignificancedeterminedatp<0.05.
CompressiveModulusofPhotoHA®:
Dynamicmechanicalanalysis(Q800,TAInstruments)wasperformedon50μLhydrogelsfabricatedin4.7mmdiametermolds.Hydrogelsweresecuredwithinafluidcupviaa0.01Npre-loadandcompressedto30%strainatarateof0.5Nmin-1.TheYoung’smodulusofeachhydrogelwascalculatedastheslopeofgeneratedstress-straincurvesbetween10%and20%strain.StatisticalcomparisonbetweenMeHAconcentrations(n=3)wasperformedviaStudentst-testwithtwo-tailedcriteriaandsignificancedeterminedatp<0.05.
ReactionBehaviorofPhotoHA®:
Rheologicaltimesweeps(AR2000stresscontrolledrheometer,TAInstruments;0.5%strain,1Hz,25°C)ofMeHAcrosslinkingwithexposuretoUVlight(=320-390nm)andinthepresenceof0.05wt%Irgacure2959(I2959).After1minute,themacromersolution(i.e.MeHAandI2959)wasexposedtoUVlight,resultingintheplateauofmodulibefore5minutes.
DirectionsforUse
DownloadthefullDirectionsforUsePDForcontinuereADIngbelow:
3DHyrdogelPreparation:
Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.
Recommendedconcentrationsare5-30mg/ml(0.5-3.0%).
Note:Thefollowingrecommendedinstructionsarefora1%hyaluronicacid(HA)methacrylatesolution.Adjustmentstothisprotocolmayberequiredforvariousconcentrations.
- Add10mlof1Xphosphatebuffersaline(PBS),waterorcellculturemediatothe100mgoflyophilizedmethacrylatedHApowder.
- Mixonashakertableorrotatorplateuntilfullysolubilized(~30to60minutes)at2-10°C.Note:SolubilizationtimesmayvarydependingonthedesiredconcentrationandvolumeofPBS,waterormediumadded,
- Calculatethevolumeofphotoinitiatortoaddbymultiplyingthevolumeofsolubilizedhyaluronicacidby0.02.Iftheresultingnumberis200ul,forexample,youwilladd200ulofLAP.
- SolubilizetherequiredamountofLAP(perstep3)ataconcentrationof17mg/mlin1XPBSorcellculturemedia.
- AddthecalculatedvolumeofphotoinitiatortotherequiredvolumeofHAmethacrylatesolutionandmixthoroughly.
- AddyourcellstotheHAmethacrylate/photoinitiatorsolution.
- DispenseyourHAmethacrylate/photoinitiator/cellsolutionintothedesiredcultureware(i.e.6-wellplate,48-wellplate).
- Forphotocrosslinking,placethehydrogelsolutiondirectlyundera405nmlightcrosslinkingsource.
ProductQ&A
Yes.TherearequiteafewpublicationscitingvarioushyaluronidaseprotocolsfordigestingPhotoHAhydrogels.Hereareafew:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460858/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4840832/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4986518/
TheHAweusecomesfrombiotechnologicalproduction,wherethehyaluronicacidisextractedfromthecellwallsofthebacteriaStreptococcuszooepidemicus.
ProductCellAssay
Humanmesenchymalstemcells(20x106/mL)wereencapsulatedin50μLhydrogels(~4.7mmx2mm).Hydrogels(1wt%PhotoHA®)werefabricatedwith0.05wt%Irgacure2959andexposureto2mW/cm2light(320-390nm)for10minutes.After24hours,encapsulatedcellswerestainedwithcalceinAMandethidiumhomodimerandsubsequentlyimagedonaLeicaSP5confocalmicroscope(usingFITC/TRITCsequentialscans).
ProductApplications
PhotoHA®MethacrylatedHyaluronicacidcanbeusedtoformhydrogelsforex-vivoengineeringofautologouscartilagetissue[1]orasamesenchymalstemcellcarrierincartilagerepair[2].
BecausethestiffnesscanbewidelyadjustedbyalteringconcentrationorUV-lightexposure,methacrylatedHAhasbeenusedtomeasuretheeffectsofmatrixstiffnessoncellphenotypeandfunction[3][4].
MethacrylatedHAcanbeusedfor3Dbioprinting(extrusion[5],inkjet[5]andphotolithographic[6])tocreatestructuresthatpromoteosteogenicdifferentiationofMSC’s[7].
ThehightunABIlityofhyaluronicacidmethacrylateallowsittobemixedwith,andreinforceothertypesofhydrogels(suchascollagen,orgelatinmethacrylate)[8].
References:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5717235/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627486/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5541838/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5447944/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615317/
https://www.ncbi.nlm.nih.gov/pubmed/21773726/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460858/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5748291/
ProductReferences
ReferencesforPhotoHA®:
Khetan,Sudhir,andOwenCorey."Maintenanceofstemcellviabilityanddifferentiationpotentialfollowingcryopreservationwithin3-dimensionalhyaluronicacidhydrogels."CryoBIOLOGy(2019).
Poldervaart,M.T.etal.3Dbioprintingofmethacrylatedhyaluronicacid(MeHA)hydrogelwithintrinsicosteogenicity.PlosOne12,(2017).
ProductCertificateofAnalysis
SafetyandDocumentation
SafetyDataSheet
CertificateofOrigin
ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
