Проект РНФ 25-19-00872 (2025-2027) «Разработка и исследование микрофлюидного устройства («Легкое на чипе») с использованием микрофлюидных, акусто- и радиоэлектронных технологий для моделирования на клеточном уровне физиологической микросреды дыхательных путей и тестирования лекарственных препаратов in vitro.» Руководитель д.ф.-м.н. Кузнецова И.Е.

2025

1. Duringtheworkusingadditivetechnologies, samples of microfluidiccellsweremadefrombiocompatiblepolymerspolydimethylsiloxane(PDMS),polymethylmethacrylate(PMMA)andpolycarbonate.Duringthetests of the cells, it wasrevealedthat the mostpromisingforfurtheruseis a cellmadebyphotopolymerprintingfromPMMA.This is due to the minimalsize of the channels, the simplicity of itsdisassemblyandtightness,comparableto a samplecellfromPDMS.
2. Duringtheplannedwork, porousmembraneswereselectedandcharacterizedforfurtheruseas a basisforgrowingepithelialcelllayers. As aresult, porousflexiblemembranes made ofpolycaprolactone,polyvinylidene fluoride andcelluloseacetatewereselectedandcharacterized. The morphology of theirsurfacewasanalyzed by electronmicroscopy. The mostpromisingmaterialfor the manufacture of membranesisPCLmodified with spidroin,which is associatedwithitshighbiocompatibilityandelasticity of the membrane. The studiedmaterialswill be usedas a basisforgrowinglayers of epithelialcellsplacedin the createdmicrofluidiccell.
3. Toanalyze the effectiveness of Caco-2epithelialcellimmobilizationonvariousporousmembranes, membranes made ofrecombinantspidroinwith the addition of polycaprolactane(PCL), polyethylene tetraphthalate(PET),acetylcelluloseandPVDFwerestudied. As aresult, it wasfoundthatpolyurethaneandPCLmembraneswith the additionofspidroin are suitableforobtaining a homogeneouscelllayer.
4. Duringtheplannedwork, the growthprocess of CaCo-2,A549, and NCI-H441 epithelialcelllineswasstudiedunderconditions of normoxiaandhypoxiaonporouspolycaprolactone-basedmembraneswith the addition of spidroin. As aresult, the growthprocess of Caco-2,A549, and NCI-H441epithelialcelllinesunderconditions of normoxiaandhypoxiawasstudiedusingconfocallasermicroscopy,atomicforcemicroscopy,scanningelectronmicroscopy, andRAMANspectroscopy.Confocallasermicroscopyis the mostpromisingmethod for monitoring the growth of celllayers,asitprovides non-invasiveobservation of cellstructureinrealtime.CombiningthismethodwithRAMANspectroscopyallowsnotonly to confirm the presence of the cellularlayer,butalso to assessitsmetabolicactivity. The methods of scanningelectronandatomicforcemicroscopyprovidethe most completedescription of the morphologyandsurfaceproperties of cellularlayers.However, theirsignificantlimitationis the need for pre-fixationanddrying of samples,whichleadstoloss of cellularactivityandexcludes the possibility of further in vitroobservations.
5. Duringtheplannedwork, the mechanicalpropertiesofpolycaprolactonemembraneswithmolecularweights of 35kDa,65kDaand100kDawith the addition of spidroinwerestudied.Astudy of the sizesand Young’s modulus of individualmembranefibers was conducted. The averagefibersizeswere70nm,77nmand110nm,and the average Young’s modulus of the fiberwas1.25MPa,1.5MPaand3.3MPaforPCLmembraneswith the abovemolecularweight,respectively. As aresultof the plannedwork, the mechanicalproperties of the membranesusedwereinvestigated in order to simulate the “breathing”effectin a two-chambermicrofluidiccell. It has been establishedthat a membranebased on polycaprolactonewith a molecularweight of 100kDapossessesoptimalelasticproperties. The parameters have been foundatwhich it is possible to achieve a periodicchange in the membranesurfaceareaby15%,i.e. to simulate the “breathing” of the membrane.
6. Duringthework, ameasuringset up was developed toprovidemeasurementsforvariousflows of liquidsin a microfluidicdevice. As aresult, a measuringset upwascreatedtoprovideandmeasurevariousmodes of fluidflowinmicrofluidicdevices.Thisstandallows you to simulate the respiration of a cellularlayerlocatedon a semi-permeableflexiblemembranemountedin a microfluidicdevice. The flowrate of liquidinto the compartments of the microfluidicdevicecan be adjustedin a widerange of values.Thecreatedstand also makes it possibletomeasure the electrophysicalcharacteristics of the celllayerusing a connectedexternalpotentiostat, agalvanostat.
7. The mostcommonmethods of sterilization of reusabledevicesinbiotechnologyareautoclaving,dryheatsterilization,ultraviolet(UV)irradiation, and chemicalsterilization.Inthissection, the methods of sterilization of the createdmicrofluidiccellwereoptimized, taking into account the need to comply with the temperatureregimedue to theuse of biocompatiblepolymers. As aresultof the work, it wasfoundthat the optimalway to sterilize a microfluidiccellis two-stagesterilization of the cellcomponentsin95%ethylalcoholsolutionandsubsequenttreatment of the assembledcellin a Sterradplasmasterilizerat120°Cfor2hours.