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Nekugadzirwa kwezvinhu zvitsva zvakapfava zvekushandisa pakurapa uye pakushandisa biomedical, kutsanangurwa kwakazara kwehunhu hwazvo hwepanyama uye hwemakanika kwakakosha uye kwakaoma. Nzira yekushandura atomic force microscopy (AFM) nanoindentation yakashandiswa kuratidza modulus yepasi zvakanyanya yelehfilcon itsva. Biomimetic silicone hydrogel contact lens yakafukidzwa nelayer yemabrashi epolymer ane brashi. Nzira iyi inobvumira kusarudzwa kwakarurama kwenzvimbo dzekubatana pasina mhedzisiro ye viscous extrusion pakuswedera kuma polymer ane brashi. Pamusoro pezvo, zvinoita kuti zvikwanisike kuona hunhu hwemakanika hwezvinhu zvebhurashi rega rega pasina mhedzisiro ye poroelasticity. Izvi zvinoitwa nekusarudza AFM probe ine dhizaini (tip size, geometry uye spring rate) yakakodzera kuyera hunhu hwezvinhu zvakapfava uye samples dzebiological. Nzira iyi inovandudza kunzwisisika uye kururama kwekuyera kwakarurama kwe lehfilcon A yezvinhu zvakapfava, ine modulus yakaderera kwazvo ye elasticity panzvimbo yepamusoro (kusvika ku 2 kPa) uye elasticity yakanyanya kwazvo munzvimbo yemukati (inenge 100%) yemvura. Zvakabuda muongororo yepamusoro hazvina kungoratidza chete hunhu hwepamusoro hwakapfava hwelens yelehfilcon A, asiwo zvakaratidza kuti modulus yemabhurashi epolymer ane branched yakafanana neye silicon-hydrogen substrate. Iyi nzira yekutarisa pamusoro inogona kushandiswa kune zvimwe zvinhu zvakapfava zvakanyanya uye zvishandiso zvekurapa.
Hunhu hwezvinhu zvakagadzirwa kuti zvibatane zvakananga nenyama mhenyu hunowanzo tsanangurwa nenzvimbo yehupenyu. Kuenderana kwakakwana kwehunhu hwezvinhu izvi kunobatsira kuwana hunhu hwekiriniki hunodiwa hwezvinhu pasina kukonzera mhinduro dzakaipa dzemasero1,2,3. Kune zvinhu zvakafanana zvakawanda, kutsanangurwa kwehunhu hwezvinhu zvemitambo kuri nyore nekuda kwekuwanikwa kwemaitiro akajairwa uye nzira dzekuyedza (semuenzaniso, microindentation4,5,6). Zvisinei, kune zvinhu zvakapfava zvakanyanya senge magel, mahydrogel, biopolymers, masero mapenyu, nezvimwewo, nzira idzi dzekuyedza hadziwanzo shandiswa nekuda kwekuganhurirwa kwekugadzirisa kuyerwa uye kusawirirana kwezvimwe zvinhu7. Mumakore apfuura, nzira dzechinyakare dzekuindentation dzakagadziriswa uye dzakagadziriswa kuti dziratidze mhando dzakasiyana dzezvinhu zvakapfava, asi nzira dzakawanda dzichiri kutambura nekusakwana kwakakomba kunoganhurira kushandiswa kwadzo8,9,10,11,12,13. Kushaikwa kwenzira dzekuyedza dzakanyanya dzinogona kuratidza hunhu hwezvinhu zve supersoft uye surface layers nenzira yakarurama uye yakavimbika inoderedza kushandiswa kwadzo mumashandisirwo akasiyana-siyana.
Mubasa redu rekare, takaunza lenzi yekubata yelehfilcon A (CL), chinhu chakapfava chakasiyana chine hunhu hwepamusoro hwakapfava hwakatorwa kubva mumagadzirirwo anogona kunge ari ebiomimetic akafuridzirwa nepamusoro pe cornea yeziso. Ichi chinhu chehupenyu chakagadzirwa nekubatanidza polymer layer ine cross-linked, cross-linked ye poly(2-methacryloyloxyethylphosphorylcholine (MPC)) (PMPC) pa silicone hydrogel (SiHy) 15 yakagadzirirwa zvishandiso zvekurapa zvichibva pa. Maitiro aya ekubatanidza anogadzira denderedzwa pamusoro rine chimiro che polymeric brash chakapfava uye chinopfava zvikuru. Basa redu rekare rakasimbisa kuti chimiro che biomimetic che lehfilcon A CL chinopa hunhu hwepamusoro hwepamusoro hwakadai sekudzivirira kunaya uye kusvibisa, kuwedzera kwemafuta, uye kuderedza kunamira kwemasero nemabhakitiriya15,16. Pamusoro pezvo, kushandiswa nekugadzirwa kwechinhu ichi che biomimetic kunoratidzawo kuwedzera kune zvimwe zvishandiso zve biomedical. Saka, zvakakosha kutsanangura hunhu hwepamusoro hwechinhu ichi chakapfava zvakanyanya uye kunzwisisa kudyidzana kwacho nemaziso kuitira kugadzira hwaro hweruzivo rwakakwana rwekutsigira budiriro nekushandiswa mune ramangwana. Magirazi ekuonana eSiHy akawanda anowanikwa mumusika anoumbwa nemusanganiswa wepolymers dzinofema uye dzinofema dzinoumba chimiro chechinhu chakafanana17. Zvidzidzo zvakati wandei zvakaitwa kuti zviongorore hunhu hwadzo hwemuchina uchishandisa nzira dzechinyakare dzekudzvanya, kudhonza uye microindentation test methods18,19,20,21. Zvisinei, dhizaini itsva yebiomimetic yelehfilcon A CL inoita kuti ive chinhu chakasiyana umo hunhu hwemuchina hwezvimiro zvemabrashi epolymer ane branched hwakasiyana zvakanyanya nehweSiHy base substrate. Nokudaro, zvakaoma kwazvo kuyera hunhu uhwu nemazvo uchishandisa nzira dzechinyakare uye dzeindentation. Nzira inovimbisa inoshandisa nzira yekuongorora nanoindentation inoshandiswa muatomic force microscopy (AFM), nzira yakashandiswa kuona hunhu hwemuchina hwezvinhu zvakapfava zveviscoelastic zvakaita semasero ebhayoloji nematishu, pamwe nemapolymers akapfava22,23,24,25. ,26,27,28,29,30. MuAFM nanoindentation, nheyo dzekuongorora nanoindentation dzinosanganiswa nekufambira mberi kwazvino mutekinoroji yeAFM kuti ipe kunzwisiswa kwakanyanya kwekuyera uye kuyedzwa kwezvinhu zvakasiyana-siyana zve supersoft31,32,33,34,35,36. Pamusoro pezvo, tekinoroji iyi inopa mamwe mabhenefiti akakosha kuburikidza nekushandisa majometri akasiyana, indenter neprobe uye mukana wekuyedza mune akasiyana liquid media.
Kuongororwa kweAFM kunogona kukamurwa kuita zvikamu zvitatu zvikuru: (1) michina (masensa, ma detector, ma probe, nezvimwewo); (2) ma paramita ekuyera (akadai sesimba, kutama, kumhanya, saizi ye ramp, nezvimwewo); (3) Kugadziriswa kwedata (kugadzirisa kwekutanga, kufungidzira kwenzvimbo yekubata, kugadzirisa data, modeling, nezvimwewo). Dambudziko guru nenzira iyi nderekuti zvidzidzo zvakati wandei mumabhuku anoshandisa AFM nanoindentation zvinoshuma mhedzisiro dzakasiyana kwazvo dzehuwandu hwemhando imwechete yesample/cell/material37,38,39,40,41. Semuenzaniso, Lekka et al. Simba reAFM probe geometry pa modulus yaYoung yakayerwa yemasampuli ehydrogel ine homogeneous mechanically uye heterogeneous cells rakadzidzwa uye rakaenzaniswa. Vanoshuma kuti modulus values ​​​​inonyanya kutsamira pakusarudzwa kwe cantilever uye chimiro chetip, ine kukosha kwakanyanya kweprobe yakaita sepiramidhi uye kukosha kwakaderera kwe42 kweprobe yakatenderera. Saizvozvowo, Selhuber-Unkel et al. Zvakaratidzwa kuti kumhanya kwe indenter, saizi ye indenter uye ukobvu hwe polyacrylamide (PAAM) samples zvinokanganisa sei Young's modulus inoyerwa ne ACM43 nanoindentation. Chimwe chinhu chinoomesa ndiko kushaikwa kwezvinhu zvekuedza zve modulus zvakajairwa zvishoma uye maitiro ekuedza emahara. Izvi zvinoita kuti zviome kuwana mhinduro dzakarurama nechivimbo. Zvisinei, nzira iyi inobatsira zvikuru pakuyera uye kuenzanisa pakati pemhando dzesamples dzakafanana, semuenzaniso kushandisa AFM nanoindentation kusiyanisa masero akajairwa kubva kumasero ekenza 44, 45.
Pakuyedza zvinhu zvakapfava neAFM nanoindentation, mutemo mukuru ndewekushandisa probe ine low spring constant (k) inoenderana ne sample modulus uye hemispherical/round tip kuitira kuti probe yekutanga isaboore pamusoro pe sample pakubata kwekutanga nezvinhu zvakapfava. Zvakakoshawo kuti chiratidzo chekutsauka chinogadzirwa ne probe chive chakasimba zvakakwana kuti chionekwe ne laser detector system24,34,46,47. Panyaya yemasero, matishu nemajeli akapfava zvakanyanya, rimwe dambudziko nderekukunda simba rekunamatira pakati pe probe ne sample surface kuti uve nechokwadi chekuyera kunodzokororwa uye kwakavimbika48,49,50. Kusvika nguva pfupi yadarika, basa rakawanda paAFM nanoindentation rakatarisana nekudzidza maitiro emakanika emasero e biological, matishu, majeli, mahydrogel, uye ma biomolecules uchishandisa ma spherical probes akakura, anowanzozivikanwa se colloidal probes (CPs). , 47, 51, 52, 53, 54, 55. Aya matips ane radius ye1 kusvika 50 µm uye anowanzogadzirwa negirazi reborosilicate, polymethyl methacrylate (PMMA), polystyrene (PS), silicon dioxide (SiO2) uye diamond-like carbon (DLC). Kunyangwe CP-AFM nanoindentation inowanzo kuve sarudzo yekutanga yekutarisa soft sample, ine matambudziko ayo uye miganhu yayo. Kushandiswa kwetips dzakakura, dzakakura micron-spherical kunowedzera nzvimbo yese yekubata muromo nemuenzaniso uye zvinoguma nekurasikirwa kukuru kwekugadzirisa kwenzvimbo. Kune sampuli dzakapfava, dzisina kufanana, uko hunhu hwemuchina hwezvinhu zvemuno hunogona kusiyana zvakanyanya neavhareji munzvimbo yakakura, CP indentation inogona kuvanza chero kusawirirana muhunhu pachiyero chemuno52. Colloidal probes dzinowanzo kugadzirwa nekubatanidza ma colloidal spheres ane saizi micron kuma cantilevers asina mativi uchishandisa epoxy adhesives. Maitiro ekugadzira pachawo ane matambudziko akawanda uye anogona kukonzera kusawirirana mumaitiro ekugadzirisa probe. Pamusoro pezvo, saizi nehuremu hwezvikamu zvecolloidal zvinokanganisa zvakananga main calibration parameters e cantilever, akadai se resonant frequency, spring stiffness, uye deflection sensitivity56,57,58. Saka, nzira dzinowanzo shandiswa dze AFM probes, dzakadai se calibration yekupisa, dzingasapa calibration chaiyo ye CP, uye dzimwe nzira dzingadiwa kuti dziite izvi kugadzirisa57, 59, 60, 61. Kuedza kwakajairika kwe CP indentation kunoshandisa large deviations cantilever kudzidza hunhu hwe soft samples, izvo zvinogadzira rimwe dambudziko pakuenzanisa maitiro asiri e linear e cantilever pakutsauka kukuru62,63,64. Nzira dzemazuva ano dze colloidal probe indentation dzinowanzo funga nezve geometry ye cantilever inoshandiswa kugadzirisa probe, asi dzinofuratira pesvedzero yezvikamu zve colloidal, izvo zvinogadzira kusava nechokwadi kwakawedzerwa mukururama kwenzira38,61. Saizvozvowo, elastic moduli inoverengwa ne contact model fitting inoenderana zvakananga ne geometry ye indentation probe, uye kusawirirana pakati petip uye sample surface characteristics kunogona kukonzera kusarongeka27, 65, 66, 67, 68. Mamwe mabasa achangobva kuitwa naSpencer et al. Zvinhu zvinofanirwa kufungwa nezvazvo pakusarudza soft polymer brushes uchishandisa CP-AFM nanoindentation method zvinoratidzwa. Vakashuma kuti kuchengetwa kwe viscous fluid muma polymer brushes sebasa rekumhanya kunoguma nekuwedzera kwemusoro wekutakura uye nokudaro kuyerwa kwakasiyana kwe speed dependent properties30,69,70,71.
Muchidzidzo chino, takatsanangura modulus yepamusoro yezvinhu zvakapfava zvakanyanya uye zvinochinjika zvakanyanya zve lehfilcon A CL tichishandisa nzira yakagadziriswa yeAFM nanoindentation. Tichifunga nezvehunhu uye chimiro chitsva chezvinhu izvi, huwandu hwekunzwa hwenzira yechinyakare yekudzika midzi hauna kukwana kuratidza modulus yezvinhu izvi zvakapfava zvakanyanya, saka zvakakosha kushandisa nzira yeAFM nanoindentation ine kunzwisisika kwakanyanya uye kunzwisisika kwakaderera. Mushure mekuongorora zvikanganiso nematambudziko emaitiro aripo ecolloidal AFM probe nanoindentation, tinoratidza kuti nei takasarudza probe diki, yakagadzirwa nemaitiro eAFM kubvisa kunzwisisika, ruzha rwekumashure, nzvimbo yekuona yekubatana, kuyera velocity modulus yezvinhu zvakapfava zvakasiyana-siyana zvakaita sekuchengetedza fluid. uye quantification chaiyo. Pamusoro pezvo, takakwanisa kuyera nemazvo chimiro nehukuru hwetip yekudzika midzi, zvichitibvumira kushandisa cone-sphere fit model kuti tizive modulus yekusimba pasina kuongorora nzvimbo yetip nezvinhu. Zvifungidziro zviviri zvisina kujeka zvinoyerwa mubasa iri ndezvekuti zvinhu zvinonyatsoshanda uye kuti modulus haina kudzika zvakanyanya sei. Tichishandisa nzira iyi, takatanga taedza zviyero zvakapfava zvakanyanya ne modulus inozivikanwa kuti tiverenge nzira iyi, uye takashandisa nzira iyi kuti titsanangure pamusoro pezvinhu zviviri zvakasiyana zvema lenzi ekuonana. Nzira iyi yekuratidza nzvimbo dzeAFM nanoindentation dzine kunzwa kwakanyanya inotarisirwa kuti ishande kune zvakasiyana-siyana zvezvinhu zve biomimetic zvakasiyana-siyana zvinogona kushandiswa mumidziyo yekurapa uye mashandisirwo emishonga.
Lehfilcon Magirazi ekuonana (Alcon, Fort Worth, Texas, USA) pamwe chete nemagirazi avo esilicone hydrogel akasarudzwa kuti ashandiswe mukuyedza kwe nanoindentation. Chigadziko che lenzi chakagadzirwa nehunyanzvi chakashandiswa mukuyedza uku. Kuti uise lenzi iyi kuti iongororwe, yakaiswa nokungwarira panzvimbo yakaita sedome, ichiita shuwa kuti hapana mabubble emhepo anopinda mukati, uye yobva yagadziriswa nemipendero. Gomba riri pamusoro pechibatiso che lenzi rinopa mukana wekupinda pakati pe lenzi kuti iongororwe ne nanoindentation uku uchibata mvura yacho. Izvi zvinochengetedza magirazi aya aine mvura yakakwana. 500 μl ye contact lens packaging solution yakashandiswa semhinduro yekuyedza. Kuti zvionekwe mhedzisiro yehuwandu, ma non-activated polyacrylamide (PAAM) hydrogels anowanikwa mumusika akagadzirwa kubva mu polyacrylamide-co-methylene-bisacrylamide composition (100 mm Petrisoft Petri dishes, Matrigen, Irvine, CA, USA), modulus inozivikanwa ye elastic ye 1 kPa. Shandisa madonhwe 4-5 (anenge 125 µl) ephosphate buffered saline (PBS kubva kuCorning Life Sciences, Tewkesbury, MA, USA) uye donhwe rimwe chete reOPTI-FREE Puremoist contact lens solution (Alcon, Vaud, TX, USA). ) paAFM hydrogel-probe interface.
Mienzaniso yeLehfilcon A CL neSiHy substrates yakatariswa uchishandisa FEI Quanta 250 Field Emission Scanning Electron Microscope (FEG SEM) system yakashongedzerwa neScanning Transmission Electron Microscope (STEM) detector. Kugadzirira mienzaniso, malenzi akatanga agezwa nemvura ochekwa kuita wedges dzakaita sepie. Kuti pave nekusiyana pakati pezvinhu zvinopisa mvura uye zvinopisa mvura zvesampuli, mhinduro yakagadzika ye0.10% yeRuO4 yakashandiswa sedhayi, umo mienzaniso yakanyudzwa kwemaminitsi makumi matatu. Kuisa lehfilcon A CL RuO4 kwakakosha kwete chete kuti pave nekusiyana kwakasiyana, asiwo kunobatsira kuchengetedza chimiro chemabhurashi epolymer ane branched muchimiro chawo chepakutanga, ayo anozoonekwa pamifananidzo yeSTEM. Akabva agezwa uye akabviswa mvura mumisanganiswa yeethanol/mvura nekuwedzera kwehuwandu hweethanol. Mienzaniso yacho yakazokandwa neEMBEd 812/Araldite epoxy, iyo yakaporeswa husiku hwese pa70°C. Mabhuroko emuenzaniso akawanikwa ne resin polymerization akachekwa ne ultramicrotome, uye zvikamu zvakatetepa zvakabuda zvakaonekwa ne STEM detector mu low vacuum mode pa acceleration voltage ye 30 kV. Iyo SEM system imwechete yakashandiswa pakuongorora zvakadzama probe ye PFQNM-LC-A-CAL AFM (Bruker Nano, Santa Barbara, CA, USA). Mifananidzo yeSEM ​​ye probe ye AFM yakawanikwa mu typical high vacuum mode ine acceleration voltage ye 30 kV. Wana mifananidzo pamakona akasiyana uye magnifications kuti unyore zvese zvechimiro nehukuru hwe AFM probe tip. Zvese zvidimbu zve tip zvinofarirwa mumifananidzo zvakayerwa ne digitally.
Maikorosikopu yesimba reatomu yeDimension FastScan Bio (Bruker Nano, Santa Barbara, CA, USA) ine "PeakForce QNM in Fluid" yakashandiswa kuona nekutarisa lehfilcon A CL, SiHy substrate, uye PAAm hydrogel samples. Pakuyedza kwemifananidzo, probe yePEAKFORCE-HIRS-FA (Bruker) ine nominal tip radius ye1 nm yakashandiswa kutora mifananidzo yepamusoro yesample pachiyero che0.50 Hz. Mifananidzo yese yakatorwa mumvura.
Kuedza kweAFM nanoindentation kwakaitwa uchishandisa probe yePFQNM-LC-A-CAL (Bruker). Probe yeAFM ine silicon tip pa nitride cantilever ine ukobvu hwe345 nm, 54 µm kureba uye 4.5 µm yakafara ine resonant frequency ye45 kHz. Yakagadzirirwa zvakananga kuratidza nekuita zviyero zve quantitative nanomechanical pa soft biological samples. Masensa anogadziriswa ega ega pafekitori nemagadzirirwo e spring akagadziriswa kare. Zvimisikidzo zve spring zve probes zvakashandiswa muchidzidzo ichi zvaive pakati pe 0.05–0.1 N/m. Kuti zvionekwe nemazvo chimiro nehukuru hwe tip, probe yakatsanangurwa zvakadzama uchishandisa SEM. Pamufananidzo. Mufananidzo 1a unoratidza high resolution, low magnification scanning electron micrograph ye PFQNM-LC-A-CAL probe, ichipa maonero akazara ekugadzira probe. Pamufananidzo 1b unoratidza maonero akakura epamusoro pe probe tip, ichipa ruzivo nezve chimiro nehukuru hwe tip. Kumucheto kwakanyanya, tsono iyi ine dhayamita inosvika 140 nm (Mufananidzo 1c). Pasi apa, muromo wacho unodzika kuita chimiro chekona, uchisvika pakureba kwakayerwa kweinenge 500 nm. Kunze kwenzvimbo inodzika, muromo wacho wakaita sedenderedzwa uye unogumira pakureba kwemuromo we1.18 µm. Ichi ndicho chikamu chikuru chinoshanda chemuromo weprobe. Pamusoro pezvo, probe hombe yepolystyrene (PS) ine dhayamita ye45 µm uye spring constant ye2 N/m yakashandiswawo kuyedza seprobe yecolloidal. neprobe yePFQNM-LC-A-CAL 140 nm yekuenzanisa.
Zvakataurwa kuti mvura inogona kuvharirwa pakati peAFM probe nepolymer brush structure panguva ye nanoindentation, iyo ichashandisa simba rekumusoro paAFM probe isati yanyatsobata pamusoro69. Iyi viscous extrusion effect nekuda kwe fluid retention inogona kuchinja nzvimbo inoonekwa yekubata, nokudaro ichikanganisa kuyerwa kwe surface modulus. Kuti tidzidze mhedzisiro ye probe geometry uye indentation speed pakuchengetwa kwemvura, indentation force curves dzakarongwa dze lehfilcon A CL samples dzichishandisa 140 nm diameter probe pa constant displacement rates ye 1 µm/s uye 2 µm/s. probe dhayamita 45 µm, fixed force setting 6 nN yakawanikwa pa 1 µm/s. Kuedza ne probe 140 nm mudhayamita kwakaitwa pa indentation speed ye 1 µm/s uye set force ye 300 pN, yakasarudzwa kugadzira contact pressure mukati me physiological range (1–8 kPa) ye upper eyelid. kudzvanywa 72. Mienzaniso yakapfava yakagadzirwa yePAA hydrogel ine kudzvanywa kwe1 kPa yakaedzwa simba rekudzika kwe50 pN nekumhanya kwe1 μm/s uchishandisa probe ine dhayamita ye140 nm.
Sezvo kureba kwechikamu chekona chemucheto wePFQNM-LC-A-CAL kuri 500 nm, kune chero kudzika kwekupinda < 500 nm zvinogona kufungidzirwa zvakachengeteka kuti geometry yeprobe panguva yekupinda icharamba iri yechokwadi kune chimiro chayo chekoni. Pamusoro pezvo, zvinofungidzirwa kuti pamusoro pechinhu chiri kuyedzwa chicharatidza mhinduro inodzoreka, iyo ichasimbiswawo muzvikamu zvinotevera. Nokudaro, zvichienderana nechimiro nehukuru hwemucheto, takasarudza modhi yekubatanidza cone-sphere yakagadzirwa naBriscoe, Sebastian naAdams, inowanikwa musoftware yemutengesi, kuti tigadzirise kuedza kwedu kweAFM nanoindentation (NanoScope). Separation data analysis software, Bruker) 73. Modhi inotsanangura hukama hweforce-displacement F(δ) yekoni ine defect yesphere. Pamufananidzo. Mufananidzo 2 unoratidza mashandisirwo ekubatana panguva yekubatana kwekoni yakasimba nemuromo wakatenderera, uko R iri radius yemuromo wakatenderera, a iri radius yekubatana, b iri radius yekubatana pamagumo emuromo wakatenderera, δ iri radius yekubatana. Kudzika kwemukati, θ ndiyo hafu yekona yekoni. Mufananidzo weSEM weprobe iyi unoratidza zvakajeka kuti muromo wakatenderera we140 nm dhayamita unosangana netambo kuita koni, saka pano b inotsanangurwa chete kuburikidza neR, kureva b = R cos θ. Software inopihwa nemutengesi inopa hukama hwekoni-sphere hwekuverenga mavalues ​​eYoung's modulus (E) kubva kudata rekuparadzanisa simba tichifunga kuti a > b. Hukama:
apo F iri simba rekupinda mukati, E iri modulus yaYoung, ν iri chiyero chaPoisson. Radius yekubatana a inogona kufungidzirwa uchishandisa:
Chimiro che geometry yekubatana kwekoni yakasimba ine muromo wakatenderera wakadzvanywa muzvinhu zveLefilcon contact lens ine pamusoro pemabhurashi epolymer ane matavi.
Kana a ≤ b, hukama hunoderera kusvika pa equation ye conventional spherical indenter;
Tinotenda kuti kudyidzana kweprobe inopinza nechimiro chebrashi rePMPC polymer kuchaita kuti radius yekubatana a ive yakakura kupfuura radius yekubatana b. Saka, pakuyera kwese kwehuwandu hwe elastic modulus kwakaitwa muchidzidzo chino, takashandisa kutsamira kwakawanikwa kwenyaya a > b.
Zvinhu zve ultrasoft biomimetic zvakadzidzwa muchidzidzo ichi zvakatorwa mifananidzo yakazara uchishandisa scanning transmission electron microscopy (STEM) ye sample cross section uye atomic force microscopy (AFM) yenzvimbo. Kutsanangurwa uku kwakadzama kwenzvimbo kwakaitwa sekuwedzera kwebasa redu rakaburitswa kare, umo takaona kuti chimiro che polymeric brush chakachinja-chinja che PMPC-modified lehfilcon A CL surface chakaratidza hunhu hwakafanana nehwe native corneal tissue 14. Nekuda kweizvi, tinotaura nezve contact lens surfaces se biomimetic materials14. Pamufananidzo 3a,b tinoratidza zvikamu zve branch PMPC polymer brush structures pamusoro pe lehfilcon A CL substrate uye SiHy substrate isina kurapwa, zvichiteerana. Nzvimbo dzemasample ese ari maviri dzakaongororwazve uchishandisa high-resolution AFM images, izvo zvakasimbisa mhedzisiro ye STEM analysis (Mufananidzo 3c, d). Kana zvabatanidzwa, mifananidzo iyi inopa hurefu hwe PMPC branch polymer brush structure pa 300–400 nm, izvo zvakakosha pakududzira zviyero zve AFM nanoindentation. Chimwe chinhu chakakosha chakawanikwa kubva mumifananidzo ndechekuti chimiro chepamusoro pezvinhu zveCL biomimetic chakasiyana pachimiro nechezvinhu zveSiHy substrate. Musiyano uyu muchimiro chavo chepamusoro unogona kuoneka panguva yekudyidzana kwavo neAFM probe inodzika uye mushure mezvo muhuwandu hwemodulus hwakayerwa.
Mifananidzo yeSTEM ine zvikamu zvakasiyana-siyana ye (a) lehfilcon A CL uye (b) SiHy substrate. Scale bar, 500 nm. Mifananidzo yeAFM yekumusoro kwe lehfilcon A CL substrate (c) uye base SiHy substrate (d) (3 µm × 3 µm).
Mapolymers anofemerwa nebio uye mabhurashi epolymer akapfava uye akadzidziswa zvakanyanya uye akashandiswa mukushandiswa kwakasiyana-siyana kwebiomedical74,75,76,77. Saka, zvakakosha kushandisa nzira yeAFM nanoindentation, iyo inogona kuyera nemazvo uye zvakavimbika hunhu hwavo hwemakanika. Asi panguva imwe chete, hunhu hwakasiyana hwezvinhu izvi zvakapfava zvakanyanya, senge modulus yakaderera zvakanyanya, huwandu hwakawanda hwemvura uye elasticity yakanyanya, zvinowanzoita kuti zviome kusarudza chinhu chakakodzera, chimiro uye chimiro cheprobe inopinza. saizi. Izvi zvakakosha kuitira kuti indenter isaboore pamusoro wakapfava wemuenzaniso, izvo zvinotungamira mukukanganisa mukuona nzvimbo yekubata nepamusoro nenzvimbo yekubata.
Kuti izvi zviitike, kunzwisisa kwakazara kwechimiro chezvinhu zvakapfava zvebiomimetic (lehfilcon A CL) kwakakosha. Ruzivo nezvehukuru uye chimiro chemabhurashi epolymer ane branched akawanikwa uchishandisa nzira yekufungidzira runopa hwaro hwekutsanangurwa kwechimiro chemuchina uchishandisa matekiniki eAFM nanoindentation. Panzvimbo pemapurobe ecolloidal ane saizi ye micron, takasarudza probe ye silicon nitride (Bruker) ine dhayamita yetip ye140 nm, yakagadzirirwa kuyera huwandu hwehunhu hwemuchina hwesamples dzebiological 78, 79, 80, 81, 82, 83, 84. Chikonzero chekushandisa mapurobe akapinza zvichienzaniswa nemapurobe ecolloidal anogona kutsanangurwa nechimiro chezvinhu zvacho. Kuenzanisa saizi yemuromo weprobe (~140 nm) nemabhurashi epolymer ane matavi ari pamusoro peCL lehfilcon A, anoratidzwa muFig. 3a, zvinogona kugumiswa kuti muromo wacho wakakura zvakakwana kuti ubatane zvakananga nemabhurashi aya, izvo zvinoderedza mukana wekuti muromo uboore nepakati pawo. Kuti tiratidze pfungwa iyi, muFig. 4 mune mufananidzo weSTEM welehfilcon A CL uye muromo weAFM probe (wakagadzirwa kuti usvike pachiyero).
Chimiro chinoratidza mufananidzo weSTEM welehfilcon A CL uye probe yekudzikisa ganda yeACM (yakagadzirwa kusvika pachiyero).
Pamusoro pezvo, saizi yetip ye140 nm idiki zvakakwana kudzivirira njodzi yechero mhedzisiro yekuwedzera inonamira yakambotaurwa nezvemabhurashi epolymer akagadzirwa neCP-AFM nanoindentation method69,71. Tinofunga kuti nekuda kwechimiro chakakosha checone-spherical uye saizi diki yetip iyi yeAFM (Mufananidzo 1), mhando ye force curve inogadzirwa ne lehfilcon A CL nanoindentation haizotsamiri pakumhanya kwe indentation kana kumhanya kwe loading/unloading. Nokudaro, haina kukanganiswa ne poroelastic effects. Kuti uedze iyi fungidziro, lehfilcon A CL samples dzakaiswa indentent pa fixed maximum force uchishandisa PFQNM-LC-A-CAL probe, asi pamatanho maviri akasiyana, uye tensile ne retract force curves zvakashandiswa kuronga simba (nN) mukupatsanurana (µm) zvinoratidzwa muMufananidzo 5a. Zviri pachena kuti simba rinokombama panguva yekurodha nekuburitsa zvinhu rinopindirana zvachose, uye hapana humbowo hwakajeka hwekuti simba rinocheka pakadzika kwe zero indentation rinowedzera nekumhanya kwe indentation mumufananidzo, zvichiratidza kuti zvinhu zvebhurashi rega rega zvakaratidzwa pasina poroelastic effect. Kusiyana neizvi, mhedzisiro yekuchengetedza mvura (viscous extrusion uye poroelasticity effects) inoonekwa kune 45 µm dhayamita AFM probe nekumhanya kumwe chete kwe indentation uye inoonekwa ne hysteresis pakati pe stretch ne retract curves, sezvakaratidzwa muMufananidzo 5b. Mhedzisiro iyi inotsigira fungidziro uye inoratidza kuti 140 nm dhayamita probes isarudzo yakanaka yekuratidza nzvimbo dzakapfava dzakadaro.
lehfilcon A CL indentation force curves uchishandisa ACM; (a) kushandisa probe ine dhayamita ye140 nm pamatanho maviri ekurodha, zvichiratidza kusavapo kwemhedzisiro ye poroelastic panguva yekuiswa pamusoro peganda; (b) kushandisa probe dzine dhayamita ye45 µm ne140 nm. s zvinoratidza mhedzisiro ye viscous extrusion uye poroelasticity yema probe makuru zvichienzaniswa nema probe madiki.
Kuti iratidze nzvimbo dzakapfava zvakanyanya, nzira dzeAFM nanoindentation dzinofanira kunge dziine probe yakanakisa yekudzidza hunhu hwechinhu chiri kudzidzwa. Pamusoro pechimiro nehukuru hwetip, kunzwisisika kweAFM detector system, kunzwisisika kwekutsauka kwetip munzvimbo yekuyedza, uye kuomarara kwe cantilever zvinoita basa rakakosha mukuona kururama uye kuvimbika kwekuyerwa kwe nanoindentation. Kune yedu AFM system, muganho wekuona wePosition Sensitive Detector (PSD) ungangoita 0.5 mV uye wakavakirwa pachiyero chechirimo chakayerwa uye kunzwisisika kwekutsauka kwemvura kwakaverengerwa kwePFQNM-LC-A-CAL probe, iyo inoenderana netheoretical load sensitivity. iri pasi pe0.1 pN. Naizvozvo, nzira iyi inobvumira kuyerwa kwesimba re indentation ≤ 0.1 pN pasina chero chikamu cheruzha rwe peripheral. Zvisinei, hazvigoneke kuti AFM system ideredze ruzha rwe peripheral kusvika padanho iri nekuda kwezvinhu zvakaita se mechanical vibration uye fluid dynamics. Zvinhu izvi zvinoderedza kunzwisisika kwese kwenzira yeAFM nanoindentation uye zvinogumawo nechiratidzo cheruzha rwekumashure cheinenge ≤ 10 pN. Pakutsanangura pamusoro, sampuro dze lehfilcon A CL neSiHy substrate dzakaiswa mukati memvura yakakwana dzichishandisa probe ye 140 nm yekutarisa SEM, uye ma force curves akabuda akaiswa pakati pesimba (pN) nekumanikidzwa. Chikamu chekuparadzanisa (µm) chinoratidzwa muMufananidzo 6a. Zvichienzaniswa neSiHy base substrate, lehfilcon A CL force curve inoratidza zvakajeka chikamu chekuchinja chinotanga panzvimbo yekubatana neforked polymer brush uye ichiguma nekuchinja kwakapinza mukubatana kweslope netip nezvinhu zviri pasi. Chikamu ichi chekuchinja kwesimba chinoratidza maitiro echokwadi elastic ebrashi repolymer rakapatsanurwa pamusoro, sezvinoratidzwa necompression curve inotevera pedyo ne tension curve uye musiyano muhunhu hwemakanika pakati pechimiro chebhurashi nezvinhu zvakakura zveSiHy. Pakuenzanisa lefilcon. Kupatsanurwa kweavhareji yehurefu hwebhurashi repolymer rine matavi mumufananidzo weSTEM wePCS (Mufananidzo 3a) uye curve yayo yekusimba iri pedyo neabscissa muMufananidzo 3a. 6a kunoratidza kuti nzira iyi inokwanisa kuona muromo nepolymer ine matavi ichisvika pamusoro chaipo penzvimbo. Kusangana pakati pezvivakwa zvebhurashi. Pamusoro pezvo, kupindirana kwepedyo kwe force curves kunoratidza kuti hapana mhedzisiro yekuchengetedza mvura. Muchiitiko ichi, hapana kubatana pakati petsono nepamusoro pemuenzaniso. Zvikamu zvepamusoro zve force curves zvemasampuro maviri aya zvinopindirana, zvichiratidza kufanana kwehunhu hwemuchina hwezvinhu zve substrate.
(a) Makomba esimba eAFM nanoindentation e lehfilcon A CL substrates neSiHy substrates, (b) makomba esimba anoratidza kufungidzira kwenzvimbo yekubatana uchishandisa nzira yekumira kweruzha rwekumashure.
Kuti tidzidze ruzivo rwakadzama rwe force curve, tension curve ye lehfilcon A CL sample inorongedzwa patsva muFig. 6b ne power yakanyanya ye 50 pN pamwe ne y-axis. Girafu iyi inopa ruzivo rwakakosha nezve ruzha rwekutanga rwekumashure. Ruzha rwuri pakati pe ±10 pN, iyo inoshandiswa kuona nemazvo nzvimbo yekubatana uye kuverenga kudzika kwe indentation. Sezvakataurwa mumabhuku, kuzivikanwa kwenzvimbo dzekubatana kwakakosha kuti tiongorore nemazvo zvinhu zvakaita se modulus85. Nzira inosanganisira kugadzirisa otomatiki data re force curve yakaratidza kuwirirana kuri nani pakati pe data fitting uye quantitative measurements yezvinhu zvakapfava86. Mubasa iri, kusarudza kwedu nzvimbo dzekubatana kuri nyore uye kwakananga, asi kune miganhu yayo. Maitiro edu ekuchengetedza ekuona nzvimbo yekubatana anogona kukonzera modulus values ​​​​dzakanyanya kufungidzirwa dzekudzika kudiki kwe indentation (< 100 nm). Kushandiswa kwe algorithm-based touchpoint detection uye automated data processing kunogona kuenderera mberi nebasa iri mune ramangwana kuti tiwedzere kuvandudza nzira yedu. Saka, kune ruzha rwemukati rwekumashure rwakaita se ±10 pN, tinotsanangura nzvimbo yekubatana senzvimbo yekutanga yedata pa x-axis muMufananidzo 6b ine kukosha kwe ≥10 pN. Zvadaro, zvichienderana nechiyero cheruzha che 10 pN, mutsetse wakamira padanho re ~0.27 µm unoratidza nzvimbo yekubatana nepamusoro, mushure mezvo curve yekutambanudza inoenderera mberi kusvika substrate yasangana nekudzika kwe indentation kwe ~270 nm. Zvinofadza kuti, zvichibva pahukuru hwe brashi re polymer rakacherwa (300–400 nm) rakayerwa uchishandisa nzira yekufungidzira, kudzika kwe indentation kwe CL lehfilcon Muenzaniso wakaonekwa uchishandisa nzira ye background noise threshold inenge 270 nm, iyo iri pedyo zvikuru nehukuru hwekuyera ne STEM. Mhedzisiro iyi inosimbisawo kuenderana uye kushanda kwechimiro nehukuru hwe AFM probe tip yekudzika kwechimiro ichi che polymer brashi chakapfava uye chakasimba. Iyi data inopawo humbowo hwakasimba hwekutsigira nzira yedu yekushandisa ruzha rwekumashure sechiyero chekutarisa nzvimbo dzekubatana. Saka, chero mhedzisiro yehuwandu inowanikwa kubva mukutevedzera kwemasvomhu uye kugadzirisa force curve inofanira kunge yakarurama.
Kuyerwa kwehuwandu nenzira dzeAFM nanoindentation kunoenderana zvachose nemamodheru emasvomhu anoshandiswa pakusarudza data uye kuongorora kwakatevera. Saka, zvakakosha kufunga nezvezvinhu zvese zvine chekuita nesarudzo ye indenter, zvinhu zvematerial uye mashandiro ekudyidzana kwavo usati wasarudza modhi yakati. Muchiitiko ichi, tip geometry yakanyatsotsanangurwa uchishandisa SEM micrographs (Mufananidzo 1), uye zvichibva pamhedzisiro, probe ye 140 nm dhayamita AFM nanoindenting ine hard cone uye spherical tip geometry isarudzo yakanaka yekutarisa lehfilcon A CL79 samples. Chimwe chinhu chakakosha chinofanira kuongororwa zvakanyatsonaka ndiko kusimba kwezvinhu zvepolymer zviri kuyedzwa. Kunyangwe data rekutanga re nanoindentation (Mifananidzo 5a na 6a) richitsanangura zvakajeka maficha ekupindirana kwema tension and compression curves, kureva, kudzoreredzwa kwakakwana kwe elastic kwezvinhu, zvakakosha kwazvo kusimbisa hunhu hwe elastic hwe contacts. Kuti izvi zviitike, kuiswa kwemasero maviri akatevedzana kwakaitwa panzvimbo imwe chete pamusoro pemuenzaniso we lehfilcon A CL pachiyero chekuiswa kwemasero che 1 µm/s pasi pemamiriro emvura akazara. Data re force curve rinobuda rinoratidzwa mumufananidzo 7 uye, sezvaitarisirwa, kuwedzera nekumanikidzwa kwema prints maviri aya kwakafanana, zvichiratidza kusimba kwakanyanya kwechimiro chebrashi re polymer rakacheneswa.
Makomba maviri esimba rekupinza panzvimbo imwe chete pamusoro pe lehfilcon A CL anoratidza kusimba kwakakodzera kwepamusoro pe lens.
Zvichibva paruzivo rwakawanikwa kubva kuSEM neSTEM mifananidzo yeprobe tip uye lehfilcon A CL surface, zvichiteerana, cone-sphere model inomiririra zvine musoro masvomhu yekudyidzana pakati peAFM probe tip nezvinhu zvakapfava zvepolymer zviri kuyedzwa. Pamusoro pezvo, kune iyi cone-sphere model, fungidziro huru pamusoro pehunhu hwe elastic hwezvinhu zvakadhindwa ndezvechokwadi kune iyi biomimetic material itsva uye inoshandiswa kuyera elastic modulus.
Mushure mekuongororwa kwakazara kwenzira yeAFM nanoindentation nezvikamu zvayo, kusanganisira hunhu hweindentation prope (chimiro, saizi, uye kuomarara kwechitubu), kunzwa (ruzha rwekumashure uye kufungidzira kwenzvimbo yekubatana), uye mamodheru ekugadzirisa data (kuyerwa kwehuwandu hwemodulus), nzira iyi yakashandiswa. ratidza masampuli akapfava anowanikwa mumusika kuti uone mhedzisiro yehuwandu. Hydrogel yekutengeserana yepolyacrylamide (PAAM) ine elastic modulus ye1 kPa yakaedzwa pasi pemamiriro emvura uchishandisa probe ye140 nm. Ruzivo rwekuyedzwa kwemodule uye kuverenga rwunopihwa muSupplementary Information. Mhedzisiro yacho yakaratidza kuti avhareji yemodulus yakayerwa yaive 0.92 kPa, uye %RSD ne percentage (%) deviation kubva kumodulus inozivikanwa zvaive pasi pe10%. Mhedzisiro iyi inosimbisa kururama uye kudzokororwa kwenzira yeAFM nanoindentation inoshandiswa mubasa iri kuyera moduli yezvinhu zve ultrasoft. Nzvimbo dzesamples dze lehfilcon A CL neSiHy base substrate dzakatsanangurwa zvakare uchishandisa nzira imwecheteyo yeAFM nanoindentation yekudzidza modulus inooneka ye ultrasoft surface sebasa re indentation depth. Indentation force separation curves yakagadzirwa kune sampuli nhatu dzerudzi rumwe nerumwe (n = 3; indentation imwe pamuenzaniso) pa force ye 300 pN, speed ye 1 µm/s, uye full hydration. Indentation force sharing curve yakafungidzirwa uchishandisa cone-sphere model. Kuti uwane modulus zvichienderana nekudzika kwe indentation, chikamu che 40 nm chakafara che force curve chakaiswa pa increment yega yega ye 20 nm kutanga kubva panzvimbo ye contact, uye kuyerwa kwe values ​​​​ye modulus padanho rega rega re force curve. Spin Cy et al. Nzira yakafanana yakashandiswa kutsanangura modulus gradient yemabhurashi epolymer epoly(lauryl methacrylate) (P12MA) uchishandisa colloidal AFM probe nanoindentation, uye zvinoenderana nedata uchishandisa Hertz contact model. Nzira iyi inopa mufananidzo we apparent contact modulus (kPa) uchienzaniswa ne indentation depth (nm), sezvakaratidzwa muMufananidzo 8, unoratidza apparent contact modulus/depth gradient. Calculated elastic modulus yeCL lehfilcon A sample iri mu range ye 2–3 kPa mukati me 100 nm yepamusoro ye sample, kupfuura iyo inotanga kuwedzera nekudzika. Kune rumwe rutivi, pakuyedza SiHy base substrate isina brush-like film pamusoro, kudzika kwakanyanya kwe indentation kunowanikwa nesimba re 300 pN kushoma pane 50 nm, uye modulus value inowanikwa kubva kudata iri inenge 400 kPa, iyo inofananidzwa ne values ​​​​dze Young's modulus yezvinhu zvakawanda.
Kuonana kwemodulus (kPa) zvichienzaniswa nekudzika kwemukati (nm) kwe lehfilcon A CL neSiHy substrates uchishandisa nzira yeAFM nanoindentation ine cone-sphere geometry kuyera modulus.
Nzvimbo yepamusoro pechimiro chitsva chebhurashi repolymer rine mabhanzi ebiomimetic inoratidza modulus yakaderera kwazvo yekusimba (2–3 kPa). Izvi zvinoenderana nemucheto wakasununguka webhurashi repolymer rakasungirirwa sezvakaratidzwa mumufananidzo weSTEM. Kunyange zvazvo paine humbowo hwekuti modulus gradient iri pamucheto wekunze weCL, substrate huru ye modulus ine simba guru. Zvisinei, 100 nm yepamusoro yepamusoro iri mukati me20% yehurefu hwese hwebhurashi repolymer rine mabhanzi, saka zvine musoro kufunga kuti zviyero zvakayerwa zvemodulus muchikamu ichi chekudzika kwe indentation zvakarurama uye hazvitsamiri zvakanyanya pamhedzisiro yechinhu chepazasi.
Nekuda kwekugadzirwa kwakasiyana kwe lehfilcon A contact lenses, ine matavi e PMPC polymer brush akasungirirwa pamusoro peSiHy substrates, zvakaoma kwazvo kutsanangura zvakavimbika hunhu hwemagadzirirwo epamusoro pemagadzirirwo avo uchishandisa nzira dzechinyakare dzekuyera. Pano tinoratidza nzira yepamusoro yeAFM nanoindentation yekutarisa nemazvo zvinhu zvakapfava zvakanyanya senge lefilcon A ine huwandu hwakawanda hwemvura uye elasticity yakanyanya. Nzira iyi yakavakirwa pakushandiswa kweAFM probe ine tip size uye geometry zvakasarudzwa zvakanyatsonaka kuti zvienderane nehukuru hwemagadzirirwo ezvimiro zvepamusoro zvakapfava zvakanyanya zvinofanira kudhindwa. Kusanganiswa uku kwehukuru pakati peprobe nechimiro kunopa kunzwa kwakawedzerwa, zvichitibvumira kuyera low modulus uye inherently elastic properties ye branched polymer brush elements, zvisinei ne poroelastic effects. Mhedzisiro yacho yakaratidza kuti mabrashes e PMPC polymer brashes akasiyana ane branches yakaderera kwazvo (kusvika ku 2 kPa) uye elasticity yakanyanya (inenge 100%) paakaedzwa munzvimbo ine mvura. Mhedzisiro yeAFM nanoindentation yakatibvumirawo kuratidza hunhu hwe contact modulus/depth gradient (30 kPa/200 nm) hwe biomimetic lens surface. Hunhu uhwu hunogona kunge hwakakonzerwa nemusiyano we modulus pakati pebrashes dze polymer dzakagadzirwa ne branched ne SiHy substrate, kana branched structure/density yema polymer brushes, kana musanganiswa wawo. Zvisinei, zvidzidzo zvakadzama zvinodiwa kuti tinzwisise hukama huripo pakati pechimiro ne properties, kunyanya mhedzisiro yebrashe branch branch pa properties dze mechanical. Kuyerwa kwakafanana kunogona kubatsira kuratidza properties dze mechanical pamusoro pezvimwe zvinhu zvakapfava zvakanyanya uye zvishandiso zvekurapa.
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