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Hamwe niterambere ryibikoresho bishya bya ultra-yoroshye kubikoresho byubuvuzi hamwe nubuzima bwa biomedical medicine, kuranga byimazeyo imiterere yumubiri nubukanishi nibyingenzi kandi biragoye.Uburyo bwa atomic force microscopie (AFM) tekinike ya nanoindentation yakoreshejwe kugirango iranga modulus yo hasi cyane ya lehfilcon Lomfilcon nshya ya biomimetike silicone hydrogel ihuza lens yometse kumurongo wububiko bwa polymer.Ubu buryo butuma hamenyekana neza aho uhurira nta ngaruka zo gukuramo ibibyimba iyo wegereye polymers ishami.Mubyongeyeho, bituma bishoboka kumenya imiterere yubukorikori bwibintu bya brush buri muntu nta ngaruka za poroelastique.Ibi bigerwaho muguhitamo probe ya AFM ifite igishushanyo (ingano yinama, geometrie nigipimo cyimpeshyi) ikwiriye cyane cyane gupima imiterere yibikoresho byoroshye hamwe nubushakashatsi bwibinyabuzima.Ubu buryo butezimbere ibyiyumvo byukuri kandi byukuri kugirango bipime neza ibintu byoroshye cyane lehfilcon A, ifite modulus nkeya cyane ya elastique ku buso (kugeza kuri 2 kPa) hamwe na elastique ikabije cyane mubidukikije byamazi (hafi 100%) .Ibyavuye mu bushakashatsi bwakozwe ntabwo byerekanaga gusa imiterere ya ultra-yoroshye yubuso bwa lehfilcon A lens, ariko yanagaragaje ko modulus yubushakashatsi bwa polymer yamashami yagereranijwe nubwa silicon-hydrogen substrate.Ubu buryo bwo kuranga tekinike burashobora gukoreshwa mubindi bikoresho bya ultra-yoroshye nibikoresho byubuvuzi.
Imiterere yubukorikori bwibikoresho byabugenewe kugirango bihuze neza na tissue nzima akenshi bigenwa nibidukikije.Ihuza ryiza ryibi bintu bifasha kugera kubintu byifuzwa byubuvuzi bitarinze gutera ibisubizo bibi bya selile1,2,3.Kubikoresho byinshi byahuje ibitsina, kuranga imiterere yubukanishi biroroshye cyane kuberako haboneka inzira zisanzwe nuburyo bwo kwipimisha (urugero, microindentation4,5,6).Nyamara, kubikoresho byoroheje cyane nka geles, hydrogel, biopolymers, selile nzima, nibindi, ubu buryo bwo kwipimisha ntabwo bukoreshwa mubisanzwe kubera imipaka yo gupima no kutagira ubumuga bwibikoresho bimwe7.Mu myaka yashize, uburyo bwa indentation gakondo bwahinduwe kandi buhuzwa kugirango buranga ibintu byinshi byoroshye, ariko uburyo bwinshi buracyafite ibibazo bikomeye bigabanya imikoreshereze yabo 8,9,10,11,12,13.Kubura uburyo bwihariye bwo kwipimisha bushobora kuranga neza kandi bwizewe kuranga imiterere yubukanishi bwibikoresho bya supersoft hamwe nubuso bwubuso bugabanya cyane imikoreshereze yabyo mubikorwa bitandukanye.
Mubikorwa byacu byabanjirije iki, twerekanye lens ya lehfilcon A (CL), ibintu byoroshye bitavanze hamwe nibintu byose bya ultra-yoroshye yubutaka bikomoka kubishushanyo mbonera bya biomimetike byahumetswe hejuru ya cornea yijisho.Iyi biomaterial yatunganijwe mugushushanya ishami rya polymer (2-methacryloyloxyethylphosphorylcholine (MPC)) (PMPC) kuri hydrogel ya silicone (SiHy) 15 yagenewe ibikoresho byubuvuzi bishingiye.Ubu buryo bwo gushushanya butera urwego hejuru rugizwe nuburyo bworoshye kandi bworoshye cyane bwishami rya polymeric brush.Ibikorwa byacu byabanje byemeje ko imiterere ya biomimetike ya lehfilcon A CL itanga imiterere isumba iyindi nko kunoza amazi meza no gukumira nabi, kongera amavuta, no kugabanya ingirabuzimafatizo na bagiteri 15,16.Byongeye kandi, imikoreshereze niterambere ryibi bikoresho bya biomimetike nabyo byerekana kurushaho kwaguka kubindi bikoresho byubuvuzi.Niyo mpamvu, ni ngombwa kuranga imiterere yubuso bwibintu byoroheje kandi byunvikana imikoreshereze yijisho ryijisho kugirango habeho ubumenyi bwuzuye bushyigikira iterambere nibikorwa.Ibicuruzwa byinshi biboneka mubucuruzi bwa SiHy bigizwe nuruvange rwimvange rwa hydrophilique na hydrophobique polymers bigize ibintu bimwe17.Ubushakashatsi bwinshi bwakozwe kugirango hakorwe iperereza ku miterere yubukanishi hakoreshejwe uburyo bwo gupima gakondo, tensile na microindentation uburyo bwo gupima 18,19,20,21.Nyamara, igitabo gishya cya biomimetike ya lehfilcon A CL ituma iba ibikoresho bidasanzwe bitandukanye aho imiterere yubukanishi bwimiterere ya polymer yamashami itandukanijwe cyane nubwa SiHy base substrate.Kubwibyo, biragoye cyane kugereranya neza iyi mitungo ukoresheje uburyo busanzwe kandi bwerekana.Uburyo butanga ikizere bukoresha uburyo bwo gupima nanoindentation bwashyizwe mubikorwa mikorosikopi yingufu za atome (AFM), uburyo bwakoreshejwe mukumenya imiterere yubukorikori bwibikoresho byoroshye bya viscoelastic nka selile biologique na tissue, hamwe na polymers yoroshye22,23,24,25 ., 26,27,28,29,30.Muri AFM nanoindentation, ibyingenzi byo kwipimisha nanoindentation byahujwe niterambere rigezweho mu ikoranabuhanga rya AFM kugirango hongerwe imbaraga zo gupima no gupima ibintu byinshi byavukijwe ibikoresho bya supersoft 31,32,33,34,35,36.Mubyongeyeho, tekinoroji itanga izindi nyungu zingenzi binyuze mugukoresha geometrie zitandukanye.indenter na probe nibishoboka byo kwipimisha mubitangazamakuru bitandukanye byamazi.
AFM nanoindentation irashobora kugabanywa mubice bitatu byingenzi: (1) ibikoresho (sensor, detector, probe, nibindi);(2) ibipimo byo gupima (nkimbaraga, kwimurwa, umuvuduko, ingano ya ramp, nibindi);(3) Gutunganya amakuru (gukosora ibyingenzi, kugereranya ingingo, kugereranya amakuru, kwerekana imiterere, nibindi).Ikibazo gikomeye hamwe nubu buryo nuko ubushakashatsi bwinshi mubuvanganzo ukoresheje raporo ya AFM nanoindentation raporo itandukanye cyane kubisubizo kubwinshi bwa sample / selile / ibikoresho 37,38,39,40,41.Kurugero, Lekka n'abandi.Ingaruka za AFM probe geometrie kuri modulus yapimwe ya Young yerekana ingero za hydrogel ya hydrogel hamwe na selile heterogeneous yakozwe kandi iragereranywa.Batangaza ko indangagaciro za modulus zishingiye cyane cyane ku guhitamo kwa kantileveri no kumiterere yinama, hamwe nagaciro gakomeye kuri piramide ishusho ya piramide nigiciro cyo hasi ya 42 kuri probe.Muri ubwo buryo, Selhuber-Unkel n'abandi.Byerekanwe uburyo umuvuduko wa indenter, ubunini bwa indenter nubunini bwa polyacrylamide (PAAM) bigira ingaruka kuri modulus ya Young yapimwe na ACM43 nanoindentation.Ikindi kintu kitoroshye nukubura ibikoresho bisanzwe byo gupima modulus hamwe nuburyo bwo gukora ibizamini kubuntu.Ibi biragoye cyane kubona ibisubizo nyabyo ufite ikizere.Nyamara, uburyo ni ingirakamaro cyane mugupima ugereranije no gusuzuma kugereranya hagati yubwoko bwikitegererezo, urugero nko gukoresha AFM nanoindentation kugirango utandukanye selile zisanzwe na selile 44, 45.
Mugihe ugerageza ibikoresho byoroshye hamwe na AFM nanoindentation, itegeko rusange ryikumwe nugukoresha probe hamwe nimpeshyi yo hasi ihoraho (k) ihuye neza nicyitegererezo modulus hamwe na hemispherical / round tip kugirango iperereza ryambere ridacengera hejuru yicyitegererezo kuri banza uhure nibikoresho byoroshye.Ni ngombwa kandi ko ibimenyetso byo gutandukana byakozwe na probe byakomera bihagije kugirango bigaragare na sisitemu yo gushakisha laser 24,34,46,47.Ku bijyanye na ultra-yoroshye ya heterogeneous selile, tissue na geles, indi mbogamizi ni ugutsinda imbaraga zifatika hagati ya probe nubuso bwikitegererezo kugirango harebwe ibipimo byororoka kandi byizewe 48,49,50.Kugeza vuba aha, imirimo myinshi kuri nanoindentation ya AFM yibanze ku kwiga ku myitwarire yimikorere ya selile biologiya, tissue, geles, hydrogels, na biomolecules ikoresheje probe nini cyane, ikunze kwitwa colloidal probes (CPs)., 47, 51, 52, 53, 54, 55. nka karubone (DLC).Nubwo CP-AFM nanoindentation aribwo buryo bwambere bwo guhitamo icyitegererezo cyoroshye, gifite ibibazo byacyo kandi bigarukira.Gukoresha inama nini, micron-nini ya spherical nama yongerera ubuso bwitumanaho bwisonga hamwe nicyitegererezo kandi bikavamo igihombo gikomeye cyo gutandukanya umwanya.Kubintu byoroheje, bidafite uburinganire, aho imiterere yubukorikori bwibintu byaho ishobora gutandukana cyane ugereranije nu kigereranyo cyagutse, CP indentation irashobora guhisha inomomogeneité iyo ari yo yose mumitungo kurwego rwaho 52.Iperereza rya colloidal mubusanzwe rikorwa muguhuza micron-nini ya colloidal sisitemu kuri cantilevers idafite tipse ukoresheje epoxy adhesives.Ibikorwa byo gukora ubwabyo byuzuyemo ibibazo byinshi kandi birashobora gutuma habaho ukudahuza mubikorwa bya kalibrasi.Byongeye kandi, ingano nubunini bwibice bya colloidal bigira ingaruka ku buryo butaziguye ibipimo nyamukuru bya kalibrasi ya cantilever, nka frequency resonant frequency, stiffness spring, hamwe na sensitivité de 56,57.58.Rero, uburyo bukoreshwa muburyo busanzwe bwa AFM, nka Calibibasi yubushyuhe, ntibishobora gutanga kalibrasi yukuri kuri CP, kandi ubundi buryo bushobora gusabwa kugirango ukosore 57, 59, 60, 61. Ubushakashatsi bwa CP bwerekana indente bukoresha gutandukana kwinshi kuri cantilever to wige imiterere yintangarugero yoroshye, itera ikindi kibazo mugihe uhindura imyitwarire itari kumurongo wa cantilever mugihe kinini ugereranije 62,63,64.Uburyo bwa kijyambere bwa colloidal probe indentation mubisanzwe hitabwa kuri geometrie ya kantileveri ikoreshwa muguhindura iperereza, ariko ukirengagiza ingaruka ziterwa nuduce duto twa colloidal, ibyo bikaba bitera ukundi gushidikanya muburyo nyabwo 38,61.Mu buryo nk'ubwo, modulike ya elastique ibarwa na moderi yo guhuza ikwiranye na geometrie ya probe ya indentation, kandi kudahuza hagati yimiterere nicyitegererezo kiranga bishobora gutuma habaho amakosa 27, 65, 66, 67, 68. Bimwe mubikorwa biherutse gukorwa na Spencer nabandi.Ibintu bigomba kwitabwaho mugihe biranga amashanyarazi yoroshye ya polymer ukoresheje uburyo bwa CP-AFM nanoindentation bwerekanwe.Batangaje ko kugumana amazi ya viscous muri brush ya polymer nkigikorwa cyihuta bituma ubwiyongere bwumutwaro bwumutwe bityo ibipimo bitandukanye byimitungo ishingiye kumuvuduko30,69,70,71.
Muri ubu bushakashatsi, twaranze modulus yubuso bwa ultra-yoroshye cyane ibikoresho byoroshye bya lehfilcon A CL dukoresheje uburyo bwa AFM bwahinduwe nanoindentation.Urebye imiterere nuburyo bushya bwibi bikoresho, urwego rwo kwiyumvisha uburyo bwa gakondo bwo kwerekana indentation biragaragara ko bidahagije kuranga modulus yibi bintu byoroshye cyane, bityo rero birakenewe ko dukoresha uburyo bwa AFM nanoindentation hamwe na sensibilité yo hejuru.urwego.Nyuma yo gusuzuma ibitagenda neza nibibazo byubuhanga busanzwe bwa AFM probe nanoindentation, twerekana impamvu twahisemo iperereza rito, ryashizweho na AFM kugirango dukureho sensibilité, urusaku rwimbere, aho duhurira, gupima modulus yihuta yibikoresho byoroshye bitavanze nko kubika amazi. kwishingikiriza.no kugereranya neza.Twongeyeho, twashoboye gupima neza imiterere nubunini bwinama ya indentation, bidufasha gukoresha moderi ya cone-sphere ikwiye kugirango tumenye modulus ya elastique tutabanje gusuzuma aho duhurira nisonga hamwe nibikoresho.Ibitekerezo bibiri bidasobanutse byagereranijwe muriyi mirimo ni ibintu byuzuye bya elastique hamwe na indentation yimbitse-yigenga.Twifashishije ubu buryo, twabanje kugerageza ultra-soft standard hamwe na modulus izwi kugirango tugereranye uburyo, hanyuma dukoresha ubu buryo kugirango turanga ubuso bwibikoresho bibiri bitandukanye byo guhuza.Ubu buryo bwo kuranga AFM nanoindentation igaragara hamwe no kwiyumvisha ibintu byitezwe ko izakoreshwa muburyo butandukanye bwibikoresho bya biomimetike ya heterogeneous ultrasoft hamwe nibishobora gukoreshwa mubikoresho byubuvuzi hamwe na biomedical medicine.
Lehfilcon Lens yo guhuza (Alcon, Fort Worth, Texas, USA) hamwe na hydrogel substrate ya silicone yatoranijwe kugirango igerageze nanoindentation.Ikigereranyo cyabugenewe cyabugenewe cyakoreshejwe mubigeragezo.Kugirango ushyireho lens kugirango ugerageze, washyizwe muburyo bwitondewe kumurongo uhagaze, urebe neza ko nta mwuka mwinshi winjiye imbere, hanyuma ugashyirwa kumpera.Umwobo uri murwego rwo hejuru ufite lens itanga uburyo bwo kugera kuri optique ya lens kugirango igerageze nanoindentation mugihe ufashe amazi mu mwanya.Ibi bituma lens zifite amazi meza.500 μl yo guhuza lens yamashanyarazi yakoreshejwe nkigisubizo cyikizamini.Kugirango hamenyekane ibisubizo byuzuye, hydrogels iboneka mubucuruzi idakoreshwa na polyacrylamide (PAAM) hydrogels yateguwe kuva polyacrylamide-co-methylene-bisacrylamide (100 mm ibyokurya bya Petrisoft Petri, Matrigen, Irvine, CA, USA), modulus izwi ya 1 ya elastique ya 1 kPa.Koresha ibitonyanga 4-5 (hafi 125)) kuri AFM hydrogel-probe.
Ingero za Lehfilcon A CL na SiHy substrates zerekanwe hakoreshejwe sisitemu ya FEI Quanta 250 yohereza ibyuka byoherezwa mu kirere Scanning Electron Microscope (FEG SEM) ifite ibikoresho bya Scanning Transmission Electron Microscope (STEM).Gutegura ibyitegererezo, lens yabanje gukaraba namazi hanyuma bayacamo imigozi imeze nka pie.Kugirango habeho itandukaniro ritandukanye hagati ya hydrophilique na hydrophobique yibigize ingero, igisubizo cya 0.10% gihamye cya RuO4 cyakoreshejwe nk'irangi, aho ingero zashizwemo iminota 30.Lehfilcon A CL RuO4 irangi ntabwo ari ngombwa kugirango igere ku itandukaniro ritandukanye gusa, ahubwo ifasha no kubungabunga imiterere y’imyanda ya polymer ishami ryayo muburyo bwumwimerere, hanyuma igaragara kumashusho ya STEM.Bahise bozwa kandi babura umwuma murukurikirane rwimvange ya Ethanol / amazi hamwe no kwiyongera kwa Ethanol.Ingero zahise ziterwa na EMBed 812 / Araldite epoxy, yakize ijoro ryose kuri 70 ° C.Icyitegererezo cyahagaritswe na resin polymerisation yaciwe na ultramicrotome, hanyuma ibice byavuyemo byerekanwe hamwe na disiketi ya STEM muburyo buke bwa vacuum kuri voltage yihuta ya 30 kV.Sisitemu imwe ya SEM yakoreshejwe mugusobanura birambuye kuranga PFQNM-LC-A-CAL AFM iperereza (Bruker Nano, Santa Barbara, CA, USA).SEM amashusho yubushakashatsi bwa AFM yabonetse muburyo busanzwe bwa vacuum hamwe na voltage yihuta ya 30 kV.Shakisha amashusho kumpande zitandukanye no gukuza kugirango wandike ibisobanuro byose byuburyo nubunini bwa AFM iperereza.Ibipimo byose byerekana inyungu mumashusho byapimwe muburyo bwa digitale.
Dimension FastScan Bio Icon ya atome yingufu za microscope (Bruker Nano, Santa Barbara, CA, USA) hamwe na "PeakForce QNM muri Fluid" yakoreshejwe mugushushanya no nanoindentate lehfilcon A CL, substrate ya SiHy, hamwe na hydrogel ya PAAm.Kubushakashatsi bwerekana amashusho, PEAKFORCE-HIRS-FA probe (Bruker) ifite numero ya nomero ya radiyo 1 nm yakoreshejwe kugirango ifate amashusho yikigereranyo cyikitegererezo kuri scan ya 0.50 Hz.Amashusho yose yafashwe mugisubizo cyamazi.
Ubushakashatsi bwa AFM nanoindentation bwakozwe hakoreshejwe PFQNM-LC-A-CAL iperereza (Bruker).Ubushakashatsi bwa AFM bufite silicon kuri cantilever ya nitride 345 nm, uburebure bwa 54 µm na 4.5 µm z'ubugari hamwe na rezonans ya 45 kHz.Yashizweho muburyo bwo kuranga no gukora ibipimo bya nanomehanike ku ngero zoroshye za biologiya.Ibyuma bifata ibyuma byahinduwe ku ruganda hamwe na progaramu ya progaramu ya progaramu ya progaramu.Isoko yimvura ya probe yakoreshejwe murubu bushakashatsi yari hagati ya 0.05–0.1 N / m.Kugirango umenye neza imiterere nubunini bwinama, iperereza ryaranzwe muburyo burambuye ukoresheje SEM.Ku mutini.Igishushanyo 1a cyerekana imiterere ihanitse, yo gukuza cyane gusikana electron micrograph ya PFQNM-LC-A-CAL iperereza, itanga icyerekezo cyuzuye cyubushakashatsi.Ku mutini.1b yerekana uburyo bwagutse bwo hejuru hejuru yubushakashatsi, butanga amakuru kubyerekeye imiterere nubunini bwinama.Ku iherezo rikabije, urushinge ni igice cy'isi hafi ya nm 140 z'umurambararo (Ishusho 1c).Munsi yibi, ibyuma bifata imashini muburyo bwa conic, bigera kuburebure bwa metero 500.Hanze y'akarere ka taping, isonga ni silindrike kandi irangirira muburebure bwa metero 1,18 µm.Iki nigice cyingenzi cyibikorwa byinama.Byongeye kandi, iperereza rinini rya polystirene (PS) (Novascan Technologies, Inc., Boone, Iowa, muri Amerika) rifite umurambararo wa diameter ya 45 µm hamwe n’isoko rihoraho rya 2 N / m naryo ryakoreshejwe mu kwipimisha nka probe ya colloidal.hamwe na PFQNM-LC-A-CAL 140 nm probe yo kugereranya.
Byavuzwe ko amazi ashobora gufatwa hagati yiperereza rya AFM nuburyo bwa polymer brush mugihe cya nanoindentation, bizakoresha imbaraga zo hejuru kuri anketi ya AFM mbere yuko ikora ku butaka69.Izi ngaruka zo gukuramo ibintu bitewe no kugumana amazi zirashobora guhindura aho zigaragara, bityo bikagira ingaruka kubipimo bya modulus.Kugira ngo bige ku ngaruka za probe geometrie n'umuvuduko wa indentation ku kugumana amazi, hateguwe umurongo w'ingufu za lendfilcon A CL ukoresheje urugero rwa 140 nm diameter ya probe ku gipimo cyimuka cya 1 µm / s na 2 µm / s.probe diameter 45 µm, imbaraga zihamye gushiraho 6 nN yagezweho kuri 1 µm / s.Ubushakashatsi bufite ipima 140 nm ya diametre bwakozwe ku muvuduko wa indentation ya 1 µm / s n'imbaraga zashyizweho na 300 pN, zahisemo gukora igitutu cyo guhura mu rwego rwa physiologique (1-8 kPa) y'ijisho ryo hejuru.igitutu 72. Ingero zoroshye ziteguye za hydrogel ya PAA ifite umuvuduko wa 1 kPa zapimwe imbaraga zo kwerekana indangururamajwi 50 pN ku muvuduko wa 1 μm / s ukoresheje iperereza rifite diameter ya 140 nm.
Kubera ko uburebure bwigice cyibice bigize isonga rya PFQNM-LC-A-CAL iperereza rigera kuri 500 nm, kubwubujyakuzimu ubwo aribwo bwose <500 nm dushobora gutekereza neza ko geometrie yubushakashatsi mugihe cyo kwerekanwa izakomeza kuba ukuri kuri yo imiterere ya cone.Mubyongeyeho, hafatwa ko ubuso bwibikoresho biri kugeragezwa bizagaragaza igisubizo cyoroshye cya elastique, nacyo kizemezwa mubice bikurikira.Kubwibyo, dukurikije imiterere nubunini bwinama, twahisemo moderi ikwiranye na cone-sphere ikwiye yakozwe na Briscoe, Sebastian na Adams, iboneka muri software y'abacuruzi, kugirango dukore ubushakashatsi bwa AFM nanoindentation (NanoScope).Porogaramu yo gusesengura amakuru yo gutandukanya, Bruker) 73. Icyitegererezo gisobanura isano-yo kwimura imbaraga F (δ) kuri cone ifite inenge ya apex.Ku mutini.Igishushanyo cya 2 cyerekana guhuza geometrie mugihe cyo gukorana kwa cone itajenjetse hamwe na sherfike, aho R ni radiyo yumutwe wa sherfike, a ni radiyo ihuza, b ni radiyo yo guhuza kumpera yumutwe wa sherfike, δ ni the vuga radiyo.ubujyakuzimu bwimbitse, θ ni kimwe cya kabiri-inguni ya cone.Ishusho ya SEM yiyi probe yerekana neza ko umurambararo wa nm 140 ya diametre uhuza mu buryo bugaragara muri cone, hano rero b isobanurwa binyuze muri R gusa, ni ukuvuga b = R cos θ.Porogaramu itangwa n'abacuruzi itanga umubano wa cone-sphere kugirango ubare indangagaciro za Modulus (E) uhereye kumibare yo gutandukanya imbaraga ukeka ko> b.Isano:
aho F ni imbaraga zo kwerekana, E ni Modulus ya Young, ν ni igipimo cya Poisson.Guhuza radiyo a irashobora kugereranywa ukoresheje:
Igishushanyo mbonera cya geometrike ya cone itajegajega hamwe nigitekerezo cya serefegitura kanda mubikoresho bya lens ya konte ya Lefilcon hamwe nubuso bwubuso bwa polymer bwashami.
Niba a ≤ b, isano igabanya kugereranya kubisanzwe bisanzwe;
Twizera ko imikoranire ya probe ya indenting hamwe nishami ryishami rya PMPC polymer brush bizatera radiyo yo guhuza kuba nini kuruta radiyo ihuza b.Kubwibyo, kubipimo byose byapimwe bya moderi ya elastique yakozwe muri ubu bushakashatsi, twakoresheje kwishingikiriza kuboneka kubibazo a> b.
Ibikoresho bya ultrasoft biomimetic byizwe muri ubu bushakashatsi byashushanijwe neza hakoreshejwe scanning yoherejwe na electron microscopi (STEM) yicyitegererezo cyambukiranya hamwe na microscopi yingufu za atome (AFM) yubuso.Iyi miterere irambuye yaranzwe no kwagura ibikorwa byacu byasohotse mbere, aho twemeje ko imiterere ya polymeric brush yamashami yimiterere ya PMPC yahinduwe na lehfilcon A CL yerekanaga imiterere yubukanishi kumubiri wa corneal 14.Kubwiyi mpamvu, twerekeza kuri lens igaragara nkibikoresho bya biomimetike14.Ku mutini.3a, b yerekana ibice byambukiranya amashami ya PMPC ya polymer brush hejuru yubuso bwa lehfilcon A CL substrate hamwe na SiHy itavuwe neza.Ubuso bwizo ngero zombi bwongeye gusesengurwa hifashishijwe amashusho akomeye ya AFM, ibyo bikaba byemeje neza ibyavuye mu isesengura rya STEM (Ishusho 3c, d).Ufatiye hamwe, aya mashusho atanga uburebure bugereranije bwa PMPC ishami rya polymer brush kuri 300-400 nm, ni ngombwa mugusobanura ibipimo bya AFM nanoindentation.Ikindi kintu cyingenzi cyakurikiranwe gikomoka ku mashusho ni uko imiterere rusange yubuso bwibikoresho bya biomimetike ya CL itandukanye na morfologiya itandukanye niyibikoresho bya SiHy substrate.Iri tandukanyirizo ryimiterere yimiterere yabo irashobora kugaragara mugihe cyo gukorana kwabo na probe ya AFM yerekana hanyuma bikagereranywa nagaciro kapimwe.
Amashusho yambukiranya amashusho ya (a) lehfilcon A CL na (b) SiHy substrate.Umwanya munini, 500 nm.AFM amashusho yubuso bwa lehfilcon A substrate ya CL (c) na base ya SiHy substrate (d) (3 µm × 3 µm).
Bioinspired polymers hamwe na polymer brush nuburyo busanzwe bworoshye kandi bwarigishijwe cyane kandi bukoreshwa muburyo butandukanye bwibinyabuzima74,75,76,77.Niyo mpamvu, ni ngombwa gukoresha uburyo bwa AFM nanoindentation, bushobora gupima neza kandi bwizewe imiterere yubukanishi.Ariko icyarimwe, imiterere yihariye yibi bikoresho byoroheje cyane, nka modulus yo hasi cyane ya elastike, ibintu byinshi byamazi menshi hamwe na elastique nyinshi, akenshi bigorana guhitamo ibikoresho, imiterere nuburyo bwa probe yerekana.ingano.Ibi nibyingenzi kugirango indenter idacumita hejuru yoroheje yicyitegererezo, byatera amakosa muguhitamo aho uhurira nubuso hamwe nubusabane bwaho.
Kubwibyo, gusobanukirwa byimazeyo morphologie yibikoresho bya ultra-yoroshye biomimetike (lehfilcon A CL) ni ngombwa.Amakuru ajyanye nubunini nuburyo bwimiterere ya polymer yamashami yabonetse hakoreshejwe uburyo bwo gufata amashusho atanga urufatiro rwo kuranga imashini ukoresheje tekinoroji ya AFM nanoindentation.Mu mwanya wa micron-nini ya spherical colloidal probe, twahisemo PFQNM-LC-A-CAL silicon nitride probe (Bruker) ifite umurambararo wa diametre 140 nm, yagenewe cyane cyane gushushanya amakarita yimiterere yibikoresho bya biologiya 78, 79, 80 , 81, 82, 83, 84 Impamvu yo gukoresha ubushakashatsi bugereranije ugereranije nubushakashatsi busanzwe bwa colloidal bushobora gusobanurwa nuburyo bwimiterere yibikoresho.Ugereranije ubunini bwa probe (~ 140 nm) hamwe nu mashami ya polymer yashami hejuru ya CL lehfilcon A, yerekanwa ku gishushanyo cya 3a, dushobora kwemeza ko inama ari nini bihagije kugirango ihure neza nuburyo bwububiko, igabanya amahirwe yo gutobora muri bo.Kugirango tugaragaze iyi ngingo, ku gishushanyo cya 4 ni ishusho ya STEM ya lehfilcon A CL hamwe nisonga ryerekana indege ya AFM (yashushanyije ku munzani).
Igishushanyo cyerekana STEM ishusho ya lehfilcon A CL hamwe na ACM yerekana indangururamajwi (yashushanyije ku munzani).
Byongeye kandi, ingano yinini ya 140 nm ni nto bihagije kugirango wirinde ingaruka ziterwa ningaruka zose zifatika zabanje kuvugwa kubushakashatsi bwa polymer bwakozwe na CP-AFM nanoindentation uburyo bwa 69,71.Dutekereza ko kubera imiterere yihariye ya cone-spherical hamwe nubunini bugereranije bwiyi nama ya AFM (Igishusho 1), imiterere yumurongo wingufu zatewe na lehfilcon A CL nanoindentation ntabwo bizaterwa numuvuduko wa indentation cyangwa umuvuduko wo gupakira / gupakurura .Kubwibyo, ntabwo byatewe ningaruka za poroelastic.Kugirango ugerageze iyi hypothesis, lehfilcon A ntangarugero ya CL yatanzwe ku mbaraga ntarengwa zagenwe hakoreshejwe iperereza rya PFQNM-LC-A-CAL, ariko ku muvuduko wa kabiri utandukanye, kandi ibisubizo bivuye ku murongo no gukuramo imbaraga byakoreshejwe mu gutegura ingufu (nN) mu gutandukana (µm) irerekanwa mu gishushanyo cya 5a.Biragaragara ko imbaraga zigoramye mugihe cyo gupakira no gupakurura byuzuye, kandi nta kimenyetso gifatika cyerekana ko ubwogosha bwimbaraga kuri zeru zeru bwiyongera hamwe n’umuvuduko wa indentation ku gishushanyo, byerekana ko buri kintu cyo gukaraba cyaranzwe nta ngaruka mbi ya poroelastique.Ibinyuranye, ingaruka zo kugumana amazi (viscous extrusion and poroelasticity effects) zigaragara kuri 45 µm diameter ya AFM iperereza kumuvuduko umwe kandi bikagaragazwa na hystereze hagati yo kurambura no gukuramo umurongo, nkuko bigaragara ku gishushanyo 5b.Ibisubizo bishyigikira hypothesis kandi byerekana ko ibipimo bya nm 140 nm ari amahitamo meza yo kuranga ibintu byoroshye.
lehfilcon A CL indentation imbaraga zigabanya umurongo ukoresheje ACM;(a) ukoresheje iperereza rifite diameter ya 140 nm ku bipimo bibiri byo gupakira, byerekana ko nta ngaruka ya poroelastique mugihe cyo kwerekana hejuru;(b) ukoresheje iperereza rifite diameter ya 45 µm na 140 nm.s herekana ingaruka zo gukuramo viscous na poroelasticitike kuri probe nini ugereranije nubushakashatsi buto.
Kuranga ultrasoft igaragara, uburyo bwa AFM nanoindentation bugomba kugira iperereza ryiza ryo kwiga imiterere yibikoresho biri kwigwa.Usibye imiterere yubunini nubunini, ibyiyumvo bya sisitemu ya detektori ya AFM, ibyiyumvo byo gutandukana kw'ibizamini mu bidukikije, hamwe no gukomera kwa cantilever bigira uruhare runini mu kumenya ukuri no kwizerwa kwa nanoindentation.ibipimo.Kuri sisitemu yacu ya AFM, Umwanya wa Sensitive Detector (PSD) ntarengwa yo gutahura ni mV 0,5 kandi ushingiye ku gipimo cyabanjirije kalibisiyoneri hamwe no kubara ibibyimba bya flux sensibilité ya PFQNM-LC-A-CAL iperereza, bihuye na umutwaro wo kwiyumvisha ibintu.ni munsi ya 0.1 pN.Kubwibyo, ubu buryo butuma igipimo cyingufu ntoya ≤ 0.1 pN nta kintu na kimwe kigize urusaku.Nubwo bimeze bityo ariko, ntibishoboka ko sisitemu ya AFM igabanya urusaku rwa peripheri kuri uru rwego bitewe nimpamvu nko kunyeganyega kwa mashini na dinamike.Izi ngingo zigabanya ibyiyumvo rusange byuburyo bwa AFM nanoindentation kandi bikavamo ibimenyetso byerekana urusaku rwerekana hafi ≤ 10 pN.Kubiranga ubuso, lehfilcon A CL hamwe na SiHy substrate ntangarugero yatanzwe mugihe cyamazi yuzuye hakoreshejwe iperereza rya nm 140 kugirango SEM iranga, kandi imirongo yavuyemo imbaraga zashyizwe hejuru yingufu (pN) nigitutu.Umugambi wo gutandukana (µm) urerekanwa mumashusho 6a.Ugereranije na SiHy base substrate, lehfilcon A CL imbaraga zigabanya umurongo werekana neza icyiciro cyinzibacyuho gitangirira aho gihurira na brush ya polymer yohasi kandi bikarangirana nimpinduka zikomeye kumurongo ugaragaza guhuza inama hamwe nibikoresho byihishe.Iki gice cyinzibacyuho yingufu zigoramye cyerekana imyitwarire yukuri ya elegitoronike ya polymer yamashami hejuru, nkuko bigaragazwa nu mugozi wo kwikuramo gukurikiranira hafi umurongo uteganijwe no gutandukanya imiterere yubukanishi hagati yimiterere ya brush nibikoresho byinshi bya SiHy.Iyo ugereranije lefilcon.Gutandukanya uburebure buringaniye bwubushakashatsi bwa polymer bwashami mumashusho ya STEM ya PCS (Igishusho 3a) hamwe nimbaraga zayo zigana umurongo kuri abscissa kumashusho 3a.6a yerekana ko uburyo bushobora kumenya inama na polymer ishami ryageze hejuru yubuso.Guhuza hagati ya brush.Mubyongeyeho, gufunga gufunga imbaraga kumurongo byerekana ko nta ngaruka zo gufata amazi.Muri iki kibazo, nta rwose gufatana hagati y'urushinge n'ubuso bw'icyitegererezo.Ibice byo hejuru byingufu zinguni zingero zombi zuzuzanya, byerekana isano ya mikoranike yibikoresho bya substrate.
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Kugirango wige amakuru arambuye yingufu zihetamye, umurongo utera umurongo wa lehfilcon A icyitegererezo cya CL wongeye gutegurwa ku gishushanyo cya 6b hamwe nimbaraga ntarengwa za 50 pN kuruhande rwa y-axis.Iyi shusho itanga amakuru yingenzi kubyerekeye urusaku rwambere.Urusaku ruri murwego rwa ± 10 pN, rukoreshwa mukumenya neza aho uhurira no kubara ubujyakuzimu.Nkuko byavuzwe mubitabo, kumenyekanisha aho uhurira ni ngombwa kugirango dusuzume neza ibintu bifatika nka modulus85.Uburyo burimo gutunganya mu buryo bwikora amakuru yingufu zerekana amakuru yerekanwe neza hagati yamakuru akwiranye nigipimo cyinshi kubikoresho byoroshye86.Muriyi mirimo, guhitamo ingingo duhuza biroroshye kandi bifite intego, ariko bifite aho bigarukira.Uburyo bwacu bwo kwibumbira hamwe kugirango tumenye aho duhurira bishobora kuvamo agaciro gake modulus indangagaciro ntoya (<100 nm).Gukoresha algorithm ishingiye kuri touchpoint yo gutahura no gutunganya amakuru mu buryo bwikora birashobora gukomeza uyu murimo mugihe kizaza kugirango turusheho kunoza uburyo bwacu.Rero, kubwurusaku rwimbere rwimbere kurutonde rwa ± 10 pN, turasobanura aho duhurira nkamakuru yambere yerekanwe kuri x-axis mu gishushanyo cya 6b gifite agaciro ka ≥10 pN.Noneho, ukurikije urusaku rwurusaku rwa 10 pN, umurongo uhagaritse kurwego rwa ~ 0.27 µm ugaragaza aho uhurira nubuso, nyuma yumurongo urambuye ukomeza kugeza substrate yujuje ubujyakuzimu bwa ~ 270 nm.Igishimishije, ukurikije ubunini bwibikoresho bya polymer bishishwa (300-400 nm) byapimwe hakoreshejwe uburyo bwo gufata amashusho, ubujyakuzimu bwa indentation ya CL lehfilcon Icyitegererezo cyagaragaye ukoresheje uburyo bw’urusaku rw’imbere ni hafi 270 nm, cyegereye cyane ingano yo gupima hamwe na STEM.Ibisubizo birashimangira kandi guhuza no gukurikizwa kumiterere nubunini bwa AFM iperereza kugirango yerekane iyi yoroshye cyane kandi yoroheje cyane ishami rya polymer brush.Aya makuru aratanga kandi ibimenyetso bifatika byo gushyigikira uburyo bwacu bwo gukoresha urusaku rwimbere nkurwego rwo kwerekana aho uhurira.Rero, ibisubizo byose byabonetse bivuye mubiharuro byerekana imibare no guhuza umurongo bikwiye bigomba kuba byukuri.
Ibipimo byuzuye byuburyo bwa AFM nanoindentation biterwa rwose nimibare yimibare ikoreshwa muguhitamo amakuru no gusesengura nyuma.Niyo mpamvu, ni ngombwa gusuzuma ibintu byose bijyanye no guhitamo indenter, ibintu bifatika hamwe nubukanishi bwimikoranire yabo mbere yo guhitamo icyitegererezo runaka.Muri iki kibazo, inama ya geometrie yaranzwe yitonze ikoresheje micrografi ya SEM (Igishusho 1), kandi ukurikije ibisubizo, iperereza rya nm 140 nm AFM nanoindenting iperereza hamwe na cone ikomeye hamwe na siporo ya geometrike ni amahitamo meza yo kuranga lehfilcon A CL79 .Ikindi kintu cyingenzi kigomba gusuzumwa neza ni elastique yibikoresho bya polymer bipimwa.Nubwo amakuru yambere ya nanoindentation (Igishusho 5a na 6a) yerekana neza ibimenyetso biranga guhuzagurika kumurongo no guhagarika umurongo, ni ukuvuga kugarura ibintu byuzuye byoroshye, ni ngombwa cyane kwemeza imiterere ya elastique gusa. .Kugira ngo ibyo bishoboke, ibyerekezo bibiri byakurikiranye byakorewe ahantu hamwe hejuru yubuso bwa lehfilcon A CL icyitegererezo ku gipimo cya 1 µm / s mugihe cyuzuye cyuzuye.Ibisubizo bivamo imbaraga zo gutondekanya amakuru byerekanwe mugitabo.7 kandi, nkuko byari byitezwe, kwaguka no kwikuramo umurongo wibicapo byombi birasa, byerekana ubuhanga bukomeye bwimiterere ya polymer brush.
Imbaraga ebyiri zo kwerekana umurongo ahantu hamwe hejuru yubuso bwa lehfilcon A CL yerekana ubworoherane bwiza bwubuso bwa lens.
Ukurikije amakuru yakuwe mu mashusho ya SEM na STEM yerekana iperereza hamwe na lehfilcon A CL hejuru, icyerekezo cya cone-sphere ni imibare yerekana imibare yerekana imikoranire hagati yiperereza rya AFM nibikoresho byoroshye bya polimeri bipimwa.Mubyongeyeho, kuri ubu buryo bwa cone-sphere, ibitekerezo byibanze kubyerekeranye na elastique yibintu byacapwe bifata ukuri kubintu bishya bya biomimetike kandi bikoreshwa mukugereranya modulus ya elastique.
Nyuma yisuzuma ryuzuye ryuburyo bwa AFM nanoindentation hamwe nibiyigize, harimo imitungo ya indentation (imiterere, ingano, hamwe no gukomera kw'isoko), sensitivite (urusaku rw'imbere no kugereranya aho uhurira), hamwe na moderi ikwiranye (gupima modulus yuzuye), uburyo bwari Byakoreshejwe.kuranga ubucuruzi buboneka ultra-yoroshye ntangarugero kugirango ugenzure ibisubizo byuzuye.Hydrogel yubucuruzi ya polyacrylamide (PAAM) hamwe na moderi ya elastike ya 1 kPa yageragejwe mubihe byamazi ikoresheje iperereza rya nm 140.Ibisobanuro birambuye byo kugerageza no kubara bitangwa mumakuru yinyongera.Ibisubizo byerekanaga ko impuzandengo ya modulus yapimwe yari 0,92 kPa, naho% RSD nijanisha (%) gutandukana na modulus izwi ntabwo byari munsi ya 10%.Ibisubizo byemeza ukuri no kubyara uburyo bwa AFM nanoindentation bwakoreshejwe muriki gikorwa cyo gupima moduli yibikoresho bya ultrasoft.Ubuso bwa lehfilcon A CL hamwe na substrate ya SiHy yarushijeho kurangwa hakoreshejwe uburyo bumwe bwa AFM nanoindentation kugirango bige modulus igaragara yo hejuru ya ultrasoft nkibikorwa byimbitse.Kwerekana imbaraga zo gutandukanya imirongo byakozwe kubintu bitatu bya buri bwoko (n = 3; indentation imwe kuri buri cyitegererezo) ku mbaraga za 300 pN, umuvuduko wa 1 µm / s, hamwe n’amazi yuzuye.Imbaraga zo kugabana umurongo zagereranijwe ukoresheje urugero rwa cone.Kugirango ubone modulus ishingiye ku burebure bwa indentation, igice cya 40 nm ubugari bwingufu zumurongo washyizweho kuri buri kwiyongera kwa 20 nm guhera aho duhurira, no gupima indangagaciro za modulus kuri buri ntambwe yingufu zingana.Kuzenguruka Cy n'abandi.Uburyo busa nabwo bwakoreshejwe mu kuranga modulus gradient ya poly (lauryl methacrylate) (P12MA) polymer brush ukoresheje colloidal AFM probe nanoindentation, kandi bihuza namakuru ukoresheje uburyo bwo guhuza Hertz.Ubu buryo butanga ikibanza kigaragara cyo guhuza modulus (kPa) nuburebure bwimbitse (nm), nkuko bigaragara ku gishushanyo cya 8, cyerekana uburyo bwo guhuza modulus / ubujyakuzimu bwa gradient.Modulus yabazwe ya elastike ya CL lehfilcon Icyitegererezo kiri murwego rwa 2-3 kPa muri 100 nm yo hejuru yicyitegererezo, hejuru yacyo gitangira kwiyongera hamwe nubujyakuzimu.Kurundi ruhande, mugihe ugerageza substrate ya SiHy idafite firime isa na brush hejuru yubuso, ubujyakuzimu bwa indentation bwagerwaho ku mbaraga za 300 pN buri munsi ya 50 nm, kandi agaciro ka modulus kabonetse mumibare ni kPa 400. , igereranywa nagaciro ka Modulus yumusore kubikoresho byinshi.
Ikigaragara cyo guhuza modulus (kPa) nuburebure bwimbitse (nm) kuri lehfilcon A CL na SiHy substrate ukoresheje uburyo bwa AFM nanoindentation hamwe na cone-sphere geometrie yo gupima modulus.
Ubuso bwo hejuru cyane bwa biomimetike ishami rya polymer brush ryerekana imiterere ya modulus yo hasi cyane (2–3 kPa).Ibi bizahuza nubusa kumanikwa yubusa ya polymer brush nkuko bigaragara mumashusho ya STEM.Mugihe hari ibimenyetso bimwe byerekana modulus ya gradient kumpera yinyuma ya CL, nyamukuru modulus substrate irakomeye cyane.Nyamara, hejuru ya 100 nm hejuru yubuso buri muri 20% yuburebure bwuburebure bwa polymer yamashami, birakwiriye rero gutekereza ko indangagaciro zapimwe za modulus muri ubu burebure bwimbitse zerekana neza kandi ntizikomeye. Biterwa n'ingaruka z'ikintu cyo hasi.
Bitewe nigishushanyo cyihariye cya biomimetike ya lehfilcon Ihuriro ryitumanaho, rigizwe nishami rya PMPC ryashushanyijemo amashanyarazi ryashizwe hejuru yubutaka bwa SiHy, biragoye cyane kuranga byimazeyo imiterere yubukorikori bwububiko bwabo hakoreshejwe uburyo bwa gakondo bwo gupima.Hano turerekana uburyo bwiza bwa AFM nanoindentation bwo kuranga neza ibikoresho bya ultra-yoroshye nka lefilcon A ifite amazi menshi kandi byoroshye cyane.Ubu buryo bushingiye ku ikoreshwa rya AFM iperereza rifite ubunini bwa geometrike hamwe na geometrie byatoranijwe neza kugirango bihuze ibipimo byimiterere ya ultra-yoroshye yubuso bugomba gucapwa.Uku guhuza ibipimo hagati yubushakashatsi nuburyo butanga ibyiyumvo byiyongera, bikadufasha gupima modulus nkeya hamwe na elastique yibintu bya elegitoronike ya polymer brush, tutitaye ku ngaruka za poroelastique.Ibisubizo byerekanaga ko amashanyarazi adasanzwe ya PMPC polymer aranga hejuru yuburinganire bwa lens yari afite modulus nkeya cyane (kugeza kuri 2 kPa) hamwe na elastique cyane (hafi 100%) mugihe yapimwe mubidukikije byamazi.Ibisubizo bya AFM nanoindentation byanadushoboje kuranga ikigaragara cyo guhuza modulus / ubujyakuzimu bwimbitse (30 kPa / 200 nm) yubuso bwa biomimetike.Iyi gradient irashobora guterwa nuburyo butandukanye bwa modulus hagati yubushakashatsi bwa polymer bwashami hamwe na SiHy substrate, cyangwa imiterere yishami / ubucucike bwimyanda ya polymer, cyangwa ikomatanya ryayo.Nyamara, ubushakashatsi bwimbitse burakenewe kugirango twumve neza isano iri hagati yimiterere nimiterere, cyane cyane ingaruka zishami rya brush kumiterere yubukanishi.Ibipimo nkibi birashobora gufasha kuranga imiterere yubukorikori bwibindi bikoresho bya ultra-yoroshye nibikoresho byubuvuzi.
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