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Site na mmepe nke ihe ọhụrụ dị nro maka ngwaọrụ ahụike yana ngwa biomedical, njirimara zuru oke nke akụrụngwa anụ ahụ na igwe ha dị mkpa ma bụrụ ihe ịma aka.Ejiri usoro nanoindentation gbanwetụrụ atomiki (AFM) iji mara modul dị oke ala nke lehfilcon ọhụrụ Akara kọntaktị silicone hydrogel biomimetic nke ejiri akwa akwa ahịhịa polymer kpuchie ya.Usoro a na-enye ohere ikpebi kpọmkwem ebe kọntaktị na-enweghị mmetụta nke extrusion viscous mgbe ọ na-abịaru nso polymers nwere alaka.Tụkwasị na nke ahụ, ọ na-eme ka o kwe omume ikpebi njirimara arụ ọrụ nke ihe ndị na-ehicha ihe n'otu n'otu na-enweghị mmetụta nke poroelasticity.A na-enweta nke a site n'ịhọrọ nyocha AFM na nhazi (ọnụọgụ ọnụ, geometry na ọnụego mmiri) nke kachasị mma maka ịlele njirimara nke ihe dị nro na ihe atụ nke ndụ.Usoro a na-eme ka ọgụgụ isi na izi ezi maka nha ziri ezi nke ihe dị nro dị nro lehfilcon A, nke nwere modul nke ngbanwe dị oke ala n'elu mpaghara (ihe ruru 2 kPa) yana ngbanwe dị oke n'ime ime (ihe fọrọ nke nta ka ọ bụrụ 100%) gburugburu mmiri. .Nsonaazụ nke ọmụmụ ihe n'elu abụghị naanị ekpughere njirimara dị nro dị nro nke lens lehfilcon A, ma gosikwara na modul nke brushes polymer nwere alaka dị ka nke mkpụrụ silicon-hydrogen.Enwere ike itinye usoro njirimara elu a na ihe ndị ọzọ dị nro na ngwaọrụ ahụike.
Ihe eji arụ ọrụ nke ihe emebere maka kọntaktị na anụ ahụ dị ndụ na-ekpebikarị site na gburugburu ebe obibi.Ngwakọta zuru oke nke ihe onwunwe ndị a na-enyere aka nweta njirimara ụlọ ọgwụ a chọrọ nke ihe ahụ n'emeghị ka nzaghachi cellular na-adịghị mma1,2,3.Maka nnukwu ihe ejikọtara ọnụ, njirimara nke akụrụngwa dị mfe n'ihi enwere usoro ọkọlọtọ na ụzọ nnwale (dịka ọmụmaatụ, microindentation4,5,6).Agbanyeghị, maka ihe ndị dị nro dị nro dị ka gels, hydrogels, biopolymers, cell ndụ, wdg, ụzọ nnwale ndị a anaghị etinye aka na ya n'ihi oke nha nha yana inhomogeneity nke ụfọdụ ihe7.N'ime afọ ndị gafeworonụ, a gbanweela ma gbanwee usoro ntinye nke ọdịnala iji mara ọtụtụ ihe dị nro, ma ọtụtụ ụzọ ka na-enwe mmejọ dị njọ nke na-egbochi ojiji ha8,9,10,11,12,13.Enweghị ụzọ nnwale pụrụ iche nke nwere ike ịkọwapụta n'ụzọ ziri ezi na ntụkwasị obi njirimara akụrụngwa nke ihe supersoft na elu elu na-egbochi oke ojiji ha na ngwa dị iche iche.
N'ime ọrụ anyị gara aga, anyị webatara oghere kọntaktị lehfilcon A (CL), ihe dị nro dị nro nke nwere ihe niile dị nro dị n'elu sitere na atụmatụ biomimetic nke sitere na elu cornea nke anya.Emepụtara biomaterial a site n'ịkwado akwa poly (2-methacryloyloxyethylphosphorylcholine (MPC)) (PMPC) nke nwere alaka, jikọtara ya na polymer na silicone hydrogel (SiHy) 15 emebere maka ngwaọrụ ahụike dabere na.Usoro grafting a na-emepụta oyi akwa n'elu nke nwere usoro ahịhịa polymeric dị nro na nke ukwuu.Ọrụ anyị gara aga ekwenyela na usoro biomimetic nke lehfilcon A CL na-enye ihe ndị dị elu dị elu dị ka mma wetting na mgbochi mmebi, ụbara mmanu, na mbelata cell na adhesion nje bacteria15,16.Na mgbakwunye, ojiji na mmepe nke ihe biomimetic a na-atụkwa aro ka mgbasawanye na ngwaọrụ ọgwụ ndị ọzọ.Ya mere, ọ dị oke mkpa iji mara njirimara elu nke ihe a dị nro ma ghọta usoro mmekọrịta ya na anya iji mepụta ihe ọmụma zuru oke iji kwado mmepe na ngwa n'ọdịnihu.Ọtụtụ lensị kọntaktị SiHy dị maka azụmahịa bụ ngwakọta nke hydrophilic na hydrophobic polymers nke na-etolite n'ụdị ihe eji eme otu17.Emeela ọtụtụ ọmụmụ iji nyochaa akụrụngwa ha site na iji mkpakọ ọdịnala, tensile na usoro nnwale microindentation18,19,20,21.Otú ọ dị, ihe ọhụrụ biomimetic imewe nke lehfilcon A CL na-eme ka ọ bụrụ ihe dị iche iche dị iche iche nke ihe eji arụ ọrụ nke ihe ọkpụkpụ polymer nwere alaka dị iche na nke SiHy base substrate.Ya mere, ọ na-esiri ike ịkọwapụta akụrụngwa ndị a n'ụzọ ziri ezi site na iji usoro omenala na ntinye.Usoro na-ekwe nkwa na-eji usoro nyocha nanoindentation emejuputa atumatu microscopy (AFM), usoro ejirila chọpụta ihe eji arụ ọrụ nke ihe viscoelastic dị nro dị ka mkpụrụ ndụ ndụ na anụ ahụ, yana polymers dị nro22,23,24,25. .,26,27,28,29,30.Na AFM nanoindentation, ihe ndị bụ isi nke nyocha nanoindentation jikọtara ya na ọganihu ọhụrụ na teknụzụ AFM iji nye mmụba nha nha na ule nke ọtụtụ ihe dị egwu supersoft31,32,33,34,35,36.Tụkwasị na nke ahụ, nkà na ụzụ na-enye uru ndị ọzọ dị mkpa site na iji geometric dị iche iche.ntinye na nyocha na ohere nke ịnwale na mgbasa ozi mmiri dị iche iche.
Enwere ike kewaa AFM nanoindentation na ọnọdụ atọ n'ime isi ihe atọ: (1) akụrụngwa (sensors, detectors, probes, etc.);(2) nha nha (dị ka ike, mbugharị, ọsọ, nha rampụ, wdg);(3) Nhazi data (ndozi ntọala, ntule ebe mmetụ aka, data dabara adaba, nhazi ihe, wdg).Nsogbu dị ịrịba ama na usoro a bụ na ọtụtụ ọmụmụ na akwụkwọ na-eji AFM nanoindentation na-akọ dị iche iche quantitative results maka otu sample / cell / ihe ụdị37,38,39,40,41.Dịka ọmụmaatụ, Leka et al.A mụrụ na atụnyere mmetụta nke AFM probe geometry na modul Young tụrụ atụ nke ihe nlere nke hydrogel na-arụkọ ọrụ ọnụ na sel dị iche iche.Ha na-akọ na ụkpụrụ modulus dabere na nhọrọ cantilever na n'ọnụ ọnụ, yana ọnụ ahịa kachasị elu maka nyocha nke nwere ụdị pyramid yana uru kacha ala nke 42 maka nyocha okirikiri.N'otu aka ahụ, Selhuber-Unkel et al.Egosipụtala ka ọsọ indenenter, nha indenenter na ọkpụrụkpụ nke ihe atụ polyacrylamide (PAAM) si emetụta modul Young nke ACM43 nanoindentation tụrụ.Ihe ọzọ na-agbagwoju anya bụ enweghị ọkọlọtọ modul ihe ule dị oke ala yana usoro nnwale efu.Nke a na-eme ka ọ sie ike iji obi ike nweta nsonaazụ ziri ezi.Otú ọ dị, usoro a bara uru nke ukwuu maka nha nhata na nyocha nke dị n'etiti ụdị ihe atụ yiri nke ahụ, dịka ọmụmaatụ site na iji AFM nanoindentation iji chọpụta mkpụrụ ndụ nkịtị site na mkpụrụ ndụ cancer 44, 45.
Mgbe ị na-anwale ihe dị nro na AFM nanoindentation, usoro isi mkpịsị aka n'ozuzu bụ iji nyocha nke nwere obere mmiri na-adịgide adịgide (k) nke dabara nke ọma na modules sample na ọnụ ọnụ hemispherical / gburugburu ka nyocha nke mbụ ghara ịmaba n'elu ihe nlele ahụ. mbụ kọntaktị na ihe dị nro.Ọ dịkwa mkpa na mgbama mgbagha nke nyocha ahụ na-eme ka ọ dị ike nke ọma ka usoro nchọpụta laser chọpụta ya24,34,46,47.N'ihe banyere sel dị iche iche dị nro dị nro, anụ ahụ na gels, ihe ịma aka ọzọ bụ imeri ike nrapado dị n'etiti nyocha na elu nlele iji hụ na nha nwere ike ịtụgharị na ntụkwasị obi48,49,50.Ruo n'oge na-adịbeghị anya, ọtụtụ ọrụ na AFM nanoindentation lekwasịrị anya n'ịmụ banyere omume nhazi nke mkpụrụ ndụ ndụ, anụ ahụ, gels, hydrogels, na biomolecules na-eji nyocha dị iche iche dị iche iche, nke a na-akpọkarị colloidal probes (CPs).. dị ka carbon (DLC).Ọ bụ ezie na CP-AFM nanoindentation na-abụkarị nhọrọ mbụ maka njirimara nlele dị nro, ọ nwere nsogbu na njedebe ya.Ojiji nke nnukwu, micron-spherical sppherical tips na-abawanye mkpokọta kọntaktị nke ọnụ ọnụ ya na ihe nlele ma na-ebute nnukwu mfu nke mkpebi oghere.Maka ihe nlere dị nro, nke na-adịghị mma, ebe akụrụngwa nke ihe ndị dị na mpaghara nwere ike ịdị iche na nkezi karịa mpaghara sara mbara, ntinye CP nwere ike zoo inhomogeneity ọ bụla na akụrụngwa na nha mpaghara52.A na-eme nyocha nke colloidal site na ijikọta spheres colloidal micron na kantilevers na-enweghị isi na-eji adhesives epoxy.Usoro nrụpụta n'onwe ya nwere ọtụtụ nsogbu ma nwee ike ibute enweghị nkwekọrịta na usoro nhazi nyocha.Na mgbakwunye, nha na oke nke ụmụ irighiri ihe nke colloidal na-emetụta kpọmkwem paramita mmezi nke cantilever, dị ka ugboro ugboro resonant, isi ike mmiri, na mmetụta mgbagha56,57,58.Ya mere, usoro ndị a na-ejikarị eme ihe maka nyocha AFM ndị a na-emekarị, dị ka nhazi okpomọkụ, nwere ike ọ gaghị enye nhazi ziri ezi maka CP, na ụzọ ndị ọzọ nwere ike ịchọrọ iji mee mgbanwe ndị a57, 59, 60, 61. Nnwale indentation CP na-ahụkarị na-eji nnukwu cantilever dị iche iche eme ihe. mụọ njirimara nke ihe atụ dị nro, nke na-emepụta nsogbu ọzọ mgbe ị na-emezi omume na-abụghị linear nke cantilever na nnukwu ọdịiche62,63,64.Ụzọ indentation colloidal nke oge a na-eburukarị n'uche geometry nke cantilever ejiri mee nyocha nyocha, mana leghara mmetụta nke ụmụ irighiri ihe colloidal anya, nke na-emepụta ejighị n'aka ọzọ na izi ezi nke usoro ahụ38,61.N'otu aka ahụ, moduli na-agbanwe agbanwe nke agbakọ site na ụdị kọntaktị dabara adaba na-adabere ozugbo na geometry nke nyocha indentation, na adabaghị n'etiti ọnụ na njirimara elu elu nwere ike iduga na-ezighị ezi27, 65, 66, 67, 68. Ụfọdụ ọrụ na-adịbeghị anya nke Spencer et al.Ihe ndị kwesịrị iburu n'uche mgbe a na-akọwapụta brushes polymer dị nro na-eji usoro nanoindentation CP-AFM pụta ìhè.Ha kwuru na njide nke mmiri viscous na brushes polymer dị ka ọrụ nke ọsọ na-ebute mmụba nke ntinye isi na ya mere nha dị iche iche nke ihe ndị na-adabere na ọsọ ọsọ30,69,70,71.
N'ime ọmụmụ ihe a, anyị ejirila usoro nanoindentation AFM gbanwetụrụ.N'iburu ihe onwunwe na nhazi ọhụrụ nke ihe a, ihe mgbagwoju anya nke usoro ntinye nke omenala bụ n'ụzọ doro anya ezughị ezu iji mara modul nke ihe a dị oke nro, n'ihi ya, ọ dị mkpa iji usoro nanoindentation AFM nwere mmetụta dị elu na mmetụta dị ala.ọkwa.Mgbe anyị nyochachara adịghị ike na nsogbu nke colloidal AFM probe nanoindentation techniques dị ugbu a, anyị na-egosi ihe mere anyị ji họrọ obere nyocha AFM emebere nke ọma iji kpochapụ mmetụta uche, mkpọtụ ndabere, akara akara kọntaktị, tụọ modul ngwa ngwa nke ihe dị iche iche dị nro dị ka njide mmiri. ndabere.na ọnụọgụgụ ziri ezi.Na mgbakwunye, anyị nwere ike ịlele n'ụzọ ziri ezi na udi na akụkụ nke indentation n'ọnụ, na-enye anyị ohere iji cone-sphere dabara nlereanya iji chọpụta modul nke elasticity na-enweghị na-atụle kọntaktị ebe nke n'ọnụ na ihe onwunwe.Echiche abụọ ahụ ezoro ezo nke agbakọtara n'ọrụ a bụ njirimara ihe na-agbanwe agbanwe na modul nke nnwere onwe dị omimi nke indentation.N'iji usoro a, anyị buru ụzọ nwalee ụkpụrụ ultra- soft na modul mara amara iji tụọ usoro ahụ, wee jiri usoro a kọwaa elu nke ihe oghere kọntaktị abụọ dị iche iche.A na-atụ anya ka usoro a iji mara AFM nanoindentation na-enwe mmetụta dị ukwuu na ọ ga-adaba n'ọtụtụ ihe dị iche iche nke biomimetic heterogeneous ultrasoft nwere ike iji na ngwaọrụ ahụike yana ngwa biomedical.
Lehfilcon A lenses kọntaktị (Alcon, Fort Worth, Texas, USA) na silicone hydrogel substrates ka ahọpụtara maka nnwale nanoindentation.A na-eji ihe mgbakwasa oghere a haziri ahazi na nnwale ahụ.Iji wụnye oghere maka ule, a na-etinye ya nke ọma n'elu ihe nkwụnye nke dome, jide n'aka na ọ dịghị ikuku ikuku batara n'ime ya, wee dozie ya na nsọtụ ya.Oghere dị na ntinye n'elu nke oghere oghere na-enye ohere ịbanye na etiti anya nke oghere maka nyocha nanoindentation ka ọ na-ejide mmiri mmiri.Nke a na-eme ka anya m hụ mmiri nke ọma.A na-eji 500 μl nke ngwọta nkwakọ lens kọntaktị mee ihe dị ka ihe nyocha.Iji nyochaa ọnụọgụ ọnụọgụgụ, a kwadebere hydrogels polyacrylamide na-adịghị arụ ọrụ nke azụmahịa (PAAM) sitere na ihe mejupụtara polyacrylamide-co-methylene-bisacrylamide (100 mm Petrisoft Petri efere, Matrigen, Irvine, CA, USA), ihe na-agbanwe agbanwe nke 1. kPa.Jiri 4-5 tụlee (ihe dị ka 125 µl) nke phosphate buffered saline (PBS si Corning Life Sciences, Tewkesbury, MA, USA) na 1 dobe nke OPTI-FREE Puremoist lens lens (Alcon, Vaud, TX, USA).) na AFM hydrogel-probe interface.
Ihe nlele nke Lehfilcon A CL na SiHy substrates ka a na-ahụ anya site na iji usoro FEI Quanta 250 Field Emission Scanning Electron Microscope (FEG SEM) nke nwere ihe nchọta Nnyefe Electron Microscope (STEM).Iji kwadebe ihe nlele ndị ahụ, a na-ebu ụzọ were mmiri saa anya anya m wee bee n'ime ibe ndị yiri achịcha.Iji nweta ọdịiche dị iche n'etiti hydrophilic na hydrophobic components nke ihe atụ, a na-eji 0.10% kwụsie ike nke RuO4 dị ka agba agba, bụ nke a na-emikpu ihe atụ maka 30 min.Ihe nkedo nke lehfilcon A CL RuO4 dị mkpa ọ bụghị naanị iji nweta ọdịiche dị iche iche dị mma, kamakwa na-enyere aka ichekwa usoro nke brushes polima nke nwere alaka n'ụdị mbụ ha, bụ nke a na-ahụ anya na ihe oyiyi STEM.Mgbe ahụ, a na-asachapụ ha ma kpochaa ha n'usoro ethanol/mmiri ngwakọta na-abawanye ụba nke ethanol.A na-eji EMBed 812/Araldite epoxy wụba ihe atụ ndị ahụ, nke gwọrọ n'abali na 70°C.A na-eji ultramicrome gbutu ihe nlere nke enwetara site na resin polymerization, wee jiri ihe nchọta STEM na-ahụta akụkụ ndị ahụ dị mkpa na ọnọdụ oghere dị ala na voltaji na-agba ọsọ nke 30 kV.Ejiri otu usoro SEM ahụ maka nkọwa zuru ezu nke nyocha PFQNM-LC-A-CAL AFM (Bruker Nano, Santa Barbara, CA, USA).E nwetara onyonyo SEM nke nyocha AFM n'ụdị oghere dị elu nke nwere oke ọkụ nke 30 kV.Nweta onyonyo n'akụkụ dị iche iche na nnukwu ihe ka ịdekọ nkọwapụta niile nke ọdịdị na nha nke ntụle nyocha AFM.A tụrụ akụkụ niile nke mmasị na onyonyo a na dijitalụ.
Ejiri microscope nke FastScan Bio Icon atomic Force (Bruker Nano, Santa Barbara, CA, USA) nwere ụdị “PeakForce QNM in Fluid” iji hụ anya wee mee nanoindentate lehfilcon A CL, SiHy substrate, na nlele PAAm hydrogel.Maka nnwale onyonyo, a na-eji nyocha PEAKFORCE-HIRS-FA (Bruker) nwere radius ọnụ ọnụ nke 1 nm iji weghara onyonyo mkpebi dị elu nke ihe nlele ahụ na ọnụ ọgụgụ nyocha nke 0.50 Hz.Ewere ihe onyonyo niile na ngwọta mmiri.
Emere nnwale nanoindentation AFM site na iji nyocha PFQNM-LC-A-CAL (Bruker).Ihe nyocha AFM nwere ntu silicon na nitride cantilever 345 nm gbara ọkpụrụkpụ, 54 µm ogologo na 4.5 µm n'obosara yana oge resonant nke 45 kHz.Emebere ya ka ọ mara ma rụpụta nha nanomechanical quantitative na nlere ihe ndụ dị nro.A na-ahazi sensọ ndị ahụ n'otu n'otu na ụlọ ọrụ mmepụta ihe nwere ntọala mmiri emebere nke ọma.Ihe nkwụsị mmiri nke nyocha ndị e ji mee ihe n'ọmụmụ ihe a dị na 0.05-0.1 N/m.Iji chọpụta n'ụzọ ziri ezi na ọdịdị na nha nke ọnụ ọnụ ahụ, a na-eji SEM mara nyocha ahụ n'ụzọ zuru ezu.Na fig.Ọnụọgụ 1a na-egosi mkpebi dị elu, nyocha micrograph eletrọn dị ala nke nyocha PFQNM-LC-A-CAL, na-enye echiche zuru oke maka nhazi nyocha.Na fig.1b na-egosi amụbawanye anya nke elu n'ọnụ nyocha, na-enye ozi gbasara ọdịdị na nha nke ọnụ ọnụ.Na njedebe dị oke egwu, agịga bụ ihe dị ka 140 nm na dayameta (Fig 1c).N'okpuru ebe a, ọnụ ya na-adaba n'ụdị conical, na-eru ogologo nke ihe dịka 500 nm.Na mpụga mpaghara tapering, ọnụ ya bụ cylindrical wee kwụsị n'ogologo ọnụ ọnụ nke 1.18 µm.Nke a bụ akụkụ bụ isi na-arụ ọrụ nke n'ọnụ nyocha.Na mgbakwunye, a na-ejikwa nnukwu nyocha polystyrene (PS) (Novascan Technologies, Inc., Boone, Iowa, USA) nke nwere dayameta n'ọnụ nke 45 µm na mmiri na-adịgide adịgide nke 2 N/m maka nnwale dị ka nyocha colloidal.ya na nyocha PFQNM-LC-A-CAL 140 nm maka ntụnyere.
A kọwapụtara na mmiri nwere ike ọnyà n'etiti nyocha AFM na usoro ahịhịa polymer n'oge nanoindentation, nke ga-eme ka ike dị elu na nyocha AFM tupu ya emetụ elu69.Mmetụta extrusion a viscous n'ihi njide mmiri nwere ike gbanwee ebe kọntaktị pụtara ìhè, si otú ahụ na-emetụta nha modulus elu.Iji mụọ mmetụta nke geometry nyocha na ọsọ indentation na njide mmiri, a kpara nkata ike indentation maka nlele lehfilcon A CL site na iji nyocha dayameta 140 nm na ọnụego mbugharị oge niile nke 1 µm/s na 2 μm/s.Dayameta nyocha 45 µm, ntọala ike kwụ ọtọ 6 nN nwetara na 1 µm/s.A na-eme nnwale na nyocha 140 nm na dayameta na ọsọ ntinye nke 1 µm / s na ike setịpụrụ 300 pN, nke a họọrọ iji mepụta nrụgide kọntaktị n'ime physiological nso (1-8 kPa) nke nkuchianaya elu.nrụgide 72. A na-anwale ihe nlele dị nro nke PAA hydrogel na nrụgide nke 1 kPa maka ike ntinye nke 50 pN na ọsọ nke 1 μm / s site na iji nyocha nke nwere dayameta nke 140 nm.
Ebe ọ bụ na ogologo akụkụ conical nke n'ọnụ nke PFQNM-LC-A-CAL nyocha bụ ihe dị ka 500 nm, maka ihe ọ bụla indentation omimi <500 nm ọ nwere ike na-eche na geometry nke nyocha n'oge indentation ga-anọgide na eziokwu na ya. ọdịdị cone.Tụkwasị na nke ahụ, a na-eche na elu nke ihe dị n'okpuru ule ga-egosipụta nzaghachi na-agbanwe agbanwe, nke a ga-ekwenyekwa na ngalaba ndị a.Ya mere, dabere na ọdịdị na nha nke ọnụ ọnụ ahụ, anyị họọrọ ụdị cone-sphere dabara adaba nke Briscoe, Sebastian na Adams mepụtara, nke dị na ngwanrọ nke onye na-ere ahịa, iji hazie nnwale nanoindentation AFM anyị (NanoScope).Akụrụngwa nyocha nkewa nkewa, Bruker) 73. Nlereanya ahụ na-akọwa mmekọrịta mmanye-nchụpụ F(δ) maka cone nwere ntụpọ sppherical apex.Na fig.Ọgụgụ 2 na-egosi geometry kọntaktị n'oge mkparịta ụka nke cone siri ike na n'ọnụ okirikiri, ebe R bụ radius nke n'ọnụ okirikiri, a bụ radius kọntaktị, b bụ radius kọntaktị na njedebe nke okirikiri, δ bụ radius kọntaktị.omimi indentation, θ bụ ọkara akụkụ nke cone.Onyonyo SEM nke nyocha a na-egosi n'ụzọ doro anya na 140 nm dayameta spherical n'ọnụ ọnụ na-ejikọta ya na cone, yabụ ebe a ka akọwara b naanị site na R, ntụgharị b = R cos θ.Akụrụngwa na-ere ahịa na-enye mmekọrịta cone-sphere iji gbakọọ ụkpụrụ nke Young's modulus (E) site na ike nkewa data na-ewere a > b.Mmekọrịta:
ebe F bụ ike indentation, E bụ modul nke Young, ν bụ oke Poisson.Enwere ike ịkọwa radius kọntaktị a site na iji:
Atụmatụ geometry kọntaktị nke cone siri ike nke nwere ọnụ ntụ ntụ ntụ ntụ nke etinyere n'ime ihe nke oghere kọntaktị Lefilcon nwere akwa elu nke brushes polymer nwere alaka.
Ọ bụrụ na ≤ b, njikọ ahụ na-ebelata na nhata maka ihe ntinye okirikiri;
Anyị kwenyere na mmekọrịta nke nyocha indenting na nhazi alaka nke PMPC polymer brush ga-eme ka radius kọntaktị dị ukwuu karịa radius kọntaktị gburugburu b.Ya mere, maka nha nha nha ọ bụla nke mgbanwe mgbanwe emere na ọmụmụ a, anyị jiri ndabere enwetara maka ikpe a> b.
Ihe onyonyo ultrasoft biomimetic a mụrụ n'ọmụmụ ihe a bụ nke egosipụtara nke ọma site na iji nyocha eletrọnịkị eletrọnịkị (STEM) nke akụkụ ihe nlere anya na atomiki microscopy (AFM) nke elu.Emere nkọwapụta nke elu elu a dị ka ndọtị nke ọrụ anyị ebipụtara na mbụ, nke anyị kpebiri na usoro ahịhịa polymeric na-agbanwe agbanwe nke PMPC-modified lehfilcon A CL elu gosipụtara ihe eji arụ ọrụ yiri ya na anụ ahụ corneal 14.N'ihi nke a, anyị na-ezo aka na oghere oghere kọntaktị dị ka ihe biomimetic14.Na fig.3a,b na-egosi akụkụ obe nke ngalaba PMPC polymer ahịhịa arụrụ arụ n'elu ala lehfilcon A CL na mkpụrụ SiHy anaghị agwọ ya, n'otu n'otu.A na-enyocha ihe ndị dị n'elu nke ihe atụ abụọ ahụ site na iji ihe oyiyi AFM dị elu, nke kwadoro nsonaazụ nke nyocha STEM (Fig 3c, d).Ejikọtara ọnụ, onyogho ndị a na-enye ihe dị ka ogologo nke PMPC branched polymer brush structure na 300-400 nm, nke dị oke mkpa maka ịkọwa nha nanoindentation AFM.Ihe ọzọ dị mkpa na-achọpụta na ihe oyiyi bụ na n'ozuzu elu Ọdịdị nke CL biomimetic ihe bụ morphologically dị iche iche na nke SiHy substrate ihe.Ọdịiche a na ọdịdị ọdịdị elu ha nwere ike ịpụta ìhè n'oge mkparịta ụka ha na nyocha AFM indenting na emesịa na ụkpụrụ modulus atụtara.
Ihe onyonyo STEM nke dị n'ofe nke (a) lehfilcon A CL na (b) mkpụrụ SiHy.Ogwe ọnụ ọgụgụ, 500 nm.Ihe oyiyi AFM nke elu nke lehfilcon A CL substrate (c) na ntọala SiHy (d) (3 µm × 3 µm).
polymers sitere n'ike mmụọ nsọ na ihe owuwu ahịhịa polymer dị nro ma bụrụ nke a mụọla ya nke ọma wee jiri ya na ngwa biomedical dị iche iche74,75,76,77.Ya mere, ọ dị mkpa iji usoro nanoindentation AFM, nke nwere ike ịlele n'ụzọ ziri ezi na ịdabere na ihe onwunwe ha.Ma n'otu oge ahụ, njirimara pụrụ iche nke ihe ndị a dị nro dị nro, dị ka modul elastic dị oke ala, nnukwu mmiri mmiri na ịdị elu dị elu, na-emekarị ka ọ sie ike ịhọrọ ihe ziri ezi, ọdịdị na ọdịdị nke nyocha indenting.nha.Nke a dị mkpa ka onye na-eme ihe ahụ ghara ịkaba elu dị nro nke ihe nlele ahụ, nke ga-eduga ná njehie n'ịchọpụta ebe kọntaktị na elu na mpaghara kọntaktị.
Maka nke a, nghọta zuru oke banyere ọdịdị ọdịdị nke ultra-soft biomimetic material (lehfilcon A CL) dị mkpa.Ozi gbasara nha na nhazi nke brushes polima nwere alaka nke enwetara site na iji usoro onyonyo na-enye ihe ndabere maka njirimara igwe nke elu site na iji usoro nanoindentation AFM.Kama nyocha nke sppherical colloidal micron, anyị họọrọ PFQNM-LC-A-CAL silicon nitride probe (Bruker) nke nwere dayameta n'ọnụ nke 140 nm, nke emebere nke ọma maka maapụ ọnụọgụ nke akụrụngwa nke ihe atụ ndu 78, 79, 80. , 81, 82, 83, 84 Enwere ike ịkọwa ebumnuche maka iji nyocha dịtụ nkọ ma e jiri ya tụnyere ihe nyocha nke colloidal a na-ahụkarị site na njirimara nhazi nke ihe ahụ.N'ịtụle nha nyocha nyocha (~ 140 nm) na brushes polymer nwere alaka n'elu CL lehfilcon A, nke egosiri na Fig. 3a, enwere ike ikwubi na ọnụ ọnụ ahụ buru ibu iji bata kpọmkwem na kọntaktị ndị a na-acha ahịhịa ndụ. na-ebelata ohere nke ọnụ na-adụpu site na ha.Iji kọwaa isi ihe a, na Fig. 4 bụ ihe oyiyi STEM nke lehfilcon A CL na ntinye ntinye nke nyocha AFM (esere na nha).
Eserese na-egosi onyonyo STEM nke lehfilcon A CL yana nyocha indentation ACM (esere na nha).
Tụkwasị na nke ahụ, ọnụ ọgụgụ nke 140 nm dị ntakịrị iji zere ihe ize ndụ nke ọ bụla n'ime mmetụta extrusion na-egbuke egbuke nke a kọrọ na mbụ maka brushes polymer mepụtara site na CP-AFM nanoindentation method69,71.Anyị na-eche na n'ihi ọdịdị cone-spherical pụrụ iche na nke dị ntakịrị nke n'ọnụ AFM a (Fig 1), ọdịdị ike nke ike na-eme site na lehfilcon A CL nanoindentation agaghị adabere na ntinye ọsọ ọsọ ma ọ bụ nbudata / nbudata ọsọ. .Ya mere, mmetụta poroelastic adịghị emetụta ya.Iji nwalee echiche a, ihe nlele lehfilcon A CL ka etinyere n'ike siri ike site na iji nyocha PFQNM-LC-A-CAL, mana n'ọsọ abụọ dị iche iche, a na-eji ihe mgbagha na mgbagha na-esi na ya pụta mee atụmatụ ike (nN) na nkewa (µm) ka egosiri na foto 5a.O doro anya na ike na-atụgharị n'oge nbudata na nbudata kpamkpam, ọ nweghịkwa ihe akaebe doro anya na ike ịkwa akwa na omimi ntinye efu na-abawanye na ọsọ ọsọ na ọnụ ọgụgụ ahụ, na-atụ aro na a na-eji ihe ndị ahụ n'otu n'otu mara ihe na-enweghị mmetụta poroelastic.N'ụzọ dị iche, mmetụta njide mmiri (mpụpụ viscous na mmetụta poroelasticity) pụtara ìhè maka nyocha 45 µm dayameta AFM n'otu ọsọ ọsọ ahụ ma na-egosipụta ya site na hysteresis n'etiti mgbatị na mgbagha, dị ka egosiri na Figure 5b.Nsonaazụ ndị a na-akwado echiche ahụ ma na-atụ aro na nyocha dayameta 140 nm bụ ezigbo nhọrọ maka ịkọwapụta ebe dị nro dị otú ahụ.
lehfilcon A CL indentation ike akụkụ iji ACM;(a) na-eji nyocha nke nwere dayameta nke 140 nm na ọnụego ntinye abụọ, na-egosi enweghị mmetụta poroelastic n'oge ntinye elu;(b) iji nyocha nwere dayameta nke 45 µm na 140 nm.s na-egosi mmetụta nke extrusion viscous na poroelasticity maka nnukwu nyocha ma e jiri ya tụnyere obere nyocha.
Iji mara elu ultrasoft, ụzọ AFM nanoindentation ga-enwerịrị nyocha kachasị mma iji mụọ njirimara nke ihe a na-amụ.Na mgbakwunye na n'ọnụ ọnụ na nha, uche nke AFM detector usoro, uche na ntughari deflection na ule gburugburu ebe obibi, na cantilever siri ike na-arụ ọrụ dị mkpa n'ịchọpụta izi ezi na ntụkwasị obi nke nanoindentation.nha.Maka usoro AFM anyị, njedebe nke nchọpụta ọnọdụ Sensitive (PSD) bụ ihe dịka 0.5 mV ma dabere na ọnụego mmiri a na-edozi tupu oge eruo na mgbagwoju anya mmiri nke PFQNM-LC-A-CAL nyocha, nke kwekọrọ na echiche ibu uche.erughị 0.1 pN.Ya mere, usoro a na-enye ohere ịlele opekempe ike indentation ≤ 0.1 pN na-enweghị akụkụ mkpọtụ ọ bụla.Otú ọ dị, ọ fọrọ nke nta ka ọ bụrụ ihe na-agaghị ekwe omume maka usoro AFM iji belata ụda gburugburu na ọkwa a n'ihi ihe ndị dị ka vibration ígwè na mmiri mmiri.Ihe ndị a na-egbochi mmetụta zuru oke nke usoro nanoindentation AFM ma na-ebutekwa ụda ụda n'azụ ihe dịka ≤ 10 pN.Maka njirimara elu, lehfilcon A CL na SiHy substrate samples ka etinyere n'okpuru ọnọdụ hydrated zuru oke site na iji nyocha 140 nm maka njirimara SEM, na ihe ndị na-akpata ike na-adabere n'etiti ike (pN) na nrụgide.Egosiri atụmatụ nkewa (µm) na eserese 6a.E jiri ya tụnyere SiHy base substrate, eriri lehfilcon A CL na-egosi n'ụzọ doro anya usoro mgbanwe na-amalite n'ebe a na-akpakọrịta ya na brush polymer forked ma na-ejedebe na mgbanwe dị nkọ na kọntaktị akara mkpọda nke ọnụ ya na ihe dị n'okpuru.Akụkụ mgbanwe a nke mgbanaka ike na-egosipụta omume ngbanwe n'ezie nke ahịhịa polima nke nwere alaka n'elu, dị ka egosipụtara site na mgbakọ mkpakọ na-esochi usoro mgbakasị ahụ na ọdịiche dị na akụrụngwa n'etiti usoro ahịhịa na nnukwu ihe SiHy.Mgbe atụnyere lefilcon.Nkewa nke nkezi ogologo nke a branched polima ahịhịa na STEM oyiyi nke PCS (Fig. 3a) na ike curve tinyere abscissa na fig. 3a.6a na-egosi na usoro ahụ na-enwe ike ịchọpụta ọnụ na polymer nwere alaka na-eru n'elu elu.Kọntaktị n'etiti ụlọ ahịhịa.Na mgbakwunye, nchikota nso nke akụkụ nke ike na-egosi enweghị mmetụta njide mmiri.N'okwu a, ọ dịghị ihe ọ bụla adhesion n'etiti agịga na elu nke sample.Akụkụ ndị kachasị elu nke ntụgharị ike maka ihe atụ abụọ ahụ na-ekpuchi, na-egosipụta myirịta nke ihe eji arụ ọrụ nke ihe ndị ahụ.
(a) AFM nanoindentation force curves maka lehfilcon A CL substrates na SiHy substrates, (b) ike usoro na-egosi ntule ebe kọntaktị site na iji usoro nbata mkpọtụ ndabere.
Iji mụọ nkọwa kachasị mma nke mgbagha ike, a na-atụgharịgharị mgbagwoju anya nke lehfilcon A CL sample na Fig. 6b na ike kachasị nke 50 pN tinyere y-axis.Eserese a na-enye ozi dị mkpa gbasara mkpọtụ ndabere izizi.Mkpọtụ ahụ dị na nso nke ± 10 pN, nke a na-eji mee ihe iji chọpụta kpọmkwem ebe kọntaktị ma gbakọọ omimi ntinye.Dịka akọwara n'akwụkwọ akụkọ ahụ, njirimara ebe kọntaktị dị oke mkpa iji nyochaa nke ọma ihe onwunwe dịka modulus85.Ụzọ metụtara nhazi akpaaka nke data mgbagharị ike egosila na ọ dabara adaba n'etiti nha data dabara na nha maka ihe dị nro86.N'ime ọrụ a, nhọrọ anyị nke isi kọntaktị dị mfe ma bụrụ nke ebumnobi, mana o nwere oke ya.Ụzọ nleba anya anyị iji chọpụta ebe a na-akpọtụrụ nwere ike ime ka ụkpụrụ modulus dị ntakịrị karịa nke ọma maka obere omimi indentation (<100 nm).Ojiji nke nchọta ihe mmetụ aka dabere na algọridim na nhazi data akpaaka nwere ike ịbụ ihe na-aga n'ihu nke ọrụ a n'ọdịnihu iji meziwanye usoro anyị.Ya mere, maka mkpọtụ ndabere dị n'ime usoro nke ± 10 pN, anyị na-akọwapụta ebe kọntaktị dị ka ebe mbụ data na x-axis na Figure 6b na uru nke ≥10 pN.Mgbe ahụ, dị ka ọnụ ọgụgụ mkpọtụ nke 10 pN si dị, ahịrị kwụ ọtọ na ọkwa nke ~ 0.27 µm na-egosi akara kọntaktị na elu, mgbe nke ahụ gasịrị, mgbatị ahụ na-agbatị na-aga n'ihu ruo mgbe mkpụrụ ahụ zutere omimi nke ~ 270 nm.N'ụzọ na-akpali mmasị, dabere na nha njirimara ahịhịa polymer nke alaka (300-400 nm) tụrụ site na iji usoro onyonyo, omimi ntinye nke CL lehfilcon Ihe nlele a hụrụ site na iji usoro mkpọtụ mkpọtụ ndabere bụ ihe dịka 270 nm, nke dị nso na ya. nha nha na STEM.Nsonaazụ ndị a na-akwadokwa ndakọrịta na ntinye nke ọdịdị na nha nke ntụle nyocha AFM maka ntinye nke usoro ahịhịa polymer a dị nro ma na-agbanwe nke ukwuu.Ihe data a na-enyekwa ihe akaebe siri ike iji kwado usoro anyị si eji mkpọtụ n'azụ mee ihe dị ka ọnụ ụzọ maka ịkọwapụta ebe kọntaktị.Ya mere, nsonaazụ ọnụọgụ ọ bụla enwetara site na nhazi mgbakọ na mwepụ yana ndochi ike kwesịrị ịbụ nke ziri ezi.
Ntụle ọnụọgụ site na ụzọ AFM nanoindentation na-adabere kpamkpam na ụdị mgbakọ na mwepụ ejiri maka nhọrọ data na nyocha na-esote.Ya mere, ọ dị mkpa ịtụle ihe niile metụtara nhọrọ nke indenter, ihe onwunwe ihe na usoro nke mmekọrịta ha tupu ịhọrọ otu ụdị.N'okwu a, ejiri nlezianya mara geometry n'ọnụ site na iji micrographs SEM (Fig 1), dabere na nsonaazụ ya, 140 nm dayameta AFM nanoindenting probe na cone siri ike na geometry okirikiri bụ nhọrọ dị mma maka ịkọwapụta lehfilcon A CL79 samples. .Ihe ọzọ dị mkpa nke kwesịrị iji nlezianya nyochaa bụ ngbanwe nke ihe polymer a na-anwale.Ọ bụ ezie na data mbụ nke nanoindentation (Figs. 5a na 6a) na-akọwapụta n'ụzọ doro anya atụmatụ nke overlapping nke esemokwu na mkpakọ akụkụ, ya bụ, mgbake na-agbanwe agbanwe nke ihe ahụ, ọ dị oke mkpa iji kwado ọdịdị na-agbanwe agbanwe nke kọntaktị. .Iji mezuo nke a, a na-eme ntinye abụọ na-aga n'ihu n'otu ebe n'elu ihe nlele lehfilcon A CL na ọnụego ntinye nke 1 µm/s n'okpuru ọnọdụ hydration zuru oke.A na-egosi data ntụgharị ike arụpụta na fig.7 na, dị ka a na-atụ anya ya, mgbasawanye na mkpakọ nke mbipụta abụọ ahụ fọrọ nke nta ka ọ bụrụ otu, na-egosipụta ịdị elu dị elu nke nhazi brush polymer nke alaka.
Ọkpụkpụ ike ntinye abụọ n'otu ebe dị n'elu lehfilcon A CL na-egosi ngbanwe dị mma nke elu oghere oghere.
Dabere na ozi sitere na SEM na ihe oyiyi STEM nke nyocha nyocha na lehfilcon A CL elu, n'otu n'otu, ụdị cone-sphere bụ ihe nnọchianya mgbakọ na mwepụ ezi uche dị na mmekọrịta dị n'etiti ọnụ ọgụgụ nyocha AFM na ihe polymer dị nro na-anwale.Na mgbakwunye, maka ụdị cone-sphere a, echiche ndị bụ isi gbasara njirimara na-agbanwe nke ihe a na-ebipụta na-ejide eziokwu maka ihe ọhụrụ biomimetic a ma jiri ya tụọ modul elastic.
Mgbe nyochachara nke ọma nke usoro nanoindentation AFM na ihe ndị mejupụtara ya, gụnyere njirimara nyocha indentation (ụdị, nha, na isi ike mmiri), uche (mkpọtụ azụ na nleba anya ebe kọntaktị), na ụdị dabara data (ọnụọgụ modulus nha), usoro ahụ bụ. eji.kọwapụta ihe nlele dị nro dị n'ahịa iji nyochaa ọnụọgụ ọnụọgụgụ.A nwalere polyacrylamide nke azụmahịa (PAAM) hydrogel nwere modul na-agbanwe nke 1 kPa n'okpuru ọnọdụ hydrated site na iji nyocha 140 nm.Enyere nkọwa nke ule modul na mgbako na ozi mgbakwunye.Nsonaazụ gosiri na nkezi modulus tụrụ bụ 0.92 kPa, na % RSD na pasentị (%) iche site na modul amara ama erughi 10%.Nsonaazụ ndị a na-akwado izi ezi na mmụgharị nke usoro AFM nanoindentation ejiri na ọrụ a tụọ moduli nke ihe ultrasoft.A kọwakwuru n'elu ala nke lehfilcon A CL sample na SiHy base substrate site na iji otu usoro nanoindentation AFM iji mụọ modul kọntaktị pụtara ìhè nke elu ultrasoft dị ka ọrụ nke omimi ntinye.A na-emepụta akụkụ nkewa nke ike indentation maka ụdị atọ nke ụdị ọ bụla (n = 3; otu ntinye n'otu ụdị) na ike nke 300 pN, ọsọ nke 1 µm / s, na hydration zuru ezu.A na-eji ụdị cone-sphere were ihe dị ka akụkụ nkesa ike ntinye aka.Iji nweta modul dabere na omimi indentation, a na-edobe akụkụ 40 nm n'obosara nke ike ike na mmụba ọ bụla nke 20 nm na-amalite site na ebe kọntaktị, wee tụọ ụkpụrụ nke modulus na nzọụkwụ ọ bụla nke ike ike.Spin Cy et al.A na-eji usoro yiri nke a iji mara modul gradient nke poly(lauryl methacrylate) (P12MA) polima brushes na-eji colloidal AFM probe nanoindentation, na ha kwekọrọ na data site na iji Hertz kọntaktị nlereanya.Ụzọ a na-enye atụmatụ nke kọntaktị modulus pụtara ìhè (kPa) na omimi indentation (nm), dị ka egosiri na eserese 8, nke na-egosipụta njirimara kọntaktị modul/ omimi gradient.Ihe ngbanwe agba agba agbagoro nke CL lehfilcon A sample dị na oke nke 2-3 kPa n'ime 100 nm nke elu nke ihe nlele ahụ, gafere nke ọ na-amalite ịbawanye na omimi.N'aka nke ọzọ, mgbe ị na-anwale mkpụrụ osisi SiHy na-enweghị ihe nkiri dị ka ahịhịa n'elu, ihe omimi kachasị elu nke a na-enweta na ike nke 300 pN bụ ihe na-erughị 50 nm, na uru modul enwetara na data bụ ihe dịka 400 kPa. , nke tụnyere ụkpụrụ nke Young's modulus maka nnukwu ihe.
Modules kọntaktị pụtara ìhè (kPa) vs. omimi indentation (nm) maka lehfilcon A CL na SiHy substrates na-eji usoro nanoindentation AFM nwere geometry cone-sphere iji tụọ modulus.
N'elu elu nke akwụkwọ akụkọ biomimetic branched polymer brush Ọdịdị na-egosipụta modul nke elasticity dị oke ala (2-3 kPa).Nke a ga-adaba na njedebe nkwudo n'efu nke brush polymer forked dị ka egosiri na foto STEM.Ọ bụ ezie na enwere ụfọdụ ihe akaebe nke gradient modul n'akụkụ mpụta nke CL, isi mkpụrụ modulus dị elu na-enwe mmetụta karịa.Otú ọ dị, elu 100 nm nke elu dị n'ime 20% nke ngụkọta ogologo nke alaka polima brush, n'ihi ya, ọ bụ ihe ezi uche dị na ya iche na a tụrụ atụ ụkpụrụ nke modulus na nke a indentation omimi nso bụ ihe ziri ezi na adịghị ike ike. dabere na mmetụta nke ihe ala.
N'ihi ihe pụrụ iche biomimetic imewe nke lehfilcon A lenses kọntaktị, nke nwere alaka PMPC polymer ahịhịa arụdo anyadokwa n'elu nke SiHy substrates, ọ na-esiri nnọọ ike iji ntụkwasị obi mara n'ibu Njirimara nke elu ihe owuwu ha site na iji omenala nha.N'ebe a, anyị na-eweta usoro nanoindentation AFM dị elu maka ịkọwapụta nke ọma ihe ndị dị nro dị ka lefilcon A nwere nnukwu mmiri na ngbanwe dị elu.Usoro a dabere na iji nyocha AFM nke a na-ahọrọ nke ọma n'ọnụ ọnụ ya na geometry ka ọ dabara na akụkụ nhazi nke njirimara elu dị nro nke a ga-ebipụta.Nchikota nke nha n'etiti nyocha na ihe owuwu na-enye mmụba dị ukwuu, na-enye anyị ohere ịlele modul dị ala na ihe na-agbanwe agbanwe nke ihe ndị na-eme ahịhịa polymer gbara agba, n'agbanyeghị mmetụta poroelastic.Nsonaazụ gosiri na prọfesọ PMPC polima nwere alaka pụrụ iche njirimara nke oghere oghere nwere modul ngbanwe dị oke ala (ruo 2kPa) yana ngbanwe dị elu (ihe fọrọ nke nta ka ọ bụrụ 100%) mgbe a nwalere ya na mpaghara mmiri.Nsonaazụ nke AFM nanoindentation nyekwara anyị ohere ịkọwapụta kọntaktị modulus / omimi gradient (30 kPa / 200 nm) nke elu lens biomimetic.gradient a nwere ike ịbụ n'ihi ọdịiche dị n'etiti brushes polima nwere alaka na mkpụrụ SiHy, ma ọ bụ nhazi/njupụta nke brushes polymer, ma ọ bụ ngwakọta ya.Agbanyeghị, ọmụmụ ihe omimi ọzọ dị mkpa iji ghọta nke ọma mmekọrịta dị n'etiti nhazi na akụrụngwa, ọkachasị mmetụta nke ahịhịa ahịhịa na akụrụngwa akụrụngwa.Ntụle ndị yiri ya nwere ike inye aka kọwaa njirimara arụ ọrụ nke elu nke ihe ndị ọzọ dị nro na ngwaọrụ ahụike.
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