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Tare da haɓaka sabbin kayan aiki masu laushi sosai don na'urorin likitanci da aikace-aikacen likitanci, cikakken fasalin halayensu na zahiri da na injiniya yana da mahimmanci kuma yana da ƙalubale. An yi amfani da dabarar nanoindentation ta atomatik mai gyara (AFM) don kwatanta yanayin saman da ke ƙasa sosai na sabon ruwan tabarau na silicone hydrogel mai launi mai launin ruwan kasa ... Sakamakon binciken saman ba wai kawai ya bayyana halayen saman ruwan tabarau na lehfilcon A mai laushi ba, har ma ya nuna cewa tsarin goge-goge na polymer mai rassan ya yi daidai da na substrate na silicon-hydrogen. Ana iya amfani da wannan dabarar siffanta saman ga wasu kayan aiki da na'urorin likitanci masu laushi sosai.
Sau da yawa ana tantance halayen injiniya na kayan da aka tsara don hulɗa kai tsaye da kyallen halitta ta hanyar yanayin halittu. Daidaiton waɗannan kaddarorin abu yana taimakawa wajen cimma halayen asibiti da ake so na kayan ba tare da haifar da mummunan martani ga ƙwayoyin halitta ba1,2,3. Ga kayan da suka yi kama da juna, halayyar halayen injiniya abu ne mai sauƙi saboda samuwar hanyoyin da aka saba da hanyoyin gwaji (misali, microindentation4,5,6). Duk da haka, ga kayan da suka yi laushi kamar gels, hydrogels, biopolymers, ƙwayoyin halitta masu rai, da sauransu, waɗannan hanyoyin gwaji gabaɗaya ba sa aiki saboda iyakokin ƙudurin aunawa da rashin daidaituwar wasu kayan7. Tsawon shekaru, an gyara hanyoyin shigar da kayan ciki na gargajiya kuma an daidaita su don siffanta nau'ikan kayan laushi iri-iri, amma hanyoyi da yawa har yanzu suna fama da ƙananan gazawa waɗanda ke iyakance amfaninsu8,9,10,11,12,13. Rashin hanyoyin gwaji na musamman waɗanda za su iya kwatanta halayen injiniya na kayan da suka yi laushi da yadudduka na saman suna da matuƙar iyakance amfani da su a aikace-aikace daban-daban.
A cikin aikinmu na baya, mun gabatar da ruwan tabarau na lehfilcon A (CL), wani abu mai laushi daban-daban tare da dukkan halayen saman mai laushi wanda aka samo daga ƙirar biomimetic da aka yi wahayi zuwa gare shi daga saman cornea na ido. An haɓaka wannan kayan halitta ta hanyar dasa wani reshe na poly(2-methacryloyloxyethylphosphorylcholine (MPC)) (PMPC) mai alaƙa da juna a kan silicone hydrogel (SiHy) 15 wanda aka tsara don na'urorin likitanci bisa ga. Wannan tsarin dasawa yana ƙirƙirar wani Layer a saman wanda ya ƙunshi tsarin goga mai laushi da reshe mai laushi. Aikinmu na baya ya tabbatar da cewa tsarin biomimetic na lehfilcon A CL yana ba da kyawawan halaye na saman kamar ingantaccen hana jika da ƙura, ƙara yawan mai, da rage mannewar ƙwayoyin cuta da ƙwayoyin cuta15,16. Bugu da ƙari, amfani da haɓaka wannan kayan biomimetic kuma yana ba da shawarar ƙarin faɗaɗawa ga sauran na'urorin biomedical. Saboda haka, yana da matuƙar muhimmanci a fayyace halayen saman wannan abu mai laushi sosai kuma a fahimci hulɗar sa ta injiniya da ido domin ƙirƙirar cikakken tushen ilimi don tallafawa ci gaba da aikace-aikace na gaba. Yawancin ruwan tabarau na SiHy da ake samu a kasuwa sun ƙunshi cakuda masu kama da juna na polymers masu hydrophilic da hydrophobic waɗanda ke samar da tsarin abu iri ɗaya17. An gudanar da bincike da yawa don bincika halayen injiniyarsu ta amfani da hanyoyin gwaji na matsewa, tensile da microindentation na gargajiya18,19,20,21. Duk da haka, sabon ƙirar biomimetic na lehfilcon A CL ya sanya shi abu na musamman wanda ke da halaye daban-daban na injiniya waɗanda halayen injiniya na tsarin goga mai reshe ya bambanta sosai da na substrate na tushen SiHy. Saboda haka, yana da matukar wahala a tantance waɗannan halaye daidai ta amfani da hanyoyin gargajiya da na indentation. Hanya mai kyau tana amfani da hanyar gwajin nanoindentation da aka aiwatar a cikin microscopy na ƙarfin atomic (AFM), hanyar da aka yi amfani da ita don tantance halayen injiniya na kayan viscoelastic masu laushi kamar ƙwayoyin halitta da kyallen takarda, da kuma polymers masu laushi22,23,24,25. ,26,27,28,29,30. A cikin nanoindentation na AFM, an haɗa muhimman abubuwan gwajin nanoindentation tare da sabbin ci gaba a cikin fasahar AFM don samar da ƙarin fahimtar ma'auni da gwaji na nau'ikan kayan aiki masu laushi iri-iri31,32,33,34,35,36. Bugu da ƙari, fasahar tana ba da wasu muhimman fa'idodi ta hanyar amfani da geometries daban-daban. indenter da probe da kuma yiwuwar gwaji a cikin kafofin watsa labarai daban-daban na ruwa.
Za a iya raba nanoindentation na AFM zuwa manyan sassa uku bisa sharaɗi: (1) kayan aiki (na'urori masu auna firikwensin, na'urori masu gano abubuwa, na'urori masu aunawa, da sauransu); (2) sigogin aunawa (kamar ƙarfi, motsawa, gudu, girman gangara, da sauransu); (3) Sarrafa bayanai (gyara tushe, kimanta wurin taɓawa, daidaita bayanai, ƙira, da sauransu). Babbar matsala game da wannan hanyar ita ce cewa bincike da yawa a cikin wallafe-wallafen ta amfani da nanoindentation na AFM sun ba da rahoton sakamako daban-daban na adadi don samfurin/tantanin halitta/nau'in abu iri ɗaya37,38,39,40,41. Misali, Lekka et al. An yi nazarin tasirin yanayin binciken AFM akan ma'aunin Young da aka auna na samfuran hydrogel mai kama da injina da ƙwayoyin halitta daban-daban. Sun ba da rahoton cewa ƙimar modulus sun dogara sosai akan zaɓin cantilever da siffar tip, tare da mafi girman ƙimar na'urar bincike mai siffar dala da mafi ƙarancin ƙimar 42 don na'urar bincike mai siffar ƙwallo. Hakazalika, Selhuber-Unkel et al. An nuna yadda saurin shigar ciki, girman shigar ciki da kauri na samfuran polyacrylamide (PAAM) ke shafar tsarin Young wanda aka auna ta hanyar ACM43 nanoindentation. Wani abu mai rikitarwa shine rashin kayan gwajin modulus masu ƙarancin inganci da hanyoyin gwaji kyauta. Wannan yana sa ya yi wuya a sami sakamako masu inganci da kwarin gwiwa. Duk da haka, hanyar tana da matukar amfani ga ma'auni na dangi da kimantawa tsakanin nau'ikan samfuran iri ɗaya, misali amfani da nanoindentation na AFM don bambance ƙwayoyin halitta na yau da kullun daga ƙwayoyin cutar kansa 44, 45.
Lokacin gwada kayan laushi tare da nanoindentation na AFM, ƙa'idar babban yatsa ita ce a yi amfani da na'urar bincike mai ƙarancin ma'aunin bazara (k) wanda ya dace da samfurin modulus da kuma ƙarshen hemispherical/zagaye don na'urar bincike ta farko ba ta huda saman samfurin ba a lokacin da ta fara hulɗa da kayan laushi. Hakanan yana da mahimmanci cewa siginar karkacewa da na'urar bincike ta samar ta kasance mai ƙarfi don a gano ta tsarin gano laser24,34,46,47. A cikin yanayin ƙwayoyin halitta masu laushi, kyallen takarda da gels, wani ƙalubale shine shawo kan ƙarfin manne tsakanin na'urar bincike da saman samfurin don tabbatar da ma'auni masu iya sake samarwa da inganci48,49,50. Har zuwa kwanan nan, yawancin aiki akan nanoindentation na AFM ya mayar da hankali kan nazarin halayen injiniya na ƙwayoyin halitta, kyallen takarda, gels, hydrogels, da biomolecules ta amfani da manyan na'urori masu siffar ƙwallo, waɗanda aka fi sani da na'urorin bincike na colloidal (CPs). , 47, 51, 52, 53, 54, 55. Waɗannan ƙananan suna da radius na 1 zuwa 50 µm kuma ana yin su ne da gilashin borosilicate, polymethyl methacrylate (PMMA), polystyrene (PS), silicon dioxide (SiO2) da carbon mai kama da lu'u-lu'u (DLC). Kodayake nanoindentation na CP-AFM sau da yawa shine zaɓi na farko don halayyar samfurin mai laushi, yana da nasa matsaloli da ƙuntatawa. Amfani da manyan ƙananan kusurwa masu girman micron yana ƙara jimlar yankin taɓawa na ƙarshen tare da samfurin kuma yana haifar da asarar ƙudurin sarari mai yawa. Ga samfuran laushi, marasa daidaituwa, inda halayen injiniya na abubuwan gida na iya bambanta sosai da matsakaicin yanki mai faɗi, CP indentation na iya ɓoye duk wani rashin daidaituwa a cikin kaddarorin akan sikelin gida52. Ana yin binciken colloidal yawanci ta hanyar haɗa ƙananan kusurwa masu girman micron zuwa cantilevers marasa tip ta amfani da manne epoxy. Tsarin kera kansa yana cike da matsaloli da yawa kuma yana iya haifar da rashin daidaito a cikin tsarin daidaita binciken. Bugu da ƙari, girma da nauyin ƙwayoyin colloidal suna shafar manyan sigogin daidaitawa na cantilever kai tsaye, kamar mitar resonant, taurin bazara, da kuma ƙarfin karkatarwa56,57,58. Don haka, hanyoyin da aka saba amfani da su don binciken AFM na gargajiya, kamar daidaita zafin jiki, ƙila ba za su samar da daidaiton daidaito ga CP ba, kuma ana iya buƙatar wasu hanyoyi don yin waɗannan gyare-gyare57, 59, 60, 61. Gwaje-gwajen shigar CP na yau da kullun suna amfani da manyan karkacewa cantilever don nazarin halayen samfuran laushi, wanda ke haifar da wata matsala yayin daidaita halayen cantilever mara layi a manyan karkacewa62,63,64. Hanyoyin shigar ƙofofin binciken colloidal na zamani galibi suna la'akari da yanayin cantilever da ake amfani da shi don daidaita binciken, amma suna watsi da tasirin ƙwayoyin colloidal, wanda ke haifar da ƙarin rashin tabbas a cikin daidaiton hanyar38,61. Hakazalika, tsarin roba da aka ƙididdige ta hanyar daidaita samfurin hulɗa ya dogara kai tsaye akan yanayin binciken shigarwa, kuma rashin daidaito tsakanin halayen saman samfurin da na iya haifar da rashin daidaito27, 65, 66, 67, 68. An haskaka wasu ayyukan da Spencer et al. Abubuwan da ya kamata a yi la'akari da su yayin siffanta goge-goge masu laushi ta amfani da hanyar nanoindentation ta CP-AFM. Sun ba da rahoton cewa riƙe ruwa mai kauri a cikin goge-goge na polymer a matsayin aikin gudu yana haifar da ƙaruwar nauyin kai kuma don haka ma'auni daban-daban na halayen da suka dogara da sauri30,69,70,71.
A cikin wannan binciken, mun bayyana yanayin saman kayan da ke da laushi sosai, wanda ke da laushi sosai, wanda ke da laushi sosai, ta amfani da hanyar AFM nanoindentation. Ganin halaye da sabon tsarin wannan kayan, kewayon yanayin yanayin da ke cikin hanyar shigar da kayan a bayyane yake bai isa ya kwatanta yanayin wannan kayan mai laushi ba, don haka ya zama dole a yi amfani da hanyar shigar da kayan a cikin AFM tare da babban ƙarfin ji da ƙarancin ƙarfin ji. Bayan duba gajarta da matsalolin dabarun shigar da kayan a cikin AFM na colloidal, mun nuna dalilin da yasa muka zaɓi ƙaramin na'urar AFM da aka tsara musamman don kawar da jin daɗi, hayaniyar bango, wurin tuntuɓar ma'auni, auna saurin kayan da ke da laushi iri-iri kamar dogaro da riƙe ruwa. da kuma ƙididdigewa daidai. Bugu da ƙari, mun sami damar auna siffar da girman tip ɗin shigar da kayan a cikin daidai, wanda ke ba mu damar amfani da samfurin dacewa da mazugi don tantance yanayin da ke cikin layin ba tare da tantance yankin da ke cikin layin tare da kayan ba. Zato biyu da ba a iya tantancewa a cikin wannan aikin sune halayen kayan da suka dace da juna da kuma modulus masu zaman kansu na zurfin shiga. Ta amfani da wannan hanyar, mun fara gwada ƙa'idodi masu laushi sosai tare da wani sanannen modulus don ƙididdige hanyar, sannan muka yi amfani da wannan hanyar don kwatanta saman kayan ruwan tabarau guda biyu daban-daban. Ana sa ran wannan hanyar siffanta saman ruwan tabarau na AFM tare da ƙarin ƙarfin fahimta za ta shafi nau'ikan kayan ultrasoft iri-iri na biomimetic tare da yuwuwar amfani da su a cikin na'urorin likitanci da aikace-aikacen biomedical.
Lehfilcon An zaɓi ruwan tabarau na hulɗa (Alcon, Fort Worth, Texas, Amurka) da kuma abubuwan da aka yi amfani da su na silicone hydrogel don gwaje-gwajen nanoindentation. An yi amfani da wani abin ɗaura ruwan tabarau na musamman a cikin gwajin. Don shigar da ruwan tabarau don gwaji, an sanya shi a hankali a kan madaurin da ke da siffar kumfa, don tabbatar da cewa babu kumfa a ciki, sannan a gyara shi da gefuna. Rami a cikin abin da aka haɗa a saman abin da aka haɗa ruwan tabarau yana ba da damar shiga tsakiyar ruwan tabarau don gwaje-gwajen nanoindentation yayin da ake riƙe ruwan a wurin. Wannan yana sa ruwan tabarau ya cika da ruwa. An yi amfani da 500 μl na maganin marufi na ruwan tabarau na hulɗa azaman maganin gwaji. Don tabbatar da sakamakon adadi, an shirya hydrogels na polyacrylamide (PAAM) waɗanda ba a kunna su ba a kasuwa daga abun da ke ciki na polyacrylamide-co-methylene-bisacrylamide (100 mm Petri dishes, Matrigen, Irvine, CA, Amurka), wani sanannen nau'in roba na 1 kPa. Yi amfani da digo 4-5 (kimanin 125 µl) na sinadarin phosphate buffered saline (PBS daga Corning Life Sciences, Tewkesbury, MA, Amurka) da digo 1 na maganin ruwan tabarau na OPTI-FREE Puremoist (Alcon, Vaud, TX, Amurka).) a mahaɗin AFM hydrogel-probe.
An yi amfani da tsarin FEI Quanta 250 Field Emission Scanning Electron Microscope (FEG SEM) wanda aka sanye da na'urar gano iskar gas ta Scanning Transmission Electron Microscope (STEM). Don shirya samfuran, an fara wanke ruwan tabarau da ruwa sannan aka yanke su zuwa gauraye masu siffar kek. Don cimma bambanci tsakanin abubuwan hydrophilic da hydrophobic na samfuran, an yi amfani da maganin RuO4 mai ƙarfi 0.10% a matsayin rini, inda aka nutsar da samfuran na tsawon minti 30. Tabon lehfilcon A CL RuO4 yana da mahimmanci ba wai kawai don cimma ingantaccen bambanci ba, har ma yana taimakawa wajen kiyaye tsarin gogayen polymer masu reshe a cikin sifar su ta asali, waɗanda a lokacin ake iya gani a hotunan STEM. Sannan aka wanke su kuma aka bushe su a cikin jerin gaurayen ethanol/ruwa tare da ƙaruwar yawan ethanol. Sannan aka jefa samfuran da EMBed 812/Araldite epoxy, wanda ya warke cikin dare a 70°C. An yanke samfuran tubalan da aka samu ta hanyar polymerization na resin da ultramicrotome, kuma an nuna sassan siririn da suka biyo baya da na'urar gano STEM a cikin yanayin injin mara iska a ƙarfin lantarki mai sauri na 30 kV. An yi amfani da tsarin SEM iri ɗaya don cikakken bayanin na'urar binciken PFQNM-LC-A-CAL AFM (Bruker Nano, Santa Barbara, CA, Amurka). An samo hotunan SEM na na'urar binciken AFM a cikin yanayin injin mai ƙarfi na yau da kullun tare da ƙarfin lantarki mai sauri na 30 kV. Sami hotuna a kusurwoyi daban-daban da girma don yin rikodin duk cikakkun bayanai na siffar da girman tip ɗin na'urar binciken AFM. An auna duk girman tip ɗin da ke da sha'awa a cikin hotunan ta hanyar dijital.
An yi amfani da na'urar hangen nesa ta Dimension FastScan Bio Icon atomic force microscope (Bruker Nano, Santa Barbara, CA, Amurka) tare da yanayin "PeakForce QNM a cikin Ruwa" don ganin da kuma nanoindentate lehfilcon A CL, SiHy substrate, da samfuran PAAm hydrogel. Don gwaje-gwajen daukar hoto, an yi amfani da na'urar bincike ta PEAKFORCE-HIRS-FA (Bruker) tare da radius na tip na 1 nm don ɗaukar hotunan samfurin masu ƙuduri mai girma a ƙimar scan na 0.50 Hz. An ɗauki duk hotunan a cikin ruwan magani.
An gudanar da gwaje-gwajen nanoindentation na AFM ta amfani da na'urar bincike ta PFQNM-LC-A-CAL (Bruker). Na'urar bincike ta AFM tana da tip ɗin silicon akan cantilever na nitride mai kauri 345 nm, tsayin µm 54 da faɗin µm 4.5 tare da mitar amsawa ta 45 kHz. An tsara shi musamman don siffantawa da kuma yin ma'aunin nanomechanical na adadi akan samfuran halittu masu laushi. Ana daidaita na'urori masu auna firikwensin daban-daban a masana'anta tare da saitunan bazara da aka riga aka daidaita. Madaidaitan maɓuɓɓugar binciken da aka yi amfani da su a cikin wannan binciken sun kasance cikin kewayon 0.05-0.1 N/m. Don tantance siffar da girman tip ɗin daidai, an bayyana na'urar dalla-dalla ta amfani da SEM. A kan siffa ta 1a yana nuna ƙaramin ƙuduri, ƙaramin girman na'urar binciken electron na PFQNM-LC-A-CAL, yana ba da cikakken ra'ayi game da ƙirar na'urar bincike. A kan siffa ta 1b yana nuna babban ra'ayi na saman tip ɗin binciken, yana ba da bayani game da siffar da girman tip ɗin. A ƙarshen ƙarshen, allurar tana da diamita kusan nm 140 (Hoto na 1c). A ƙasa da wannan, ƙarshen yana raguwa zuwa siffar mazugi, yana kaiwa tsawon da aka auna na kimanin nm 500. A wajen yankin da ke jujjuyawa, ƙarshen yana da silinda kuma yana ƙarewa da jimillar tsawon ƙarshen 1.18 µm. Wannan shine babban ɓangaren aikin tip ɗin bincike. Bugu da ƙari, an yi amfani da babban polystyrene mai siffar ƙwallo (PS) mai diamita na tip na 45 µm da kuma madaurin bazara na 2 N/m don gwaji azaman gwajin colloidal. tare da PFQNM-LC-A-CAL 140 nm binciken don kwatantawa.
An ruwaito cewa ruwa zai iya makale tsakanin na'urar binciken AFM da tsarin goga na polymer yayin nanoindentation, wanda zai yi ƙarfin sama akan na'urar binciken AFM kafin ya taɓa saman69. Wannan tasirin extrusion mai kauri saboda riƙe ruwa zai iya canza wurin da aka gani, wanda hakan zai shafi ma'aunin yanayin saman. Don nazarin tasirin yanayin binciken da saurin shigarwa akan riƙe ruwa, an zana lanƙwasa ƙarfin shigarwa don samfuran lehfilcon A CL ta amfani da na'urar binciken diamita 140 nm a cikin ƙimar ƙaura akai-akai na 1 µm/s da 2 µm/s. diamita na binciken 45 µm, saitin ƙarfi mai ƙarfi 6 nN an cimma shi a 1 µm/s. An gudanar da gwaje-gwaje tare da na'urar binciken 140 nm a diamita a saurin shigarwa na 1 µm/s da ƙarfin saita na 300 pN, wanda aka zaɓa don ƙirƙirar matsin lamba a cikin kewayon ilimin halittar jiki (1-8 kPa) na fatar ido ta sama. Matsi 72. An gwada samfuran PAA hydrogel masu laushi waɗanda aka shirya tare da matsin lamba na 1 kPa don ƙarfin shiga na 50 pN a gudun 1 μm/s ta amfani da na'urar bincike mai diamita na 140 nm.
Tunda tsawon ɓangaren mazugi na ƙarshen binciken PFQNM-LC-A-CAL yana da kusan nm 500, ga kowane zurfin mazugi < 500 nm, ana iya ɗauka lafiya cewa yanayin binciken yayin mazugi zai kasance daidai da siffar mazugi. Bugu da ƙari, ana ɗauka cewa saman kayan da ake gwadawa zai nuna amsawar roba mai juyawa, wanda kuma za a tabbatar a cikin sassan masu zuwa. Saboda haka, dangane da siffar da girman tip ɗin, mun zaɓi samfurin dacewa da mazugi wanda Briscoe, Sebastian da Adams suka haɓaka, wanda ke samuwa a cikin software na mai siyarwa, don aiwatar da gwaje-gwajen nanoindentation na AFM (NanoScope). Software na nazarin bayanai na rabuwa, Bruker) 73. Samfurin ya bayyana dangantakar motsi da motsi F(δ) don mazugi mai lahani na kololuwa mai siffar ƙwallo. A kan siffa Siffa ta 2 tana nuna yanayin hulɗa yayin hulɗar mazugi mai tauri tare da ƙarshen zagaye, inda R shine radius na ƙarshen zagaye, a shine radius na hulɗa, b shine radius na hulɗa a ƙarshen zagaye, δ shine radius na hulɗa. Zurfin shiga, θ shine rabin kusurwar mazugi. Hoton SEM na wannan binciken ya nuna a sarari cewa ƙarshen zagaye mai diamita 140 nm yana haɗuwa ta hanyar tangentially zuwa mazugi, don haka a nan b an bayyana shi ne kawai ta hanyar R, watau b = R cos θ. Manhajar da mai siyarwa ke bayarwa tana ba da alaƙar mazugi-sphere don ƙididdige ƙimar modulus (E) na Young daga bayanan rabuwar ƙarfi da ake ɗauka a > b. Dangantaka:
inda F shine ƙarfin shigar da ƙofa, E shine modulus na Young, ν shine rabon Poisson. Ana iya kimanta radius ɗin hulɗar a ta amfani da:
Tsarin yanayin hulɗa na mazugi mai tauri tare da ƙarshen zagaye wanda aka matse a cikin kayan ruwan tabarau na Lefilcon tare da saman goge-goge masu reshe.
Idan ≤ b, dangantakar ta ragu zuwa ga daidaiton mai shiga zagaye na al'ada;
Mun yi imanin cewa hulɗar binciken da ke shiga da tsarin reshe na goga polymer na PMPC zai sa radius ɗin hulɗar a ya fi radius ɗin hulɗar mai siffar ƙwallo b. Saboda haka, ga duk ma'aunin adadi na modulus na roba da aka yi a cikin wannan binciken, mun yi amfani da dogaro da aka samu don yanayin a > b.
An yi cikakken hoton kayan aikin biomimetic na ultrasoft da aka yi nazari a kansu a cikin wannan binciken ta amfani da na'urar daukar hoto ta na'urar daukar hoto ta na'urar daukar hoto ta na'urar daukar hoto (STEM) ta samfurin giciye sashe da kuma na'urar daukar hoto ta atomic force microscopy (AFM) ta saman. An yi wannan cikakken bayanin yanayin saman a matsayin fadada aikin da muka buga a baya, inda muka gano cewa tsarin goga mai rassa na polymeric na saman lehfilcon A CL da PMPC ta gyara ya nuna irin wannan halayen injiniya ga nama na corneal na asali 14. Saboda wannan dalili, muna kiran saman ruwan tabarau na lamba a matsayin kayan biomimetic14. A hoto na 3a,b yana nuna sassan giciye na tsarin goga mai reshe na PMPC polymer a saman substrate na lehfilcon A CL da kuma substrate na SiHy da ba a yi masa magani ba, bi da bi. An kara yin nazari kan saman samfuran biyu ta amfani da hotunan AFM masu inganci, wanda ya kara tabbatar da sakamakon binciken STEM (Hoto na 3c, d). Idan aka hada su, wadannan hotunan suna ba da kimanin tsawon tsarin goga mai reshe na PMPC a 300-400 nm, wanda yake da mahimmanci don fassara ma'aunin nanoindentation na AFM. Wani muhimmin abin lura da aka samo daga hotunan shine cewa tsarin saman gaba ɗaya na kayan biomimetic na CL ya bambanta da na kayan substrate na SiHy a fannin siffa. Wannan bambanci a cikin yanayin saman su na iya bayyana a lokacin hulɗar injiniya da binciken AFM mai shiga ciki da kuma daga baya a cikin ƙimar modulus da aka auna.
Hotunan STEM masu sassaka na (a) lehfilcon A CL da (b) SiHy substrate. Madaurin sikelin, 500 nm. Hotunan AFM na saman substrate na lehfilcon A CL (c) da tushen substrate na SiHy (d) (3 µm × 3 µm).
Tsarin polymers da aka yi wahayi zuwa gare su da kuma tsarin goga na polymer suna da laushi a zahiri kuma an yi nazari sosai kuma an yi amfani da su sosai a aikace-aikacen likitanci daban-daban74,75,76,77. Saboda haka, yana da mahimmanci a yi amfani da hanyar nanoindentation ta AFM, wacce za ta iya auna halayen injinan su daidai kuma cikin aminci. Amma a lokaci guda, keɓantattun kaddarorin waɗannan kayan masu laushi, kamar ƙarancin modulus na roba, babban abun ciki na ruwa da babban sassauci, sau da yawa suna sa ya zama da wahala a zaɓi kayan da suka dace, siffa da siffar girman binciken da ke shiga. Wannan yana da mahimmanci don kada indenter ya huda saman laushi na samfurin, wanda zai haifar da kurakurai wajen tantance wurin da aka taɓa da saman da yankin da aka taɓa.
Don haka, cikakken fahimtar yanayin kayan biomimetic masu laushi (lehfilcon A CL) yana da mahimmanci. Bayani game da girma da tsarin gogayen polymer da aka samu ta hanyar amfani da hanyar daukar hoto yana samar da tushen fasalin injiniya na saman ta amfani da dabarun nanoindentation na AFM. Maimakon binciken colloidal mai siffar micron, mun zaɓi binciken silicon nitride na PFQNM-LC-A-CAL (Bruker) mai diamita na tip na 140 nm, wanda aka tsara musamman don taswirar adadi na halayen injiniya na samfuran halittu 78, 79, 80, 81, 82, 83, 84 Za a iya bayyana dalilin amfani da binciken kaifi idan aka kwatanta da binciken colloidal na gargajiya ta hanyar fasalin kayan. Idan aka kwatanta girman tip ɗin binciken (~140 nm) da gogayen polymer masu reshe a saman CL lehfilcon A, wanda aka nuna a Hoto na 3a, za a iya kammala da cewa tip ɗin ya isa ya shiga kai tsaye da waɗannan tsarin goga, wanda hakan ke rage damar huda tip ɗin ta cikinsu. Don kwatanta wannan batu, a Hoto na 4 akwai hoton STEM na lehfilcon A CL da kuma tip ɗin binciken AFM (wanda aka zana zuwa sikelin).
Tsarin da ke nuna hoton STEM na lehfilcon A CL da kuma binciken shigar ACM (wanda aka zana zuwa sikelin).
Bugu da ƙari, girman ƙarshen 140 nm ya isa ya guji haɗarin duk wani tasirin fitar da manne da aka ruwaito a baya ga goge-goge na polymer da aka samar ta hanyar nanoindentation na CP-AFM69,71. Muna ɗauka cewa saboda siffar musamman mai siffar mazugi da ƙaramin girman wannan tip ɗin AFM (Hoto na 1), yanayin lanƙwasa ƙarfi da lehfilcon A CL nanoindentation ya samar ba zai dogara da saurin shigar da shi ko saurin ɗaukar kaya/saukewa ba. Saboda haka, tasirin poroelastic ba ya shafar shi. Don gwada wannan hasashe, an sanya samfuran lehfilcon A CL a cikin matsakaicin ƙarfi mai ƙarfi ta amfani da binciken PFQNM-LC-A-CAL, amma a cikin gudu biyu daban-daban, kuma an yi amfani da lanƙwasa ƙarfi da ja da baya da aka samu don zana ƙarfin (nN) a cikin rabuwa (µm) a cikin Hoto na 5a. A bayyane yake cewa lanƙwasa ƙarfin yayin lodawa da saukewa gaba ɗaya suna haɗuwa, kuma babu wata shaida bayyananna cewa yankewar ƙarfi a zurfin shigarwar sifili yana ƙaruwa tare da saurin shigarwa a cikin hoton, yana nuna cewa an siffanta abubuwan goga ɗaya ba tare da tasirin poroelastic ba. Sabanin haka, tasirin riƙe ruwa (fitar da ruwa mai kauri da tasirin poroelasticity) a bayyane yake ga binciken AFM mai diamita 45 µm a daidai saurin shigarwa kuma an haskaka su ta hanyar hysteresis tsakanin lanƙwasa mai shimfiɗawa da ja da baya, kamar yadda aka nuna a Hoto na 5b. Waɗannan sakamakon suna goyan bayan hasashen kuma suna nuna cewa binciken diamita 140 nm kyakkyawan zaɓi ne don siffanta irin waɗannan saman masu laushi.
lehfilcon Lanƙwasa ƙarfin CL mai shiga ta amfani da ACM; (a) amfani da na'urar bincike mai diamita na 140 nm a ƙimar lodi biyu, yana nuna rashin tasirin poroelastic yayin shigar saman; (b) amfani da na'urori masu diamita na 45 µm da 140 nm. s yana nuna tasirin fitar da viscous extrusion da poroelasticity ga manyan na'urori idan aka kwatanta da ƙananan na'urori.
Don kwatanta saman da ke da laushi sosai, hanyoyin nanoindentation na AFM dole ne su sami mafi kyawun bincike don nazarin halayen kayan da ake nazari a kansu. Baya ga siffar da girman tip, ƙarfin tsarin gano AFM, ƙarfin juyewar tip a cikin yanayin gwaji, da kuma taurin cantilever suna taka muhimmiyar rawa wajen tantance daidaito da amincin ma'aunin nanoindentation. Ga tsarin AFM ɗinmu, iyakar gano Matsayi Mai Sensitive Detector (PSD) shine kimanin 0.5 mV kuma ya dogara ne akan ƙimar bazara da aka riga aka daidaita da kuma ƙimar juyewar ruwa da aka ƙididdige na binciken PFQNM-LC-A-CAL, wanda ya dace da ƙimar nauyi na ka'ida. ƙasa da 0.1 pN. Saboda haka, wannan hanyar tana ba da damar auna mafi ƙarancin ƙarfin shiga ≤ 0.1 pN ba tare da wani ɓangaren hayaniyar gefe ba. Duk da haka, kusan ba zai yiwu ba ga tsarin AFM ya rage hayaniyar gefe zuwa wannan matakin saboda dalilai kamar girgizar injiniya da ƙarfin ruwa. Waɗannan abubuwan suna iyakance jimillar ƙarfin hanyar AFM nanoindentation kuma suna haifar da siginar hayaniyar bango na kusan ≤ 10 pN. Don siffanta saman, an sanya samfuran substrate lehfilcon A CL da SiHy a cikin yanayin da ya cika da ruwa ta amfani da na'urar bincike ta 140 nm don siffanta SEM, kuma an haɗa lanƙwasa ƙarfin da ya haifar tsakanin ƙarfi (pN) da matsin lamba. An nuna taswirar rabuwar (µm) a cikin Hoto na 6a. Idan aka kwatanta da substrate na tushen SiHy, lanƙwasa ƙarfin lehfilcon A CL a bayyane yana nuna matakin canji wanda ya fara daga wurin da aka taɓa goga polymer mai yankan ya ƙare da canji mai kaifi a cikin alamar gangara ta gefen tare da kayan da ke ƙarƙashinsa. Wannan ɓangaren canji na lanƙwasa ƙarfi yana nuna halayen roba na goga polymer mai reshe a saman, kamar yadda aka tabbatar ta lanƙwasa matsewa kusa da lanƙwasa mai ƙarfi da bambancin halayen injiniya tsakanin tsarin goga da babban kayan SiHy. Lokacin kwatanta lefilcon. Raba matsakaicin tsawon goga mai reshe na polymer a cikin hoton STEM na PCS (Hoto na 3a) da kuma lanƙwasa ƙarfinsa tare da abscissa a Hoto na 3a. 6a ya nuna cewa hanyar tana iya gano ƙarshen da kuma polymer mai reshe da ke kaiwa saman saman. Hulɗa tsakanin tsarin goga. Bugu da ƙari, haɗuwa kusa da lanƙwasa ƙarfi yana nuna babu wani tasirin riƙe ruwa. A wannan yanayin, babu mannewa gaba ɗaya tsakanin allura da saman samfurin. Sassan sama na lanƙwasa ƙarfi don samfuran biyu suna haɗuwa, suna nuna kamannin halayen injiniya na kayan substrate.
(a) Lanƙwasa ƙarfin nanoindentation na AFM don substrates na lehfilcon A CL da substrates na SiHy, (b) lanƙwasa ƙarfi da ke nuna kimanta wurin hulɗa ta amfani da hanyar iyakar hayaniyar bango.
Domin yin nazarin cikakkun bayanai game da lanƙwasa ƙarfi, an sake tsara lanƙwasa matsin lamba na samfurin lehfilcon A CL a cikin Hoto na 6b tare da matsakaicin ƙarfin pN 50 tare da axis na y. Wannan jadawalin yana ba da mahimman bayanai game da hayaniyar asali. Hayaniyar tana cikin kewayon ±10 pN, wanda ake amfani da shi don tantance wurin hulɗa daidai da ƙididdige zurfin lanƙwasa. Kamar yadda aka ruwaito a cikin wallafe-wallafen, gano wuraren hulɗa yana da mahimmanci don tantance halayen kayan kamar modulus85 daidai. Hanyar da ta shafi sarrafa bayanai ta atomatik na lanƙwasa ƙarfi ta nuna ingantacciyar dacewa tsakanin daidaita bayanai da ma'aunin adadi don kayan laushi86. A cikin wannan aikin, zaɓin wuraren hulɗarmu yana da sauƙi kuma mai ma'ana, amma yana da iyakokinta. Hanyarmu mai ra'ayin mazan jiya don tantance wurin hulɗa na iya haifar da ƙimar modulus da aka ƙima kaɗan don ƙananan zurfin lanƙwasa (< 100 nm). Amfani da gano wurin taɓawa bisa algorithm da sarrafa bayanai ta atomatik na iya zama ci gaba da wannan aikin a nan gaba don ƙara inganta hanyarmu. Saboda haka, ga hayaniyar bango ta ciki akan tsari na ±10 pN, mun ayyana wurin hulɗa a matsayin wurin bayanai na farko akan axis na x a cikin Hoto na 6b tare da ƙimar ≥10 pN. Sannan, daidai da iyakar hayaniyar 10 pN, layi a tsaye a matakin ~0.27 µm yana nuna wurin hulɗa da saman, bayan haka lanƙwasa mai shimfiɗawa ya ci gaba har sai substrate ya cika zurfin shigarwa na ~270 nm. Abin sha'awa, bisa ga girman siffofin goga mai reshe na polymer (300–400 nm) wanda aka auna ta amfani da hanyar hoto, zurfin shigarwa na CL lehfilcon Samfurin da aka lura ta amfani da hanyar iyakar hayaniyar bango yana kusan 270 nm, wanda yake kusa da girman ma'auni tare da STEM. Waɗannan sakamakon sun ƙara tabbatar da dacewa da amfani da siffa da girman tip ɗin binciken AFM don shigar da wannan tsarin goga mai laushi da reshe mai laushi. Wannan bayanan kuma yana ba da shaida mai ƙarfi don tallafawa hanyarmu ta amfani da hayaniyar bango azaman ma'auni don gano wuraren hulɗa. Saboda haka, duk wani sakamako na adadi da aka samu daga ƙirar lissafi da kuma daidaita lanƙwasa ƙarfi ya kamata ya zama daidai.
Ma'aunin ƙididdiga ta hanyoyin nanoindentation na AFM ya dogara gaba ɗaya akan samfuran lissafi da ake amfani da su don zaɓar bayanai da kuma nazarin da ke gaba. Saboda haka, yana da mahimmanci a yi la'akari da duk abubuwan da suka shafi zaɓin indenter, halayen kayan aiki da kuma hanyoyin hulɗarsu kafin zaɓar wani takamaiman samfuri. A wannan yanayin, an yi amfani da micrographs na SEM a hankali, kuma bisa ga sakamakon, binciken nanoindenting na AFM mai diamita 140 nm tare da mazugi mai tauri da kuma siffar tip mai zagaye kyakkyawan zaɓi ne don siffanta samfuran lehfilcon A CL79. Wani muhimmin abu da ke buƙatar a tantance shi da kyau shine sassaucin kayan polymer da ake gwadawa. Kodayake bayanan farko na nanoindentation (Figures. 5a da 6a) sun bayyana fasalin haɗuwar lanƙwasa na matsi da matsi, watau, cikakken dawo da robar kayan, yana da matuƙar mahimmanci a tabbatar da yanayin robar kawai na lambobin sadarwa. Don haka, an yi ƙoƙon shiga guda biyu a jere a wuri ɗaya a saman samfurin lehfilcon A CL a ƙimar shiga ta 1 µm/s a ƙarƙashin cikakken yanayin ruwa. Bayanan lanƙwasa ƙarfi da aka samu an nuna su a cikin fig. 7 kuma, kamar yadda aka zata, lanƙwasa faɗaɗawa da matsewa na kwafi biyu kusan iri ɗaya ne, suna nuna babban sassaucin tsarin goga mai reshe.
Lanƙwasa guda biyu na ƙarfin shiga a wuri ɗaya a saman lehfilcon A CL suna nuna cikakkiyar sassaucin saman ruwan tabarau.
Dangane da bayanan da aka samu daga hotunan SEM da STEM na tip ɗin bincike da kuma surface ɗin lehfilcon A CL, bi da bi, samfurin cone-sphere wakilci ne mai ma'ana na lissafi na hulɗar da ke tsakanin tip ɗin binciken AFM da kayan polymer mai laushi da ake gwadawa. Bugu da ƙari, ga wannan samfurin cone-sphere, ainihin zato game da kaddarorin roba na kayan da aka buga suna da gaskiya ga wannan sabon kayan biomimetic kuma ana amfani da su don ƙididdige modulus na roba.
Bayan cikakken kimantawa na hanyar AFM nanoindentation da abubuwan da ke cikinta, gami da halayen binciken shigarwa (siffa, girma, da taurin bazara), jin daɗi (hayaniyar bango da kimanta wurin tuntuɓar), da samfuran daidaita bayanai (ma'aunin modulus na adadi), an yi amfani da hanyar. An kwatanta samfuran taushi masu yawa da ake samu a kasuwa don tabbatar da sakamakon adadi. An gwada hydrogel na polyacrylamide (PAAM) na kasuwanci tare da modulus na roba na 1 kPa a ƙarƙashin yanayin ruwa ta amfani da binciken nm 140. An bayar da cikakkun bayanai game da gwajin module da lissafi a cikin Ƙarin Bayani. Sakamakon ya nuna cewa matsakaicin modulus da aka auna shine 0.92 kPa, kuma karkacewar %RSD da kashi (%) daga modulus da aka sani sun kasance ƙasa da 10%. Waɗannan sakamakon sun tabbatar da daidaito da sake haifar da hanyar AFM nanoindentation da aka yi amfani da ita a cikin wannan aikin don auna moduli na kayan ultrasoft. An ƙara siffanta saman samfuran lehfilcon A CL da kuma tushen SiHy ta amfani da wannan hanyar nanoindentation ta AFM don nazarin yanayin hulɗar da ke bayyane na saman mai laushi a matsayin aikin zurfin shiga. An samar da lanƙwasa na rabuwar ƙarfin shiga ga samfura uku na kowane nau'i (n = 3; shigarwa ɗaya a kowane samfuri) a ƙarfin 300 pN, saurin 1 µm/s, da cikakken ruwa. An kimanta lanƙwasa raba ƙarfin shiga ta amfani da samfurin mazugi. Don samun modulus dangane da zurfin shiga, an saita wani yanki mai faɗi na 40 nm na lanƙwasa ƙarfi a kowane ƙaruwa na 20 nm fara daga wurin haɗuwa, kuma an auna ƙimar modulus a kowane mataki na lanƙwasa ƙarfi. Spin Cy et al. An yi amfani da irin wannan hanyar don kwatanta yanayin modulus na gogayen polymer poly(lauryl methacrylate) (P12MA) ta amfani da nanoindentation na colloidal AFM probe, kuma sun yi daidai da bayanai ta amfani da samfurin lamba na Hertz. Wannan hanyar tana ba da labarin yanayin lamba a bayyane (kPa) da zurfin shiga (nm), kamar yadda aka nuna a Hoto na 8, wanda ke nuna yanayin lamba a bayyane/zurfin shiga. Tsarin roba da aka ƙididdige na samfurin CL lehfilcon A yana cikin kewayon 2-3 kPa a cikin saman 100 nm na samfurin, wanda bayan haka ya fara ƙaruwa da zurfi. A gefe guda kuma, lokacin gwada tushen SiHy ba tare da fim mai kama da goga a saman ba, matsakaicin zurfin shiga da aka samu a ƙarfin 300 pN bai wuce 50 nm ba, kuma ƙimar modulus da aka samu daga bayanan shine kusan 400 kPa, wanda yayi daidai da ƙimar modulus na Young don kayan da yawa.
Bayyanannen tsarin hulɗa (kPa) idan aka kwatanta da zurfin shigarwa (nm) don substrates na lehfilcon A CL da SiHy ta amfani da hanyar nanoindentation ta AFM tare da tsarin cone-sphere don auna modulus.
Saman saman sabon tsarin goga mai reshe na biomimetic yana nuna ƙarancin modulus na sassauci (2-3 kPa). Wannan zai dace da ƙarshen rataye kyauta na goga mai fork kamar yadda aka nuna a hoton STEM. Duk da cewa akwai wasu shaidu na canjin modulus a gefen waje na CL, babban substrate mai girma modulus ya fi tasiri. Duk da haka, saman 100 nm na saman yana cikin 20% na jimlar tsawon goga mai reshe na polymer, don haka yana da kyau a ɗauka cewa ƙimar da aka auna na modulus a cikin wannan kewayon zurfin shigarwa daidai ne kuma ba su dogara sosai kan tasirin abin ƙasa ba.
Saboda ƙirar biomimetic ta musamman ta ruwan tabarau na lahfilcon A, wanda ya ƙunshi tsarin buroshin polymer na PMPC da aka dasa a saman substrates na SiHy, yana da matuƙar wahala a iya siffanta halayen injiniya na tsarin saman su ta amfani da hanyoyin aunawa na gargajiya. A nan mun gabatar da wata hanyar nanoindentation ta AFM mai ci gaba don tantance kayan da suka yi laushi kamar lefilcon A daidai tare da yawan ruwa da kuma yawan sassauci. Wannan hanyar ta dogara ne akan amfani da na'urar bincike ta AFM wacce aka zaɓi girmanta da yanayinta a hankali don dacewa da girman tsarin fasalin saman mai laushi da za a buga. Wannan haɗin girma tsakanin na'urar bincike da tsari yana ba da ƙarin hankali, yana ba mu damar auna ƙananan modulus da halayen roba na abubuwan goga polymer masu reshe, ba tare da la'akari da tasirin poroelastic ba. Sakamakon ya nuna cewa goge-goga na polymer na PMPC na musamman waɗanda ke da alaƙa da saman ruwan tabarau suna da ƙarancin modulus na roba (har zuwa 2 kPa) da kuma babban sassauci (kusan 100%) lokacin da aka gwada su a cikin yanayi mai ruwa. Sakamakon nanoindentation na AFM ya kuma ba mu damar kwatanta yanayin hulɗa/zurfin haske (30 kPa/200 nm) na saman ruwan tabarau na biomimetic. Wannan yanayin haske na iya kasancewa saboda bambancin modulus tsakanin gogayen polymer masu reshe da substrate na SiHy, ko kuma tsarin/yawan gogayen polymer, ko haɗuwa da su. Duk da haka, ana buƙatar ƙarin bincike mai zurfi don fahimtar cikakkiyar alaƙar da ke tsakanin tsari da halaye, musamman tasirin reshe na goga akan halayen injiniya. Irin waɗannan ma'aunai na iya taimakawa wajen kwatanta halayen injiniya na saman wasu kayan aiki masu laushi da na'urorin likitanci.
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