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I te whanaketanga o ngā rauemi tino-ngohengohe hou mō ngā taputapu hauora me ngā tono hauora, he mea nui, he wero hoki te whakaahua whānui o ō rātou āhuatanga ā-tinana me ā-pūkaha. I whakamahia he tikanga nanoindentation microscopy kaha ngota (AFM) whakarerekētia hei whakaahua i te modulus mata tino iti o te karaihe whakapā silicone hydrogel biomimetic lehfilcon A hou kua pania ki te paparanga o ngā hanganga paraihe polymer peka. Mā tēnei tikanga ka taea te whakatau tika i ngā pūwāhi whakapā me te kore e pā ki te extrusion matotoru ina whakatata atu ki ngā polymer peka. Hei tāpiri, ka taea te whakatau i ngā āhuatanga ā-pūkaha o ngā huānga paraihe takitahi me te kore e pā ki te poroelasticity. Ka tutuki tēnei mā te whiriwhiri i tētahi probe AFM me te hoahoa (rahi pito, āhuahanga me te tere puna) e tino pai ana mō te ine i ngā āhuatanga o ngā rauemi ngohengohe me ngā tauira koiora. Ka whakapai ake tēnei tikanga i te aro me te tika mō te ine tika o te rauemi tino ngohengohe lehfilcon A, he tino iti te modulus o te elasticity i runga i te horahanga mata (tae atu ki te 2 kPa) me te tino elasticity i roto i te taiao wai o roto (tata ki te 100%). Kāore i te whakaatu noa ngā hua o te rangahau mata i ngā āhuatanga mata tino ngawari o te arotahi lehfilcon A, engari i whakaatu anō hoki he rite te modulus o ngā paraihe polymer manga ki te papa silicon-hauwai. Ka taea te whakamahi i tēnei tikanga tautuhi mata ki ētahi atu rauemi tino ngawari me ngā taputapu hauora.
Ko ngā āhuatanga ā-pūkaha o ngā rauemi i hangaia mō te whakapā tika ki te kiko ora ka whakatauhia e te taiao koiora. Ko te ōritetanga tino pai o ēnei āhuatanga rauemi ka āwhina ki te whakatutuki i ngā āhuatanga haumanu e hiahiatia ana o te rauemi me te kore e puta he urupare kino ki te pūtau1,2,3. Mō ngā rauemi papatipu ōrite, he ngāwari noa te tautuhi i ngā āhuatanga ā-pūkaha nā te wātea o ngā tikanga paerewa me ngā tikanga whakamātautau (hei tauira, te microindentation4,5,6). Heoi, mō ngā rauemi tino ngawari pērā i ngā gels, hydrogels, biopolymers, pūtau ora, me ētahi atu, ko ēnei tikanga whakamātautau kāore e tika ana te whakamahi nā te mea he iti te taumira ine me te kore ōrite o ētahi rauemi7. I roto i ngā tau, kua whakarerekētia, kua urutauhia hoki ngā tikanga indentation tuku iho hei tautuhi i te whānuitanga o ngā rauemi ngohengohe, engari he maha tonu ngā tikanga e pāngia ana e ngā ngoikoretanga nui e whakawhāiti ana i tā rātou whakamahinga8,9,10,11,12,13. Ko te kore o ngā tikanga whakamātautau motuhake ka taea te tautuhi tika me te pono i ngā āhuatanga ā-pūkaha o ngā rauemi tino ngawari me ngā paparanga mata e tino whakawhāiti ana i tā rātou whakamahinga i roto i ngā tono maha.
I roto i tā mātou mahi o mua, i whakaurua e mātou te karaihe whakapā lehfilcon A (CL), he rauemi ngohengohe rerekē me ngā āhuatanga mata tino ngawari katoa i ahu mai i ngā hoahoa koiora-whakamimetic pea i ahu mai i te mata o te cornea o te kanohi. I whakawhanakehia tēnei rauemi koiora mā te hono i tētahi paparanga polymer peka, whakawhiti-hono o te poly(2-methacryloyloxyethylphosphorylcholine (MPC)) (PMPC) ki runga i tētahi silicone hydrogel (SiHy) 15 i hangaia mō ngā taputapu hauora i runga i. Mā tēnei tukanga hono ka hangaia he paparanga ki runga i te mata he hanganga paraihe polymeric peka tino ngohengohe me te tino ngāwari. Kua whakaūhia e tā mātou mahi o mua ko te hanganga koiora-whakamimetic o te lehfilcon A CL e whakarato ana i ngā āhuatanga mata pai ake pērā i te whakapai ake i te aukati i te makuku me te poke, te whakanui ake i te hinu, me te whakaiti i te piri o te pūtau me te huakita15,16. Hei tāpiri, ko te whakamahinga me te whanaketanga o tēnei rauemi koiora-whakamimetic e kī ana hoki he whānui ake ki ētahi atu taputapu hauora. Nō reira, he mea nui kia whakaahuahia ngā āhuatanga o te mata o tēnei rauemi tino ngohengohe, kia mārama hoki ki tana taunekeneke ā-pūkaha ki te kanohi kia taea ai te hanga i tētahi pūtake mātauranga whānui hei tautoko i ngā whanaketanga me ngā tono ā muri ake nei. Ko te nuinga o ngā karaihe whakapā SiHy e wātea ana i te hokohoko he mea tito mai i tētahi ranunga ōrite o ngā pūmua hydrophilic me hydrophobic e hanga ana i tētahi hanganga rauemi ōrite17. He maha ngā rangahau kua mahia hei tirotiro i ō rātou āhuatanga ā-pūkaha mā te whakamahi i ngā tikanga whakamātautau kōpeketanga, tensile me te microindentation tuku iho18,19,20,21. Heoi, ko te hoahoa biomimetic hou o te lehfilcon A CL e hanga ana hei rauemi ahurei heterogeneous e rerekē ana ngā āhuatanga ā-pūkaha o ngā hanganga paraihe pūmua peka mai i ērā o te papa turanga SiHy. Nō reira, he tino uaua ki te ine tika i ēnei āhuatanga mā te whakamahi i ngā tikanga tuku iho me ngā tikanga kōpeketanga. Ko tētahi tikanga whai hua e whakamahi ana i te tikanga whakamātautau nanoindentation kua whakatinanahia i roto i te atomic force microscopy (AFM), he tikanga kua whakamahia hei whakatau i ngā āhuatanga ā-pūkaha o ngā rauemi viscoelastic ngohengohe pērā i ngā pūtau me ngā kiko koiora, tae atu ki ngā pūmua ngohengohe22,23,24,25. ,26,27,28,29,30. I roto i te AFM nanoindentation, ka honoa ngā kaupapa matua o te whakamātautau nanoindentation me ngā whanaketanga hou o te hangarau AFM hei whakarato i te tairongo ine me te whakamātautau i te whānuitanga o ngā rauemi ngohengohe rawa31,32,33,34,35,36. Hei tāpiri, ka tukuna e te hangarau ētahi atu painga nui mā te whakamahi i ngā āhuahanga rerekē. te indenter me te probe me te āheinga o te whakamātautau i roto i ngā momo wai.
Ka taea te wehewehe i te nanoindentation AFM kia toru ngā wāhanga matua: (1) ngā taputapu (ngā pūoko, ngā kite, ngā probe, me ētahi atu); (2) ngā tawhā ine (pēnei i te kaha, te nekehanga, te tere, te rahi o te arawhata, me ētahi atu); (3) te tukatuka raraunga (te whakatikatika turanga, te whakatau tata i te pūwāhi pa, te whakauru raraunga, te whakatauira, me ētahi atu). Ko tētahi raruraru nui o tēnei tikanga ko te maha o ngā rangahau i roto i ngā tuhinga e whakamahi ana i te nanoindentation AFM e whakaatu ana i ngā hua tau rerekē rawa atu mō te momo tauira/pūtau/rauemi kotahi37,38,39,40,41. Hei tauira, i akohia, i whakaritea hoki e Lekka et al. te awe o te āhua o te probe AFM ki te modulus Young i inehia o ngā tauira o ngā pūtau hydrogel me ngā pūtau heterogeneous he rite tonu te āhua. E ai ki a rātou he tino whakawhirinaki ngā uara modulus ki te kōwhiringa cantilever me te āhua o te pito, me te uara teitei rawa atu mō te probe āhua-piramita me te uara iti rawa o te 42 mō te probe porowhita. Waihoki, i akohia e Selhuber-Unkel et al. Kua whakaaturia te pānga o te tere o te whakaurunga, te rahi o te whakaurunga me te matotoru o ngā tauira polyacrylamide (PAAM) ki te modulus a Young i inehia e te nanoindentation ACM43. Ko tētahi atu āhuatanga uaua ko te kore o ngā rauemi whakamātautau modulus tino iti paerewa me ngā tikanga whakamātautau kore utu. Nā tēnei ka tino uaua ki te whiwhi hua tika me te maia. Heoi, he tino whai hua te tikanga mō ngā inenga whanaunga me ngā aromatawai whakatairite i waenga i ngā momo tauira rite, hei tauira te whakamahi i te nanoindentation AFM hei wehewehe i ngā pūtau noa mai i ngā pūtau matepukupuku 44, 45.
I te whakamātautau i ngā rauemi ngohengohe me te nanoindentation AFM, ko te ture whānui ko te whakamahi i tētahi ine me te pūmau puna iti (k) e ōrite ana ki te modulus tauira me te pito porowhita/hemispherical kia kore ai te ine tuatahi e wero i ngā mata tauira i te pa tuatahi ki ngā rauemi ngohengohe. He mea nui hoki kia kaha te tohu whakapēhanga i hangaia e te ine kia kitea e te pūnaha kite laser24,34,46,47. Mō ngā pūtau, kiko me ngā tiēre tino ngohengohe, ko tētahi atu wero ko te wikitoria i te kaha piri i waenga i te ine me te mata tauira kia taea ai te whakaputa anō me te pono o ngā inenga48,49,50. Tae noa ki ēnei wā tata nei, ko te nuinga o ngā mahi i runga i te nanoindentation AFM kua arotahi ki te ako i te whanonga miihini o ngā pūtau koiora, ngā kiko, ngā tiēre, ngā hydrogel, me ngā biomolecules mā te whakamahi i ngā ine porowhita nui, e kiia nei ko ngā ine colloidal (CP). , 47, 51, 52, 53, 54, 55. Ko te whānui o ēnei pito he 1 ki te 50 µm, ā, he mea hanga noa mai i te karāhe borosilicate, te polymethyl methacrylate (PMMA), te polystyrene (PS), te silicon dioxide (SiO2) me te waro rite-taimana (DLC). Ahakoa ko te nanoindentation CP-AFM te mea tuatahi hei whiriwhiri mā te whakaahua i ngā tauira ngohengohe, he raruraru me ngā herenga anō ōna. Mā te whakamahi i ngā pito porowhita nui, he rite ki te micron te rahi, ka nui ake te horahanga o te whakapā o te pito ki te tauira, ā, ka ngaro te taumira ā-wāhi. Mō ngā tauira ngohengohe, kāore i te ōrite, ka rerekē pea ngā āhuatanga ā-pūkaha o ngā huānga ā-rohe i te toharite i runga i te rohe whānui, ka taea e te indentation CP te huna i te kore ōrite o ngā āhuatanga i runga i te tauine ā-rohe52. Ko ngā tirotiro colloidal ka hangaia mā te tāpiri i ngā porowhita colloidal rahi-micron ki ngā cantilevers kore pito mā te whakamahi i ngā piripiri epoxy. He maha ngā raruraru o te tukanga hanga, ā, ka arahi pea ki ngā rerekētanga i roto i te tukanga whakatikatika i te tirotiro. Hei tāpiri, ko te rahi me te papatipu o ngā matūriki colloidal e pā tika ana ki ngā tawhā whakatikatika matua o te cantilever, pērā i te auau oro, te pakari o te puna, me te aro ki te piko56,57,58. Nō reira, ko ngā tikanga e whakamahia whānuitia ana mō ngā tirotiro AFM tuku iho, pērā i te whakatikatika pāmahana, kāore pea e whakarato i te whakatikatika tika mō te CP, ā, me whakamahi pea ētahi atu tikanga hei mahi i ēnei whakatikatika57, 59, 60, 61. Ko ngā whakamātautau whakatō CP noa e whakamahi ana i ngā rerekētanga nui o te cantilever hei ako i ngā āhuatanga o ngā tauira ngohengohe, ka puta he raruraru anō i te wā e whakatikatika ana i te whanonga kore-raina o te cantilever i ngā rerekētanga nui62,63,64. Ko ngā tikanga whakatō whakatō hou o te tirotiro colloidal e whai whakaaro ana ki te āhua o te cantilever e whakamahia ana hei whakatikatika i te tirotiro, engari kāore e aro ki te awe o ngā matūriki colloidal, ka puta he koretake anō i roto i te tika o te tikanga38,61. Waihoki, ko ngā moduli rapa e tatauhia ana mā te taunga tauira whakapā e whakawhirinaki tika ana ki te āhua o te tirotiro kōpiko, ā, ko te kore e ōrite i waenga i ngā āhuatanga o te mata o te pito me te tauira ka arahi pea ki ngā hapa27, 65, 66, 67, 68. Kua whakamāramahia ētahi mahi hou a Spencer et al. Ngā āhuatanga me whai whakaaro i te wā e whakaahua ana i ngā paraihe polymer ngohengohe mā te whakamahi i te tikanga nanoindentation CP-AFM. I kōrero rātou ko te pupuri i te wai matotoru i roto i ngā paraihe polymer hei mahi a te tere ka hua ake te pikinga o te utaina o te upoko, ā, nō reira he rerekē ngā inenga o ngā āhuatanga e whakawhirinaki ana ki te tere30,69,70,71.
I roto i tēnei rangahau, kua whakaahuahia e mātou te modulus mata o te rauemi tino ngawari, tino ngāwari, lehfilcon A CL mā te whakamahi i te tikanga nanoindentation AFM kua whakarerekētia. I runga i ngā āhuatanga me te hanganga hou o tēnei rauemi, kāore i te rawaka te whānuitanga o te aro o te tikanga indentation tuku iho ki te whakaahua i te modulus o tēnei rauemi tino ngawari, nō reira me whakamahi i te tikanga nanoindentation AFM me te taumata aro teitei ake me te taumata aro iti ake. Whai muri i te arotake i ngā ngoikoretanga me ngā raruraru o ngā tikanga nanoindentation probe AFM colloidal o nāianei, ka whakaaturia e mātou te take i whiriwhiria ai e mātou he probe AFM iti ake, he mea hanga motuhake hei whakakore i te aro, te haruru papamuri, te tohu i te pūwāhi whakapā, te ine i te modulus tere o ngā rauemi ngohengohe rerekē pērā i te whakawhirinaki ki te pupuri wai, me te ine tika. Hei tāpiri, i taea e mātou te ine tika i te āhua me ngā inenga o te pito indentation, e āhei ai mātou ki te whakamahi i te tauira taunga cone-sphere hei whakatau i te modulus o te elasticity me te kore e aromatawai i te horahanga whakapā o te pito ki te rauemi. Ko ngā whakaaro huna e rua e inehia ana i roto i tēnei mahi ko ngā āhuatanga rauemi tino ngāwari me te modulus hohonu-kore indentation. Mā te whakamahi i tēnei tikanga, i whakamatautauria tuatahitia e mātou ngā paerewa tino ngohengohe me te modulus e mōhiotia ana hei ine i te tikanga, kātahi ka whakamahia tēnei tikanga hei whakaahua i ngā mata o ngā rauemi arotahi whakapā rerekē e rua. Ko tēnei tikanga whakaahua i ngā mata nanoindentation AFM me te tairongo nui ake e tika ana kia whakamahia ki te whānuitanga o ngā rauemi tino ngohengohe kanorau koiora me te whakamahinga pea i roto i ngā taputapu hauora me ngā tono koiora.
I whiriwhiria ngā karāhe whakapā Lehfilcon A (Alcon, Fort Worth, Texas, USA) me ā rātou papa silicone hydrogel mō ngā whakamātautau nanoindentation. I whakamahia he whakairinga karāhe i hangaia mō te whakamātautau. Hei tāuta i te karāhe mō te whakamātautau, i ata whakatakotoria ki runga i te tūnga āhua-poupou, me te whakarite kia kore he mirumiru hau e uru ki roto, kātahi ka whakapirihia ki ngā taha. He kōhao i roto i te hononga i te tihi o te pupuri karāhe e whakarato ana i te urunga ki te pokapū whatu o te karāhe mō ngā whakamātautau nanoindentation me te pupuri i te wai ki tōna wāhi. Mā tēnei ka noho makuku rawa ngā karāhe. 500 μl o te otinga takai karāhe whakapā i whakamahia hei otinga whakamātautau. Hei manatoko i ngā hua tauanga, i whakaritea ngā hydrogel polyacrylamide kore-whakahohe (PAAM) e wātea ana i te hokohoko mai i te hanganga polyacrylamide-co-methylene-bisacrylamide (100 mm Petrisoft Petri dishes, Matrigen, Irvine, CA, USA), he modulus elastic e mōhiotia ana o te 1 kPa. Whakamahia kia 4-5 pata (tata ki te 125 µl) o te tote phosphate buffered (PBS mai i Corning Life Sciences, Tewkesbury, MA, USA) me te 1 pata o te otinga OPTI-FREE Puremoist contact lens (Alcon, Vaud, TX, USA). ) ki te atanga AFM hydrogel-probe.
I tirohia ngā tauira o ngā papa Lehfilcon A CL me SiHy mā te whakamahi i tētahi pūnaha FEI Quanta 250 Field Emission Scanning Electron Microscope (FEG SEM) e mau ana i tētahi taputapu tirotiro Scanning Transmission Electron Microscope (STEM). Hei whakarite i ngā tauira, i horoia tuatahitia ngā karaihe ki te wai, ā, i tapahia kia rite ki ngā poro porotaka. Hei whakatutuki i te rerekētanga rerekē i waenga i ngā wāhanga hydrophilic me hydrophobic o ngā tauira, i whakamahia he otinga 0.10% o RuO4 kua whakapūmautia hei tae, ā, i rumakina ngā tauira mō te 30 meneti. He mea nui te tae lehfilcon A CL RuO4, ehara i te mea hei whakatutuki i te rerekētanga rerekētanga pai ake, engari hei āwhina hoki ki te tiaki i te hanganga o ngā paraihe polymer peka i roto i tō rātou āhua taketake, ka kitea i runga i ngā whakaahua STEM. Kātahi ka horoia, ka whakamaroketia ki roto i tētahi raupapa ranunga etanoru/wai me te pikinga o te kukū etanoru. Kātahi ka makahia ngā tauira ki te epoxy EMBed 812/Araldite, i whakamaroketia i te pō i te 70°C. I tapahia ngā poraka tauira i whiwhi mā te whakapūhui kapia ki te ultramicrotome, ā, i tirohia ngā wāhanga angiangi i puta mai mā te whakamahi i te taputapu kite STEM i te aratau korehau iti i te ngaohiko whakatere o te 30 kV. I whakamahia te pūnaha SEM kotahi mō te whakaahua taipitopito o te pūwero PFQNM-LC-A-CAL AFM (Bruker Nano, Santa Barbara, CA, USA). I ​​whiwhihia ngā whakaahua SEM o te pūwero AFM i te aratau korehau teitei noa me te ngaohiko whakatere o te 30 kV. Tikina ngā whakaahua i ngā koki me ngā whakanuinga rerekē hei tuhi i ngā taipitopito katoa o te āhua me te rahi o te pito pūwero AFM. I inehia ngā āhuatanga katoa o te pito e hiahiatia ana i roto i ngā whakaahua mā te mamati.
I whakamahia he karu hiko ā-atomika Dimension FastScan Bio Icon (Bruker Nano, Santa Barbara, CA, USA) me te aratau "PeakForce QNM in Fluid" hei tiro me te whaka-nano i ngā tauira lehfilcon A CL, te papa SiHy, me te PAAm hydrogel. Mō ngā whakamātautau whakaahua, i whakamahia he probe PEAKFORCE-HIRS-FA (Bruker) me te radius pito ingoa o te 1 nm hei hopu i ngā whakaahua taumira teitei o te tauira i te tere matawai o te 0.50 Hz. I tangohia ngā whakaahua katoa i roto i te wairewa.
I whakahaerehia ngā whakamātautau nanoindentation AFM mā te whakamahi i tētahi pūwero PFQNM-LC-A-CAL (Bruker). He pito silicon kei runga i tētahi cantilever nitride he 345 nm te matotoru, he 54 µm te roa, he 4.5 µm te whānui, me te auau resonant o te 45 kHz. He mea hanga motuhake hei whakaahua me te mahi i ngā inenga nanomechanical tauanga i runga i ngā tauira koiora ngohengohe. Ka whakatikatikaina takitahitia ngā pūoko i te wheketere me ngā tautuhinga puna kua whakatikatikaina i mua. Ko ngā pūmau puna o ngā pūwero i whakamahia i tēnei rangahau i roto i te awhe o te 0.05–0.1 N/m. Hei whakatau tika i te āhua me te rahi o te pito, i whakaahuahia te pūwero i roto i ngā taipitopito mā te whakamahi i te SEM. I runga i te pikitia. E whakaatu ana te Pikitia 1a i tētahi kauwhata hiko matawai teitei, iti te whakanui o te pūwero PFQNM-LC-A-CAL, e whakarato ana i tētahi tirohanga whānui o te hoahoa pūwero. I runga i te pikitia. E whakaatu ana te pikitia. 1b i tētahi tirohanga whakanuia o te tihi o te pito pūwero, e whakarato ana i ngā mōhiohio e pā ana ki te āhua me te rahi o te pito. I te pito rawa, he taha tuawhā te ngira, he 140 nm te whānui (Pikitia 1c). I raro iho i tēnei, ka whakawhāiti te pito ki te āhua kōniko, ka eke ki te roa ine tata ki te 500 nm. I waho o te rohe whakawhāiti, he porotaka te pito, ā, ka mutu ki te roa katoa o te pito o te 1.18 µm. Koinei te wāhanga mahi matua o te pito o te mata o te ngira. Hei tāpiri, i whakamahia hoki he mata nui porowhita polystyrene (PS) (Novascan Technologies, Inc., Boone, Iowa, USA) me te whānui o te pito o te 45 µm me te pūmau puna o te 2 N/m hei whakamātautau hei mata kōrori. me te mata PFQNM-LC-A-CAL 140 nm hei whakataurite.
Kua pūrongohia ka taea te mau i te wai i waenganui i te pūwero AFM me te hanganga paraihe polymer i te wā o te nanoindentation, ka puta he kaha ki runga i te pūwero AFM i mua i te pa atu ki te mata69. Ka taea e tēnei pānga extrusion matotoru nā te pupuri wai te whakarerekē i te pūwāhi whakapā kitea, ā, ka pā ki ngā inenga modulus mata. Hei ako i te pānga o te āhua o te pūwero me te tere o te kōpiko ki te pupuri wai, i tuhia ngā ānau kaha kōpiko mō ngā tauira lehfilcon A CL mā te whakamahi i tētahi pūwero diameter 140 nm i ngā tere nekehanga pumau o te 1 µm/s me te 2 µm/s. Ko te diameter o te pūwero he 45 µm, ko te tautuhinga kaha pumau 6 nN i tutuki i te 1 µm/s. I whakahaerehia ngā whakamātautau me tētahi pūwero he 140 nm te diameter i te tere kōpiko o te 1 µm/s me te kaha kua whakaritea o te 300 pN, i whiriwhiria hei hanga i tētahi pēhanga whakapā i roto i te awhe ā-tinana (1–8 kPa) o te kamo o runga. pēhanga 72. I whakamatautauria ngā tauira ngohengohe kua oti te hanga o te PAA hydrogel me te pēhanga o te 1 kPa mō te kaha kōpiko o te 50 pN i te tere o te 1 μm/s mā te whakamahi i tētahi ine he diameter o te 140 nm.
I te mea ko te roa o te wāhanga kōniko o te pito o te pūwero PFQNM-LC-A-CAL he tata ki te 500 nm, mō tētahi hōhonutanga kōpiko < 500 nm ka taea te kī haumaru ka noho tonu te āhua o te pūwero i te wā kōpiko ki tōna āhua kōniko. Hei tāpiri, e kīia ana ka whakaatu te mata o te rauemi e whakamatautauria ana i tētahi urupare whakapūmau ka taea te huri, ka whakaūtia hoki i ngā wāhanga e whai ake nei. Nō reira, i runga i te āhua me te rahi o te pito, i whiriwhiria e mātou te tauira whakauru kōniko-porowhita i whakawhanakehia e Briscoe, Sebastian me Adams, e wātea ana i roto i te pūmanawa a te kaihoko, hei tukatuka i ā mātou whakamātautau nanoindentation AFM (NanoScope). Pūmanawa tātari raraunga wehenga, Bruker) 73. E whakaahua ana te tauira i te whanaungatanga kaha-nekehanga F(δ) mō tētahi kōniko me te hapa pito porowhita. I runga i te pikitia. E whakaatu ana te Pikitia 2 i te āhua whakapā i te wā e taunekeneke ana te koeko mārō me te pito porowhita, ko R te pūtoro o te pito porowhita, ko a te pūtoro whakapā, ko b te pūtoro whakapā i te pito o te pito porowhita, ko δ te pūtoro whakapā. te hohonutanga o te kōhao, ko θ te haurua-koki o te koeko. E whakaatu mārama ana te ahua SEM o tēnei tirotiro ka hanumi te pito porowhita 140 nm te whānui ki roto i te koeko, nō reira i konei ka tautuhia a b mā R anake, arā, ko b = R cos θ. Ka whakaratohia e te pūmanawa i tukuna mai e te kaihoko he whanaungatanga koeko-porowhita hei tatau i ngā uara modulus a Young (E) mai i ngā raraunga wehenga kaha me te whakaaro ko a > b. Te whanaungatanga:
ko F te kaha whakatōtō, ko E te modulus a Young, ko ν te ōwehenga a Poisson. Ka taea te whakatau tata i te radius whakapā a mā te whakamahi i:
Kauwhata o te āhua whakapā o tētahi koeko mārō me te pito porowhita kua pēhia ki roto i te rauemi o tētahi karāhe whakapā Lefilcon me tētahi paparanga mata o ngā paraihe pūmua peka.
Mena ko a ≤ b, ka heke te whanaungatanga ki te whārite mō te kōpeketanga porowhita tikanga;
E whakapono ana mātou mā te taunekeneke o te pūwero whakatōtō me te hanganga peka o te paraihe pūwero PMPC ka nui ake te pūtoro whakapā a i te pūtoro whakapā porowhita b. Nō reira, mō ngā inenga tauanga katoa o te modulus rapa i mahia i roto i tēnei rangahau, i whakamahia e mātou te whakawhirinakitanga i whiwhihia mō te take a > b.
Ko ngā rauemi biomimetic tino ngohengohe i akohia i roto i tēnei rangahau i whakaahuahia whānuitia mā te whakamahi i te karu hiko tuku matawai (STEM) o te wāhanga whakawhiti tauira me te karu kaha atomika (AFM) o te mata. I mahia tēnei whakaahua mata taipitopito hei toronga o tā mātou mahi kua whakaputaina i mua, i roto i tēnei i whakatauhia ai e mātou ko te hanganga paraihe polymeric peka hihiri o te mata lehfilcon A CL kua whakarerekētia e te PMPC he rite ngā āhuatanga miihini ki te kiko corneal taketake 14. Mō tēnei take, ka kiia e mātou ngā mata arotahi whakapā he rauemi biomimetic14. I te pikitia 3a, b e whakaatu ana i ngā wāhanga whakawhiti o ngā hanganga paraihe polymer PMPC peka i runga i te mata o te papa lehfilcon A CL me te papa SiHy kāore i rongoatia. I tātarihia anō ngā mata o ngā tauira e rua mā te whakamahi i ngā whakaahua AFM taumira teitei, i whakaū anō i ngā hua o te tātari STEM (Pikitia 3c, d). Ina tangohia ngātahi, ka hoatu e ēnei whakaahua he roa tata o te hanganga paraihe polymer peka PMPC i te 300–400 nm, he mea nui mō te whakamaori i ngā inenga nanoindentation AFM. Ko tētahi atu kitenga matua i ahu mai i ngā whakaahua ko te hanganga mata whānui o te rauemi koiora-koiora CL he rerekē te āhua o te hanganga mai i te rauemi papa SiHy. Ka kitea pea tēnei rerekētanga i roto i te āhua o te mata i roto i tā rātou taunekeneke ā-pūkaha me te pūwero AFM e whakatōtō ana, ā, muri iho i roto i ngā uara modulus i inehia.
Ngā whakaahua STEM whakawhiti-wāhanga o (a) te papa lehfilcon A CL me (b) te papa SiHy. Pae tauine, 500 nm. Ngā whakaahua AFM o te mata o te papa lehfilcon A CL (c) me te papa SiHy turanga (d) (3 µm × 3 µm).
He ngohengohe ngā pūmua koiora me ngā hanganga paraihe pūmua, ā, kua whānuitia te rangahau me te whakamahi i roto i ngā tono hauora maha74,75,76,77. Nō reira, he mea nui te whakamahi i te tikanga nanoindentation AFM, ka taea te ine tika me te pono i ō rātou āhuatanga miihini. Engari i te wa ano, ko ngā āhuatanga ahurei o ēnei rauemi tino ngohengohe, pērā i te modulus tino iti o te elasticity, te nui o te wai me te teitei o te elasticity, he maha ngā wā ka uaua ki te whiriwhiri i te rauemi tika, te āhua me te āhua o te probe indentation. rahi. He mea nui tēnei kia kore ai te indenter e wero i te mata ngohengohe o te tauira, ka arahi pea ki ngā hapa i te whakatau i te pūwāhi e pa ana ki te mata me te rohe e pa ana.
Mō tēnei, he mea nui te māramatanga whānui ki te āhua o ngā rauemi koiora tino ngohengohe (lehfilcon A CL). Ko ngā mōhiohio e pā ana ki te rahi me te hanganga o ngā paraihe pūmua peka i whiwhihia mā te whakamahi i te tikanga whakaahua e whakarato ana i te pūtake mō te whakaahua ā-pūkaha o te mata mā te whakamahi i ngā tikanga nanoindentation AFM. Engari i ngā probes colloidal porowhita-rahi-micron, i whiriwhiria e mātou te probe silicon nitride PFQNM-LC-A-CAL (Bruker) me te diameter pito o te 140 nm, i hangaia mō te mahere tauanga o ngā āhuatanga ā-pūkaha o ngā tauira koiora 78, 79, 80, 81, 82, 83, 84. Ka taea te whakamārama i te take mō te whakamahi i ngā probes koi ki te whakataurite ki ngā probes colloidal tikanga mā ngā āhuatanga hanganga o te rauemi. Mā te whakataurite i te rahi o te pito o te ine (~140 nm) ki ngā paraihe pūmua peka i runga i te mata o te CL lehfilcon A, e whakaaturia ana i te Pikitia 3a, ka taea te whakatau he nui te pito kia pā tika atu ki ēnei hanganga paraihe, ka whakaiti i te tūponotanga o te pito kia werohia mā roto i a rātou. Hei whakaatu i tēnei kōrero, kei te Pikitia 4 he whakaahua STEM o te lehfilcon A CL me te pito kōkuhu o te ine AFM (kua tuhia ki te tauine).
He hoahoa e whakaatu ana i te ahua STEM o te lehfilcon A CL me te inenga kōpiko ACM (kua tuhia ki te tauine).
Hei tāpiritanga, he iti te rahi o te pito o te 140 nm hei karo i te mōrearea o tētahi o ngā pānga whakapiripiri i kōrerotia i mua mō ngā paraihe polymer i hangaia e te tikanga nanoindentation CP-AFM69,71. E whakaaro ana mātou nā te āhua porowhita-kōne motuhake me te iti o te rahi o tēnei pito AFM (Pikitia 1), ko te āhua o te pihi kaha i hangaia e te nanoindentation lehfilcon A CL kāore e whakawhirinaki ki te tere o te kōpeketanga, ki te tere uta/tango rānei. Nō reira, kāore e pāngia e ngā pānga poroelastic. Hei whakamātautau i tēnei whakapae, i kōpeketia ngā tauira lehfilcon A CL ki tētahi kaha mōrahi pumau mā te whakamahi i tētahi tirotiro PFQNM-LC-A-CAL, engari i ngā tere rerekē e rua, ā, ko ngā pihi kaha kume me te whakahoki i puta mai i whakamahia hei tuhi i te kaha (nN) i te wehenga (µm) e whakaaturia ana i te Pikitia 5a. He mārama kei te taupatupatu ngā pihi kaha i te wā e utaina ana, e tangohia ana hoki, ā, kāore he taunakitanga mārama e whakaatu ana ka piki ake te kutikuti kaha i te hohonutanga kore o te kōhao me te tere kōhao i roto i te ahua, e kī ana i whakaahuatia ngā wāhanga paraihe takitahi me te kore he pānga poroelastic. He rerekē, e kitea ana ngā pānga pupuri wai (ngā pānga extrusion viscous me te poroelasticity) mō te probe AFM diameter 45 µm i te tere kōhao kotahi, ā, e whakamāramahia ana e te hysteresis i waenga i ngā pihi totoro me ngā pihi hoki whakamuri, e whakaaturia ana i te Pikitia 5b. E tautoko ana ēnei hua i te whakapae, ā, e kī ana he pai ngā probe diameter 140 nm mō te whakaahua i aua mata ngohengohe.
Ngā kōpiko kaha whakatōtō lehfilcon A CL mā te whakamahi i te ACM; (a) te whakamahi i tētahi inenga he whānui o te 140 nm i ngā tere uta e rua, e whakaatu ana i te korenga o tētahi pānga poroelastic i te wā e whakatōtō ana te mata; (b) te whakamahi i ngā inenga he whānui o te 45 µm me te 140 nm. E whakaatu ana ngā s i ngā pānga o te extrusion viscous me te poroelastic mō ngā inenga nui ki te whakataurite ki ngā inenga iti ake.
Hei whakaahua i ngā mata tino ngohengohe, me whai i ngā tikanga nanoindentation AFM te tirotiro pai rawa atu hei ako i ngā āhuatanga o te rauemi e akohia ana. Haunga te āhua me te rahi o te pito, he mea nui te aro o te pūnaha kite AFM, te aro ki te piko o te pito i roto i te taiao whakamātautau, me te pakari o te cantilever ki te whakatau i te tika me te pono o ngā inenga nanoindentation. Mō tā mātou pūnaha AFM, ko te rohe kite o te Position Sensitive Detector (PSD) he tata ki te 0.5 mV, ā, e hangai ana ki te tere puna kua whakatikatikaina i mua me te aro o te piko wai kua tatauhia o te tirotiro PFQNM-LC-A-CAL, e rite ana ki te aro o te kawenga ariā. he iti iho i te 0.1 pN. Nō reira, ka taea e tēnei tikanga te ine i te kaha piko iti rawa ≤ 0.1 pN me te kore he wāhanga haruru taha. Heoi, he tata kore e taea e te pūnaha AFM te whakaiti i te haruru taha ki tēnei taumata nā ngā āhuatanga pēnei i te wiri miihini me te hihiri wai. Ka whakawhāitihia e ēnei āhuatanga te aro whānui o te tikanga nanoindentation AFM, ā, ka hua ake he tohu haruru papamuri tata ki te ≤ 10 pN. Mō te whakaahuatanga o te mata, i whakatōkia ngā tauira papa lehfilcon A CL me SiHy i raro i ngā āhuatanga tino makuku mā te whakamahi i tētahi tirotiro 140 nm mō te whakaahuatanga SEM, ā, ko ngā ānau kaha i puta mai i whakakīia i waenga i te kaha (pN) me te pēhanga. Kei te whakaatuhia te kauwhata wehenga (µm) i te Pikitia 6a. Ki te whakatauritea ki te papa turanga SiHy, ka whakaatu mārama te ānau kaha lehfilcon A CL i tētahi wāhanga whakawhiti e tīmata ana i te wāhi e pā ana ki te paraihe polymer kua marau, ā, ka mutu me te huringa koi o te hononga tohu pari o te pito ki te rauemi kei raro. E whakaatu ana tēnei wāhanga whakawhiti o te ānau kaha i te whanonga tino ngāwari o te paraihe polymer peka i runga i te mata, e whakaatuhia ana e te ānau pēhitanga e whai tata ana i te ānau kume me te rerekētanga o ngā āhuatanga miihini i waenga i te hanganga paraihe me te rauemi SiHy matotoru. I te whakataurite i te lefilcon. Ko te wehenga o te roa toharite o te paraihe pūmua peka i roto i te ahua STEM o te PCS (Pikitia 3a) me tōna pihi kaha i te taha o te abscissa i roto i te Pikitia 3a. 6a e whakaatu ana ka taea e te tikanga te kite i te pito me te pūmua peka e tae ana ki te tihi rawa o te mata. Te pa i waenga i ngā hanganga paraihe. Hei tāpiri, ko te taupatupatu tata o ngā pihi kaha e tohu ana kāore he pānga pupuri wai. I tēnei take, kāore rawa he piri i waenga i te ngira me te mata o te tauira. Ko ngā wāhanga o runga rawa o ngā pihi kaha mō ngā tauira e rua e taupatupatu ana, e whakaata ana i te ōrite o ngā āhuatanga ā-pūkaha o ngā rauemi papa.
(a) Ngā ānau kaha nanoindentation AFM mō ngā papa lehfilcon A CL me ngā papa SiHy, (b) ngā ānau kaha e whakaatu ana i te whakatau tataitanga o te pūwāhi whakapā mā te whakamahi i te tikanga paepae haruru papamuri.
Hei ako i ngā taipitopito pai ake o te pihi kaha, ka tuhia anō te pihi kume o te tauira lehfilcon A CL ki te Pikitia 6b me te kaha mōrahi o te 50 pN i te tuaka-y. Ka whakaratohia e tēnei kauwhata he kōrero nui mō te haruru papamuri taketake. Kei roto i te awhe o te ±10 pN te haruru, e whakamahia ana hei whakatau tika i te pūwāhi whakapā me te tatau i te hohonutanga o te kōhao. E ai ki ngā tuhinga, he mea nui te tautuhi i ngā pūwāhi whakapā hei aromatawai tika i ngā āhuatanga rauemi pēnei i te modulus85. Kua whakaatuhia e tētahi huarahi e uru ana ki te tukatuka aunoa i ngā raraunga pihi kaha he pai ake te taunga i waenga i te taunga raraunga me ngā inenga tauanga mō ngā rauemi ngohengohe86. I roto i tēnei mahi, he māmā noa iho, he whāinga hoki tā mātou whiriwhiri i ngā pūwāhi whakapā, engari he iti iho ōna herenga. Tērā pea ka hua ake i tā mātou huarahi matawhānui ki te whakatau i te pūwāhi whakapā he nui rawa ngā uara modulus mō ngā hohonutanga kōhao iti ake (< 100 nm). Ko te whakamahinga o te kimi i ngā pūwāhi pā e hangai ana ki te rauropi me te tukatuka raraunga aunoa he haere tonu pea o tēnei mahi ā muri ake nei hei whakapai ake i tā mātou tikanga. Nō reira, mō te haruru papamuri ā-roto i te raupapa o te ±10 pN, ka tautuhia e mātou te pūwāhi whakapā hei pūwāhi raraunga tuatahi i runga i te tuaka-x i te Pikitia 6b me te uara o te ≥10 pN. Kātahi, i runga anō i te paepae haruru o te 10 pN, ka tohua he rārangi poutū i te taumata o te ~0.27 µm te pūwāhi whakapā ki te mata, ā, muri iho ka haere tonu te pihi totoro kia tae rā anō te papa ki te hohonutanga o te ~270 nm. He mea whakamīharo, i runga i te rahi o ngā āhuatanga paraihe polymer manga (300–400 nm) i inehia mā te whakamahi i te tikanga whakaahua, ko te hohonutanga o te tauira CL lehfilcon i kitea mā te whakamahi i te tikanga paepae haruru papamuri he tata ki te 270 nm, he tino tata ki te rahi ine me te STEM. Ka whakaū anō ēnei hua i te hototahitanga me te whakamahinga o te āhua me te rahi o te pito tirotiro AFM mō te whakaurunga o tēnei hanganga paraihe polymer manga tino ngohengohe me te tino ngāwari. Ka whakaratohia hoki e ēnei raraunga he taunakitanga kaha hei tautoko i tā mātou tikanga whakamahi i te haruru papamuri hei paepae mō te tohu i ngā pūwāhi whakapā. Nō reira, ko ngā hua tauanga katoa e puta mai ana i te whakatauira pāngarau me te taurite i te pihi kaha me tino tika.
Ko ngā inenga tauanga mā ngā tikanga nanoindentation AFM e tino whakawhirinaki ana ki ngā tauira pāngarau e whakamahia ana mō te whiriwhiri raraunga me te tātari i muri mai. Nō reira, he mea nui kia whakaarohia ngā āhuatanga katoa e pā ana ki te whiriwhiringa o te indenter, ngā āhuatanga rauemi me te tikanga o tā rātou taunekeneke i mua i te whiriwhiri i tētahi tauira. I tēnei take, i āta whakaahuahia te āhua o te pito mā te whakamahi i ngā whakaahua SEM (Pikitia 1), ā, i runga i ngā hua, ko te probe nanoindenting AFM 140 nm te whānui me te koeko mārō me te āhua o te pito porowhita he kōwhiringa pai mō te whakaahua i ngā tauira lehfilcon A CL79. Ko tētahi atu āhuatanga nui me āta arotake ko te ngāwari o te rauemi polymer e whakamatautauria ana. Ahakoa e whakamārama mārama ana ngā raraunga tuatahi o te nanoindentation (Pikitia 5a me te 6a) i ngā āhuatanga o te taupatupatu o ngā pihi kume me te kōpeketanga, arā, te whakaoranga katoa o te rauemi, he mea tino nui kia whakaūhia te āhua ngāwari o ngā hononga. Hei whakatutuki i tēnei, e rua ngā kōhao i mahia i te wāhi kotahi i runga i te mata o te tauira lehfilcon A CL i te tere kōhao o te 1 µm/s i raro i ngā āhuatanga whakamakuku katoa. Ko ngā raraunga o te pihi kaha i puta mai ka whakaaturia i te pikitia 7, ā, e ai ki te whakaaro, he tata rite tonu ngā pihi whakawhānui me te kōpeketanga o ngā tānga e rua, e whakaatu ana i te ngāwari teitei o te hanganga paraihe polymer peka.
E rua ngā ānau kaha kōpiko i te wāhi kotahi i runga i te mata o te lehfilcon. E tohu ana te A CL i te ngāwari pai o te mata o te arotahi.
I runga i ngā mōhiohio i whiwhi mai i ngā whakaahua SEM me STEM o te pito o te pūwero me te mata lehfilcon A CL, ko te tauira porowhita-kōne he whakaaturanga pāngarau whaitake o te taunekeneke i waenga i te pito o te pūwero AFM me te rauemi polymer ngohengohe e whakamatautauria ana. Hei tāpiri, mō tēnei tauira porowhita-kōne, ko ngā whakapae taketake mō ngā āhuatanga rapa o te rauemi kua tāia he pono mō tēnei rauemi biomimetic hou, ā, e whakamahia ana hei ine i te modulus rapa.
I muri i te aromatawai whānui o te tikanga nanoindentation AFM me ōna wāhanga, tae atu ki ngā āhuatanga o te probe indentation (āhua, rahi, me te pakari o te puna), te aro (te haruru papamuri me te whakatau tataitanga), me ngā tauira taunga raraunga (ngā inenga modulus tauanga), i whakamahia te tikanga. whakaahuahia ngā tauira tino ngawari e wātea ana i te hokohoko hei manatoko i ngā hua tauanga. I whakamatautauria he hydrogel polyacrylamide arumoni (PAAM) me te modulus elastic o te 1 kPa i raro i ngā āhuatanga wai mā te whakamahi i te probe 140 nm. Kei roto i ngā Mōhiohio Tāpiri ngā taipitopito o te whakamātautau me ngā tataunga o te kōwae. I whakaatuhia e ngā hua ko te modulus toharite i inehia he 0.92 kPa, ā, ko te %RSD me te ōrau (%) te rerekētanga mai i te modulus e mōhiotia ana he iti iho i te 10%. E whakaū ana ēnei hua i te tika me te taea te whakaputa anō o te tikanga nanoindentation AFM i whakamahia i roto i tēnei mahi hei ine i ngā moduli o ngā rauemi tino ngawari. I whakaahuahia anō ngā mata o ngā tauira lehfilcon A CL me te papa turanga SiHy mā te whakamahi i te tikanga nanoindentation AFM kotahi hei ako i te modulus whakapā kitea o te mata tino ngohengohe hei mahi a te hohonutanga o te indentation. I hangaia ngā pihi wehenga kaha indentation mō ngā tauira e toru o ia momo (n = 3; kotahi te indentation mō ia tauira) i te kaha o te 300 pN, te tere o te 1 µm/s, me te makuku katoa. I whakatatahia te pihi tiritiri kaha indentation mā te whakamahi i te tauira porowhita-kōne. Hei whiwhi i te modulus e whakawhirinaki ana ki te hohonutanga o te indentation, i whakatakotoria he wāhanga whānui 40 nm o te pihi kaha i ia pikinga o te 20 nm mai i te pūwāhi whakapā, ā, i inehia ngā uara o te modulus i ia taahiraa o te pihi kaha. Spin Cy et al. Kua whakamahia he huarahi rite hei whakaahua i te gradient modulus o ngā paraihe polymer poly(lauryl methacrylate) (P12MA) mā te whakamahi i te nanoindentation probe AFM colloidal, ā, he rite tonu ki ngā raraunga e whakamahi ana i te tauira whakapā Hertz. Mā tēnei huarahi ka whakaratohia he kauwhata o te modulus whakapā kitea (kPa) me te hohonutanga kōpiko (nm), e whakaaturia ana i te Pikitia 8, e whakaatu ana i te modulus whakapā kitea/te rōnaki hohonu. Ko te modulus elastic i tatauhia o te tauira CL lehfilcon A kei roto i te awhe o te 2–3 kPa i roto i te 100 nm o runga o te tauira, ā, ki tua atu i tēnā ka tīmata te piki haere me te hohonu. I tētahi atu taha, i te whakamātautau i te papa turanga SiHy me te kore he kiriata rite ki te paraihe i runga i te mata, ko te hohonutanga kōpiko mōrahi i tutukihia i te kaha o te 300 pN he iti iho i te 50 nm, ā, ko te uara modulus i whiwhi mai i ngā raraunga he tata ki te 400 kPa, he rite ki ngā uara o te modulus a Young mō ngā rauemi papatipu.
Te modulus whakapā āhua (kPa) vs. te hohonutanga kōhao (nm) mō ngā papa lehfilcon A CL me SiHy mā te whakamahi i te tikanga nanoindentation AFM me te āhua porowhita-kōne hei ine i te modulus.
Ko te mata o runga rawa o te hanganga paraihe pūmua peka biomimetic hou e whakaatu ana i te modulus o te ngāwari (2–3 kPa). Ka ōrite tēnei ki te pito iri kore utu o te paraihe pūmua peka e whakaaturia ana i te ahua STEM. Ahakoa he taunakitanga o te gradient modulus i te taha o waho o te CL, ko te papa matua modulus teitei te mea nui ake. Heoi, ko te 100 nm o runga o te mata kei roto i te 20% o te roa katoa o te paraihe pūmua peka, nō reira he tika ki te whakaaro he tika ngā uara ine o te modulus i roto i tēnei awhe hohonutanga o te kōhao, ā, kāore e tino whakawhirinaki ki te pānga o te mea o raro.
Nā te hoahoa koiora ahurei o ngā karāhe whakapā lehfilcon A, he mea hanga mai i ngā hanganga paraihe pūmua PMPC peka kua honoa ki te mata o ngā papa SiHy, he tino uaua ki te tautuhi pono i ngā āhuatanga miihini o ō rātou hanganga mata mā te whakamahi i ngā tikanga ine tuku iho. I konei ka whakaatuhia e mātou he tikanga nanoindentation AFM matatau hei tautuhi tika i ngā rauemi tino ngawari pērā i te lefilcon A he nui te wai me te tino ngāwari. Ko tēnei tikanga e hangai ana ki te whakamahinga o tētahi pūwero AFM kua whiriwhiria marietia te rahi me te āhua o te pito kia ōrite ki ngā āhuatanga hanganga o ngā āhuatanga mata tino ngawari hei tā. Mā tēnei huinga o ngā āhuatanga i waenga i te pūwero me te hanganga ka nui ake te aro, ka taea ai e mātou te ine i te modulus iti me ngā āhuatanga ngāwari ā-roto o ngā huānga paraihe pūmua peka, ahakoa ngā pānga poroelastic. I whakaatuhia e ngā hua he tino iti te modulus ngāwari o ngā paraihe pūmua PMPC peka ahurei e āhua ana ki te mata o te karāhe (tae atu ki te 2 kPa) me te tino ngāwari (tata ki te 100%) i te wā i whakamatautauria ai i roto i te taiao wai. Nā ngā hua o te nanoindentation AFM i āhei ai mātou ki te whakaahua i te rerekētanga modulus/hohonutanga whakapā kitea (30 kPa/200 nm) o te mata arotahi biomimetic. Tērā pea nā te rerekētanga modulus i waenga i ngā paraihe polymer peka me te papa SiHy tēnei rerekētanga, nā te hanganga/kiato peka rānei o ngā paraihe polymer, nā te huinga rānei o ēnei. Heoi anō, me nui ake ngā rangahau hōhonu hei mārama katoa ki te whanaungatanga i waenga i te hanganga me ngā āhuatanga, inā koa te pānga o te peka o te paraihe ki ngā āhuatanga hangarau. Ka taea e ngā inenga rite te āwhina i te whakaahua i ngā āhuatanga hangarau o te mata o ētahi atu rauemi tino ngohengohe me ngā taputapu hauora.
Kei te wātea mai i ngā kaituhi takitahi ngā huinga raraunga i hangaia, i tātarihia rānei i te wā o te rangahau o nāianei, mēnā he tika te tono.
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