Rau cov neeg mob uas tau phais orthopedic implants, kab mob thiab kab mob ntawm lub cev tiv thaiv kab mob yog ib txwm ua rau muaj kev phom sij rau lub neej.Cov khoom siv tshuaj lom neeg muaj feem cuam tshuam rau cov kab mob lom lom, uas ua rau cov kab mob nkag mus rau qhov chaw raug mob thiab ua rau muaj kab mob tom qab phais.Yog li ntawd, nws yog ib qho tsim nyog yuav tsum tau ua kom muaj kev tiv thaiv kab mob thiab tiv thaiv kab mob tiv thaiv kab mob rau kev cog hniav orthopedic.Ntawm no, peb tau tsim ib qho kev hloov kho saum npoo av rau orthopedic implants hu ua Lubricated Orthopedic Implant Surface (LOIS), uas tau txais kev tshoov siab los ntawm lub ntsej muag du ntawm pitcher cog pitchers.LOIS muaj qhov ua tau ntev thiab muaj zog ua kua rau ntau yam kua thiab lom (xws li cov hlwb, cov protein, calcium thiab cov kab mob).Tsis tas li ntawd, peb tau lees paub qhov kev ua haujlwm ruaj khov tiv thaiv khawb thiab kho lub zog los ntawm kev simulating qhov kev puas tsuaj uas tsis tuaj yeem tshwm sim thaum lub sijhawm phais hauv vitro.Tus luav pob txha pob txha inflammatory femoral fracture qauv tau siv los ua kom zoo kawm txog kev tiv thaiv kab mob lom thiab tiv thaiv kab mob ntawm LOIS.Peb xav tias LOIS, uas muaj cov khoom siv los tiv thaiv biofouling thiab kev siv tshuab ua haujlwm ntev, yog ib kauj ruam tom ntej hauv kev phais mob tsis muaj kab mob orthopedic.
Niaj hnub no, vim kev laus tag nrho, tus naj npawb ntawm cov neeg mob uas muaj kab mob orthopedic (xws li cov neeg laus pob txha, cov kab mob sib koom ua ke, thiab pob txha pob txha) tau nce ntau (1, 2).Yog li ntawd, cov tsev kho mob muab qhov tseem ceeb rau kev phais orthopedic, nrog rau orthopedic implants ntawm cov ntsia hlau, daim hlau, rau tes thiab cov pob qij txha (3, 4).Txawm li cas los xij, cov tshuaj orthopedic implants tau raug tshaj tawm tias muaj kev cuam tshuam rau cov kab mob adhesion thiab biofilm tsim, uas tuaj yeem ua rau qhov chaw phais mob (SSI) tom qab phais (5, 6).Thaum cov biofilm tau tsim nyob rau saum npoo ntawm orthopedic implant, kev tshem tawm ntawm biofilm yuav nyuaj heev txawm tias siv cov tshuaj tua kab mob loj.Yog li, nws feem ntau ua rau mob hnyav tom qab phais mob (7, 8).Vim yog cov teeb meem saum toj no, kev kho cov kab mob cog qoob loo yuav tsum muaj kev rov ua haujlwm dua, suav nrog kev tshem tawm tag nrho cov cog thiab cov ntaub so ntswg ib puag ncig;Yog li ntawd, tus neeg mob yuav raug mob hnyav thiab qee qhov kev pheej hmoo (9, 10).
Txhawm rau daws qee qhov teeb meem no, cov tshuaj siv tshuaj tua kab mob orthopedic tau tsim los tiv thaiv kev kis kab mob los ntawm kev tshem tawm cov kab mob uas txuas rau saum npoo (11, 12).Txawm li cas los xij, lub tswv yim tseem qhia ntau yam kev txwv.Nws tau raug tshaj tawm tias kev cog qoob loo ntev ntev ntawm kev cog cov tshuaj eluting tau ua rau muaj kev puas tsuaj rau cov ntaub so ntswg ib puag ncig thiab ua rau mob, uas tuaj yeem ua rau necrosis (13, 14).Tsis tas li ntawd, cov kuab tshuaj organic uas yuav muaj nyob tom qab tsim cov txheej txheem ntawm kev siv tshuaj tua kab mob orthopedic implants, uas tau txwv nruj los ntawm US Food and Drug Administration, yuav tsum tau ua kom huv ntxiv kom ua tau raws li nws cov qauv (15).Kev cog cov tshuaj eluting yog qhov nyuaj rau kev tswj cov tshuaj tso tawm, thiab vim lawv txwv cov tshuaj thauj khoom, kev siv tshuaj mus sij hawm ntev tsis ua tau (16).
Lwm lub tswv yim zoo tshaj plaws yog txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawmPiv txwv li, zwitterionic polymers tau nyiam cov xim vim lawv cov khoom tsis yog nplaum thaum sib cuag nrog cov plasma proteins, hlwb, thiab cov kab mob.Txawm li cas los xij, nws muaj qee qhov kev txwv ntsig txog kev ruaj ntseg mus sij hawm ntev thiab kev siv tshuab ua haujlwm ntev, uas cuam tshuam nws txoj kev siv rau hauv orthopedic implants, tshwj xeeb tshaj yog vim cov tshuab scraping thaum cov txheej txheem phais (18, 19).Tsis tas li ntawd, vim nws cov biocompatibility siab, tsis xav tau kev phais tshem tawm, thiab cov khoom ntxuav saum npoo los ntawm corrosion, orthopedic implants ua los ntawm cov ntaub ntawv biodegradable tau siv (20, 21).Thaum lub sij hawm corrosion, cov tshuaj bonds ntawm lub polymer matrix yog tawg thiab detached los ntawm qhov chaw, thiab cov adherents ntxuav lub nto.Txawm li cas los xij, tshuaj tiv thaiv kab mob fouling los ntawm kev ntxuav saum npoo yog siv tau rau lub sijhawm luv luv.Tsis tas li ntawd, cov ntaub ntawv absorbable feem ntau suav nrog poly (lactic acid-glycolic acid copolymer) (PLGA), polylactic acid (PLA) thiab magnesium-raws li alloys yuav ua rau tsis sib xws biodegradation thiab yaig hauv lub cev, uas yuav cuam tshuam tsis zoo rau txhua yam kev ruaj ntseg.( nees nkaum ob).Tsis tas li ntawd, biodegradable phaj fragments muab ib qho chaw rau cov kab mob mus txuas, uas ua rau kom muaj feem kis kab mob mus ntev.Qhov kev pheej hmoo ntawm kev degradation thiab kab mob no txwv tsis pub siv cov kev phais yas (23).
Superhydrophobic (SHP) qhov chaw uas ua raws li cov qauv hierarchical ntawm lotus nplooj tau dhau los ua ib qho kev daws teeb meem rau qhov chaw tiv thaiv fouling (24, 25).Thaum SHP nto yog raus rau hauv cov kua, huab cua npuas yuav raug daig, yog li tsim cov hnab tshos cua thiab tiv thaiv cov kab mob adhesion (26).Txawm li cas los xij, cov kev tshawb fawb tsis ntev los no tau pom tias SHP nto muaj qhov tsis zoo cuam tshuam txog kev siv tshuab ua haujlwm ntev thiab kev ruaj ntseg mus ntev, uas cuam tshuam rau nws daim ntawv thov hauv kev kho mob cog.Tsis tas li ntawd, cov hnab ntim huab cua yuav yaj thiab poob lawv cov khoom tiv thaiv fouling, yog li ua rau cov kab mob dav dua vim qhov loj ntawm SHP nto (27, 28).Tsis ntev los no, Aizenberg thiab cov npoj yaig tau qhia txog txoj hauv kev tshiab ntawm kev tiv thaiv biofouling nto txheej los ntawm kev txhim kho qhov chaw du los ntawm Nepenthes pitcher cog (29, 30).Qhov saum npoo du qhia tau tias muaj kev ruaj ntseg ntev nyob rau hauv hydraulic tej yam kev mob, yog tsis tshua muaj kua repellent rau cov kua roj ntsha, thiab muaj nws tus kheej-repairing zog.Txawm li cas los xij, tsis muaj ib txoj hauv kev los siv cov txheej txheem rau kev cog qoob loo zoo li kev kho mob, thiab tsis muaj pov thawj los txhawb txoj kev kho ntawm cov ntaub so ntswg puas tom qab cog.
Ntawm no, peb qhia txog lub lubricated orthopedic implant nto (LOIS), ib tug micro / nano-structured orthopedic implant nto thiab nruj nreem ua ke nrog ib tug nyias lubricant txheej los tiv thaiv nws los ntawm kev txuam nrog yas phais kab mob, xws li pob txha fixation.Vim tias cov fluorine-functionalized micro/nano-theem qauv ruaj khov kho cov roj nplua nyeem ntawm cov qauv, cov tsim LOIS tuaj yeem ua rau tag nrho cov adhesion ntawm ntau cov kua thiab tswj kev ua haujlwm zoo rau lub sijhawm ntev.LOIS coatings tuaj yeem siv rau cov ntaub ntawv ntawm ntau yam duab tsim rau cov pob txha synthesis.Cov khoom zoo heev los tiv thaiv biofouling ntawm LOIS tiv thaiv cov kab mob biofilm [Pseudomonas aeruginosa thiab methicillin-resistant Staphylococcus aureus (MRSA)] thiab cov tshuaj lom (cells, proteins thiab calcium) tau lees paub hauv vitro.Lub adhesion tus nqi ntawm qhov uas nws kim heev adhesion rau lub substrate yog tsawg tshaj li 1%.Tsis tas li ntawd, txawm tias tom qab txhua yam kev ntxhov siab xws li khawb qhov chaw tshwm sim, kev kho tus kheej los ntawm kev nkag mus rau lubricant pab tswj nws cov khoom tiv thaiv fouling.Cov txiaj ntsig ntawm kev ntsuas kev ua haujlwm tau zoo qhia tau tias txawm tias tom qab kev hloov kho cov qauv thiab tshuaj lom neeg, tag nrho lub zog yuav tsis txo qis.Tsis tas li ntawd, ib qho kev sim hauv vitro uas simulates cov neeg kho tshuab kev ntxhov siab hauv qhov chaw phais mob tau ua pov thawj tias LOIS tuaj yeem tiv taus ntau yam kev ntxhov siab uas tshwm sim thaum phais yas.Thaum kawg, peb siv tus luav-raws li nyob rau hauv vivo femoral fracture qauv, uas ua pov thawj tias LOIS muaj superior antibacterial zog thiab biocompatibility.Radiological thiab histological cov txiaj ntsig tau lees paub tias kev coj tus cwj pwm ruaj khov thiab cov khoom siv biofouling tsis pub dhau 4 lub lis piam tom qab cog qoob loo tuaj yeem ua tiav kev tiv thaiv kab mob thiab tiv thaiv kev tiv thaiv kev ua haujlwm yam tsis muaj kev ncua sijhawm ntawm cov pob txha kho.
Daim duab 1A qhia ib daim duab schematic ntawm LOIS tsim, uas yog cog nrog micro / nano-scale qauv hauv luav femoral fracture qauv kom paub meej tias nws zoo heev los tiv thaiv kab mob fouling thiab tiv thaiv kab mob.Ib txoj kev biomimetic yog ua los simulate saum npoo ntawm lub lauj kaub dej, thiab tiv thaiv biofouling los ntawm kev sib xyaw cov roj nplua nyeem hauv cov qauv micro / nano ntawm qhov chaw.Qhov chaw txhaj tshuaj nrog roj nplua nyeem tuaj yeem txo qhov kev sib cuag ntawm cov tshuaj lom neeg thiab qhov chaw.Yog li ntawd, vim yog kev tsim cov tshuaj ruaj khov rau ntawm qhov chaw, nws muaj kev ua haujlwm zoo los tiv thaiv fouling thiab kev ruaj ntseg mus ntev.Yog li ntawd, cov khoom los tiv thaiv biofouling ntawm lubricating nto tso cai rau ntau yam kev siv hauv kev tshawb fawb biomedical.Txawm li cas los xij, kev tshawb fawb dav dav txog seb qhov chaw tshwj xeeb no cuam tshuam li cas hauv lub cev tseem tsis tau tiav.Los ntawm kev sib piv LOIS nrog liab qab substrates hauv vitro siv cov kab mob albumin thiab biofilm, qhov tsis-adhesiveness ntawm LOIS tuaj yeem lees paub (Daim duab 1B).Tsis tas li ntawd, los ntawm dov tawm cov tee dej ntawm lub inclined liab qab substrate thiab LOIS substrate (Daim duab S1 thiab Movie S1), qhov kev ua tau zoo ntawm cov kab mob lom tuaj yeem ua rau pom tau.Raws li tau pom nyob rau hauv daim duab fluorescence microscope, cov substrate raug incubated nyob rau hauv ib tug ncua kev kawm ntawv ntawm cov protein thiab cov kab mob pom ib tug loj npaum li cas ntawm cov khoom lom neeg adhering rau saum npoo.Txawm li cas los xij, vim nws cov khoom tiv thaiv biofouling zoo heev, LOIS tsis tshua pom muaj fluorescence.Txhawm rau kom paub meej tias nws cov khoom tiv thaiv biofouling thiab tiv thaiv kab mob, LOIS tau siv rau saum npoo ntawm orthopedic implants rau cov pob txha synthesis (cov phaj thiab cov ntsia hlau) thiab muab tso rau hauv cov qauv luav pob txha.Ua ntej implantation, lub liab qab orthopedic implant thiab LOIS tau incubated nyob rau hauv ib tug kab mob ncua kev kawm ntawv rau 12 teev.Lub pre-incubation xyuas kom meej tias ib tug biofilm yog tsim nyob rau saum npoo ntawm qhov chaw cog rau kev sib piv.Daim duab 1C qhia ib daim duab ntawm qhov chaw tawg 4 lub lis piam tom qab cog.Nyob rau sab laug, ib tug luav nrog ib tug liab qab orthopedic implants pom ib tug loj theem ntawm o vim yog tsim ntawm ib tug biofilm nyob rau saum npoo ntawm cog.Cov txiaj ntsig zoo sib xws tau pom nyob rau hauv cov luav cog nrog LOIS, uas yog, cov ntaub so ntswg ib puag ncig ntawm LOIS tsis pom muaj kab mob lossis cov tsos mob ntawm qhov mob.Tsis tas li ntawd, cov duab kho qhov muag ntawm sab laug qhia txog qhov chaw phais ntawm tus luav nrog rau qhov chaw cog qoob loo, qhia tias tsis muaj ntau cov nplaum uas muaj nyob rau saum npoo ntawm cov khoom cog tau pom nyob rau saum npoo ntawm LOIS.Qhov no qhia tau hais tias LOIS muaj kev ruaj ntseg mus sij hawm ntev thiab muaj peev xwm tswj tau nws cov khoom tiv thaiv kab mob lom thiab tiv thaiv adhesion.
(A) Schematic daim duab ntawm LOIS thiab nws implantation nyob rau hauv ib tug luav femoral fracture qauv.(B) Fluorescence microscopy duab ntawm cov protein thiab cov kab mob biofilm ntawm liab qab thiab LOIS substrate.4 lub lis piam tom qab cog, (C) daim duab duab ntawm qhov chaw tawg thiab (D) daim duab X-ray (teeb pom los ntawm daim duab plaub liab).Yees duab courtesy: Kyomin Chae, Yonsei University.
Cov kab mob sterilized, raug pom tsis zoo implants luav tau pom cov txheej txheem kho pob txha zoo yam tsis muaj cov tsos mob ntawm o lossis kab mob.Ntawm qhov tod tes, SHP implants pre-incubated nyob rau hauv ib tug kab mob ncua kev kawm ntawv muaj ib tug kab mob uas muaj feem xyuam rau o ntawm cov ntaub so ntswg nyob ib ncig ntawm.Qhov no tuaj yeem raug ntaus nqi vim nws tsis muaj peev xwm tiv thaiv kab mob adhesion rau lub sijhawm ntev (Daim duab S2).Txhawm rau ua pov thawj tias LOIS tsis cuam tshuam rau cov txheej txheem kho, tab sis inhibits kev kis kab mob cuam tshuam nrog kev cog qoob loo, X-ray duab ntawm qhov pom zoo matrix thiab LOIS ntawm qhov chaw tawg tau muab piv (Daim duab 1D).Daim duab X-ray ntawm qhov tsis pom zoo cog tau pom cov kab mob osteolysis tsis tu ncua, qhia tias cov pob txha tsis zoo tag nrho.Qhov no qhia tau hais tias cov txheej txheem ntawm cov pob txha rov qab tuaj yeem ncua ntev heev vim muaj kev kis kab mob.Ntawm qhov tsis sib xws, nws pom tau tias cov luav cog nrog LOIS tau kho thiab tsis pom muaj qhov pom tseeb tawg qhov chaw.
Txhawm rau txhim kho cov kev cog lus kho mob nrog kev ruaj ntseg ntev thiab kev ua haujlwm (nrog rau kev tiv thaiv biofouling), ntau qhov kev siv zog tau ua.Txawm li cas los xij, muaj ntau yam tshuaj lom neeg thiab cov kev ua haujlwm ntawm cov ntaub so ntswg adhesion txwv txoj kev loj hlob ntawm lawv cov kev kho mob txhim khu kev qha.Txhawm rau kom kov yeej cov kev tsis txaus no, peb tau tsim cov qauv micro / nano txheej thiab cov tshuaj hloov pauv, uas yog qhov ua tau zoo vim muaj zog capillary siab thiab tshuaj lom neeg affinity kom lub smoothest lubricant mus rau qhov loj tshaj plaws.Daim duab 2A qhia tag nrho cov txheej txheem tsim khoom ntawm LOIS.Ua ntej, npaj ib qho kev kho mob qib stainless hlau (SS) 304 substrate.Qhov thib ob, cov qauv micro / nano yog tsim los ntawm SS substrate los ntawm cov tshuaj etching siv hydrofluoric acid (HF) tov.Txhawm rau kho qhov corrosion kuj ntawm SS, nitric acid (HNO3) tov (31) yog siv los ua cov txheej txheem etched substrate.Passivation txhim khu kev tiv thaiv corrosion ntawm SS substrate thiab ua kom qeeb ntawm cov txheej txheem corrosion uas yuav txo tau tag nrho cov kev ua tau zoo ntawm LOIS.Tom qab ntawd, los ntawm kev tsim tus kheej-sib dhos monolayer (SAM) nrog 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane (POTS), qhov saum npoo yog hloov kho tshuaj los txhim kho cov tshuaj sib cuam tshuam ntawm qhov chaw thiab cov roj nplua nyeem Affinity.Qhov kev hloov kho saum npoo ua rau txo qis lub zog saum npoo ntawm cov khoom tsim micro / nano-scale qauv, uas sib phim lub zog saum npoo ntawm cov roj nplua nyeem.Qhov no tso cai rau cov roj nplua nyeem kom ntub tag nrho, yog li tsim cov roj nplaum ruaj khov rau ntawm qhov chaw.Hloov kho qhov chaw nthuav tawm txhim kho hydrophobicity.Cov txiaj ntsig tau pom tias cov roj nplua nyeem ua rau muaj tus cwj pwm ruaj khov ntawm LOIS vim yog cov tshuaj lom neeg siab thiab cov capillary quab yuam los ntawm cov qauv micro / nano (32, 33).Kev hloov kho qhov muag ntawm qhov chaw ntawm SS tom qab kev hloov kho saum npoo thiab cov roj nplua nyeem tau kawm.Cov txheej txheem micro / nano txheej tsim rau ntawm qhov chaw tuaj yeem ua rau pom kev hloov pauv thiab ua kom tsaus rau saum npoo.Qhov tshwm sim no raug ntaus nqi los ntawm kev txhim kho lub teeb tawg rau ntawm qhov chaw ntxhib, uas ua rau kom muaj kev cuam tshuam ntawm kev cuam tshuam los ntawm lub teeb ntxiab mechanism (34).Tsis tas li ntawd, tom qab cov roj nplua nyeem tau txhaj, LOIS yuav tsaus dua.Lub lubricating txheej ua rau lub teeb tsawg dua los ntawm lub substrate, yog li ua rau lub LOIS darkening.Txhawm rau txhawm rau txhim kho microstructure / nanostructure los qhia lub kaum sab xis me tshaj plaws (SA) kom ua tiav kev ua haujlwm ntawm biofouling, scanning electron microscopy (SEM) thiab atomic khub tau siv los ua HF etching lub sijhawm sib txawv (0, 3)., 15 thiab 60 feeb) Force Microscope (AFM) (Daim duab 2B).SEM thiab AFM cov duab qhia tau hais tias tom qab lub sij hawm luv luv ntawm etching (3 feeb ntawm etching), lub substrate liab qab tau tsim uneven nano-scale roughness.Qhov saum npoo roughness hloov nrog lub sij hawm etching (Daim duab S3).Lub sij hawm-hloov nkhaus qhia tau hais tias qhov nto roughness tseem nce ntxiv thiab nce mus txog qhov siab tshaj ntawm 15 feeb ntawm etching, thiab tom qab ntawd tsuas yog txo qis me ntsis ntawm tus nqi roughness ntawm 30 feeb ntawm etching.Nyob rau ntawm lub sijhawm no, nano-theem roughness yog etched tam sim ntawd, thaum lub micro-qib roughness loj hlob, ua rau cov roughness hloov ntau ruaj khov.Tom qab etching rau ntau tshaj 30 feeb, pom qhov nce ntxiv hauv qhov ntxhib, uas tau piav qhia hauv qab no: SS yog tsim los ntawm cov hlau, alloyed nrog cov ntsiab lus xws li hlau, chromium, npib tsib xee, molybdenum thiab ntau lwm yam.Ntawm cov ntsiab lus no, hlau, chromium thiab molybdenum ua lub luag haujlwm tseem ceeb hauv kev tsim micron / nano-scale roughness ntawm SS los ntawm HF etching.Nyob rau theem pib ntawm corrosion, hlau thiab chromium feem ntau yog corroded vim molybdenum muaj corrosion kuj ntau dua li molybdenum.Raws li etching zuj zus, cov tshuaj etching mus txog hauv zos oversaturation, tsim fluorides thiab oxides tshwm sim los ntawm etching.Fluoride thiab oxide precipitate thiab nws thiaj li redeposit rau ntawm qhov chaw, ua ib tug nto roughness nyob rau hauv lub micron / nano ntau yam (31).Qhov no micro / nano-level roughness plays lub luag haujlwm tseem ceeb hauv kev kho tus kheej ntawm LOIS.Lub dual nplai nto ua rau muaj kev sib koom ua ke, ua kom lub zog capillary zoo heev.Qhov tshwm sim no tso cai rau cov roj nplua nyeem nkag mus rau qhov chaw ruaj khov thiab ua rau nws tus kheej zoo (35).Qhov tsim ntawm roughness nyob ntawm lub sij hawm etching.Nyob rau hauv 10 feeb ntawm etching, qhov saum npoo tsuas muaj nano-scale roughness, uas yog tsis txaus los tuav txaus lubricant kom muaj biofouling kuj (36).Ntawm qhov tod tes, yog tias lub sij hawm etching tshaj 30 feeb, nano-scale roughness tsim los ntawm redeposition ntawm hlau thiab chromium yuav ploj mus, thiab tsuas yog micro-scale roughness yuav nyob twj ywm vim molybdenum.Lub over-etched nto tsis muaj nano-scale roughness thiab poob lub synergistic nyhuv ntawm ob-theem roughness, uas cuam tshuam rau tus kheej-kho tus yam ntxwv ntawm LOIS.SA ntsuas tau ua nyob rau hauv substrates nrog sib txawv etching lub sij hawm los ua pov thawj anti-fouling kev ua tau zoo.Ntau hom kua tau raug xaiv raws li viscosity thiab lub zog saum npoo, suav nrog deionized (DI) dej, ntshav, ethylene glycol (EG), ethanol (EtOH) thiab hexadecane (HD) (Daim duab S4).Lub sij hawm sib txawv etching qauv qhia tau hais tias rau ntau yam kua nrog txawv zog nto thiab viscosities, SA ntawm LOIS tom qab 15 feeb ntawm etching yog qhov qis tshaj.Yog li ntawd, LOIS yog optimized rau etch rau 15 feeb los tsim micron thiab nano-scale roughness, uas yog haum rau zoo tswj lub durability ntawm lubricant thiab zoo heev anti-fouling zog.
(A) Schematic daim duab ntawm plaub-kauj ruam kev tsim khoom ntawm LOIS.Lub inset qhia SAM tsim ntawm lub substrate.(B) SEM thiab AFM cov duab, siv los txhim kho cov qauv micro/nano ntawm cov substrate nyob rau hauv txawv etching lub sij hawm.X-ray photoelectron spectroscopy (XPS) spectra ntawm (C) Cr2p thiab (D) F1s tom qab nto passivation thiab SAM txheej.aw, arbitrary unit.(E) Cov duab sawv cev ntawm cov tee dej ntawm liab qab, etched, SHP thiab LOIS substrates.(F) Lub kaum sab xis ntawm kev sib cuag (CA) thiab SA ntsuas cov kua nrog qhov sib txawv ntawm qhov sib txawv ntawm SHP thiab LOIS.Cov ntaub ntawv raug qhia raws li txhais tau tias ± SD.
Tom qab ntawd, txhawm rau txhawm rau paub meej qhov kev hloov pauv ntawm cov khoom siv tshuaj ntawm qhov chaw, X-ray photoelectron spectroscopy (XPS) tau siv los kawm txog kev hloov pauv ntawm cov tshuaj muaj pes tsawg leeg ntawm substrate nto tom qab txhua txheej txheej.Daim duab 2C qhia txog XPS ntsuas qhov tshwm sim ntawm HF etched nto thiab HNO 3 kho nto.Ob lub ntsiab peaks ntawm 587.3 thiab 577.7 eV tuaj yeem raug ntaus nqi rau Cr-O daim ntawv cog lus uas muaj nyob rau hauv chromium oxide txheej, uas yog qhov txawv ntawm HF etched nto.Qhov no feem ntau yog vim kev noj cov hlau thiab chromium fluoride ntawm qhov chaw los ntawm HNO3.HNO3-raws li etching tso cai rau chromium los tsim ib txheej txheej oxide passivating rau saum npoo, uas ua rau etched SS dua resistant rau corrosion.Nyob rau hauv daim duab 2D, XPS spectra tau txais los xyuas kom meej tias fluorocarbon-raws li silane tau tsim rau saum npoo tom qab SAM txheej, uas muaj cov kua ua kua ntau heev txawm rau EG, ntshav thiab EtOH.SAM txheej yog ua tiav los ntawm reacting silane functional pawg nrog hydroxyl pawg tsim los ntawm kev kho ntshav plasma.Raws li qhov tshwm sim, qhov nce tseem ceeb hauv CF2 thiab CF3 peaks tau pom.Lub zog khi ntawm 286 thiab 296 eV qhia tias kev hloov kho tshuaj tau ua tiav tiav los ntawm SAM txheej.SHP qhia tau hais tias loj heev CF2 (290.1 ​​​​eV) thiab CF3 (293.3 ​​eV) peaks, uas yog tshwm sim los ntawm fluorocarbon-raws li silane tsim nyob rau saum npoo.Daim duab 2E qhia cov duab sawv cev kho qhov muag ntawm kev sib cuag lub kaum sab xis (CA) rau cov pab pawg sib txawv ntawm cov dej deionized nyob rau hauv kev sib cuag nrog liab qab, etched, SHP, thiab LOIS.Cov duab no qhia tau hais tias qhov etched nto ua hydrophilic vim lub micro / nano qauv tsim los ntawm cov tshuaj etching kom cov dej deionized absorbed rau hauv cov qauv.Txawm li cas los xij, thaum lub substrate yog coated nrog SAM, lub substrate muaj zog tiv thaiv dej, yog li ib tug nto SHP yog tsim thiab qhov chaw sib cuag ntawm dej thiab saum npoo yog me me.Thaum kawg, qhov txo qis hauv CA tau pom nyob rau hauv LOIS, uas tuaj yeem raug ntaus nqi rau kev nkag mus ntawm cov roj nplua nyeem rau hauv microstructure, yog li ua rau thaj chaw sib cuag.Txhawm rau ua pov thawj tias qhov chaw muaj cov kua dej zoo heev thiab cov khoom tsis yog nplaum, LOIS tau muab piv nrog SHP substrate los ntawm kev ntsuas CA thiab SA siv ntau yam kua (Daim duab 2F).Ntau hom kua tau raug xaiv raws li viscosity thiab lub zog saum npoo, suav nrog dej deionized, ntshav, EG, EtOH thiab HD (Daim duab S4).Kev ntsuas CA qhia tau tias thaum CA nyhav rau HD, qhov txo qis ntawm CA, qhov twg CA muaj lub zog qis tshaj plaws.Tsis tas li ntawd, LOIS ntawm tag nrho CA yog tsawg.Txawm li cas los xij, qhov ntsuas SA qhia tau qhov tshwm sim sib txawv kiag li.Tsuas yog cov dej ionized, tag nrho cov kua dej ua raws li SHP substrate yam tsis tau swb.Ntawm qhov tod tes, LOIS qhia SA tsawg heev, qhov twg thaum tag nrho cov kua yog qaij ntawm lub kaum sab xis qis dua 10 ° txog 15 °, tag nrho cov kua yuav yob tawm.Qhov no qhia tau hais tias tsis yog-nplaum ntawm LOIS zoo dua li ntawm SHP nto.Tsis tas li ntawd, LOIS coatings kuj tau siv rau ntau hom ntaub ntawv, suav nrog titanium (Ti), polyphenylsulfone (PPSU), polyoxymethylene (POM), polyether ether ketone (PEEK) thiab bioabsorbable polymers (PLGA), Lawv yog cov khoom siv orthopedic implantable (Daim duab S5)).Cov duab ua ntu zus ntawm cov tee dej ntawm cov khoom kho los ntawm LOIS qhia tias cov khoom tiv thaiv biofouling ntawm LOIS yog tib yam ntawm txhua qhov substrates.Tsis tas li ntawd, qhov ntsuas ntsuas ntawm CA thiab SA qhia tau tias cov khoom tsis yog nplaum ntawm LOIS tuaj yeem siv rau lwm cov ntaub ntawv.
Txhawm rau kom paub meej tias cov khoom tiv thaiv fouling ntawm LOIS, ntau hom substrates (xws li liab qab, etched, SHP thiab LOIS) tau incubated nrog Pseudomonas aeruginosa thiab MRSA.Ob tus kab mob no tau raug xaiv los ua tus neeg sawv cev hauv tsev kho mob cov kab mob, uas tuaj yeem ua rau muaj cov kab mob biofilms, ua rau SSI (37).Daim duab 3 (A thiab B) qhia txog fluorescence microscope cov duab thiab cov colony forming unit (CFU) kev ntsuas ntawm cov substrates incubated nyob rau hauv cov kab mob ncua sij hawm luv luv (12 teev) thiab ntev (72 teev), feem.Nyob rau hauv ib lub sij hawm luv luv, cov kab mob yuav tsim pawg thiab loj hlob tuaj, npog lawv tus kheej nrog cov mucus zoo li tshuaj thiab tiv thaiv lawv tshem tawm.Txawm li cas los xij, thaum lub sij hawm 72-teev incubation, cov kab mob yuav loj hlob tuaj thiab yooj yim rau disperse mus tsim ntau colonies los yog pawg.Yog li, nws tuaj yeem txiav txim siab tias 72-teev incubation yog lub sij hawm ntev thiab yog lub sij hawm tsim nyog incubation los tsim ib tug muaj zog biofilm rau saum npoo (38).Nyob rau hauv ib lub sij hawm luv luv, etched nto thiab nto ntawm SHP nthuav tawm cov kab mob adhesion, uas tau txo los ntawm 25% mus rau 50% piv rau cov liab qab substrate.Txawm li cas los xij, vim nws qhov kev ua tau zoo los tiv thaiv biofouling thiab kev ruaj ntseg, LOIS tsis pom cov kab mob biofilm adhesion nyob rau lub sijhawm luv thiab ntev.Daim duab schematic (Daim duab 3C) piav qhia txog kev tiv thaiv kab mob fouling mechanism ntawm etching tov, SHP thiab LOIS.Qhov kev xav yog tias etched substrate nrog hydrophilic zog yuav muaj thaj tsam loj dua li qhov liab qab substrate.Yog li ntawd, ntau cov kab mob adhesion yuav tshwm sim ntawm etched substrate.Txawm li cas los xij, piv nrog rau qhov liab qab substrate, etched substrate muaj tsawg dua biofilm tsim rau saum npoo.Qhov no yog vim cov dej molecules khi ruaj khov rau ntawm qhov chaw hydrophilic thiab ua cov roj nplua nyeem rau dej, yog li cuam tshuam nrog cov kab mob adhesion hauv lub sijhawm luv (39).Txawm li cas los xij, txheej ntawm cov dej molecules yog nyias thiab soluble hauv cov kab mob suspensions.Yog li ntawd, cov dej molecular txheej ploj mus rau lub sijhawm ntev, ua rau muaj cov kab mob adhesion thiab proliferation.Rau SHP, vim nws lub sijhawm luv luv uas tsis yog ntub dej, cov kab mob adhesion yog inhibited.Qhov txo cov kab mob adhesion tuaj yeem raug ntaus nqi rau cov hnab ntim huab cua uas nyob rau hauv cov txheej txheem txheej thiab lub zog qis dua, yog li txo qis kev sib cuag ntawm cov kab mob ncua kev kawm ntawv thiab qhov chaw.Txawm li cas los xij, cov kab mob loj heev tau pom nyob rau hauv SHP vim nws poob nws cov khoom tiv thaiv fouling ntev.Qhov no feem ntau yog vim qhov ploj ntawm cov hnab ris hauv huab cua vim yog hydrostatic siab thiab kev sib cais ntawm huab cua hauv dej.Qhov no feem ntau yog vim qhov ploj ntawm cov hnab ris huab cua vim yog kev sib cais thiab cov txheej txheem txheej uas muab thaj chaw loj dua rau adhesion (27, 40).Tsis zoo li ob lub substrates no uas muaj qhov cuam tshuam tseem ceeb rau kev ruaj ntseg mus sij hawm ntev, lub lubricating lubricant muaj nyob rau hauv LOIS yog txhaj rau hauv micro / nano qauv thiab yuav tsis ploj txawm nyob rau lub sij hawm ntev.Lubricants uas muaj cov qauv micro/nano yog qhov ruaj khov thiab muaj zog txaus nyiam rau saum npoo vim lawv cov tshuaj lom neeg muaj zog, yog li tiv thaiv kab mob adhesion rau lub sijhawm ntev.Daim duab S6 qhia tau hais tias ib qho kev xav txog confocal microscope duab ntawm cov roj nplaum-infused substrate rau hauv phosphate buffered saline (PBS).Cov duab txuas ntxiv qhia tau hais tias txawm tias tom qab 120 teev ntawm kev co me ntsis (120 rpm), cov roj nplaum txheej ntawm LOIS tseem tsis hloov pauv, qhia txog kev ruaj ntseg mus ntev hauv cov dej ntws.Qhov no yog vim muaj tshuaj lom neeg muaj txiaj ntsig ntawm cov tshuaj fluorine-based SAM txheej thiab perfluorocarbon-based lubricant, kom cov roj nplua nyeem ruaj khov tuaj yeem tsim.Yog li ntawd, qhov kev ua haujlwm tiv thaiv fouling yog khaws cia.Tsis tas li ntawd, lub substrate tau sim tawm tsam cov neeg sawv cev ntawm cov proteins (albumin thiab fibrinogen), uas muaj nyob rau hauv plasma, cov hlwb muaj feem xyuam nrog kev tiv thaiv kab mob (macrophages thiab fibroblasts), thiab cov uas cuam tshuam txog kev tsim cov pob txha.Cov ntsiab lus ntawm calcium yog siab heev.(Daim duab 3D, 1 thiab 2, thiab daim duab S7) (41, 42).Tsis tas li ntawd, fluorescence microscope dluab ntawm adhesion test rau fibrinogen, albumin thiab calcium pom txawv adhesion yam ntxwv ntawm txhua pawg substrate (Daim duab S8).Thaum lub sij hawm tsim cov pob txha, cov pob txha tshiab thiab cov txheej txheem calcium tuaj yeem nyob ib puag ncig ntawm cov pob txha orthopedic, uas tsis tsuas yog ua rau kev tshem tawm nyuaj, tab sis kuj tseem yuav ua rau muaj kev phom sij rau tus neeg mob thaum lub sijhawm tshem tawm.Yog li ntawd, tsis tshua muaj calcium deposits ntawm cov pob txha daim hlau thiab cov ntsia hlau yog qhov zoo rau kev phais orthopedic uas yuav tsum tau muab tshem tawm.Raws li qhov muaj nuj nqis ntawm thaj chaw txuas nrog raws li kev siv fluorescence thiab cov xov tooj ntawm tes, peb tau lees paub tias LOIS qhia tau zoo heev los tiv thaiv biofouling rau txhua yam tshuaj lom neeg piv nrog rau lwm cov substrates.Raws li cov txiaj ntsig ntawm kev sim hauv vitro, cov tshuaj tiv thaiv kab mob fouling LOIS tuaj yeem siv rau orthopedic implants, uas tuaj yeem tsis tsuas yog cuam tshuam cov kab mob los ntawm cov kab mob biofilm, tab sis kuj txo qhov mob tshwm sim los ntawm lub cev tiv thaiv kab mob.
(A) Fluorescence microscope dluab ntawm txhua pab pawg (liab qab, etched, SHP thiab LOIS) incubated nyob rau hauv Pseudomonas aeruginosa thiab MRSA ncua kev kawm ntawv rau 12 thiab 72 teev.(B) Tus naj npawb ntawm adherent CFU ntawm Pseudomonas aeruginosa thiab MRSA nyob rau saum npoo ntawm txhua pab pawg.(C) Schematic daim duab ntawm anti-biological fouling mechanism ntawm lub sij hawm luv thiab ntev etching, SHP thiab LOIS.(D) (1) Tus naj npawb ntawm fibroblasts adhered rau txhua lub substrate thiab fluorescence microscope cov duab ntawm cov hlwb adhered rau liab qab thiab LOIS.(2) Adhesion test of immune-related proteins, albumin and calcium uas koom nrog hauv cov txheej txheem kho pob txha (* P <0.05, ** P <0.01, *** P <0.001 thiab **** P <0.0001).ns, tsis tseem ceeb.
Nyob rau hauv cov ntaub ntawv ntawm unavoidable concentrated kev nyuaj siab, mechanical durability yeej ib txwm yog lub ntsiab kev sib tw rau daim ntawv thov ntawm antifouling txheej.Cov txheej txheem los tiv thaiv dej phwj gel yog raws li cov polymers uas tsis tshua muaj dej solubility thiab fragility.Yog li ntawd, lawv feem ntau raug rau cov neeg kho tshuab kev nyuaj siab hauv kev siv biomedical.Yog li ntawd, mechanically check antifouling coatings tseem yog ib qho nyuaj rau kev siv xws li orthopedic implants (43, 44).Daim duab 4A(1) qhia txog ob hom kev ntxhov siab uas siv rau orthopedic implants, nrog rau khawb (shear stress) thiab compression nrog cov duab kho qhov muag ntawm cov khoom cog uas puas tsim los ntawm forceps.Piv txwv li, thaum tus ntsia hlau yog nruj nreem ntsia hlau ntsia hlau, los yog thaum tus kws phais tuav cov pob txha phaj nruj nrog tweezers thiab siv compressive quab yuam, cov pob txha yas yuav puas thiab khawb ntawm ob lub macro thiab micro / nano nplai (Daim duab 4A, 2).Txhawm rau kuaj seb LOIS cov khoom tsim puas tuaj yeem tiv taus cov kev puas tsuaj no thaum lub sijhawm phais yas, nanoindentation tau ua los sib piv cov tawv tawv ntawm cov tawv nqaij liab qab thiab LOIS ntawm micro / nano nplai los kawm cov khoom siv ntawm micro / nano qauv cuam tshuam (Daim duab 4B).Daim duab schematic qhia qhov sib txawv deformation tus cwj pwm ntawm LOIS vim muaj cov qauv micro/nano.Ib qho kev quab yuam-yav dhau los tau kos raws li cov txiaj ntsig ntawm nanoindentation (Daim duab 4C).Cov duab xiav sawv cev rau qhov liab qab substrate, uas pom tsuas yog me ntsis deformation, raws li pom los ntawm qhov siab tshaj plaws indentation qhov tob ntawm 0.26-μm.Ntawm qhov tod tes, qhov maj mam nce hauv nanoindentation quab yuam thiab kev hloov pauv tau pom hauv LOIS (liab nkhaus) tuaj yeem pom cov cim ntawm cov khoom siv hluav taws xob txo qis, ua rau muaj qhov tob nanoindentation ntawm 1.61μm.Qhov no yog vim hais tias cov qauv micro / nano tam sim no nyob rau hauv LOIS muab qhov chaw sib sib zog nqus mus rau qhov kawg ntawm nanoindenter, yog li nws deformation ntau dua li ntawm qhov liab qab substrate.Konsta-Gdoutos et al.(45) ntseeg hais tias vim muaj cov nanostructures, nanoindentation thiab micro/nano roughness ua rau tsis xwm yeem nanoindentation nkhaus.Qhov chaw ntxoov ntxoo sib raug rau qhov tsis xwm yeem deformation nkhaus ntaus nqi los ntawm nanostructure, thaum lub cheeb tsam tsis-shaded yog ntaus nqi rau microstructure.Qhov deformation no tuaj yeem ua rau lub microstructure / nanostructure ntawm cov roj nplaum tuav thiab ua rau tsis zoo rau nws cov kev ua haujlwm tiv thaiv fouling.Txhawm rau kawm txog qhov cuam tshuam ntawm kev puas tsuaj ntawm LOIS, kev puas tsuaj uas tsis tuaj yeem cuam tshuam rau micro / nano cov qauv tau rov ua dua hauv lub cev thaum phais yas.Los ntawm kev siv cov ntshav thiab cov protein adhesion kuaj, kev ruaj ntseg ntawm cov khoom tiv thaiv biofouling ntawm LOIS tom qab hauv vitro tuaj yeem txiav txim siab (Daim duab 4D).Cov duab kho qhov muag pom tau pom qhov kev puas tsuaj uas tshwm sim ze ntawm qhov ntawm txhua lub substrate.Kev kuaj ntshav adhesion tau ua los ua kom pom qhov cuam tshuam ntawm kev puas tsuaj ntawm cov txheej txheem biofouling (Daim duab 4E).Zoo li SHP, cov khoom tiv thaiv fouling ploj vim kev puas tsuaj, thiab LOIS nthuav tawm cov khoom tiv thaiv fouling zoo heev los ntawm kev rov ua cov ntshav.Qhov no yog vim hais tias, vim hais tias lub zog saum npoo yog tsav los ntawm cov capillary nkaus uas npog thaj chaw puas, cov dej ntws hauv microstructured lubricant lubricant restores cov khoom tiv thaiv fouling (35).Tib qhov sib txawv tau pom nyob rau hauv kev kuaj protein adhesion siv albumin.Nyob rau hauv qhov chaw puas, qhov adhesion ntawm cov protein nyob rau saum npoo ntawm SHP yog dav pom, thiab los ntawm kev ntsuas nws cheeb tsam, nws muaj peev xwm muab xam raws li ib nrab ntawm cov adhesion theem ntawm cov liab qab substrate.Ntawm qhov tod tes, LOIS khaws nws cov khoom tiv thaiv biofouling yam tsis ua rau adhesion (Daim duab 4, F thiab G).Tsis tas li ntawd, qhov saum npoo ntawm cov ntsia hlau feem ntau raug rau cov neeg kho tshuab muaj zog, xws li drilling, yog li peb tau kawm txog lub peev xwm ntawm LOIS txheej kom nyob twj ywm ntawm cov ntsia hlau hauv vitro.Daim duab 4H qhia cov duab kho qhov muag ntawm cov ntsia hlau sib txawv, suav nrog liab qab, SHP thiab LOIS.Cov duab plaub liab sawv cev rau thaj chaw lub hom phiaj uas muaj zog txhua yam kev ntxhov siab tshwm sim thaum cog pob txha.Zoo ib yam li cov protein adhesion test ntawm lub phaj, fluorescence microscope yog siv los ntsuas cov protein adhesion thiab ntsuas thaj tsam kev pab cuam los ua pov thawj qhov kev ncaj ncees ntawm LOIS txheej, txawm tias muaj zog mechanical stress (Daim duab 4, I thiab J).Lub LOIS-kho cov ntsia hlau muaj qhov ua tau zoo los tiv thaiv fouling, thiab yuav luag tsis muaj protein ntau adheres rau saum npoo.Ntawm qhov tod tes, protein adhesion tau pom nyob rau hauv liab qab screws thiab SHP screws, qhov twg thaj tsam ntawm SHP screws yog ib feem peb ntawm cov liab qab screws.Tsis tas li ntawd, lub orthopedic implant siv rau fixation yuav tsum tau mechanically muaj zog los tiv thaiv kev nyuaj siab siv rau qhov chaw tawg, raws li qhia nyob rau hauv daim duab 4K.Yog li ntawd, ib qho kev sim khoov tau ua los txiav txim siab qhov cuam tshuam ntawm kev hloov kho tshuaj ntawm cov khoom siv kho tshuab.Tsis tas li ntawd, qhov no yog ua los tswj cov kev ntxhov siab ruaj khov los ntawm kev cog qoob loo.Siv cov tshuab ntsug ntsug kom txog rau thaum cov cog cog tau tag nrho folded thiab tau txais ib qho kev ntxhov siab-strain nkhaus (Daim duab 4L, 1).Ob lub zog suav nrog Young's modulus thiab flexural zog tau muab piv ntawm liab qab thiab LOIS substrates raws li qhov taw qhia ntawm lawv lub zog siv tshuab (Daim duab 4L, 2 thiab 3).Young's modulus qhia txog lub peev xwm ntawm cov khoom siv los tiv thaiv kev hloov kho tshuab.Cov Young's modulus ntawm txhua lub substrate yog 41.48 ± 1.01 thiab 40.06 ± 0.96 GPa, feem;qhov pom qhov sib txawv yog li ntawm 3.4%.Tsis tas li ntawd, nws tau tshaj tawm tias lub zog dabtsi yog khoov, uas txiav txim siab qhov nyuaj ntawm cov khoom, yog 102.34 ± 1.51 GPa rau cov khoom qab zib liab qab thiab 96.99 ± 0.86 GPa rau SHP.Lub substrate liab qab yog kwv yees li 5.3% siab dua.Kev txo qis me ntsis ntawm cov khoom siv kho tshuab tuaj yeem tshwm sim los ntawm qhov cuam tshuam.Nyob rau hauv cov nyhuv notch, lub micro / nano roughness yuav ua raws li ib tug txheej ntawm notches, ua rau lub zos kev nyuaj siab concentration thiab cuam tshuam rau lub mechanical zog ntawm lub cog (46).Txawm li cas los xij, raws li qhov tseeb tias qhov tawv nqaij ntawm tib neeg cov pob txha cortical tau tshaj tawm tias nyob nruab nrab ntawm 7.4 thiab 31.6 GPa, thiab ntsuas LOIS modulus tshaj li ntawm tib neeg cov pob txha cortical (47), LOIS txaus los txhawb kev puas tsuaj thiab nws tag nrho. mechanical zog yog me ntsis cuam tshuam los ntawm kev hloov kho saum npoo.
(A) Schematic daim duab ntawm (1) cov neeg kho tshuab kev nyuaj siab siv rau orthopedic implant thaum lub sij hawm ua hauj lwm, thiab (2) kho qhov muag duab ntawm lub puas orthopedic implant.(B) Schematic daim duab ntawm nano-mechanical zog ntsuas los ntawm nanoindentation thiab LOIS ntawm qhov liab qab.(C) Nanoindentation force-displacement nkhaus ntawm liab qab nto thiab LOIS.(D) Tom qab kev sim hauv vitro, simulate cov duab kho qhov muag ntawm ntau hom orthopedic daim hlau (qhov chaw puas yog qhov tseem ceeb nrog lub duab plaub liab) los simulate cov neeg kho tshuab kev ntxhov siab thaum lub sijhawm ua haujlwm.(E) Kev kuaj ntshav adhesion thiab (F) protein adhesion test ntawm pawg pawg orthopedic puas.(G) Ntsuas thaj tsam ntawm cov protein adhering rau lub phaj.(H) Cov duab kho qhov muag ntawm ntau hom orthopedic screws tom qab kev sim hauv vitro.(I) Protein adhesion test los kawm txog kev ncaj ncees ntawm cov txheej txheem sib txawv.(J) Ntsuas thaj tsam ntawm cov protein ua raws li cov ntsia hlau.(K) Kev txav ntawm tus luav yog npaj los tsim kom muaj kev ntxhov siab ruaj khov ntawm cov pob txha tawg.(L) (1) Khoov cov txiaj ntsig kuaj thiab cov duab kho qhov muag ua ntej thiab tom qab khoov.Qhov txawv ntawm (2) Young's modulus thiab (3) dabtsi yog khoov lub zog ntawm liab qab implant thiab SHP.Cov ntaub ntawv raug nthuav tawm raws li txhais tau tias ± SD (*P<0.05, **P<0.01, ***P<0.001 thiab ****P<0.0001).Yees duab courtesy: Kyomin Chae, Yonsei University.
Hauv cov xwm txheej kho mob, feem ntau cov kab mob sib cuag nrog cov khoom siv roj ntsha thiab qhov chaw mob los ntawm cov kab mob loj, cov kab mob loj (48).Yog li ntawd, Tebchaws Asmeskas Lub Chaw Tiv Thaiv thiab Tiv Thaiv Kab Mob tau kwv yees tias 65% ntawm tag nrho cov tib neeg kis tau tus mob muaj feem xyuam rau biofilms (49).Nyob rau hauv cov ntaub ntawv no, nws yog ib qhov tsim nyog los muab ib tug nyob rau hauv vivo kev sim tsim uas muab zoo ib yam biofilm tsim nyob rau saum npoo ntawm cog.Yog li ntawd, peb tau tsim ib tus luav femoral fracture qauv nyob rau hauv uas orthopedic implants twb pre-incubated nyob rau hauv ib tug kab mob ncua kev kawm ntawv thiab ces implanted nyob rau hauv luav femurs los kawm cov anti-fouling zog ntawm LOIS nyob rau hauv vivo.Vim yog peb qhov tseeb tseem ceeb hauv qab no, kab mob kis tau los ntawm kev coj noj coj ua ua ntej es tsis yog kev txhaj tshuaj ntawm cov kab mob ncaj qha: (i) Lub cev tiv thaiv kab mob ntawm luav yog ib txwm muaj zog dua li ntawm tib neeg;Yog li ntawd, kev txhaj tshuaj ntawm cov kab mob tshem tawm thiab cov kab mob planktonic yog ua tau Nws tsis muaj kev cuam tshuam rau kev tsim cov biofilms.(Ii) Cov kab mob Planktonic yog cov tshuaj tua kab mob ntau dua, thiab cov tshuaj tua kab mob feem ntau yog siv tom qab phais;Thaum kawg, (iii) cov kab mob planktonic raug tshem tawm yuav raug diluted los ntawm tus tsiaj lub cev kua (50).Los ntawm kev cog qoob loo ua ntej cog rau hauv cov kab mob raug tshem tawm ua ntej cog, peb tuaj yeem tshawb xyuas qhov teeb meem ntawm cov kab mob thiab txawv teb chaws lub cev cov tshuaj tiv thaiv (FBR) ntawm cov txheej txheem kho pob txha.Cov luav tau muab txi rau 4 lub lis piam tom qab cog, vim hais tias osseointegration tseem ceeb rau cov txheej txheem kho pob txha yuav ua tiav hauv 4 lub lis piam.Tom qab ntawd, cov implants raug tshem tawm los ntawm cov luav rau kev kawm downstream.Daim duab 5A qhia txog kev loj hlob ntawm cov kab mob.Tus kab mob orthopedic implant yog nkag mus rau hauv lub cev.Raws li qhov tshwm sim ntawm qhov ua ntej incubation nyob rau hauv cov kab mob ncua, rau ntawm rau ntawm rau tus luav implants nrog liab qab implants tau kis tus kab mob, thaum tsis muaj ib tug luav implants nrog LOIS-kho implants tau kis.Cov kab mob kis mus rau hauv peb kauj ruam, suav nrog kev loj hlob, kev loj hlob thiab tawg (51).Ua ntej, cov kab mob uas txuas nrog tsim tawm thiab loj hlob ntawm qhov chaw, thiab tom qab ntawd cov kab mob tsim cov kab mob biofilm thaum lawv tso tawm cov khoom siv polymer (EPS), amyloid thiab extracellular DNA.Biofilm tsis tsuas yog cuam tshuam nrog kev nkag mus ntawm cov tshuaj tua kab mob, tab sis kuj txhawb kev sib sau ntawm cov tshuaj tua kab mob-degrading enzymes (xws li β-lactamase) (52).Thaum kawg, biofilm kis cov kab mob loj hlob mus rau hauv cov ntaub so ntswg.Yog li ntawd, kis tau tshwm sim.Tsis tas li ntawd, thaum lub cev txawv teb chaws nkag mus rau hauv lub cev, tus kab mob uas tuaj yeem ua rau lub cev tiv thaiv kab mob tuaj yeem ua rau mob hnyav, mob, thiab txo qis kev tiv thaiv.Daim duab 5B muab cov ntsiab lus ntawm FBR tshwm sim los ntawm kev tso cov tshuaj orthopedic implants, es tsis yog lub cev tiv thaiv kab mob los ntawm kab mob.Lub cev tiv thaiv kab mob lees paub qhov muab tso rau hauv lub cev raws li lub cev txawv teb chaws, thiab tom qab ntawd ua rau cov hlwb thiab cov ntaub so ntswg ua rau lub cev txawv teb chaws (53).Nyob rau hauv thaum ntxov hnub ntawm FBR, cov khoom matrix tau tsim nyob rau saum npoo ntawm orthopedic implants, uas ua rau lub adsorption ntawm fibrinogen.Tus adsorbed fibrinogen tom qab ntawd ua ib qho tuab tuab siv lub network, uas txhawb nqa cov leukocytes (54).Thaum cov fibrin network tsim, mob mob yuav tshwm sim vim yog infiltration ntawm neutrophils.Hauv cov kauj ruam no, ntau yam cytokines xws li qog necrosis factor-α (TNF-α), interleukin-4 (IL-4) thiab IL-β raug tso tawm, thiab monocytes pib nkag mus rau qhov chaw cog qoob loo thiab sib txawv rau hauv cov hlwb loj.Phaj (41, 55, 56).Txo FBR ib txwm yog qhov nyuaj vim tias FBR ntau dhau tuaj yeem ua rau mob hnyav thiab mob ntev, uas tuaj yeem ua rau muaj teeb meem tuag taus.Txhawm rau ntsuas qhov cuam tshuam ntawm cov kab mob kab mob hauv cov ntaub so ntswg nyob ib puag ncig ntawm kev cog qoob loo thiab LOIS, hematoxylin thiab eosin (H&E) thiab Masson trichrome (MT) staining tau siv.Rau cov luav cog nrog liab qab substrates, cov kab mob loj zuj zus tuaj, thiab H&E cov ntaub so ntswg ua kom pom tseeb pom abscesses thiab necrosis tshwm sim los ntawm o.Ntawm qhov tod tes, qhov muaj zog heev los tiv thaiv biofouling nto LOIS inhibits cov kab mob adhesion, yog li nws tsis pom muaj kab mob thiab txo qhov mob (Daim duab 5C).Cov txiaj ntsig ntawm MT staining pom zoo li qub.Txawm li cas los xij, MT staining kuj pom edema hauv cov luav cog nrog LOIS, qhia tias kev rov qab los yuav tshwm sim (Daim duab 5D).Txhawm rau kawm txog qib ntawm kev tiv thaiv kab mob, immunohistochemical (IHC) staining tau ua tiav siv cytokines TNF-α thiab IL-6 ntsig txog kev tiv thaiv kab mob.Ib qho kev cog qoob loo tsis zoo uas tsis raug cov kab mob tau muab piv nrog LOIS uas tau kis rau cov kab mob tab sis tsis kis kab mob los kawm txog kev kho kom zoo thaum tsis muaj kab mob.Daim duab 5E qhia cov duab kho qhov muag ntawm IHC swb uas qhia txog TNF-α.Qhov chaw xim av sawv cev rau lub cev tiv thaiv kab mob, qhia tias kev tiv thaiv kab mob hauv LOIS tau txo qis me ntsis.Tsis tas li ntawd, qhov kev qhia ntawm IL-6 hauv LOIS tau tsawg dua qhov kev qhia tsis zoo ntawm kev tsis muaj menyuam liab qab (Daim duab 5F).Qhov kev qhia ntawm cytokine tau raug ntsuas los ntawm kev ntsuas thaj tsam ntawm cov tshuaj tiv thaiv kab mob sib txuas nrog cytokine (Daim duab 5G).Piv nrog rau cov luav tau pom qhov kev cog lus tsis zoo, cov lus qhia theem ntawm cov luav cog nrog LOIS tau qis dua, qhia qhov sib txawv.Kev txo qis hauv cytokine qhia tau hais tias lub sijhawm ntev, ruaj khov los tiv thaiv fouling ntawm LOIS tsis yog tsuas yog cuam tshuam nrog kev cuam tshuam ntawm cov kab mob, tab sis kuj txo qis FBR, uas yog induced los ntawm macrophages adhering rau lub substrate (53, 57, 58).Yog li, qhov txo qis hauv lub cev tiv thaiv kab mob vim yog lub zog tiv thaiv kab mob ntawm LOIS tuaj yeem daws cov kev mob tshwm sim tom qab implantation, xws li kev tiv thaiv kab mob ntau dhau tom qab phais yas.
(A) Daim duab schematic ntawm cov txheej txheem ntawm biofilm tsim thiab kis rau ntawm qhov chaw ntawm tus kab mob orthopedic cog.eDNA, extracellular DNA.(B) Schematic daim duab ntawm lub cev tiv thaiv kab mob tom qab orthopedic implant insertion.(C) H&E staining thiab (D) MT staining ntawm cov ntaub so ntswg ncig ntawm orthopedic implants nrog liab qab zoo thiab LOIS.IHC ntawm kev tiv thaiv kab mob cytokines (E) TNF-α thiab (F) IL-6 yog stained dluab ntawm liab qab-tsis zoo thiab LOIS-implanted luav.(G) Quantification ntawm cytokine qhia los ntawm cheeb tsam kev ntsuas (** P <0.01).
Lub biocompatibility ntawm LOIS thiab nws cov txiaj ntsig ntawm cov txheej txheem kho pob txha raug kuaj hauv vivo siv kev kuaj pom [x-ray thiab micro-computed tomography (CT)] thiab osteoclast IHC.Daim duab 6A qhia txog cov txheej txheem kho pob txha uas muaj peb theem sib txawv: o, kho, thiab kho dua tshiab.Thaum muaj pob txha tawg, cov kab mob inflammatory thiab fibroblasts yuav nkag mus rau hauv cov pob txha tawg thiab pib loj hlob mus rau hauv cov ntaub so ntswg.Thaum lub sij hawm kho, qhov ingrowth ntawm vascular cov ntaub so ntswg kis nyob ze ntawm qhov chaw tawg.Vascular cov ntaub so ntswg muab cov as-ham rau kev tsim cov pob txha tshiab, uas yog hu ua callus.Cov theem kawg ntawm cov txheej txheem kho pob txha yog qhov hloov kho, uas qhov loj ntawm callus raug txo mus rau qhov loj ntawm cov pob txha nrog kev pab los ntawm kev nce qib ntawm activated osteoclasts (59).Peb-dimensional (3D) rov tsim kho ntawm qhov chaw tawg tau ua tiav siv micro-CT scans los soj ntsuam qhov sib txawv ntawm qib ntawm callus tsim hauv txhua pab pawg.Saib xyuas qhov hla ntawm lub femur kom pom qhov tuab ntawm callus nyob ib puag ncig cov pob txha tawg (Daim duab 6, B thiab C).X-rays kuj tau siv los tshuaj xyuas qhov chaw tawg ntawm txhua pab pawg txhua lub lis piam los soj ntsuam cov txheej txheem sib txawv ntawm cov pob txha hauv txhua pab pawg (Daim duab S9).Callus thiab cov pob txha paub tab yog pom hauv xiav / ntsuab thiab ivory, feem.Feem ntau cov ntaub so ntswg yog lim tawm nrog lub preset pib.Nude zoo thiab SHP tau lees paub qhov tsim ntawm me me ntawm callus nyob ib ncig ntawm qhov chaw tawg.Ntawm qhov tod tes, qhov cuam tshuam tsis zoo ntawm LOIS thiab qhov chaw tawg yog ib puag ncig los ntawm cov callus tuab.Micro-CT cov duab qhia tau hais tias kev tsim ntawm callus tau cuam tshuam los ntawm cov kab mob thiab cov kab mob cuam tshuam txog kev mob.Qhov no yog vim lub cev tiv thaiv kab mob tseem ceeb rau kev kho mob ntawm cov kab mob septic uas tshwm sim los ntawm kev kis kab mob, tsis yog cov pob txha rov qab (60).IHC thiab Tartrate-resistant Acid Phosphatase (TRAP) staining tau ua los soj ntsuam kev ua haujlwm ntawm osteoclast thiab pob txha resorption (Daim duab 6D) (61).Tsuas yog ob peb lub tshuab ua haujlwm osteoclasts stained ntshav tau pom nyob rau hauv liab qab zoo thiab SHP.Ntawm qhov tod tes, ntau cov activated osteoclasts tau pom nyob ze ntawm cov pob txha liab qab zoo thiab paub tab ntawm LOIS.Qhov tshwm sim no qhia tau hais tias nyob rau hauv lub xub ntiag ntawm osteoclasts, lub callus nyob ib ncig ntawm lub pob txha tawg yog nyob rau hauv ib tug nruj kev hloov kho txheej txheem (62).Cov pob txha ntim thiab osteoclast qhia cheeb tsam ntawm callus tau ntsuas los sib piv cov theem ntawm callus tsim nyob ib ncig ntawm qhov chaw tawg hauv txhua pab pawg, txhawm rau txhawm rau txheeb xyuas qhov micro-CT scan thiab IHC cov txiaj ntsig (Daim duab 6E, 1 thiab 2).Raws li qhov xav tau, qhov liab qab tsis zoo thiab kev tsim callus hauv LOIS tau nce siab dua li lwm pab pawg, qhia tias kev kho pob txha zoo tshwm sim (63).Daim duab S10 qhia pom cov duab kho qhov muag ntawm qhov chaw phais, MT staining tshwm sim ntawm cov ntaub so ntswg khaws ze ntawm cov ntsia hlau, thiab TRAP staining tshwm sim qhia txog cov ntsia hlau-pob txha interface.Nyob rau hauv qhov liab qab substrate, muaj zog callus thiab fibrosis tsim tau pom, thaum lub LOIS-kho implant pom ib tug kuj unadhered nto.Ib yam li ntawd, piv rau cov liab qab tsis zoo, qis fibrosis tau pom nyob rau hauv luav cog nrog LOIS, raws li qhia los ntawm cov xub dawb.Tsis tas li ntawd, qhov ruaj khov edema (xiav xub) tuaj yeem raug ntaus nqi rau lub cev tiv thaiv kab mob ntawm LOIS, yog li txo qhov mob hnyav.Qhov uas tsis yog-stick nto nyob ib ncig ntawm qhov cog thiab txo cov fibrosis qhia tias cov txheej txheem tshem tawm yooj yim dua, uas feem ntau ua rau lwm cov pob txha los yog mob.Cov txheej txheem kho pob txha tom qab tshem tawm cov ntsia hlau tau soj ntsuam los ntawm kev ua haujlwm osteoclast ntawm cov ntsia hlau-pob txha interface.Ob leeg pob txha liab qab thiab LOIS cog lus cuam tshuam tau txais cov qib zoo sib xws ntawm osteoclasts kom kho pob txha ntxiv, qhia tias LOIS txheej tsis muaj qhov tsis zoo rau cov pob txha kho lossis tiv thaiv kab mob.Txhawm rau kom paub meej tias qhov kev hloov kho saum npoo ntawm LOIS tsis cuam tshuam nrog cov txheej txheem kho pob txha, kev kuaj X-ray tau siv los sib piv cov pob txha kho ntawm cov luav nrog cov ions tsis zoo thiab 6 lub lis piam ntawm LOIS implantation (Daim duab 6F).Cov txiaj ntsig tau pom tias piv nrog cov pab pawg tsis muaj kab mob zoo nkauj, LOIS tau pom zoo tib yam ntawm cov pob txha kho, thiab tsis muaj cov cim qhia pom tseeb ntawm pob txha (nruam osteolysis kab) hauv ob pawg.
(A) Schematic daim duab ntawm cov pob txha kho cov txheej txheem tom qab tawg.(B) Qhov sib txawv ntawm qhov degree ntawm callus tsim ntawm txhua pawg saum npoo thiab (C) cov duab hla ntawm qhov chaw tawg.(D) TRAP staining kom pom kev ua haujlwm osteoclast thiab pob txha resorption.Raws li kev ua TRAP, qhov tsim ntawm sab nraud callus ntawm cov pob txha cortical tau soj ntsuam ntau los ntawm (E) (1) micro-CT thiab (2) osteoclast kev ua haujlwm.(F) 6 lub lis piam tom qab cog, X-ray duab ntawm cov pob txha tawg ntawm qhov pom qhov tsis zoo (tshwj xeeb los ntawm cov duab plaub liab liab) thiab LOIS (highlighted los ntawm xiav dashed duab plaub).Kev txheeb xyuas txheeb cais tau ua los ntawm ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA).* P <0.05.** P <0.01.
Nyob rau hauv luv luv, LOIS muab ib hom tshiab ntawm kev tiv thaiv kab mob kev tiv thaiv kab mob thiab tiv thaiv kev tiv thaiv txheej rau orthopedic implants.Pa orthopedic implants nrog SHP functionalization nthuav tawm lub sij hawm luv luv los tiv thaiv biofouling, tab sis tsis tuaj yeem tuav lawv cov khoom rau lub sijhawm ntev.Lub superhydrophobicity ntawm lub substrate ntxiab huab cua npuas ntawm cov kab mob thiab cov substrate, yog li tsim cov huab cua hnab ris, yog li tiv thaiv kab mob.Txawm li cas los xij, vim muaj huab cua sib txawv, cov hnab ntim cua no tau yooj yim tshem tawm.Ntawm qhov tod tes, LOIS tau ua pov thawj zoo nws lub peev xwm los tiv thaiv kab mob biofilm.Yog li ntawd, vim yog cov khoom tiv thaiv kev tsis lees paub ntawm cov roj nplaum txheej txhaj rau hauv cov txheej txheem micro / nano txheej txheej, cov kab mob ntsig txog kev mob tuaj yeem tiv thaiv.Ntau yam kev ua cim suav nrog SEM, AFM, XPS thiab CA ntsuas yog siv los ua kom zoo dua LOIS kev tsim khoom.Tsis tas li ntawd, LOIS tseem tuaj yeem siv rau ntau yam khoom siv roj ntsha uas feem ntau siv hauv cov khoom siv kho mob, xws li PLGA, Ti, PE, POM thiab PPSU.Tom qab ntawd, LOIS tau sim hauv vitro los ua pov thawj nws cov khoom tiv thaiv biofouling tawm tsam cov kab mob thiab cov tshuaj lom neeg muaj feem xyuam rau lub cev tiv thaiv kab mob.Cov txiaj ntsig tau pom tias nws muaj cov tshuaj tua kab mob zoo heev thiab cov tshuaj tiv thaiv biofouling piv rau cov cog ntoo liab qab.Tsis tas li ntawd, LOIS qhia tau hais tias muaj zog txhua yam txawm tias tom qab siv cov kev ntxhov siab, uas tsis tuaj yeem zam tau hauv kev phais yas.Vim nws txoj kev kho tus kheej ntawm cov roj nplua nyeem nyob rau saum npoo ntawm micro / nano qauv, LOIS tau ua tiav nws cov khoom tiv thaiv kab mob fouling.Txhawm rau kawm txog biocompatibility thiab antibacterial thaj chaw ntawm LOIS hauv vivo, LOIS tau cog rau hauv luav femur rau 4 lub lis piam.Tsis pom muaj kab mob kab mob hauv cov luav cog nrog LOIS.Tsis tas li ntawd, kev siv IHC tau ua pov thawj txo qis ntawm kev tiv thaiv kab mob hauv zos, qhia tias LOIS tsis cuam tshuam cov txheej txheem kho pob txha.LOIS nthuav tawm cov tshuaj tua kab mob zoo heev thiab tiv thaiv kab mob, thiab tau raug pov thawj los tiv thaiv kev tsim biofilm ua ntej thiab thaum lub sij hawm phais orthopedic, tshwj xeeb tshaj yog rau cov pob txha synthesis.Los ntawm kev siv tus luav pob txha pob txha inflammatory femoral fracture qauv, cov nyhuv ntawm biofilm-hais txog kab mob ntawm cov pob txha kho txheej txheem induced los ntawm pre-incubated implants tau kawm tob.Raws li kev kawm yav tom ntej, yuav tsum muaj tus qauv tshiab hauv vivo los kawm txog kev kis kab mob tom qab cog qoob loo kom nkag siab thiab tiv thaiv kab mob biofilm thaum lub sijhawm kho tag nrho.Tsis tas li ntawd, osteoinduction tseem yog ib qho kev sib tw uas tsis tau daws teeb meem hauv kev koom ua ke nrog LOIS.Kev tshawb fawb ntxiv yog xav tau los muab cov kev xaiv adhesion ntawm osteoinductive hlwb los yog cov tshuaj regenerative nrog LOIS kom kov yeej qhov kev sib tw.Zuag qhia tag nrho, LOIS sawv cev rau kev cog lus orthopedic implant txheej nrog txhua yam robustness thiab zoo heev anti-biofouling zog, uas yuav txo tau SSI thiab tiv thaiv kab mob.
Ntxuav 15mm x 15mm x 1mm 304 SS substrate (Dong Kang M-Tech Co., Kauslim) hauv acetone, EtOH thiab DI dej rau 15 feeb kom tshem tawm cov kab mob.Txhawm rau tsim cov qauv micro / nano-theem rau ntawm qhov chaw, cov txheej txheem huv si yog immersed hauv 48% mus rau 51% HF tov (DUKSAN Corp., South Kauslim) ntawm 50 ° C.Lub sij hawm etching txawv ntawm 0 mus rau 60 feeb.Tom qab ntawd, cov etched substrate raug ntxuav nrog dej deionized thiab muab tso rau hauv 65% HNO3 (Korea DUKSAN Corp.) daws ntawm 50 ° C rau 30 feeb los tsim cov chromium oxide passivation txheej ntawm qhov chaw.Tom qab passivation, lub substrate yog ntxuav nrog dej deionized thiab qhuav kom tau ib tug substrate nrog ib txheej txheej.Tom ntej no, lub substrate raug rau oxygen plasma (100 W, 3 feeb), thiab tam sim ntawd raus rau hauv ib tug tov ntawm 8.88 mM POTS (Sigma-Aldrich, lub teb chaws Yelemees) nyob rau hauv toluene ntawm chav tsev kub rau 12 teev.Tom qab ntawd, lub substrate coated nrog POTS yog ntxuav nrog EtOH, thiab annealed ntawm 150 ° C rau 2 teev kom tau ib tug ntom POTS SAM.Tom qab SAM txheej, ib txheej lubricant tau tsim nyob rau hauv lub substrate los ntawm kev siv ib tug perfluoropolyether lubricant (Krytox 101; DuPont, USA) nrog ib tug loading ntim ntawm 20 μm / cm 2. Ua ntej siv, lim lub lubricant los ntawm ib tug 0.2 micron lim.Tshem tawm cov roj nplua nyeem ntau dhau los ntawm tilting ntawm lub kaum sab xis 45 ° rau 15 feeb.Tib txoj kev tsim khoom siv rau orthopedic implants ua los ntawm 304 SS (locking phaj thiab cortical locking ntsia hlau; Dong Kang M-Tech Co., Kauslim).Tag nrho cov orthopedic implants yog tsim los kom haum lub geometry ntawm luav femur.
Qhov chaw morphology ntawm lub substrate thiab orthopedic implants tau soj ntsuam los ntawm teb emission SEM (Inspect F50, FEI, USA) thiab AFM (XE-100, Park Systems, South Kauslim).Qhov saum npoo roughness (Ra, Rq) yog ntsuas los ntawm kev sib npaug ntawm thaj tsam ntawm 20 μm los ntawm 20 μm (n = 4).Ib qho XPS (PHI 5000 VersaProbe, ULVAC PHI, Nyiv) qhov system nruab nrog Al Kα X-ray qhov chaw nrog qhov chaw loj ntawm 100μm2 tau siv los txheeb xyuas cov tshuaj lom neeg hauv qab.Lub CA ntsuas qhov nruab nrog lub koob yees duab dynamic duab ntes (SmartDrop, FEMTOBIOMED, ​​South Kauslim) tau siv los ntsuas cov kua CA thiab SA.Rau txhua qhov kev ntsuas, 6 mus rau 10 μl ntawm cov tee dej (deionized dej, ntshav nees, EG, 30% ethanol, thiab HD) muab tso rau saum npoo los ntsuas CA.Thaum lub kaum sab xis inclination ntawm lub substrate nce ntawm qhov ceev ntawm 2 ° / s (n = 4), SA yog ntsuas thaum lub tee ntog.
Pseudomonas aeruginosa [American Type Culture Collection (ATCC) 27853] thiab MRSA (ATCC 25923) tau yuav los ntawm ATCC (Manassas, Virginia, USA), thiab cov kab lis kev cai tau khaws cia ntawm -80 ° C.Ua ntej siv, cov kab lis kev cai khov tau muab tso rau hauv trypsin-thawed soybean broth (Komed, Kauslim) ntawm 37 ° C rau 18 teev thiab tom qab ntawd hloov ob zaug kom qhib nws.Tom qab incubation, kab lis kev cai tau centrifuged ntawm 10,000 rpm rau 10 feeb ntawm 4 ° C thiab ntxuav ob zaug nrog PBS (pH 7.3) tov.Cov kab lis kev cai centrifuged yog ces subcultured ntawm ntshav agar daim hlau (BAP).MRSA thiab Pseudomonas aeruginosa tau npaj thaum hmo ntuj thiab coj mus rau hauv Luria-Bertani broth.Qhov concentration ntawm Pseudomonas aeruginosa thiab MRSA nyob rau hauv lub inoculum tau txiav txim ntau npaum li cas los ntawm CFU ntawm kev ncua nyob rau hauv serial dilutions ntawm agar.Tom qab ntawd, kho cov kab mob siab rau 0.5 McFarland tus qauv, uas yog sib npaug rau 108 CFU / ml.Tom qab ntawd dilute cov kab mob ua haujlwm 100 zaug rau 106 CFU / ml.Txhawm rau kuaj cov kab mob ua kom cov kab mob adhesion, lub substrate tau sterilized ntawm 121 ° C rau 15 feeb ua ntej siv.Tom qab ntawd, lub substrate tau pauv mus rau 25 ml ntawm cov kab mob raug tshem tawm thiab incubated ntawm 37 ° C nrog lub zog co (200 rpm) rau 12 thiab 72 teev.Tom qab incubation, txhua lub substrate raug tshem tawm ntawm lub incubator thiab ntxuav 3 zaug nrog PBS kom tshem tawm cov kab mob uas ntab saum npoo.Txhawm rau soj ntsuam biofilm ntawm lub substrate, biofilm tau kho nrog methanol thiab stained nrog 1 ml ntawm crimidine txiv kab ntxwv rau 2 feeb.Tom qab ntawd siv lub tshuab hluav taws xob fluorescence (BX51TR, Olympus, Nyiv) tau siv los thaij duab ntawm biofilm stained.Txhawm rau kom muaj nuj nqis biofilm ntawm lub substrate, cov hlwb txuas tau raug cais tawm ntawm lub substrate los ntawm txoj kev hlaws vortex, uas tau suav hais tias yog txoj kev tsim nyog tshaj plaws los tshem tawm cov kab mob txuas nrog (n = 4).Siv sterile forceps, tshem tawm cov substrate los ntawm qhov nruab nrab txoj kev loj hlob thiab coj mus rhaub lub phaj dej kom tshem tawm cov kua ntau dhau.Loosely txuas cov hlwb raug tshem tawm los ntawm kev ntxuav ob zaug nrog tsis muaj PBS.Txhua lub substrate raug xa mus rau lub raj kuaj tsis muaj menyuam uas muaj 9 ml ntawm 0.1% protein ept saline (PSW) thiab 2 g ntawm 20 mus rau 25 sterile iav hlaws dai (0.4 mus rau 0.5 mm inch).Tom qab ntawd nws tau vortexed rau 3 feeb kom tshem tawm cov hlwb ntawm cov qauv.Tom qab vortexing, lub ncua kev kawm ntawv yog serially diluted 10-fold nrog 0.1% PSW, thiab ces 0.1 ml ntawm txhua dilution yog inoculated rau BAP.Tom qab 24 teev ntawm incubation ntawm 37 ° C, CFU raug suav manually.
Rau cov hlwb, nas fibroblasts NIH/3T3 (CRL-1658; American ATCC) thiab nas macrophages RAW 264.7 (TIB-71; American ATCC) tau siv.Siv Dulbecco's modified Eagle medium (DMEM; LM001-05, Welgene, Kauslim) rau kab lis kev cai nas fibroblasts thiab ntxiv nrog 10% calf serum (S103-01, Welgene) thiab 1% penicillin-streptomycin (PS; LS202-02, Welgene (Welgene) ). Cov hlwb raug incubated thaum hmo ntuj ntawm 37 ° C thiab 5% CO2 rau cell staining, cov hlwb raug kho nrog 4% paraformaldehyde rau 20 feeb thiab muab tso rau hauv 0.5% Triton X Incubate rau 5 feeb hauv -100 raus substrate hauv 50nM tetramethylrhodamine ntawm 37 ° C rau 30 feeb Tom qab cov txheej txheem incubation, siv cov substrate nrog 4′,6-diamino-2-phenylindole (H -1200, Vector Laboratories, UK) VECTASHIELD fixation nruab nrab (n = 4 ib cell). , fluorescein, fluorescein isothiocyanate-albumin (A9771, Sigma-Aldrich, Lub teb chaws Yelemees) thiab tib neeg plasma Lub Alexa Fluor 488-conjugated fibrinogen (F13191, Invitrogen, USA) tau yaj hauv PBS (10 mM, pH 7.4).Qhov siab ntawm albumin thiab fibrinogen yog 1 thiab 150 μg / ml, raws li.Tom qab lub substrate Ua ntej immersing nyob rau hauv cov protein tov, yaug lawv nrog PBS kom rehydrate saum npoo.Tom qab ntawd muab tag nrho cov substrates rau hauv ib lub phaj rau lub qhov dej uas muaj cov tshuaj protein thiab incubate ntawm 37 ° C rau 30 thiab 90 feeb.Tom qab incubation, Lub substrate yog ces muab tshem tawm los ntawm cov protein tov, ntxuav maj mam nrog PBS 3 zaug, thiab tsau nrog 4% paraformaldehyde (n = 4 rau txhua protein).Rau calcium, sodium chloride (0.21 M) thiab potassium phosphate (3.77 mM) tau yaj hauv dej deionized.Cov pH ntawm cov tshuaj tau hloov mus rau 2.0 los ntawm kev ntxiv cov tshuaj hydrochloride (1M).Tom qab ntawd calcium chloride (5.62 mM) tau yaj hauv cov tshuaj.Los ntawm kev ntxiv 1M tris (hydroxymethyl)-amino Methane kho cov pH ntawm cov tshuaj rau 7.4.Raus tag nrho cov substrates nyob rau hauv ib tug rau-zoo phaj ntim nrog 1.5 × calcium phosphate tov thiab tshem tawm los ntawm cov tshuaj tom qab 30 feeb.Rau staining, 2 g Alizarin Liab S (CI 58005) Sib tov nrog 100 ml ntawm dej deionized.Tom qab ntawd, siv 10% ammonium hydroxide los kho cov pH mus rau 4. dye lub substrate nrog Alizarin Red tov rau 5 feeb, thiab ces co tawm cov ntau dye thiab blot.Tom qab cov txheej txheem co, tshem tawm cov substrate.Cov khoom yog lub cev qhuav dej, ces muab tso rau hauv acetone rau 5 feeb, ces muab tso rau hauv acetone-xylene (1: 1) tov rau 5 feeb, thiab thaum kawg ntxuav nrog xylene (n = 4).Fluorescence microscope (Axio Imager) nrog × 10 thiab × 20 lub hom phiaj lo ntsiab muag yog siv..A2m, Zeiss, Lub teb chaws Yelemees) duab tag nrho cov substrates.ImageJ/FIJI (https://imagej.nih.gov/ij/) tau siv los txheeb xyuas cov ntaub ntawv adhesion ntawm cov tshuaj lom neeg ntawm txhua pab pawg ntawm plaub qhov chaw sib txawv.Hloov tag nrho cov duab rau binary dluab nrog rau qhov chaw ruaj khov rau kev sib piv substrate.
Zeiss LSM 700 confocal microscope tau siv los saib xyuas kev ruaj ntseg ntawm cov roj nplua nyeem hauv PBS hauv hom kev xav.Cov fluorine-raws li SAM-coated iav qauv nrog cov txheej txheej lubricating tau muab tso rau hauv PBS tov, thiab sim siv lub orbital shaker (SHO-1D; Daihan Scientific, South Kauslim) nyob rau hauv me me co co (120 rpm).Tom qab ntawd coj tus qauv thiab saib xyuas qhov poob ntawm cov roj nplua nyeem los ntawm kev ntsuas qhov poob ntawm lub teeb pom kev.Txhawm rau kom tau txais cov duab fluorescence hauv hom kev xav, cov qauv raug xa mus rau 633 nm laser thiab tom qab ntawd sau, vim tias lub teeb yuav rov qab los ntawm cov qauv.Cov qauv ntsuas tau ntsuas ntawm lub sijhawm ntawm 0, 30, 60, thiab 120 teev.
Txhawm rau txiav txim siab qhov cuam tshuam ntawm cov txheej txheem hloov kho saum npoo ntawm nanomechanical zog ntawm orthopedic implants, nanoindenter (TI 950 TriboIndenter, Hysitron, USA) nruab nrog peb sab pyramid-shaped Berkovich pob zeb diamond taub tau siv los ntsuas nanoindenedione.Lub ncov load yog 10 mN thiab cheeb tsam yog 100μmx 100μm.Rau txhua qhov kev ntsuas, lub sijhawm thauj khoom thiab lub sijhawm thauj khoom yog 10 s, thiab lub sijhawm tuav nyob rau hauv qhov siab tshaj plaws indentation load yog 2 s.Siv kev ntsuas los ntawm tsib qhov chaw sib txawv thiab coj qhov nruab nrab.Txhawm rau ntsuas qhov kev ua tau zoo ntawm cov khoom siv dag zog hauv kev thauj khoom, kev ntsuas peb-taw tes dabtsi yog khoov tau ua tiav siv lub tshuab ntsuas thoob ntiaj teb (Instron 5966, Instron, USA).Lub substrate yog compressed ntawm tus nqi tas li ntawm 10 N / s nrog kev nce ntxiv.Lub Bluehill Universal software program (n = 3) tau siv los xam cov flexural modulus thiab qhov siab tshaj plaws compressive.
Txhawm rau simulate cov txheej txheem ua haujlwm thiab cuam tshuam txog kev puas tsuaj thaum lub sijhawm ua haujlwm, cov txheej txheem ua haujlwm tau ua hauv vitro.Cov femurs tau sau los ntawm tua New Zealand dawb luav.Lub femur tau ntxuav thiab kho hauv 4% paraformaldehyde rau 1 lub lis piam.Raws li tau piav qhia nyob rau hauv txoj kev sim tsiaj, qhov ruaj khov femur tau raug phais.Tom qab kev ua haujlwm, kev cog hniav orthopedic tau muab tso rau hauv cov ntshav (nees ntshav, KISAN, Kaus Lim Kauslim) rau 10 s kom paub meej tias cov ntshav adhesions tshwm sim tom qab kev raug mob (n = 3).
Tag nrho ntawm 24 tus txiv neej New Zealand dawb luav (qhov hnyav 3.0 txog 3.5kg, hnub nyoog nruab nrab 6 lub hlis) tau muab faib ua plaub pawg: liab qab tsis zoo, liab qab zoo, SHP thiab LOIS.Tag nrho cov txheej txheem cuam tshuam nrog cov tsiaj tau ua raws li cov qauv kev coj ua ntawm Pawg Saib Xyuas Tsiaj thiab Siv Tshuaj (IAACUC pom zoo, KOREA-2017-0159).Lub orthopedic implant muaj ib tug xauv phaj nrog tsib qhov (ntev 41 hli, dav 7 hli thiab thickness 2 hli) thiab cortical locking screws (ntev 12 hli, txoj kab uas hla 2.7 hli) rau fracture fixation.Tshwj tsis yog rau cov phaj thiab cov ntsia hlau siv hauv cov pab pawg tsis pom zoo, tag nrho cov phaj thiab cov ntsia hlau tau muab tso rau hauv MRSA ncua kev kawm (106 CFU / ml) rau 12 teev.Cov pab pawg liab qab-tsis zoo (n = 6) tau kho nrog liab qab qhov chaw cog qoob loo yam tsis muaj cov kab mob raug tshem tawm, raws li kev tswj tsis zoo rau kev kis kab mob.Cov pab pawg neeg liab qab zoo (n = 6) tau kho nrog lub pob zeb liab qab uas raug rau cov kab mob ua rau muaj kev tiv thaiv kab mob zoo.SHP pab pawg (n = 6) tau kho nrog cov kab mob SHP cog.Thaum kawg, pab pawg LOIS tau kho nrog cov kab mob kis tau LOIS implants (n = 6).Txhua tus tsiaj tau khaws cia rau hauv lub tawb, thiab muab zaub mov thiab dej ntau.Ua ntej kev ua haujlwm, cov luav tau yoo mov rau 12 teev.Cov tsiaj tau muab tshuaj loog los ntawm kev txhaj tshuaj intramuscular ntawm xylazine (5mg / kg) thiab intravenously txhaj ntawm paclitaxel (3mg / kg) rau induction.Tom qab ntawd, xa 2% isoflurane thiab 50% mus rau 70% kev kho mob cov pa oxygen (kwj tus nqi 2 L / min) los ntawm kev ua pa kom tswj tau cov tshuaj loog.Nws yog implanted los ntawm ib tug ncaj qha mus rau lub lateral femur.Tom qab tshem tawm cov plaub hau thiab povidone-iodine tshuaj tua kab mob ntawm daim tawv nqaij, ib qho incision txog 6 cm ntev tau ua rau sab nraud ntawm sab laug nruab nrab femur.Los ntawm qhib qhov sib txawv ntawm cov nqaij ntshiv npog lub femur, lub femur tau nthuav tawm tag nrho.Muab lub phaj rau pem hauv ntej ntawm femoral ncej thiab kho nws nrog plaub screws.Tom qab kho, siv cov saw hniav (1 hli tuab) los tsim cov pob txha tawg hauv thaj tsam ntawm lub qhov thib ob thiab lub qhov thib plaub.Thaum kawg ntawm kev ua haujlwm, lub qhov txhab raug ntxuav nrog saline thiab kaw nrog sutures.Txhua tus luav tau txhaj tshuaj subcutaneously nrog enrofloxacin (5 mg / kg) diluted ib feem peb hauv saline.Postoperative X-rays ntawm lub femur tau coj nyob rau hauv tag nrho cov tsiaj (0, 7, 14, 21, 28, thiab 42 hnub) kom paub meej tias cov pob txha osteotomy.Tom qab siv tshuaj loog sib sib zog nqus, tag nrho cov tsiaj raug tua los ntawm kev txhaj tshuaj KCl (2 mmol / kg) ntawm 28 thiab 42 hnub.Tom qab ua tiav, lub femur tau kuaj xyuas los ntawm micro-CT los soj ntsuam thiab sib piv cov txheej txheem kho pob txha thiab cov pob txha tshiab ntawm plaub pawg.
Tom qab ua tiav, cov ntaub so ntswg uas nyob rau hauv kev sib cuag ncaj qha nrog orthopedic implants tau sau.Cov ntaub so ntswg tau kho nyob rau hauv 10% nruab nrab buffered formalin thaum hmo ntuj thiab tom qab ntawd lub cev qhuav dej hauv EtOH.Cov ntaub so ntswg qhuav tau muab tso rau hauv paraffin thiab muab faib rau ntawm qhov tuab ntawm 40 μm siv microtome (400CS; EXAKT, Lub teb chaws Yelemees).Txhawm rau pom tus kab mob, H&E staining thiab MT staining tau ua.Txhawm rau txheeb xyuas cov lus teb tus tswv, cov ntaub so ntswg tau muab tso rau hauv cov kab mob TNF-α thawj cov tshuaj tiv thaiv kab mob (AB6671, Abcam, USA) thiab luav anti-IL-6 (AB6672; Abcam, USA), thiab tom qab ntawd kho nrog horseradish.Oxidase.Siv cov avidin-biotin complex (ABC) staining system rau cov ntu raws li cov chaw tsim khoom cov lus qhia.Yuav kom tshwm sim raws li cov tshuaj tiv thaiv xim av, 3,3-diaminobenzidine tau siv rau txhua qhov chaw.Lub tshuab luam ntawv swb digital (Pannoramic 250 Flash III, 3DHISTECH, Hungary) tau siv los ua kom pom tag nrho cov hlais, thiab tsawg kawg plaub lub substrates hauv txhua pab pawg tau soj ntsuam los ntawm ImageJ software.
X-ray duab tau muab coj los rau hauv txhua tus tsiaj tom qab kev phais thiab txhua lub lim tiam los saib xyuas kev kho pob txha (n = 6 ib pab).Tom qab ua tiav, kev daws teeb meem siab micro-CT tau siv los suav cov tsim ntawm callus nyob ib ncig ntawm lub femur tom qab kho.Cov femur tau raug ntxuav, tsau hauv 4% paraformaldehyde rau 3 hnub, thiab lub cev qhuav dej hauv 75% ethanol.Cov pob txha qhuav dej tau raug kuaj xyuas los ntawm kev siv micro-CT (SkyScan 1173, Brooke Micro-CT, Kandy, Belgium) los tsim cov duab 3D voxel (2240 ​​× 2240 pixels) ntawm cov qauv pob txha.Siv 1.0 hli Al lim kom txo cov suab nrov thiab siv cov kev daws teeb meem siab rau txhua qhov kev soj ntsuam (E = 133 kVp, I = 60 μA, lub sijhawm sib xyaw = 500 ms).Nrecon software (version 1.6.9.8, Bruker microCT, Kontich, Belgium) tau siv los ua kom muaj 3D ntim ntawm cov qauv scanned los ntawm qhov tau txais 2D lateral projection.Rau kev tsom xam, 3D reconstructed duab muab faib ua 10mm × 10mm × 10mm cubes raws li qhov chaw tawg.Xam cov callus sab nraum cov pob txha cortical.DataViewer (version 1.5.1.2; Bruker microCT, Kontich, Belgium) software tau siv los digitally redirect lub scanned pob txha ntim, thiab CT-Analyzer (version 1.14.4.1; Bruker microCT, Kontich, Belgium) software tau siv los tshuaj xyuas.Cov txheeb ze x-ray nqus coefficients nyob rau hauv cov pob txha laus thiab callus yog qhov txawv ntawm lawv qhov ntom, thiab tom qab ntawd qhov ntim ntawm callus yog quantified (n = 4).Txhawm rau kom paub meej tias biocompatibility ntawm LOIS tsis ncua cov txheej txheem kho pob txha, ntxiv X-ray thiab micro-CT tsom xam tau ua nyob rau hauv ob lub luav: cov liab qab-tsis zoo thiab LOIS pawg.Ob pawg tau raug tua nyob rau lub lim tiam thib 6.
Cov femurs los ntawm cov tsiaj txia tau sau thiab kho hauv 4% paraformaldehyde rau 3 hnub.Tom qab ntawd lub orthopedic implant yog ua tib zoo tshem tawm ntawm lub femur.Tus femur tau decalcified rau 21 hnub los ntawm kev siv 0.5 M EDTA (EC-900, National Diagnostics Corporation).Tom qab ntawd lub decalcified femur tau muab tso rau hauv EtOH kom nws lub cev qhuav dej.Lub cev qhuav dej femur raug tshem tawm hauv xylene thiab muab tso rau hauv paraffin.Tom qab ntawd cov qauv raug muab txiav nrog lub tshuab tsis siv neeg teb microtome (Leica RM2255, Leica Biosystems, Lub teb chaws Yelemees) nrog lub thickness ntawm 3 μm.Rau TRAP staining (F6760, Sigma-Aldrich, Lub teb chaws Yelemees), cov qauv uas tau muab faib tau deparaffinized, rehydrated thiab incubated hauv TRAP reagent ntawm 37 ° C rau 1 teev.Cov duab tau txais los ntawm kev siv tus swb scanner (Pannoramic 250 Flash III, 3DHISTECH, Hungary) thiab ntsuas qhov ntsuas ntawm thaj tsam ntawm thaj chaw stained.Hauv txhua qhov kev sim, tsawg kawg plaub lub substrates hauv txhua pab pawg tau txheeb xyuas los ntawm ImageJ software.
Kev txheeb xyuas qhov tseem ceeb tau ua los ntawm kev siv GraphPad Prism (GraphPad Software Inc., USA).Unpaired t-test thiab one-way analysis of variance (ANOVA) tau siv los ntsuas qhov sib txawv ntawm pawg ntsuas.Cov theem tseem ceeb yog qhia hauv daim duab hauv qab no: *P<0.05, **P<0.01, ***P<0.001 thiab ****P<0.0001;NS, tsis muaj qhov sib txawv tseem ceeb.
Rau cov ntaub ntawv ntxiv rau tsab xov xwm no, thov mus saib http://advances.sciencemag.org/cgi/content/full/6/44/eabb0025/DC1
Qhov no yog ib tsab xov xwm qhib tau muab faib raws li cov ntsiab lus ntawm Creative Commons Attribution-Non-Commercial License, uas tso cai rau siv, faib thiab luam tawm nyob rau hauv ib qho nruab nrab, tsuav yog siv tsis yog rau kev lag luam nce thiab qhov chaw yog qhov qub. ua haujlwm yog lawm.Kev siv.
Nco tseg: Peb tsuas yog hais kom koj muab email chaw nyob kom tus neeg koj pom zoo rau nplooj ntawv paub tias koj xav kom lawv pom email thiab email tsis yog spam.Peb yuav tsis ntes tej email chaw nyob.
Lo lus nug no yog siv los ntsuas seb koj puas yog tib neeg tuaj xyuas thiab tiv thaiv tsis siv neeg spam xa tawm.
Choe Kyung Min, Oh Young Jang, Park Jun Joon, Lee Jin Hyuk, Kim Hyun Cheol, Lee Kyung Moon, Lee Chang Kyu, Lee Yeon Taek, Lee Sun-uck, Jeong Morui
Cov tshuaj tua kab mob thiab tiv thaiv kab mob tiv thaiv kab mob ntawm cov tshuaj orthopedic tuaj yeem txo cov kab mob thiab tiv thaiv kab mob los ntawm kev kis kab mob.
Choe Kyung Min, Oh Young Jang, Park Jun Joon, Lee Jin Hyuk, Kim Hyun Cheol, Lee Kyung Moon, Lee Chang Kyu, Lee Yeon Taek, Lee Sun-uck, Jeong Morui
Cov tshuaj tua kab mob thiab tiv thaiv kab mob tiv thaiv kab mob ntawm cov tshuaj orthopedic tuaj yeem txo cov kab mob thiab tiv thaiv kab mob los ntawm kev kis kab mob.
© 2021 American Association rau Advancement of Science.tag nrho cov cai.AAAS yog tus khub ntawm HINARI, AGORA, OARE, CHORUS, CLOCKSS, CrossRef thiab COUNTER.ScienceAdvances ISSN 2375-2548.