Kwizigulane ezenza utyando lokufakela amathambo, usulelo lwebhaktheriya kunye neempendulo ze-immune ezibangelwa yintsholongwane ziye zahlala ziyingozi ebomini.Izinto eziqhelekileyo zebhayoloji zichaphazeleka kungcoliseko lwebhayoloji, obangela ukuba ibhaktheriya ihlasele indawo eyonzakeleyo kwaye ibangele usulelo emva kokuhlinzwa.Ngoko ke, kukho imfuneko engxamisekileyo yokuphuhlisa i-anti-infection kunye ne-immune yokubalekela i-immune implants ye-orthopedic implants.Apha, siphuhlise itekhnoloji ephuculweyo yokulungiswa komphezulu wokufakelwa kwamathambo okubizwa ngokuba yiLubricated Orthopedic Implant Surface (LOIS), ephefumlelwe ngumphezulu ogudileyo weengqayi zezityalo zepitcher.I-LOIS inolwelo oluhlala ixesha elide nolunamandla olugxotha ulwelo olwahlukeneyo kunye nezinto zebhayoloji (kubandakanywa iiseli, iiproteni, icalcium kunye neebhaktheriya).Ukongeza, siqinisekise ukuqina komatshini ngokuchasene nemikrwelo kunye nokulungisa amandla ngokufanisa umonakalo ongenakuthintelwa ngexesha lotyando lwe-in vitro.Umongo wethambo lomvundla wasetyenziselwa ukuphonononga ngokucokisekileyo isikali esichasene nebhayoloji kunye nesakhono sokuchasa ukosuleleka kweLOIS.Sicinga ukuba i-LOIS, eneempawu zokuchasana ne-biofouling kunye nokuqina koomatshini, linyathelo eliya phambili kutyando lwamathambo olungenalo usulelo.
Namhlanje, ngenxa yokuguga ngokubanzi, inani lezigulane ezinezifo ze-orthopedic (ezifana neefractures ezikhulile, izifo ezidibeneyo ezidibeneyo, kunye ne-osteoporosis) ziye zanda kakhulu (1, 2).Ngoko ke, amaziko ezonyango afaka ukubaluleka okukhulu kwi-orthopedic utyando, kubandakanywa ukufakelwa kwe-orthopedic ye-screws, iipleyiti, izikhonkwane kunye namalungu okwenziwa (3, 4).Nangona kunjalo, i-orthopedic implants yendabuko iye yaxelwa ukuba ichaphazeleka kwi-bacterial adhesion kunye nokwakheka kwe-biofilm, enokubangela ukusuleleka kwendawo yokuhlinzwa (SSI) emva kokuhlinzwa (5, 6).Nje ukuba i-biofilm yenziwe kumphezulu wokufakelwa kwe-orthopedic, ukususwa kwe-biofilm kuba nzima kakhulu nangona kusetyenziswa iidosi ezinkulu ze-antibiotics.Ngoko ke, ngokuqhelekileyo kukhokelela kwizifo ezinzima emva kokuhlinzwa (7, 8).Ngenxa yeengxaki ezingentla apha, unyango lokufakelwa okusulelekileyo kufuneka lubandakanye ukusebenza kwakhona, kubandakanywa nokususwa kwazo zonke ii-implants kunye nezicubu ezijikelezileyo;ngoko ke, isigulane siya kuba neentlungu ezibuhlungu kunye neengozi ezithile (9, 10).
Ukusombulula ezinye zezi ngxaki, iziyobisi-eluting implanti wamathambo ziye zaphuhliswa ukuthintela usulelo ngokuphelisa iibhaktheriya ezincanyathiselwe kumphezulu (11, 12).Nangona kunjalo, isicwangciso sisabonisa imida emininzi.Kuye kwaxelwa ukuba ukufakelwa kwexesha elide lokufakelwa kweziyobisi kubangele umonakalo kwizicubu ezijikelezayo kwaye kubangele ukuvuvukala, oku kunokukhokelela kwi-necrosis (13, 14).Ukongeza, i-solvents ye-organic enokuthi ibekho emva kwenkqubo yokwenziwa kweziyobisi-eluting i-orthopedic implants, ezithintelwe ngokungqongqo yi-US Food and Drug Administration, zifuna amanyathelo okucoca okongezelelweyo ukuhlangabezana nemigangatho yayo (15).Ukufakelwa kweziyobisi eluting kungumngeni ekukhutshweni okulawulwayo kwamachiza, kwaye ngenxa yokulayisha kwawo okunyiniweyo, ukusetyenziswa kwexesha elide kwechiza akunakwenzeka (16).
Esinye isicwangciso esiqhelekileyo kukugquma ukufakelwa ngepolymer echasene ne-antifouling ukukhusela umcimbi webhayoloji kunye neebhaktheriya ekubambeleleni kumphezulu (17).Umzekelo, iipolima zezwitterionic zitsale umdla ngenxa yokungancamatheli kwazo xa zidibana neeproteni zeplasma, iiseli kunye neebhaktheriya.Nangona kunjalo, inemida ethile ehambelana nokuzinza kwexesha elide kunye nokuqina komatshini, okuthintela ukusetyenziswa kwayo ngokufanelekileyo kwi-orthopedic implants, ngakumbi ngenxa yokutshiza ngomatshini ngexesha lotyando (18, 19).Ukongezelela, ngenxa ye-biocompatibility ephezulu, ukungabikho kwesidingo sokuhlinzwa kokususwa, kunye neepropati zokucoca indawo ngokusebenzisa i-corrosion, i-orthopedic implants eyenziwe ngezinto eziphathekayo ezisetyenzisiweyo zisetyenzisiwe (20, 21).Ngexesha lokubola, iibhondi zeekhemikhali phakathi kwe-matrix ye-polymer zichithwa kwaye zikhutshiwe ukusuka phezulu, kwaye abalandeli bahlambulula umphezulu.Nangona kunjalo, ukungcola okuchasene nebhayoloji ngokucoca umphezulu kusebenza ngexesha elifutshane.Ukongeza, uninzi lwezixhobo ezithathayo ezibandakanya i-poly (lactic acid-glycolic acid copolymer) (PLGA), i-polylactic acid (PLA) kunye ne-magnesium-based alloys ziya kuba ne-biodegradation engalinganiyo kunye nokukhukuliseka komzimba, okuya kuchaphazela kakubi ukuzinza komatshini.(amashumi anambini).Ukongeza, amaqhekeza epleyiti anokubola abonelela ngendawo yokuncamathela ibhaktheriya, nto leyo eyandisa ithuba lokusuleleka kwithuba elide.Lo mngcipheko wokuthotywa komatshini kunye nosulelo luthintela ukusetyenziswa kokuhlinzwa kweplastiki (23).
Iimpawu ze-Superhydrophobic (SHP) ezifanisa isakhiwo se-hierarchical samagqabi e-lotus ziye zaba sisisombululo esinokubakho kwiindawo ezichasayo (24, 25).Xa umphezulu we-SHP untywiliselwa kulwelo, amaqamza omoya aya kubanjiswa, ngaloo ndlela enze iipokotho zomoya kunye nokuthintela ukunamathela kwebhaktheriya (26).Nangona kunjalo, uphando olutshanje luye lwabonisa ukuba i-SHP surface inezinto ezingalunganga ezinxulumene nokuqina komatshini kunye nokuzinza kwexesha elide, elithintela ukusetyenziswa kwayo kwiimpawu zonyango.Ngapha koko, iipokotho zomoya ziya kunyibilika kwaye ziphulukane neempawu zazo ezichasayo, oko kukhokelela ekunamatheleni okubanzi kwebhaktheriya ngenxa yendawo enkulu yomphezulu we-SHP (27, 28).Kutshanje, i-Aizenberg kunye noogxa bakhe bazise indlela entsha yokugquma umphezulu we-anti-biofouling ngokuphuhlisa indawo egudileyo ephefumlelwe sisityalo se-Nepenthes pitcher (29, 30).Umphezulu ogudileyo ubonisa uzinzo lwexesha elide phantsi kweemeko zehydraulic, lugxotha ngokugqithisileyo ulwelo kulwelo lwebhayoloji, kwaye uneempawu zokuzilungisa.Nangona kunjalo, akukho ndlela yokufaka i-coating kwi-complex-shample-shaped implant, kwaye akukho bungqina bokuxhasa inkqubo yokuphilisa izicubu ezonakalisiweyo emva kokufakelwa.
Apha, sazisa i-lubricated orthopedic implant surface (LOIS), i-micro/nano-structured orthopedic implant surface kwaye idityaniswe ngokuqinileyo kunye ne-lubricant layer ukuze ithintele ukuba ingadibanisi notyando lweplastiki Usulelo lwebhaktheriya, njengokulungiswa kwe-fracture.Ngenxa yokuba i-fluorine-functionalized micro / nano-level structure ilungisa ngokuqinileyo i-lubricant kwisakhiwo, i-LOIS ephuhlisiwe inokugxotha ngokupheleleyo ukunamathela kweentlobo ezahlukeneyo ze-liquids kunye nokugcina ukusebenza kokuchasana nokungcola ixesha elide.Iingubo ze-LOIS zinokusetyenziswa kwizinto ezahlukeneyo zeemilo ezijoliswe ekuhlanganiseni amathambo.Iipropathi ezigqwesileyo zokulwa ne-biofouling ze-LOIS ngokuchasene nebhaktiriya ye-biofilm [Pseudomonas aeruginosa kunye ne-methicillin-resistant Staphylococcus aureus (MRSA)] kunye nezinto zebhayoloji (iiseli, iiproteni kunye ne-calcium) ziqinisekisiwe kwi-vitro.Umlinganiselo wokubambelela wokubambelela ngokubanzi kwi-substrate ungaphantsi kwe-1%.Ukongeza, nasemva koxinzelelo lomatshini olufana nokukrwempa komphezulu kwenzeka, ukuziphilisa okubangelwa yi-lubricant yokungena kunceda ukugcina iipropathi zayo ezichasayo.Iziphumo zovavanyo lokuqina koomatshini zibonisa ukuba nasemva kokuguqulwa kolwakhiwo kunye neekhemikhali, amandla ewonke awayi kuncitshiswa kakhulu.Ukongeza, uvavanyo lwe-in vitro olulinganisa uxinzelelo lomatshini kwindawo yotyando lwenziwa ukubonisa ukuba i-LOIS inokumelana noxinzelelo olwahlukeneyo lomatshini olwenzeka ngexesha lotyando lweplastiki.Okokugqibela, sasebenzisa imodeli yomvundla esekwe kwi-vivo femoral fracture, engqina ukuba i-LOIS ineempawu eziphezulu zokulwa ne-antibacterial kunye ne-biocompatibility.Iziphumo ze-radiological kunye ne-histological ziqinisekisile ukuba ukuziphatha okuzinzileyo kwe-lubricant kunye ne-anti-biofouling iipropathi kwiiveki ezi-4 emva kokufakelwa kunokufikelela kwi-anti-infection esebenzayo kunye nokusebenza komzimba wokuzivikela ngaphandle kokulibazisa inkqubo yokuphulukisa amathambo.
Umzobo we-1A ubonisa umzobo we-skim ye-LOIS ephuhlisiwe, efakwe kunye nezakhiwo ezincinci / ze-nano-scale kwimodeli yomvundla we-femoral fracture ukuqinisekisa ukuba i-anti-biological fouling egqwesileyo kunye ne-anti-infection properties.Indlela ye-biomimetic iqhutyelwa ukulinganisa umphezulu wesityalo sembiza yamanzi, kunye nokukhusela i-biofouling ngokubandakanya i-lubricant layer ngaphakathi kwesakhiwo se-micro / nano yomhlaba.Umphezulu otofwe ngesithambiso unokunciphisa unxibelelwano phakathi kwezinto zebhayoloji nomphezulu.Ngoko ke, ngenxa yokuqulunqwa kweebhondi zeekhemikhali ezizinzile kumphezulu, unomsebenzi obalaseleyo wokuchasana nokuxhatshazwa kunye nokuzinza kwexesha elide.Ngenxa yoko, iipropathi ezichasene ne-biofouling zomgangatho wokuthambisa zivumela ukusetyenziswa kwezinto ezahlukeneyo kuphando lwebhayoloji.Nangona kunjalo, uphando olubanzi malunga nendlela le ndawo ekhethekileyo idibana ngayo emzimbeni ayikagqitywa.Ngokuthelekisa i-LOIS kunye ne-substrates ehamba ze-in vitro usebenzisa i-albumin kunye nebhaktheriya ye-biofilm, i-non-adhesiveness ye-LOIS inokuqinisekiswa (Umfanekiso 1B).Ukongezelela, ngokukhupha amaconsi amanzi kwi-substrate engenanto ethathiweyo kunye ne-LOIS substrate (Umfanekiso we-S1 kunye ne-Movie S1), ukusebenza kokungcola kwezinto eziphilayo kunokubonakaliswa.Njengoko kubonisiwe kumfanekiso wemicroscope ye-fluorescence, i-substrate evezwayo efakwe kwi-suspension yeprotheni kunye neebhaktheriya ibonise ubuninzi bezinto zebhayoloji ezibambelela kumphezulu.Nangona kunjalo, ngenxa yeempawu zayo ezibalaseleyo zokulwa ne-biofouling, i-LOIS ayifane ibonise nayiphi na i-fluorescence.Ukuze kuqinisekiswe iipropathi zayo zokulwa ne-biofouling kunye ne-anti-infection, i-LOIS isetyenziswe kumphezulu we-orthopedic implants for bone synthesis (iipleyiti kunye ne-screws) kwaye ifakwe kwimodeli yokuphuka komvundla.Ngaphambi kokufakelwa, ukufakelwa kwe-orthopedic ehamba ze kunye ne-LOIS zafakwa kwi-bacterial suspension iiyure ezili-12.I-pre-incubation iqinisekisa ukuba i-biofilm yenziwa kumphezulu wokufakelwa okuveziweyo ukuze kuthelekiswe.Umzobo we-1C ubonisa ifoto yendawo yokuphuka kwiiveki ezi-4 emva kokufakelwa.Ngakwesobunxele, umvundla kunye nokufakelwa kwe-orthopedic engenanto kubonisa inqanaba elibi lokuvuvukala ngenxa yokuqulunqwa kwe-biofilm ebusweni bokufakelwa.Isiphumo esichaseneyo sabonwa kwimivundla efakwe kwi-LOIS, oko kukuthi, izicubu ezijikelezileyo ze-LOIS azizange zibonise iimpawu zosulelo okanye iimpawu zokudumba.Ukongezelela, umfanekiso we-optical ngakwesobunxele ubonisa indawo yokuhlinzwa yomvundla kunye nokufakelwa okubonakalayo, ebonisa ukuba akukho zixhobo ezininzi zokubambelela ezikhoyo kwindawo yokufakelwa okubonakalayo kufunyenwe phezu kwe-LOIS.Oku kubonisa ukuba i-LOIS inozinzo lwexesha elide kwaye inamandla okugcina i-anti-biological fouling kunye ne-anti-adhesion properties.
(A) Idayagram ecwangcisiweyo yeLOIS kunye nokufakelwa kwayo kwimodeli yomvundla yokwaphuka kwebhinqa.(B) Umfanekiso we-Fluorescence microscopy weprotheyini kunye nebhaktiriya biofilm kumphezulu ongenanto kunye ne-LOIS substrate.Iiveki ezi-4 emva kokufakelwa, (C) umfanekiso wesithombe sendawo yokuphuka kunye (D) umfanekiso we-X-ray (uphawulwe ngoxande obomvu).Umfanekiso ngoncedo: Kyomin Chae, Yonsei University.
Imivundla efakwe inzala, evezwe kakubi ibonise inkqubo eqhelekileyo yokuphilisa amathambo ngaphandle kweempawu zokudumba okanye usulelo.Kwelinye icala, i-SHP ifakela i-pre-incubated kwi-bacterial suspension ibonisa ukudumba okunxulumene nosulelo kwiithishu ezijikelezileyo.Oku kunokuthi kubangelwa ukungakwazi ukuvimbela ukunamathela kwebhaktheriya ixesha elide (Umfanekiso S2).Ukuze kuqinisekiswe ukuba i-LOIS ayichaphazeli inkqubo yokuphulukisa, kodwa inqanda ukusuleleka okunokwenzeka okunxulumene nokufakelwa, imifanekiso ye-X-ray ye-matrix evezwe kakuhle kunye ne-LOIS kwindawo yokuphuka yafaniswa (Umfanekiso 1D).Umfanekiso we-X-reyi wokufakelwa kwezinto ezilungileyo ubonise imigca ye-osteolysis eqhubekayo, ebonisa ukuba ithambo alizange liphile ngokupheleleyo.Oku kuphakamisa ukuba inkqubo yokubuyisela amathambo inokulibaziseka kakhulu ngenxa yokuvuvukala okunxulumene nosulelo.Ngokuchaseneyo noko, yabonisa ukuba imivundla eyayifakwe kwi-LOIS yaphila kwaye ayizange ibonise nasiphi na isiza esicacileyo sokwaphuka.
Ukuze kuphuhliswe ukufakelwa kwezonyango kunye nokuzinza kwexesha elide kunye nokusebenza (kubandakanywa nokuchasana ne-biofouling), ezininzi iinzame zenziwe.Nangona kunjalo, ubukho bezinto ezahlukeneyo zebhayoloji kunye nokuguquguquka kokuncamathela kwezicubu kunciphisa uphuhliso lweendlela zabo ezithembekileyo zeklinikhi.Ukuze sinqobe ezi ntsilelo, siye saqulunqa isakhiwo se-micro / nano layered kunye ne-chemical modified surface, ephuculwe ngenxa yamandla aphezulu e-capillary kunye nokudibanisa kweekhemikhali ukugcina i-lubricant egudileyo ukuya kwinqanaba elikhulu.Umzobo 2A ubonisa inkqubo yokwenziwa kwe-LOIS iyonke.Okokuqala, lungiselela ibakala lezonyango insimbi engenasici (SS) 304 substrate.Okwesibini, i-micro / nano isakhiwo senziwe kwi-substrate ye-SS nge-chemical etching usebenzisa isisombululo se-hydrofluoric acid (HF).Ukuze ubuyisele ukuxhathisa ukubola kwe-SS, isisombululo se-nitric acid (HNO3) (31) sisetyenziselwa ukucubungula i-substrate edibeneyo.I-Passivation iphucula ukuxhathisa kwe-corrosion ye-SS substrate kwaye icotha kakhulu inkqubo ye-corrosion enokunciphisa ukusebenza ngokubanzi kwe-LOIS.Emva koko, ngokwenza i-monolayer edibeneyo (i-SAM) ene-1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane (i-POTS), umphezulu uguqulwa ngokwemichiza ukuze uphucule ukusebenzisana kweekhemikhali phakathi kwendawo kunye ne-lubricant Affinity egudileyo.Ukuguqulwa komphezulu kunciphisa kakhulu amandla omhlaba owenziweyo we-micro / nano-scale-scale structured surface, ehambelana namandla angaphezulu kwe-lubricant egudileyo.Oku kuvumela i-lubricant ukuba imanziswe ngokupheleleyo, ngaloo ndlela yenza umaleko we-lubricant ozinzileyo phezu komhlaba.Umphezulu olungisiweyo ubonisa i-hydrophobicity eyandisiweyo.Iziphumo zibonisa ukuba i-lubricant slippery ibonisa ukuziphatha okuzinzile kwi-LOIS ngenxa yokubambisana kweekhemikhali eziphezulu kunye ne-capillary force ebangelwa yi-micro / nano structure (32, 33).Utshintsho lwamehlo kumphezulu we-SS emva kokuguqulwa komphezulu kunye nenaliti ye-lubricant yafundwa.I-micro / nano layered structure eyenziwe phezu komhlaba ingabangela utshintsho olubonakalayo kwaye lube mnyama ebusweni.Esi siganeko sibangelwa yimpembelelo yokusasazwa kokukhanya kwindawo erhabaxa, eyandisa ukubonakaliswa okubangelwa yindlela yokubambisa ukukhanya (34).Ukongeza, emva kokuba i-lubricant ifakwe, i-LOIS iba mnyama.Umaleko wokuthambisa ubangela ukuba ukukhanya okuncinci kubonakaliswe kwi-substrate, ngaloo ndlela kube mnyama iLOIS.Ukuze kuphuculwe i-microstructure/nanostructure ukubonisa eyona angle incinci sliding (SA) ukuphumeza ukusebenza kwe-anti-biofouling, ukuskena i-electron microscopy (SEM) kunye neeperi zeathom zisetyenziselwa ukwenza amaxesha ahlukeneyo e-HF etching (0, 3)., I-15 kunye nemizuzu ye-60) Force Microscope (AFM) (Figure 2B).Imifanekiso ye-SEM kunye ne-AFM ibonisa ukuba emva kwexesha elifutshane le-etching (imizuzu ye-3 ye-etching), i-substrate engenanto yenze i-nano-scale roughness engalinganiyo.Uburhabaxa bomphezulu butshintsha kunye nexesha le-etching (Figure S3).Ijika eliguquguqukayo lexesha libonisa ukuba uburhabaxa bomphezulu buqhubeka bukhula kwaye bufikelele kwincopho kwimizuzu eyi-15 yokucofa, kwaye ke ukuhla okuncinci nje kwexabiso loburhalarhume kubonwa kwimizuzu engama-30 ye-etching.Ngeli xesha, i-nano-level roughness ikhutshwe, ngelixa i-micro-level roughness ikhula ngamandla, okwenza ukuba ukuguquka kuguquke kuzinzile.Emva kwe-etching ngaphezu kwemizuzu engama-30, ukwanda okungaphezulu koburhalarhume kubonwa, okuchazwa ngokweenkcukacha ngolu hlobo lulandelayo: I-SS yenziwe ngentsimbi, exutywe ngezinto ezibandakanya intsimbi, i-chromium, i-nickel, i-molybdenum kunye nezinye izinto ezininzi.Phakathi kwezi zinto, isinyithi, i-chromium kunye ne-molybdenum idlala indima ebalulekileyo ekwenzeni i-micron / nano-scale roughness kwi-SS nge-HF etching.Kumanqanaba okuqala omhlwa, intsimbi kunye nechromium zidla ngokutya ngenxa yokuba imolybdenum inokumelana nomhlwa okuphezulu kunemolybdenum.Njengoko i-etching iqhubela phambili, isisombululo se-etching sifikelela kwi-oversaturation yendawo, senza i-fluorides kunye ne-oxides ebangelwa ukubetha.I-Fluoride kunye ne-oksidi zigqithise kwaye ekugqibeleni ziphinde zigcinwe phezulu, zenze uburhabaxa bomphezulu kuluhlu lwe-micron/nano (31).Le micro/nano-level roughness idlala indima ebalulekileyo kwiipropati zokuziphilisa ze-LOIS.Umphezulu we-double scale surface uvelisa umphumo we-synergistic, ukwandisa kakhulu amandla e-capillary.Lo mcimbi uvumela i-lubricant ukuba ingene kumphezulu ngokuzinzileyo kwaye igalelo kwiipropati zokuziphilisa (35).Ukubunjwa koburhabaxa kuxhomekeke kwixesha le-etching.Ngaphantsi kwemizuzu eli-10 yokuchongwa, umphezulu uqulethe uburhabaxa be-nano-scale kuphela, obungonelanga ukubamba i-lubricant eyaneleyo ukuba ibe nokuchasana kwe-biofouling (36).Ngakolunye uhlangothi, ukuba ixesha lokubetha lidlula imizuzu engama-30, uburhabaxa be-nano-scale obuqulunqwe yi-redeposition yentsimbi kunye ne-chromium buya kunyamalala, kwaye kuphela i-micro-scale roughness iya kuhlala ngenxa ye-molybdenum.Umphezulu ogqithisiweyo awunayo i-nano-scale roughness kwaye ulahlekelwa yimpembelelo ye-synergistic ye-rough-stage roughness, echaphazela kakubi iimpawu zokuziphilisa ze-LOIS.Imilinganiselo yoMzantsi Afrika yenziwa kwii-substrates ezinamaxesha ahlukeneyo e-etching ukubonisa ukusebenza okuchasayo.Iindidi ezahlukahlukeneyo zolwelo zikhethwe ngokusekelwe kwi-viscosity kunye namandla angaphezulu, kubandakanywa amanzi e-deionized (DI), igazi, i-ethylene glycol (EG), i-ethanol (EtOH) kunye ne-hexadecane (HD) (Figure S4).Ixesha elitshintshayo lepatheni ye-etching libonisa ukuba kulwelo olwahlukeneyo olunamandla ahlukeneyo omphezulu kunye ne-viscosities, i-SA ye-LOIS emva kwemizuzu eyi-15 yokuchongwa yeyona iphantsi.Ngoko ke, i-LOIS ilungiselelwe ukuba i-etch imizuzu ye-15 ukwenza i-micron kunye ne-nano-scale roughness, efanelekile ukugcina ngokufanelekileyo ukuqina kwe-lubricant kunye neempawu ezibalaseleyo zokuchasana nokungcola.
(A) Umzobo wesicwangciso senkqubo yokwenziwa kwamanyathelo amane eLOIS.I-inset ibonisa i-SAM eyenziwe kwi-substrate.(B) Imifanekiso ye-SEM kunye ne-AFM, esetyenziselwa ukunyusa i-micro / nano isakhiwo se-substrate phantsi kwamaxesha ahlukeneyo etching.I-X-ray ye-photoelectron spectroscopy (XPS) i-spectra ye-(C) ye-Cr2p kunye (i-D) ne-F1s emva kokugqithiswa komphezulu kunye nokwaleka kwe-SAM.okanye, iyunithi engenasizathu.(E) Imifanekiso emele amathontsi amanzi kwindawo engenanto, echotshiweyo, ye-SHP kunye ne-LOIS.(F) I-engile yoqhagamshelwano (CA) kunye nomlinganiselo we-SA wolwelo olunoxinzelelo olwahlukileyo lomphezulu kwi-SHP nakwi-LOIS.Idatha ichazwa njengentsingiselo ± SD.
Emva koko, ukuze kuqinisekiswe utshintsho kwiimpawu zeekhemikhali zomphezulu, i-X-ray photoelectron spectroscopy (XPS) yayisetyenziselwa ukufundisisa utshintsho kwimichiza ye-substrate surface emva kokugubungela umphezulu ngamnye.Umzobo we-2C ubonisa iziphumo zokulinganisa i-XPS ye-HF etched surface kunye ne-HNO 3 ephathwayo.Iinqununu ezimbini eziphambili kwi-587.3 kunye ne-577.7 eV zinokuthi zifakwe kwi-bond ye-Cr-O ekhoyo kwi-chromium oxide layer, eyona nto ibalulekileyo kwi-HF etched surface.Oku kungenxa yokusetyenziswa kwe-iron kunye ne-chromium fluoride kumphezulu yi-HNO3.I-etching esekelwe kwi-HNO3 ivumela i-chromium ukuba yenze umaleko we-oxide odlulayo kumphezulu, okwenza ukuba i-SS iphinde imelane nokubola.Kumfanekiso we-2D, i-spectra ye-XPS yafunyanwa ukuze kuqinisekiswe ukuba i-silane esekwe kwi-fluorocarbon yenziwa kumphezulu emva kokugquma kwe-SAM, enokugxotha ulwelo oluphezulu kakhulu nakwi-EG, igazi kunye ne-EtOH.Ukugquma kwe-SAM kugqitywe ngokuphendula amaqela asebenzayo e-silane kunye namaqela e-hydroxyl eyenziwe ngonyango lwe-plasma.Ngenxa yoko, ukwanda okubonakalayo kweCF2 kunye neencopho zeCF3 kwabonwa.Amandla okubopha phakathi kwe-286 kunye ne-296 eV abonisa ukuba ukulungiswa kweekhemikhali kugqitywe ngempumelelo yi-SAM coating.I-SHP ibonisa iincopho ze-CF2 (290.1 ​​​​eV) kunye ne-CF3 (293.3 eV) ezinkulu, ezibangelwa yi-silane esekwe kwi-fluorocarbon eyenziwe phezulu.Umzobo we-2E ubonisa imifanekiso emele i-optical angle ye-contact angle (CA) yemilinganiselo yamaqela ahlukeneyo amanzi adibeneyo adityaniswe ne-bare, etched, SHP, kunye ne-LOIS.Le mifanekiso ibonisa ukuba i-etched surface iba yi-hydrophilic ngenxa yesakhiwo se-micro / nano esakhiwe yi-chemical etching ukuze amanzi adibeneyo afakwe kwisakhiwo.Nangona kunjalo, xa i-substrate ihlanganiswe ne-SAM, i-substrate ibonisa ukunyanzeliswa kwamanzi okunamandla, ngoko ke i-SHP yomhlaba wonke yenziwe kwaye indawo yokudibanisa phakathi kwamanzi kunye nomhlaba omncinci.Ekugqibeleni, ukuhla kwe-CA kwabonwa kwi-LOIS, enokuthi ifakwe ekungeneni kwe-lubricant kwi-microstructure, ngoko kwandisa indawo yoqhagamshelwano.Ukuze kuqinisekiswe ukuba umphezulu uneempawu ezigqwesileyo zokugxotha ulwelo kunye neepropati ezingabambeleliyo, i-LOIS yathelekiswa ne-SHP substrate ngokulinganisa i-CA kunye ne-SA kusetyenziswa ulwelo olwahlukeneyo (Figure 2F).Iindidi ezahlukeneyo zolwelo zakhethwa ngokusekelwe kwi-viscosity kunye namandla angaphezulu, kubandakanywa namanzi adibeneyo, igazi, i-EG, i-EtOH kunye ne-HD (Umfanekiso we-S4).Iziphumo zokulinganisa i-CA zibonisa ukuba xa i-CA ithandeka kwi-HD, ixabiso lokunciphisa i-CA, apho i-CA inamandla aphantsi komhlaba.Ukongeza, i-LOIS ye-CA iyonke iphantsi.Nangona kunjalo, umlinganiselo woMzantsi Afrika ubonisa into eyahlukileyo ngokupheleleyo.Ngaphandle kwamanzi ane-ionized, zonke izinto ezimanzi zibambelela kwi-substrate ye-SHP ngaphandle kokuphuma.Ngakolunye uhlangothi, i-LOIS ibonisa i-SA ephantsi kakhulu, apho xa lonke ulwelo luthambekele kwi-engile engaphantsi kwe-10 ° ukuya kwi-15 °, lonke ulwelo luya kuqengqeleka.Oku kubonisa ngamandla ukuba ukunganamatheli kwe-LOIS kungcono kunomphezulu we-SHP.Ukongeza, iingubo ze-LOIS zikwasetyenziswa kwiintlobo ezahlukeneyo zezixhobo, ezibandakanya i-titanium (Ti), i-polyphenylsulfone (PPSU), i-polyoxymethylene (i-POM), i-polyether ether ketone (PEEK) kunye ne-bioabsorbable polymers (PLGA), Zizinto ezifakelwayo zamathambo (Figure S5)).Imifanekiso elandelelanayo yamaconsi kwizinto eziphathwa yi-LOIS ibonisa ukuba iimpawu ze-anti-biofouling ze-LOIS ziyafana kuzo zonke ii-substrates.Ukongezelela, iziphumo zokulinganisa ze-CA kunye ne-SA zibonisa ukuba iipropathi ezingabambeleli ze-LOIS zinokusetyenziswa kwezinye izinto.
Ukuze kuqinisekiswe iipropathi ezichasayo ze-LOIS, iintlobo ezahlukeneyo ze-substrates (kubandakanywa i-bare, etched, i-SHP kunye ne-LOIS) zifakwe kwi-Pseudomonas aeruginosa kunye ne-MRSA.Ezi bhaktheriya zimbini zikhethwe njengeebhaktheriya zesibhedlele ezimele, ezinokukhokelela ekubunjweni kwe-biofilms, ekhokelela kwi-SSI (37).Umzobo we-3 (A kunye no-B) ubonisa imifanekiso ye-microscope ye-fluorescence kunye ne-colon forming unit (CFU) iziphumo zokulinganisa ii-substrates ezifakwe kwi-bacterial suspension ixesha elifutshane (iiyure ze-12) kunye nexesha elide (ii-72 iiyure), ngokulandelanayo.Ngexesha elifutshane, iibhaktheriya ziya kwenza amaqoqo kwaye zikhule ngobukhulu, zizigubungele ngezinto ezinjenge-mucus kunye nokuthintela ukususwa kwazo.Nangona kunjalo, ngexesha le-72 yeeyure ze-incubation, iibhaktheriya ziya kuvuthwa kwaye kube lula ukusabalalisa ukuze zenze iikholoni okanye amaqela amaninzi.Ngoko ke, kunokuqwalaselwa ukuba i-incubation yeeyure ezingama-72 yithuba elide kwaye lixesha elifanelekileyo lokufakelwa ukwenza i-biofilm eqinile phezu komhlaba (38).Ngexesha elifutshane, indawo edibeneyo kunye ne-SHP ibonise ukunamathela kwebhaktheriya, eyancitshiswa malunga ne-25% ukuya kwi-50% xa kuthelekiswa ne-substrate engenanto.Nangona kunjalo, ngenxa yokusebenza kwayo kakuhle kwe-anti-biofouling kunye nokuzinza, i-LOIS ayizange ibonise ukunamathela kwebhaktheriya yebhaktheriya kwixesha elifutshane kunye nexesha elide.Umzobo wesikimu (Umfanekiso 3C) uchaza inkcazo yendlela yokuchasana ne-biological yesisombululo se-etching, i-SHP kunye ne-LOIS.Ingcinga yokuba i-substrate edibeneyo eneempawu ze-hydrophilic iya kuba nommandla omkhulu kune-substrate engenanto.Ke ngoko, ukunamathela ngakumbi kwebhaktheriya kuya kwenzeka kwi-substrate edibeneyo.Nangona kunjalo, xa kuthelekiswa ne-substrate engenanto, i-substrate ehlanganisiweyo ine-biofilm encinci kakhulu eyenziwe phezulu.Oku kungenxa yokuba iimolekyuli zamanzi zibophelela ngokuqinileyo kumphezulu we-hydrophilic kwaye zisebenze njenge-lubricant yamanzi, ngaloo ndlela ziphazamisa ukunamathela kweebhaktheriya kwixesha elifutshane (39).Nangona kunjalo, umaleko weemolekyuli zamanzi zincinci kakhulu kwaye zinyibilika ekumisweni kwebhaktheriya.Ngoko ke, i-molecular layer yamanzi iyanyamalala ixesha elide, ekhokelela ekunamatheleni okubanzi kwebhaktheriya kunye nokwanda.Kwi-SHP, ngenxa yeempawu zayo zexesha elifutshane ezingenamanzi, ukunamathela kwebhaktheriya kunqatshelwe.Ukuncamathisela okuncitshisiweyo kwebhaktheriya kunokunxulunyaniswa neepokotho zomoya ezivaleleke kwisakhiwo esenziweyo kunye namandla angaphantsi komhlaba, ngaloo ndlela kuncitshiswe ukunxibelelana phakathi kokumiswa kwebhaktheriya kunye nomphezulu.Nangona kunjalo, ukunamathela okubanzi kwebhaktheriya kwabonwa kwi-SHP kuba ilahlekelwe ziipropathi zayo zokuchasana nokungcola ixesha elide.Oku ikakhulu ngenxa yokunyamalala kweepokotho zomoya ngenxa yoxinzelelo lwe-hydrostatic kunye nokuchithwa komoya emanzini.Oku kubangelwa ikakhulu ngenxa yokunyamalala kweepokotho zomoya ngenxa yokuchithwa kunye nesakhiwo esine-layer esinika indawo enkulu yokubambelela (27, 40).Ngokungafani nala ma-substrates amabini anempembelelo ebalulekileyo ekuzinzeni kwexesha elide, i-lubricant lubricant equlethwe kwi-LOIS ifakwe kwi-micro / nano structure kwaye ayiyi kunyamalala nakwixesha elide.I-Lubricants ezaliswe ngezakhiwo ezincinci / ze-nano zizinzile kakhulu kwaye zikhanga kakhulu kumphezulu ngenxa yokubambisana kwazo kweekhemikhali, ngaloo ndlela zikhusela ukunamathela kwebhaktheriya ixesha elide.Umzobo S6 ubonisa umfanekiso we-confocal microscope we-lubricant-efakwe kwi-substrate entywiliselwe kwi-phosphate buffered saline (PBS).Imifanekiso eqhubekayo ibonisa ukuba nangemva kweeyure ze-120 zokugubha kancinci (120 rpm), i-lubricant layer kwi-LOIS ihlala ingatshintshi, ebonisa ukuzinza kwexesha elide phantsi kweemeko zokuhamba.Oku kungenxa yobudlelwane obuphezulu beekhemikhali phakathi kwe-SAM esekelwe kwi-fluorine kunye ne-lubricant esekelwe kwi-perfluorocarbon, ukwenzela ukuba i-lubricant layer ingenziwa.Ngoko ke, ukusebenza okuchasene nokungcola kuyagcinwa.Ukongezelela, i-substrate yavavanywa ngokuchasene neeprotheni ezimele (i-albumin kunye ne-fibrinogen), ezikwi-plasma, iiseli ezinxulumene ngokusondeleyo nokusebenza komzimba (i-macrophages kunye ne-fibroblasts), kunye nezo zihambelana nokubunjwa kwamathambo.Umxholo we-calcium uphezulu kakhulu.(Umfanekiso we-3D, i-1 kunye ne-2, kunye ne-Figure S7) (41, 42).Ukongezelela, imifanekiso ye-fluorescence microscope yovavanyo lokubambelela kwi-fibrinogen, i-albumin kunye ne-calcium ibonise iimpawu zokubambelela ezahlukeneyo zeqela ngalinye le-substrate (Umfanekiso we-S8).Ngethuba lokubunjwa kwamathambo, amathambo asanda kwakhiwa kunye neengqimba ze-calcium zinokujikeleza i-orthopedic implant, engenzi nje ukususwa kube nzima, kodwa kunokubangela ingozi engalindelekanga kwisigulane ngexesha lokususwa.Ngoko ke, amanqanaba aphantsi e-calcium ediphozithi kwiipleyiti zethambo kunye nezikrufu ziluncedo kutyando lwamathambo olufuna ukususwa kokufakelwa.Ngokusekwe kubungakanani bendawo eqhotyoshelweyo ngokusekwe kukuqina kwe-fluorescence kunye nokubalwa kweeseli, siqinisekisile ukuba i-LOIS ibonisa iimpawu ezigqwesileyo zokulwa ne-biofouling kuzo zonke izinto zebhayoloji xa kuthelekiswa nezinye ii-substrates.Ngokweziphumo zovavanyo lwe-in vitro, i-anti-biological fouling i-LOIS inokusetyenziswa kukufakelwa kwe-orthopedic, enokuthi ithintele kuphela usulelo olubangelwa yibhaktiriya ye-biofilm, kodwa nokunciphisa ukudumba okubangelwa yinkqubo yokuzikhusela yomzimba esebenzayo.
(A) Imifanekiso ye-Fluorescence microscope yeqela ngalinye (ze, ifakwe, i-SHP kunye ne-LOIS) efakwe kwi-Pseudomonas aeruginosa kunye ne-MRSA yokunqunyanyiswa kwe-12 kunye ne-72 iiyure.(B) Inani le-CFU ebambeleyo ye-Pseudomonas aeruginosa kunye ne-MRSA kumphezulu weqela ngalinye.(C) Umzobo weSchematic we-anti-biological foiling mechanism ye-etching yexesha elifutshane kunye nexesha elide, i-SHP kunye ne-LOIS.(D) (1) Inani le-fibroblasts linamathele kwi-substrate nganye kunye ne-fluorescence microscope imifanekiso yeeseli ezinamathele kwi-bare kunye ne-LOIS.(2) Uvavanyo lokubambelela kwiiprotheni ezinxulumene nomzimba, i-albumin kunye ne-calcium ebandakanyekayo kwinkqubo yokuphulukisa amathambo (* P <0.05, ** P <0.01, *** P <0.001 kunye **** P <0.0001).ns, ayibalulekanga.
Kwimeko yoxinzelelo olugxininisiweyo olungenakuthintelwa, ukuqina komatshini kuye kwaba ngumngeni oyintloko wokusetyenziswa kweengubo ezichasene ne-antifouling.Iindlela ze-gel ezichasene ne-sewage zisekelwe kwi-polymers kunye ne-solubility encinci yamanzi kunye ne-fragility.Ke ngoko, bahlala bechaphazeleka kuxinzelelo lomatshini kwizicelo ze-biomedical.Ngoko ke, iingubo ezichasene ne-antifouling zihlala zingumngeni kwizicelo ezifana nokufakelwa kwamathambo (43, 44).Umzobo we-4A (1) ubonisa iindidi ezimbini eziphambili zoxinzelelo ezisetyenziselwa ukufakelwa kwe-orthopedic implants, kubandakanywa ukukrazula (uxinzelelo lwe-shear) kunye nokunyanzeliswa ngomfanekiso we-optical we-implant eyonakalisiweyo eyenziwa yi-forceps.Ngokomzekelo, xa i-screw iqiniswe nge-screwdriver, okanye xa ugqirha ebambe i-plate yethambo ngokuqinileyo kunye ne-tweezers kwaye esebenzisa amandla oxinzelelo, ipleyiti yamathambo eplastiki iya konakaliswa kwaye ikrwelwe kuzo zombini izikali ze-macro kunye ne-micro / nano (Umfanekiso 4A, 2) .Ukuze uvavanye ukuba i-LOIS eyenziweyo inokumelana nale monakalo ngexesha lotyando lweplastiki, i-nanoindentation yenziwa ukuthelekisa ubunzima be-substrate engenanto kunye ne-LOIS kwi-micro / nano scale ukufunda iimpawu zomatshini we-micro / nano structure Impact (Figure 4B).Umzobo weskimu ubonisa ukuziphatha okungafaniyo kwe-LOIS ngenxa yobukho bezakhiwo ezincinci / ze-nano.I-curve-force-displacement curve yatsalwa ngokusekelwe kwiziphumo ze-nanoindentation (Umfanekiso 4C).Umfanekiso ohlaza okwesibhakabhaka ubonisa i-substrate engenanto, ebonisa i-deformation encinci kuphela, njengoko ibonwa ngobuninzi bobunzulu be-indentation ye-0.26-μm.Ngakolunye uhlangothi, ukunyuka ngokuthe ngcembe kwi-nanoindentation force kunye nokufuduka okubonwa kwi-LOIS (i-curve ebomvu) kunokubonisa iimpawu zokunciphisa izakhiwo zomatshini, okubangelwa ubunzulu be-nanoindentation ye-1.61μm.Oku kungenxa yokuba i-micro / nano structure ekhoyo kwi-LOIS inika indawo enzulu yokuqhubela phambili kwi-tip ye-nanoindenter, ngoko ke i-deformation yayo inkulu kune-substrate engenanto.UKonsta-Gdoutos et al.(45) ikholelwa ukuba ngenxa yobukho be-nanostructures, i-nanoindentation kunye ne-micro / nano roughness ikhokelela kwii-curves ze-nanoindentation ezingaqhelekanga.Indawo enomthunzi ihambelana ne-curve ye-deformation engaqhelekanga echazwe kwi-nanostructure, ngelixa indawo engabonakaliyo ibonakaliswe kwi-microstructure.Olu tshintsho lungonakalisa i-microstructure / nanostructure ye-lubricant yokubamba kwaye ichaphazele kakubi ukusebenza kwayo okuchasayo.Ukuze ufunde impembelelo yomonakalo kwi-LOIS, umonakalo ongenakuphepheka kwizakhiwo ezincinci / ze-nano ziphindwe emzimbeni ngexesha lotyando lweplastiki.Ngokusebenzisa iimvavanyo zegazi kunye neprotheyini yokubambelela, ukuzinza kweempawu ze-anti-biofouling ze-LOIS emva kwe-in vitro kunokumiselwa (Umfanekiso we-4D).Uluhlu lwemifanekiso ye-optical lubonisa umonakalo owenzekileyo kufuphi nemingxuma ye-substrate nganye.Uvavanyo lokunamathela kwegazi lwenziwa ukubonisa umphumo wokulimala komatshini kwi-anti-biofouling coating (Umfanekiso 4E).Njenge-SHP, iipropathi ezichasene ne-fouling zilahlekile ngenxa yomonakalo, kwaye i-LOIS ibonisa iimpawu ezibalaseleyo zokuchasa ukungcola ngokugxotha igazi.Oku kungenxa yokuba, ngenxa yokuba amandla angaphezulu aqhutywa linyathelo le-capillary eligubungela indawo eyonakeleyo, ukuhamba kwi-lubricant ye-lubricant ye-microstructured ibuyisela iipropati ezichasayo (35).Umkhwa ofanayo wabonwa kuvavanyo lokunamathela kwiprotheni kusetyenziswa ialbumin.Kwindawo eyonakalisiweyo, ukunamathela kweprotheni ebusweni be-SHP kubonwa ngokubanzi, kwaye ngokulinganisa indawo yayo, inokulinganiswa njengesiqingatha senqanaba lokubambelela kwi-substrate engenanto.Ngakolunye uhlangothi, i-LOIS igcine iimpawu zayo zokulwa ne-biofouling ngaphandle kokubangela ukunamathela (Umfanekiso 4, F kunye no-G).Ukongeza, umphezulu we-screw uhlala uphantsi koxinzelelo olomeleleyo lomatshini, njengokugrunjwa, ngoko ke sifunde isakhono se-LOIS yokwambathisa ukuze sihlale sinjalo kwi-screw in vitro.Umzobo we-4H ubonisa imifanekiso ye-optical ye-screws eyahlukeneyo, kuquka i-bare, i-SHP kunye ne-LOIS.Uxande olubomvu lumele indawo ekujoliswe kuyo apho ukuxinzezeleka komatshini okunamandla kwenzeka ngexesha lokufakelwa kwamathambo.Ngokufana novavanyo lokubambelela kwiprotheyini yepleyiti, i-microscope ye-fluorescence isetyenziselwa umfanekiso weprotheyini yokubambelela kunye nokulinganisa indawo yokugubungela ukubonisa ukunyaniseka kwe-LOIS yokugqoka, naphantsi koxinzelelo oluqinileyo lomatshini (Umfanekiso 4, I kunye no-J).Izikrufu eziphathwe nge-LOIS zibonisa ukusebenza kakuhle kokuchasana nokungcolisa, kwaye phantse akukho protein ibambelela kumphezulu.Kwelinye icala, ukuncamathela kwiprotheyini kwabonwa kwizikrufu ezingenanto kunye nezikrufu ze-SHP, apho indawo yokugqunywa kwezikrufu ze-SHP yayisisinye kwisithathu sezikrufu ezingenanto.Ukongezelela, i-orthopedic implant esetyenziselwa ukulungiswa kufuneka ibe namandla ngoomatshini ukumelana noxinzelelo olusetyenziswe kwindawo yokuphuka, njengoko kuboniswe kuMzobo 4K.Ngoko ke, uvavanyo lokugoba lwenziwa ukufumanisa umphumo wokuguqulwa kweekhemikhali kwiipropati zomatshini.Ukongezelela, oku kwenzelwa ukugcina uxinzelelo olusisigxina kwi-implant.Faka isicelo samandla omatshini omileyo kuze kube yilapho ukufakelwa kugoqwe ngokupheleleyo kwaye i-curve yoxinzelelo-strain ifunyenwe (Umfanekiso 4L, 1).Iipropati ezimbini ezibandakanya i-Young's modulus kunye namandla aguquguqukayo zifaniswa phakathi kwe-bare kunye ne-LOIS substrates njengezibonakaliso zamandla abo omatshini (Umfanekiso we-4L, i-2 kunye ne-3).Imodyuli encinci ibonisa ukukwazi kwezinto eziphathekayo ukumelana nokutshintsha komatshini.Imodyuli yoMncinci kwisubstrate nganye yi-41.48±1.01 kunye ne-40.06±0.96 GPa, ngokulandelelanayo;Umahluko oboniweyo umalunga ne-3.4%.Ukongezelela, kuxelwa ukuba amandla okugoba, amisela ukuqina kwezinto eziphathekayo, yi-102.34±1.51 GPa ye-substrate engenanto kunye ne-96.99±0.86 GPa ye-SHP.I-substrate engenanto imalunga ne-5.3% ephezulu.Ukuhla okuncinci kwiipropati zoomatshini kunokubangelwa yimpembelelo yenotshi.Kwisiphumo senotshi, uburhabaxa be-micro/nano bunokusebenza njengeseti yeenotshi, ezikhokelela ekugxininiseni koxinzelelo lwendawo kunye nokuchaphazela iipropathi zomatshini zokufakelwa (46).Nangona kunjalo, ngokusekelwe kwinto yokuba ukuqina kwethambo le-cortical yomntu kubikwa ukuba phakathi kwe-7.4 kunye ne-31.6 GPa, kwaye imodyuli ye-LOIS elinganisiweyo idlula ithambo lomntu we-cortical (47), i-LOIS yanele ukuxhasa ukuphuka kunye neyonke. iimpawu zomatshini zichaphazeleka kancinci ngokuguqulwa komphezulu.
(A) Umzobo we-Schematic (1) uxinzelelo lomatshini olusetyenziswa kwi-orthopedic implant ngexesha lomsebenzi, kunye (2) nomfanekiso we-optical we-orthopedic implant eyonakalisiweyo.(B) I-Schematic diagram ye-nano-mechanical properties ukulinganisa nge-nanoindentation kunye ne-LOIS kwindawo engenanto.(C) I-Nanoindentation force-displacement curve yendawo engenanto kunye ne-LOIS.(D) Emva kovavanyo lwe-in vitro, linganisa imifanekiso ye-optical yeentlobo ezahlukeneyo zamacwecwe amathambo (indawo eyonakalisiweyo igxininiswe ngoxande olubomvu) ukulinganisa uxinzelelo lomatshini olubangelwa ngexesha lokusebenza.(E) Uvavanyo lokubambelela kwegazi kunye (F) novavanyo lweprotheyini yokubambelela kwiqela leplate yamathambo eyonakalisiweyo.(G) Linganisa ummandla weprotheyini ebambelela kwipleyiti.(H) Imifanekiso ye-Optical yeentlobo ezahlukeneyo zezikrufu zamathambo emva komfuniselo we-in vitro.(I) Uvavanyo lokubambelela kwiprotheyini yokufunda ukuthembeka kweengubo ezahlukeneyo.(J) Ukulinganisa indawo yeprotheyini ebambelela kwisikrufu.(K) Intshukumo yomvundla yenzelwe ukuvelisa uxinzelelo olusisigxina kwithambo eliqhekekileyo.(L) (1) Goba iziphumo zovavanyo kunye nemifanekiso yamehlo ngaphambi nasemva kokugoba.Umahluko (2) kwimodyuli yoLutsha kunye (3) namandla okugoba phakathi kofakelo olungenanto kunye ne-SHP.Idatha ichazwa njengentsingiselo ± SD (*P <0.05, **P <0.01, ***P <0.001 kunye ****P <0.0001).Umfanekiso ngoncedo: Kyomin Chae, Yonsei University.
Kwiimeko zeklinikhi, unxibelelwano oluninzi lwebhaktheriya kunye nezixhobo zebhayoloji kunye neendawo zamanxeba zivela kwii-biofilms ezivuthiweyo, ezivuthiweyo (48).Ngoko ke, amaZiko e-US oLawulo lweSifo kunye noThintelo aqikelela ukuba i-65% yazo zonke izifo zabantu zihlobene ne-biofilms (49).Kule meko, kuyimfuneko ukubonelela ngoyilo lovavanyo lwe-vivo olubonelela ngokwenziwa kwe-biofilm engaguqukiyo kumphezulu wokufakelwa.Ngoko ke, siye savelisa imodeli yomvundla we-femoral fracture apho i-orthopedic implants ifakwe ngaphambili kwi-bacterial suspension kwaye emva koko ifakwe kwi-rabbit femurs ukufunda iimpawu ezichasayo ze-LOIS kwi-vivo.Ngenxa yezi ngongoma zintathu zilandelayo, usulelo lwebhaktiriya lubangelwa yinkcubeko yangaphambili endaweni yokutofwa ngokuthe ngqo kokunqunyanyiswa kwebhaktiriya: (i) Amajoni omzimba emivundla anamandla ngokwendalo kunowabantu;ngoko ke, i-injection yokumiswa kwebhaktheriya kunye neebhaktheriya zeplankonic zinokwenzeka Akunampembelelo ekubunjweni kwe-biofilms.(Ii) Iibhaktheriya zePlanktonic zichaphazeleka ngakumbi kwi-antibiotics, kwaye i-antibiotics idla ngokusetyenziswa emva kokuhlinzwa;okokugqibela, (iii) ukunqunyanyiswa kwebhaktheriya yeplanktonic kunokuxutywa ngamanzi asemzimbeni wesilwanyana (50).Ngokucwangciswa kwangaphambili kokufakelwa kwi-bacterial suspension ngaphambi kokufakwa, sinokufunda ngokucokisekileyo iziphumo ezinobungozi bosulelo lwebhaktheriya kunye nokusabela komzimba wangaphandle (FBR) kwinkqubo yokuphulukisa amathambo.Imivundla yabingelelwa kwiiveki ezi-4 emva kokufakelwa, kuba i-osseointegration ebalulekileyo kwinkqubo yokuphulukisa amathambo iya kugqitywa kwiiveki ezi-4.Emva koko, ukufakelwa kwasuswa kwimivundla kwizifundo ezisezantsi.Umzobo 5A ubonisa indlela yokwandisa ibhaktheriya.Ufakelo lwamathambo olosulelekileyo lungeniswa emzimbeni.Njengesiphumo sokufukanyelwa kwangaphambili kwibhaktiriya yokunqunyanyiswa, imivundla emithandathu kwisithandathu efakwe ngofakelo lweze yosulelekile, lo gama kungekho namnye umvundla owafakelwe ukufakwa kwe-LOIS owasuleleka.Usulelo lwebhaktheriya luqhubeka ngamanyathelo amathathu, kubandakanywa ukukhula, ukuvuthwa kunye nokusasazwa (51).Okokuqala, iibhaktheriya eziqhotyoshelweyo zivelisa kwaye zikhule phezu komhlaba, kwaye iibhaktheriya zenza i-biofilm xa zikhupha i-polymer extracellular (EPS), i-amyloid kunye ne-extracellular DNA.I-Biofilm ayiphazamisi kuphela ukungena kwe-antibiotics, kodwa ikhuthaza nokuqokelela i-antibiotic-degrading enzymes (njenge-β-lactamase) (52).Ekugqibeleni, i-biofilm isasaza ibhaktheriya evuthiweyo kwizicubu ezijikelezileyo.Ngoko ke, ukusuleleka kwenzeka.Ukongezelela, xa umzimba wangaphandle ungena emzimbeni, usulelo olunokubangela ukusabela okunamandla komzimba kunokubangela ukuvuvukala okukhulu, intlungu, kunye nokunciphisa i-immunity.Umzobo we-5B unikezela ngombono we-FBR obangelwa ukufakwa kwe-orthopedic implant, kunokuba impendulo ye-immune ebangelwa yintsholongwane yebhaktheriya.Amajoni omzimba ayakubona ukufakwa okufakiweyo njengomzimba wasemzini, kwaye emva koko kubangele ukuba iiseli kunye nezicubu zisabele ukugquma umzimba wasemzini (53).Kwiintsuku zokuqala ze-FBR, i-matrix yokubonelela yaqulunqwa kumphezulu wokufakelwa kwamathambo, okubangele i-adsorption ye-fibrinogen.I-adsorbed fibrinogen ke yenza inethiwekhi ye-fibrin eninzi kakhulu, ekhuthaza ukudibanisa kwe-leukocytes (54).Emva kokuba inethiwekhi ye-fibrin yenziwe, ukuvuvukala okukrakra kuya kwenzeka ngenxa yokungena kwe-neutrophils.Kule nyathelo, iindidi ze-cytokines ezifana ne-tumor necrosis factor-α (TNF-α), i-interleukin-4 (IL-4) kunye ne-IL-β zikhululwa, kwaye i-monocytes iqala ukungena kwindawo yokufakelwa kwaye ihluke kwiiseli ezinkulu.Phage (41, 55, 56).Ukunciphisa i-FBR ibisoloko ingumngeni kuba i-FBR egqithisileyo inokubangela ukudumba okubukhali nokungapheliyo, okunokukhokelela kwiingxaki ezibulalayo.Ukuze kuhlolwe impembelelo yosulelo lwebhaktheriya kwiithishu ezijikeleze ukufakelwa okungenanto kunye ne-LOIS, i-hematoxylin kunye ne-eosin (H & E) kunye ne-Masson trichrome (MT) staining yasetyenziswa.Kwimivundla efakwe kwi-substrates engenanto, usulelo olunzima lwebhaktheriya luye lwaqhubela phambili, kwaye izilayidi ze-H & E ze-tissue zabonisa ngokucacileyo amathumba kunye ne-necrosis ebangelwa kukuvuvukala.Kwelinye icala, indawo eyomeleleyo ye-anti-biofouling i-LOIS inqanda ukunamathela kwebhaktiriya, ngoko ke ayibonisi zimpawu zosulelo kwaye yehlisa ukudumba (Figure 5C).Iziphumo ze-MT staining zibonise umkhwa ofanayo.Nangona kunjalo, i-MT staining ibonise i-edema kwimivundla efakwe kwi-LOIS, ebonisa ukuba ukuchacha sekuza kwenzeka (Figure 5D).Ukuze ufunde iqondo lokuphendula kwe-immune, i-immunohistochemical (IHC) i-staining yenziwa ngokusebenzisa i-cytokines TNF-α kunye ne-IL-6 ehambelana nokuphendula komzimba.Ukufakelwa kwe-negative enqunu engazange ibonakale kwiibhaktheriya yafaniswa ne-LOIS eyayivezwe kwiibhaktheriya kodwa ingenalo intsholongwane ukuze ifunde inkqubo yokuphilisa ngokungabikho kosulelo lwebhaktheriya.Umzobo 5E ubonisa umfanekiso obonakalayo wesilayidi se-IHC ebonisa i-TNF-α.Ummandla omdaka ubonisa impendulo yokuzivikela komzimba, ebonisa ukuba impendulo yokuzivikela kwi-LOIS iyancipha kancinci.Ukongeza, inkcazo ye-IL-6 kwi-LOIS yayingaphantsi kakhulu kunokubonakaliswa okungalunganga kwe-Nzalo enqunu (Umfanekiso 5F).Ukubonakaliswa kwe-cytokine kwalinganiswa ngokulinganisa indawo ye-antibody staining ehambelana ne-cytokine (Umfanekiso 5G).Xa kuthelekiswa nemivundla evezwe kwizinto ezifakelweyo ezingalunganga, amanqanaba okuchaza imivundla etyalwe ngeLOIS ebesezantsi, ebonisa umahluko onentsingiselo.Ukuncipha kwintetho ye-cytokine kubonisa ukuba ixesha elide, iipropathi ezizinzileyo zokuchasana ne-LOIS azihambelani kuphela nokuthintela usulelo lwebhaktheriya, kodwa kunye nokuncipha kwe-FBR, eyenziwa yi-macrophages ebambelele kwi-substrate (53), 57 , 58).Ke ngoko, ukuncitshiswa kwempendulo yokuzivikela komzimba ngenxa yepropathi yokuphepha i-immune ye-LOIS inokusombulula iziphumo ebezingalindelekanga emva kokufakelwa, njengokuphendula ngokugqithisileyo komzimba emva kotyando lweplastiki.
(A) Umzobo oqingqiweyo wendlela yokwenza i-biofilm kunye nokusasazeka kumphezulu we-orthopedic implant.I-eDNA, i-extracellular DNA.(B) Umzobo we-Schematic wempendulo ye-immune emva kokufakwa kwe-orthopedic implant.(C) I-H&E staining kunye (D) ne-MT staining yezicubu ezijikelezileyo ze-orthopedic implants nge-bare positive kunye ne-LOIS.I-IHC ye-cytokines enxulumene nomzimba (E) i-TNF-α kunye (F) i-IL-6 yimifanekiso engcolileyo yomvundla ohamba ze-negative kunye ne-LOIS efakwe.(G) Ubungakanani benkcazo ye-cytokine ngomlinganiselo wendawo yokugubungela (** P <0.01).
I-biocompatibility ye-LOIS kunye nefuthe layo kwinkqubo yokuphulukisa amathambo yavavanywa kwi-vivo ngokusebenzisa i-imaging yokuxilonga [i-x-ray kunye ne-micro-computed tomography (CT)] kunye ne-osteoclast IHC.Umzobo we-6A ubonisa inkqubo yokuphulukisa amathambo ebandakanya izigaba ezintathu ezahlukeneyo: ukuvuvukala, ukulungiswa kunye nokulungiswa kwakhona.Xa ukuphuka kwenzeka, iiseli ezivuthayo kunye ne-fibroblasts ziya kungena kwithambo eliqhekekileyo kwaye ziqale ukukhula zibe yi-vascular tissue.Ngethuba lesigaba sokulungisa, i-ingrowth ye-vascular tissue isasazeka kufuphi nendawo yokuphuka.Izicubu ze-Vascular zibonelela ngezondlo zokwenziwa kwethambo elitsha, elibizwa ngokuba yi-callus.Isigaba sokugqibela senkqubo yokuphulukisa amathambo yinqanaba lokuhlaziya, apho ubukhulu be-callus buncitshiswe kubukhulu bethambo eliqhelekileyo ngoncedo lokunyuka kwinqanaba le-osteoclast esebenzayo (59).Ukuhlaziywa kwe-3-dimensional (3D) ngokutsha kwendawo yokuphuka kwenziwa ngokusebenzisa i-micro-CT scans ukujonga ukungafani kwinqanaba lokubunjwa kwefowuni kwiqela ngalinye.Qaphela i-cross-section ye-femur ukujonga ubukhulu be-callus ejikeleze ithambo eliqhekekileyo (Umfanekiso 6, B kunye no-C).I-X-rays nayo yayisetyenziselwa ukuhlolisisa iindawo zokuphuka kwawo onke amaqela ngeveki ukujonga iinkqubo ezahlukeneyo zokuvuselela amathambo kwiqela ngalinye (Umfanekiso S9).I-Callus kunye namathambo avuthiweyo aboniswa kwi-blue / eluhlaza kunye neendlovu, ngokulandelanayo.Uninzi lwezicubu ezithambileyo zihluzwa ngaphandle ngomda osetwe kwangaphambili.I-Nude positive kunye ne-SHP iqinisekisile ukubunjwa kwenani elincinci le-callus ejikeleze indawo yokuphuka.Kwelinye icala, i-negative eveziweyo ye-LOIS kunye nendawo yokwaphuka ijikelezwe yi-callus eshinyeneyo.Imifanekiso ye-Micro-CT ibonise ukuba ukubunjwa kwe-callus kwakuthintelwe yintsholongwane yebhaktheriya kunye nokuvuvukala okunxulumene nosulelo.Oku kungenxa yokuba i-immune system ibeka phambili ukuphiliswa kokulimala kwe-septic okubangelwa ukuvuvukala okunxulumene nosulelo, kunokuba kubuyiselwe amathambo (60).I-IHC kunye ne-Tartrate-resistant Acid Phosphatase (TRAP) staining yenziwa ukujonga umsebenzi we-osteoclast kunye ne-bone resorption (Figure 6D) (61).Zimbalwa kuphela ii-osteoclasts ezisebenzayo ezinebala elimfusa ezifunyenwe kwii-positives ze-ze kunye ne-SHP.Kwelinye icala, ii-osteoclasts ezininzi ezisebenzayo zabonwa kufutshane namathambo amahle kunye naqolileyo e-LOIS.Esi siganeko sibonisa ukuba phambi kwee-osteoclasts, i-callus ejikeleze indawo yokuphuka iphantsi kwenkqubo yokuvuselela ubundlobongela (62).Umthamo wamathambo kunye nommandla we-osteoclast we-osteoclast ye-callus yalinganiswa ukuthelekisa inqanaba lokubunjwa kwe-callus ejikeleze indawo yokuphuka kuwo onke amaqela, ukwenzela ukuba kuqikelelwe i-micro-CT scan kunye neziphumo ze-IHC (Umfanekiso 6E, 1 kunye ne-2).Njengoko kulindelekile, ii-negatives ze-negative kunye ne-callus formation kwi-LOIS yayiphezulu kakhulu kunamanye amaqela, ebonisa ukuba ukulungiswa kwamathambo aphilileyo kwenzeka (63).Umzobo we-S10 ubonisa umfanekiso obonakalayo wendawo yotyando, isiphumo se-MT esingcolisayo se-tissue eqokelelwe kufuphi ne-screw, kunye nesiphumo se-TRAP sibalaselisa i-screw-bone interface.Kwi-substrate engenanto, i-callus enamandla kunye nokwakheka kwe-fibrosis kwabonwa, ngelixa ukufakwa kwe-LOIS-treatment implant kubonisa indawo engahambelaniyo.Ngokufanayo, xa kuthelekiswa nee-negatives ze-negatives, i-fibrosis ephantsi yabonwa kwimivundla efakwe kwi-LOIS, njengoko kuboniswe ngeentolo ezimhlophe.Ukongeza, i-edema eqinile (utolo oluhlaza okwesibhakabhaka) ingabalelwa kwiipropathi zokukhusela i-immune ye-LOIS, ngaloo ndlela inciphisa ukuvuvukala okukhulu.Umphezulu ongenanto ojikeleze ukufakelwa kunye nokunciphisa i-fibrosis ibonisa ukuba inkqubo yokususa ilula, edla ngokubangela ezinye iifractures okanye ukuvuvukala.Inkqubo yokuphilisa ithambo emva kokususwa kwesikrufu yavavanywa ngumsebenzi we-osteoclast kwi-screw-bone interface.Zombini ithambo elingenanto kunye ne-LOIS implant interface ifunxa amanqanaba afanayo e-osteoclasts ukuqhubela phambili ukuphiliswa kwamathambo, ebonisa ukuba i-LOIS yokwambathisa ayinayo impembelelo embi ekuphiliseni amathambo okanye ukuphendula komzimba.Ukuze kuqinisekiswe ukuba ukuguqulwa komhlaba okwenziwa kwi-LOIS akuphazamisi inkqubo yokuphulukisa amathambo, ukuhlolwa kwe-X-ray kusetyenziswe ukuthelekisa ukuphulukiswa kwamathambo oonogwaja kunye nee-ion ezingabonakaliyo eziveziweyo kunye neeveki ze-6 zokufakelwa kwe-LOIS (Umfanekiso 6F).Iziphumo zibonise ukuba xa kuthelekiswa neqela elingabonakaliyo elinqunu, i-LOIS ibonise iqondo elifanayo lokuphulukisa amathambo, kwaye akukho zibonakaliso ezicacileyo zokuphuka (umgca we-osteolysis oqhubekayo) kumaqela omabini.
(A) Umzobo weSchematic wenkqubo yokuphulukisa amathambo emva kokuphuka.(B) Umahluko kwiqondo lokubunjwa kwefowuni yeqela ngalinye lomphezulu kunye (C) nomfanekiso wecandelo elinqamlezayo lendawo yokuphuka.(D) I-TRAP staining ukujonga umsebenzi we-osteoclast kunye ne-bone resorption.Ngokusekelwe kumsebenzi we-TRAP, ukubunjwa kwe-callus yangaphandle ye-cortical bone ihlalutywe ngokobungakanani (E) (1) i-micro-CT kunye (2) nomsebenzi we-osteoclast.(F) Iiveki ezi-6 emva kokufakelwa, imifanekiso ye-X-ray yethambo eliqhekekileyo le-negative evezwe (ephawulwe ngoxande oludayiweyo olubomvu) kunye ne-LOIS (ephawulwe nge-blue dashed rectangle).Uhlalutyo lwamanani lwenziwa ngohlalutyo lwendlela enye yokwahluka (ANOVA).* P <0.05.** P <0.01.
Ngamafutshane, i-LOIS ibonelela ngohlobo olutsha lwesicwangciso sosulelo lwe-antibacterial kunye ne-immune yokuphunyuka kwe-immune implants ye-orthopedic.Ukufakelwa kwamathambo okuqhelekileyo kunye nokusebenza kwe-SHP kubonisa iimpawu zexesha elifutshane zokulwa ne-biofouling, kodwa azikwazi ukugcina iimpawu zazo ixesha elide.I-superhydrophobicity ye-substrate ibamba amaqamza omoya phakathi kwebhaktheriya kunye ne-substrate, ngaloo ndlela yenze iipokotho zomoya, ngaloo ndlela ikhusela usulelo lwebhaktheriya.Nangona kunjalo, ngenxa yokusasazwa komoya, ezi pokotho zomoya zisuswa ngokulula.Kwelinye icala, i-LOIS ibonakalise amandla ayo okuthintela usulelo olunxulumene ne-biofilm.Ngoko ke, ngenxa yeempawu ezichasayo zokuchaswa kwe-lubricant layer injected in layered micro / nano structure surface, ukuvutha okunxulumene nokusuleleka kunokuthintelwa.Iindlela ezahlukeneyo zokubonisa iimpawu ezibandakanya i-SEM, i-AFM, i-XPS kunye nemilinganiselo ye-CA zisetyenziselwa ukunyusa iimeko zokuvelisa i-LOIS.Ukongeza, i-LOIS inokusetyenziswa kwizinto ezahlukeneyo zebhayoloji eziqhele ukusetyenziswa kwizixhobo zokulungisa amathambo, njenge-PLGA, Ti, PE, POM kunye nePPSU.Ke, i-LOIS yavavanywa kwi-vitro ukubonisa iipropathi zayo zokulwa ne-biofouling ngokuchasene neebhaktheriya kunye nezinto zebhayoloji ezinxulumene nokuphendula komzimba.Iziphumo zibonisa ukuba ineempembelelo ezibalaseleyo ze-antibacterial kunye ne-anti-biofouling xa kuthelekiswa nokufakelwa okungenanto.Ukongeza, i-LOIS ibonisa amandla omatshini nangemva kokusebenzisa uxinzelelo lomatshini, olungenakuphepheka kwi-plastic surgery.Ngenxa yeempawu zokuziphilisa ze-lubricant ebusweni besakhiwo se-micro / nano, i-LOIS igcine ngempumelelo i-anti-biological fouling properties.Ukuze ufunde i-biocompatibility kunye ne-antibacterial properties ye-LOIS kwi-vivo, i-LOIS yafakwa kwi-femur yomvundla kwiiveki ezi-4.Alukho usulelo lwebhaktiriya luye lwabonwa kwimivundla etyalwe ngeLOIS.Ukongezelela, ukusetyenziswa kwe-IHC kubonise izinga elincitshisiweyo lempendulo ye-immune yendawo, ebonisa ukuba i-LOIS ayivimbeli inkqubo yokuphulukisa amathambo.I-LOIS ibonisa iimpawu ezibalaseleyo zokulwa ne-antibacterial kunye ne-immune evasion properties, kwaye ibonakaliswe ukuba ithintele ngokufanelekileyo ukubunjwa kwe-biofilm ngaphambi nangexesha lotyando lwamathambo, ngakumbi ukudibanisa amathambo.Ngokusebenzisa imodeli yomvundla wethambo lomvundla oluvuthayo, umphumo wezifo ezinxulumene ne-biofilm kwinkqubo yokuphulukisa ithambo eyenziwa yi-pre-incubated implants yafundwa ngokunzulu.Njengophononongo lwexesha elizayo, imodeli entsha ye-vivo iyadingeka ukufunda ukusuleleka okunokwenzeka emva kokufakelwa ukuqonda ngokupheleleyo kunye nokukhusela izifo ezinxulumene ne-biofilm ngexesha lenkqubo yonke yokuphilisa.Ukongeza, i-osteoinduction isengumngeni ongasonjululwanga ekudityanisweni kwe-LOIS.Uphando olongezelelweyo luyafuneka ukudibanisa ukunamathela okukhethiweyo kweeseli ze-osteoinductive okanye iyeza lokuvuselela kunye ne-LOIS ukoyisa umngeni.Ngokubanzi, i-LOIS imele isithembiso sokufakelwa kwamathambo kunye nokomelela koomatshini kunye neepropathi ezigqwesileyo zokulwa ne-biofouling, ezinokunciphisa i-SSI kunye neziphumo ebezingalindelekanga ze-immune.
Geza i-15mm x 15mm x 1mm 304 SS substrate (i-Dong Kang M-Tech Co., Korea) kwi-acetone, i-EtOH kunye ne-DI yamanzi imizuzu eyi-15 ukususa ukungcola.Ukuze wenze isakhiwo se-micro / nano-level kwi-surface, i-substrate ecocekileyo ifakwe kwi-48% ukuya kwi-51% isisombululo se-HF (i-DUKSAN Corp., i-South Korea) kwi-50 ° C.Ixesha le-etching lihluka ukusuka kwi-0 ukuya kwi-60 imizuzu.Emva koko, i-substrate edibeneyo ihlanjululwe ngamanzi adibeneyo kwaye ifakwe kwi-65% ye-HNO3 (Korea DUKSAN Corp.) isisombululo kwi-50 ° C imizuzu engama-30 ukwenza i-chromium oxide passivation layer phezu komhlaba.Emva kokugqithisa, i-substrate ihlanjwe ngamanzi adibeneyo kwaye yomiswe ukuze ifumane i-substrate enesakhiwo esine-layer.Emva koko, i-substrate ibonakaliswe kwi-oxygen plasma (i-100 W, i-3 imizuzu), kwaye ngokukhawuleza ifakwe kwisisombululo se-8.88 mM POTS (i-Sigma-Aldrich, eJamani) kwi-toluene kwiqondo lokushisa kweeyure ze-12.Emva koko, i-substrate ehlanganiswe ne-POTS ihlanjululwe nge-EtOH, kwaye ifakwe kwi-150 ° C kwiiyure ze-2 ukufumana i-POTS SAM eninzi.Emva kokugquma kwe-SAM, i-lubricant layer yenziwe kwi-substrate ngokusebenzisa i-lubricant perfluoropolyether (Krytox 101; DuPont, USA) kunye nomthamo wokulayisha we-20 μm / cm 2. Ngaphambi kokusetyenziswa, hluza i-lubricant nge-0.2 micron filter.Susa i-lubricant egqithisileyo ngokutyekela kwi-engile engama-45° imizuzu eli-15.Inkqubo efanayo yokuvelisa yayisetyenziselwa ukufakelwa kwe-orthopedic eyenziwe nge-304 SS (ipleyiti yokutshixa kunye ne-cortical locking screw; i-Dong Kang M-Tech Co., Korea).Zonke izifakelo ze-orthopedic zenzelwe ukulingana nejometri ye-rabbit femur.
I-morphology ye-surface ye-substrate kunye ne-orthopedic implants ihlolwe yi-SEM ye-field emission (Hlola i-F50, i-FEI, i-USA) kunye ne-AFM (XE-100, i-Park Systems, eMzantsi Korea).Uburhabaxa bomphezulu (Ra, Rq) bulinganiswa ngokuphinda-phinda indawo ye-20 μm ngama-20 μm (n=4).Inkqubo ye-XPS (PHI 5000 VersaProbe, ULVAC PHI, Japan) ixhotyiswe ngomthombo we-Al Kα X-ray kunye nobukhulu bendawo ye-100μm2 yayisetyenziselwa ukuhlalutya ukubunjwa kweekhemikhali zomhlaba.Inkqubo yokulinganisa i-CA enekhamera yokuthatha umfanekiso oguqukayo (i-SmartDrop, i-FEMTOBIOMED, ​​i-South Korea) isetyenziselwe ukulinganisa ulwelo lwe-CA kunye ne-SA.Kumlinganiselo ngamnye, i-6 ukuya kwi-10 μl yamathontsi (amanzi adiyiniweyo, igazi lehashe, i-EG, i-ethanol engama-30%, kunye ne-HD) zibekwe phezu komhlaba ukulinganisa i-CA.Xa i-angle inclination ye-substrate ikhula ngesantya se-2 ° / s (n = 4), i-SA ilinganiswa xa i-droplet iwa.
I-Pseudomonas aeruginosa [I-American Type Culture Collection (ATCC) 27853] kunye ne-MRSA (ATCC 25923) yathengwa kwi-ATCC (eManassas, eVirginia, eU.SA), kwaye inkcubeko yesitokhwe yagcinwa kwi -80 ° C.Phambi kokusetyenziswa, inkcubeko engumkhenkce yayifukanyelwa kwi-trypsin-thawed soya broth (i-Komed, Korea) kwi-37°C ngeeyure ze-18 kwaye emva koko idluliselwe kabini ukuze isebenze.Emva kwe-incubation, inkcubeko i-centrifuged kwi-10,000 rpm imizuzu eyi-10 kwi-4 ° C kwaye ihlanjwe kabini nge-PBS (pH 7.3) isisombululo.Inkcubeko ye-centrifuged ke i-subcultured kwi-blood agar plates (BAP).I-MRSA kunye nePseudomonas aeruginosa zalungiswa ngobusuku kwaye zakhuliswa kwi-Luria-Bertani broth.Ukuxinwa kwe-Pseudomonas aeruginosa kunye ne-MRSA kwi-inoculum kunqunywe ngokobungakanani be-CFU yokumiswa kwi-serial dilutions kwi-agar.Emva koko, lungisa i-bacterial concentration kwi-0.5 McFarland standard, elingana ne-108 CFU / ml.Emva koko nciphisa ukumiswa kwebhaktheriya esebenzayo ngamaxesha angama-100 ukuya kwi-106 CFU / ml.Ukuvavanya iimpawu zokubambelela kwi-antibacterial, i-substrate yahlanjululwa kwi-121 ° C imizuzu eyi-15 ngaphambi kokusetyenziswa.I-substrate yabe idluliselwa kwi-25 ml yokumiswa kwebhaktheriya kwaye ifakwe kwi-37 ° C kunye nokugubha ngamandla (200 rpm) kwiiyure ze-12 kunye ne-72.Emva kokufukanyiswa, i-substrate nganye ikhutshwe kwi-incubator kwaye ihlanjwe ngamaxesha e-3 nge-PBS ukususa nayiphi na ibhaktheriya edadayo phezu komhlaba.Ukuze kugcinwe i-biofilm kwi-substrate, i-biofilm yayilungiswe nge-methanol kwaye ihlanjwe nge-1 ml ye-orange crimidine imizuzu emi-2.Emva koko i-microscope ye-fluorescence (BX51TR, Olympus, Japan) yayisetyenziselwa ukuthatha imifanekiso ye-biofilm enebala.Ukuze ulinganise i-biofilm kwi-substrate, iiseli eziqhotyoshelweyo zahlulwa kwi-substrate ngendlela ye-bead vortex, eyayibhekwa njengeyona ndlela ifanelekileyo yokususa ibhaktheriya eqhotyoshelweyo (n = 4).Ukusebenzisa i-forceps engenazintsholongwane, susa i-substrate kwindawo yokukhula kwaye ucofe ipleyiti yomthombo ukususa ulwelo olugqithisileyo.Iiseli ezincanyathiselwe ngokukhululekileyo zisuswe ngokuhlanjwa kabini nge-PBS enyumba.I-substrate nganye idluliselwe kwi-tube yovavanyo oluyinyumba equkethe i-9 ml ye-0.1% ye-protein ept saline (PSW) kunye ne-2 g ye-20 ukuya kwi-25 i-glass beads eyinyumba (0.4 ukuya kwi-0.5 mm ububanzi).Emva koko i-vortexed imizuzu ye-3 ukukhupha iiseli kwisampuli.Emva kwe-vortexing, ukumiswa kwakuhlanjululwe ngokuphindaphindiweyo kwi-10-fold kunye ne-0.1% ye-PSW, kwaye emva koko i-0.1 ml ye-dilution nganye ifakwe kwi-BAP.Emva kweeyure ezingama-24 zokufukamela kwi-37°C, i-CFU yabalwa ngesandla.
Kwiiseli, i-fibroblasts ye-mouse i-NIH / 3T3 (CRL-1658; i-American ATCC) kunye ne-mouse macrophages RAW 264.7 (TIB-71; American ATCC) yasetyenziswa.Sebenzisa i-Dulbecco's modified Eagle medium (DMEM; LM001-05, Welgene, Korea) kwinkcubeko ye-fibroblasts ye-mouse kunye nokuncedisa nge-10% ithole serum (S103-01, Welgene) kunye ne-1% penicillin-streptomycin (PS; LS202-02, Welgene (Welgene) Sebenzisa i-DMEM kwinkcubeko ye-macrophages ye-mouse, eyongezwa nge-10% ye-fetal bovine serum (S001-01, Welgene) kunye ne-1% PS Beka i-substrate kwipleyiti yenkcubeko ye-cell-well , Kwaye ugobe iiseli kwiiseli ze-105 / cm2. Iiseli zifakwe ngobusuku kwi-37 ° C kunye ne-5% ye-CO2 ye-cell staining, iiseli zilungiswe nge-4% ye-paraformaldehyde imizuzu ye-20 kwaye ifakwe kwi-0.5% ye-Triton X Incubate kwi-5 imizuzu kwi-100 kwi-37 ° C imizuzu engama-30 emva kwenkqubo yokufukamela, sebenzisa i-substrate ene-4′, 6-diamino-2-phenylindole (H -1200, i-Vector Laboratories, UK) i-VECTASHIELD yokulungisa i-medium (n = 4 ngeseli). , i-fluorescein, i-fluorescein isothiocyanate-albumin (A9771, Sigma-Aldrich, eJamani) kunye neplasma yabantu I-Alexa Fluor 488-conjugated fibrinogen (F13191, Invitrogen, USA) yachithwa kwi-PBS (10 mM, pH 7.4).Ukugxininiswa kwe-albumin kunye ne-fibrinogen kwakuyi-1 kunye ne-150 μg / ml, ngokulandelanayo.Emva kwe-substrate Ngaphambi kokuntywiliselwa kwisisombululo seprotheni, zihlambulule nge-PBS ukuze ubuyisele amanzi kwindawo.Emva koko ucwilise zonke ii-substrates kwisitya esinemithombo emithandathu enesisombululo seprotheni kwaye ufukamele kwi-37 ° C kwi-30 kunye ne-90 imizuzu.Emva kokufakwa kwe-incubation, i-substrate yasuswa kwisisombululo seprotheni, ihlanjwe ngobumnene kunye namaxesha e-PBS e-3, kwaye igxininiswe nge-4% ye-paraformaldehyde (n = 4 kwiprotheni nganye).I-calcium, i-sodium chloride (0.21 M) kunye ne-potassium phosphate (3.77 mM) ) Yachithwa emanzini adibeneyo.I-pH yesisombululo yahlengahlengiswa kwi-2.0 ngokongeza isisombululo se-hydrochloride (1M).Emva koko i-calcium chloride (5.62 mM) yachithwa kwisisombululo.Ngokudibanisa i-1M tris (hydroxymethyl) -amino iMethane ilungisa i-pH yesisombululo kwi-7.4.Gxila zonke ii-substrates kwisitya esinemithombo emithandathu egcwele isisombululo se-1.5 × calcium phosphate kwaye ususe kwisisombululo emva kwemizuzu engama-30.Ukucoca, i-2 g Alizarin Red S (CI 58005) Hlanganisa kunye ne-100 ml yamanzi adibeneyo.Emva koko, sebenzisa i-10% ye-ammonium hydroxide ukulungisa i-pH kwi-4. Daya i-substrate nge-Alizarin Red isisombululo semizuzu emi-5, uze ugubungele idayi engaphezulu kunye ne-blot.Emva kwenkqubo yokugubha, susa i-substrate.Izinto eziphathekayo ziphelelwe ngamanzi, emva koko zifakwe kwi-acetone imizuzu emi-5, emva koko zifakwe kwisisombululo se-acetone-xylene (1: 1) imizuzu emi-5, kwaye ekugqibeleni ihlanjwe nge-xylene (n = 4).Imakroskopu ye-Fluorescence (i-Axio Imager) ene-×10 kunye ne-× 20 iilensi zenjongo isetyenziswa..A2m, Zeiss, Germany) imifanekiso zonke substrates.I-ImageJ/FIJI (https://imagej.nih.gov/ij/) yayisetyenziselwa ukulinganisa idatha yokubambelela kwizinto zebhayoloji kwiqela ngalinye leendawo ezine ezahlukeneyo zokucinga.Guqula yonke imifanekiso ibe yimifanekiso yokubini enemigangatho emiselweyo yokuthelekisa i-substrate.
I-microscope ye-Zeiss LSM 700 ye-confocal yayisetyenziselwa ukubeka esweni uzinzo lwe-lubricant layer kwi-PBS kwimodi yokubonakalisa.Isampulu yeglasi efakwe kwi-fluorine efakwe kwi-SAM ene-lubricating layer ifakwe kwisisombululo se-PBS, kwaye yavavanywa kusetyenziswa i-orbital shaker (i-SHO-1D; i-Daihan Scientific, i-South Korea) phantsi kweemeko zokugubha kancinci (120 rpm).Emva koko thatha isampuli kwaye ubeke iliso ekulahlekeni kwe-lubricant ngokulinganisa ilahleko yokukhanya okubonakalisiweyo.Ukufumana imifanekiso ye-fluorescence kwimodi yokubonakalisa, isampuli ibonakaliswe kwi-laser ye-633 nm kwaye emva koko iqokelelwe, kuba ukukhanya kuya kubonakaliswa emva kwesampuli.Iisampuli zilinganiswe ngexesha lexesha le-0, i-30, i-60, kunye ne-120 iiyure.
Ukuze unqume impembelelo yenkqubo yokuguqulwa komhlaba kwiipropati ze-nanomechanical zokufakelwa kwamathambo, i-nanoindenter (TI 950 TriboIndenter, Hysitron, USA) exhotywe ngecala le-diamond ye-diamond ye-piramidi ye-Berkovich enecala elithathu yayisetyenziselwa ukulinganisa i-nanoindenedione.Umthwalo ophakamileyo yi-10 mN kwaye indawo yi-100μmx 100μm.Kuyo yonke imilinganiselo, ixesha lokulayishwa kunye nokulayishwa li-10, kwaye ixesha lokubamba phantsi komthwalo we-peak indentation ngu-2 s.Thatha imilinganiselo kwiindawo ezintlanu ezahlukeneyo kwaye uthathe umndilili.Ukuze uvavanye ukusebenza kwamandla omatshini phantsi komthwalo, uvavanyo lokugoba olunqamlezayo lwamanqaku amathathu lwenziwa kusetyenziswa umatshini wokuvavanya jikelele (Instron 5966, Instron, USA).I-substrate ixinzelelwe kwinqanaba eliqhubekayo le-10 N / s kunye nomthwalo owandisiweyo.Inkqubo yesoftware yeBluehill Universal (n = 3) yasetyenziselwa ukubala imodyuli eguquguqukayo kunye noxinzelelo olukhulu loxinzelelo.
Ukwenzela ukulinganisa inkqubo yokusebenza kunye nomonakalo onxulumene noomatshini owenziwe ngexesha lokusebenza, inkqubo yokusebenza yenziwa kwi-vitro.Iifemurs zaqokelelwa kwimivundla emhlophe yaseNew Zealand eyabulawa.I-femur yahlanjululwa kwaye yalungiswa kwi-4% ye-paraformaldehyde ye-1 iveki.Njengoko kuchazwe kwindlela yovavanyo lwezilwanyana, i-femur esisigxina yenziwa utyando.Emva kokusebenza, i-orthopedic implant ifakwe kwigazi (igazi lehashe, i-KISAN, iKorea) kwi-10 s ukuqinisekisa ukuba ukunamathela kwegazi kwenzeka emva kokulimala komatshini (n = 3).
Iyonke imivundla emhlophe yaseNew Zealand eyinkunzi engama-24 (ubunzima obuyi-3.0 ukuya kwi-3.5kg, i-avareji yeenyanga ezi-6) yahlulwa ngokungenamkhethe yangamaqela amane: i-nude negative, nude positive, i-SHP ne-LOIS.Zonke iinkqubo ezibandakanya izilwanyana zenziwa ngokuhambelana nemigangatho yokuziphatha ye-Institutional Animal Care and Use Committee (IACUC evunyiweyo, KOREA-2017-0159).I-orthopedic implant iqukethe ipleyiti yokutshixa enemingxuma emihlanu (ubude obuyi-41 mm, ububanzi be-7 mm kunye nobukhulu be-2 mm) kunye ne-cortical locking screws (ubude obuyi-12 mm, ububanzi obuyi-2.7 mm) ukulungiswa kwe-fracture.Ngaphandle kwezo pleyiti kunye nezikrufu ezisetyenziswe kwiqela elingenanto-negative, zonke iipleyiti kunye nezikrufu zazifakwe kwi-MRSA ukumiswa (106 CFU / ml) kwiiyure ze-12.Iqela elihamba ze-negative (n = 6) liphathwe ngokufakela umphezulu oze ngaphandle kokuvezwa kwi-bacterial suspension, njengolawulo olubi losulelo.Iqela elingenanto elilungileyo (n = 6) liphathwe ngokufakelwa kwendawo engenanto evezwe kwiibhaktheriya njengolawulo olulungileyo losulelo.Iqela le-SHP (n = 6) liphathwe ngofakelo lwe-SHP oluvezwe yibhaktiriya.Ekugqibeleni, iqela le-LOIS liphathwe nge-bacterial-exposed LOIS implants (n = 6).Zonke izilwanyana zigcinwa ehokweni, kwaye kukho ukutya okuninzi namanzi.Phambi koqhaqho, imivundla yayizila ukutya iiyure ezili-12.Izilwanyana zenziwa i-anesthetized nge-injection ye-intramuscular ye-xylazine (5mg / kg) kunye ne-injection ye-intravenous ye-paclitaxel (3mg / kg) yokungeniswa.Emva koko, hambisa i-2% isoflurane kunye ne-50% ukuya kwi-70% ye-oksijeni yonyango (izinga lokuhamba kwe-2 L / min) ngokusebenzisa inkqubo yokuphefumula ukugcina i-anesthesia.Ifakwe ngendlela ethe ngqo kwi-femur esecaleni.Emva kokususwa kweenwele kunye ne-povidone-iodine yokubulala iintsholongwane eluswini, ukusika malunga ne-6 cm ubude kwenziwa ngaphandle kwe-femur ephakathi ekhohlo.Ngokuvula i-gap phakathi kwezihlunu ezigubungela i-femur, i-femur ibonakaliswe ngokupheleleyo.Beka ipleyiti phambi kwe-femoral shaft kwaye uyilungise nge-screws ezine.Emva kokulungiswa, sebenzisa i-saw blade (1 mm ubukhulu) ukuze wenze ngokusemthethweni ukuphuka kwindawo phakathi komngxuma wesibini kunye nomgodi wesine.Ekupheleni kotyando, inxeba lihlanjwe nge-saline kwaye livalwe ngeesutures.Umvundla ngamnye wawutofwe nge-enrofloxacin (5 mg/kg) exutywe isinye kwisithathu kwi-saline.I-X-rays ye-postoperative ye-femur ithathwe kuzo zonke izilwanyana (0, 7, 14, 21, 28, kunye neentsuku ze-42) ukuqinisekisa i-osteotomy yethambo.Emva kwe-anesthesia enzulu, zonke izilwanyana zabulawa yi-KCl (2 mmol / kg) nge-28 kunye neentsuku ze-42.Emva kokubulawa, i-femur ihlolwe yi-micro-CT ukujonga nokuthelekisa inkqubo yokuphulukisa amathambo kunye nokwakhiwa kwamathambo amatsha phakathi kwamaqela amane.
Emva kokubulawa, izicubu ezithambileyo ezidibene ngokuthe ngqo kunye nokufakelwa kwamathambo ziqokelelwe.I-tissue yalungiswa kwi-10% engathathi hlangothi i-formalin engathathi hlangothi ngobusuku kwaye yachithwa kwi-EtOH.I-tissue ye-dehydrated ifakwe kwiparafini kwaye yahlulwe kwi-thickness ye-40 μm usebenzisa i-microtome (400CS; EXAKT, eJamani).Ukuze ube nombono wosulelo, i-H&E staining kunye ne-MT staining yenziwa.Ukuze ukhangele impendulo ye-host host, i-tissue edibeneyo ifakwe kunye ne-rabbit anti-TNF-α primary antibody (AB6671, Abcam, USA) kunye ne-rabbit anti-IL-6 (AB6672; Abcam, USA), kwaye emva koko iphathwa nge-horseradish.Oxidase.Faka i-avidin-biotin complex (ABC) inkqubo yokungcolisa kumacandelo ngokwemiyalelo yomenzi.Ukuze kubonakale njengemveliso ye-brown reaction, i-3,3-diaminobenzidine isetyenziswe kuzo zonke iindawo.I-digital slide scanner (i-Pannoramic 250 Flash III, 3DHISTECH, Hungary) yayisetyenziselwa ukujonga zonke izilayi, kwaye ubuncinane ii-substrates ezine kwiqela ngalinye zahlalutywa yi-software ye-ImageJ.
Imifanekiso ye-X-ray ithathwe kuzo zonke izilwanyana emva kokuhlinzwa kunye neveki nganye ukujonga ukuphulukiswa kwe-fracture (n = 6 ngeqela ngalinye).Emva kokubulawa, i-high-resolution micro-CT yayisetyenziselwa ukubala ukubunjwa kwe-callus ejikeleze i-femur emva kokuphilisa.I-femur efunyenweyo yahlanjululwa, igxininiswe kwi-4% ye-paraformaldehyde kwiintsuku ze-3, kwaye iyancipha kwi-ethanol ye-75%.Amathambo aphelelwe ngumzimba aye ahlolwa ngokusebenzisa i-micro-CT (SkyScan 1173, Brooke Micro-CT, Kandy, Belgium) ukuvelisa imifanekiso ye-3D ye-voxel (2240 ​​× 2240 pixels) yesampuli yamathambo.Sebenzisa i-1.0 mm i-Al filter ukunciphisa ingxolo yesignali kwaye usebenzise isisombululo esiphezulu kuzo zonke izikena (E = 133 kVp, I = 60 μA, ixesha lokudibanisa = 500 ms).I-software ye-Nrecon (inguqulo ye-1.6.9.8, i-Bruker microCT, i-Kontich, eBelgium) isetyenziselwe ukuvelisa umthamo we-3D wesampuli eskeniweyo kwi-2D efunyenweyo ye-lateral projection.Ukuhlalutya, umfanekiso owenziwe ngokutsha we-3D uhlukaniswe kwii-cubes ze-10mm × 10mm × 10mm ngokwendawo yokuphuka.Bala i-callus ngaphandle kwethambo lecortical.I-DataViewer (inguqulo ye-1.5.1.2; i-Bruker microCT, i-Kontich, i-Belgium) isofthiwe yayisetyenziselwa ukuqondisa kwakhona i-digital umthamo wethambo, kunye ne-CT-Analyzer (inguqulo ye-1.14.4.1; i-Bruker microCT, i-Kontich, i-Belgium) isofthiwe isetyenziselwa ukuhlalutya.I-x-ray ye-absorption coefficients ehambelanayo kwithambo elivuthiweyo kunye ne-callus iyahlukaniswa ngobuninzi babo, kwaye ke umthamo we-callus ulinganiswe (n = 4).Ukuze kuqinisekiswe ukuba i-biocompatibility ye-LOIS ayilibazisi inkqubo yokuphulukisa amathambo, uhlalutyo olongezelelweyo lwe-X-ray kunye ne-micro-CT lwenziwa kwiimvundla ezimbini: i-naked-negative kunye namaqela e-LOIS.Omabini amaqela abulawa kwiveki ye-6.
Iifemu ezivela kwizilwanyana ezibingelwayo ziqokelelwe kwaye zilungiswe kwi-4% ye-paraformaldehyde kwiintsuku ze-3.I-orthopedic implant isuswa ngononophelo kwi-femur.I-femur yachithwa iintsuku ze-21 ngokusebenzisa i-0.5 M EDTA (EC-900, i-National Diagnostics Corporation).Emva koko i-femur ye-decalcified yacwiliswa kwi-EtOH ukuze ikhuphe amanzi.I-femur ephelelwe ngamanzi emzimbeni yasuswa kwi-xylene kwaye yafakwa kwiparafini.Emva koko isampuli yasikwa nge-microtome ejikelezayo ezenzekelayo (iLeica RM2255, iLeica Biosystems, eJamani) enobunzima be-3 μm.I-TRAP staining (i-F6760, i-Sigma-Aldrich, eJamani), iisampulu ezicatshulweyo zachithwa, zihlaziywa kwaye zifakwe kwi-reagent ye-TRAP kwi-37 ° C ngeyure eli-1.Imifanekiso ifunyenwe kusetyenziswa iskena sesilayidi (i-Pannoramic 250 Flash III, 3DHISTECH, Hungary) yaza yalinganiswa ngokulinganisa indawo egqunyiweyo yendawo.Kuvavanyo ngalunye, ubuncinci ii-substrates ezine kwiqela ngalinye zihlalutywe ngesoftware ye-ImageJ.
Uhlalutyo lokubaluleka kwamanani lwenziwa kusetyenziswa iGraphPad Prism (iGraphPad Software Inc., USA).Uvavanyo lwe-t olungabhangqwanga kunye nohlalutyo lwendlela enye yokwahluka (ANOVA) zisetyenziselwe ukuvavanya umahluko phakathi kwamaqela ovavanyo.Inqanaba lokubaluleka liboniswa kwinani ngolu hlobo lulandelayo: *P<0.05, **P<0.01, ***P<0.001 kunye ****P<0.0001;NS, akukho mahluko ubalulekileyo.
Ukufumana imathiriyeli eyongezelelweyo yeli nqaku, nceda ubone http://advances.sciencemag.org/cgi/content/full/6/44/eabb0025/DC1
Eli linqaku elivulekileyo lokufikelela elisasazwa phantsi kwemigaqo ye-Creative Commons Attribution-Non-Commercial License, evumela ukusetyenziswa, ukusabalalisa kunye nokuveliswa kwakhona kuyo nayiphi na imidiya, nje ukuba ukusetyenziswa kungekhona inzuzo yorhwebo kwaye isiseko kukuba i-original umsebenzi ulungile.Isalathiso.
Qaphela: Sikucela kuphela ukuba usinike idilesi ye-imeyile ukuze umntu omcebisayo kwiphepha azi ukuba ufuna ukubona i-imeyile kwaye i-imeyile ayingospam.Asiyi kubamba naziphi na iidilesi ze-imeyile.
Lo mbuzo usetyenziselwa ukuvavanya ukuba ngaba undwendwe lomntu kunye nokuthintela ukuthunyelwa kwe-spam okuzenzekelayo.
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
I-antibacterial kunye ne-immune yokubalekela i-implants implants ye-orthopedic implants inokunciphisa usulelo kunye neempendulo ze-immune ezibangelwa zizifo.
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
I-antibacterial kunye ne-immune yokubalekela i-implants implants ye-orthopedic implants inokunciphisa usulelo kunye neempendulo ze-immune ezibangelwa zizifo.
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