Kune varwere vari kuvhiyiwa orthopedic implant oparesheni, hutachiona hwehutachiona uye hutachiona-induced immune immune mhinduro dzagara dziri njodzi dzinotyisa.Zvinhu zvakajairwa zvipenyu zvinokonzeresa kusvibiswa kwebiological, izvo zvinoita kuti mabhakitiriya apinde munzvimbo yakakuvara uye kukonzera hutachiona hwepashure.Nokudaro, pane chido chekukurumidzira chekugadzira anti-infection uye immune kupukunyuka coatings kune orthopedic implants.Pano, takagadzira tekinoroji yepamberi yekugadziridza tekinoroji yemabhonzo anodyarwa anonzi Lubricated Orthopedic Implant Surface (LOIS), iyo inofemerwa neyakatsetseka pamusoro pepitcher plant pitchers.LOIS ine inogara kwenguva refu uye yakasimba mvura inodzingira kune zvakasiyana siyana zvinwiwa uye biological zvinhu (kusanganisira masero, mapuroteni, calcium uye mabhakitiriya).Pamusoro pezvo, isu takasimbisa kusimba kwemechani pakatarisana nekukwenya uye kugadzirisa simba nekutevedzera kukuvara kusingadzivisike panguva yekuvhiya in vitro.Tsuro bone mwongo wekuzvimba femoral fracture model yakashandiswa kunyatsodzidza anti-biological scaling uye anti-infection kugona kweLOIS.Isu tinofungidzira kuti LOIS, iyo ine anti-biofouling zvimiro uye kusimba kwemagetsi, inhanho yekumberi mukuvhiya-pasina hutachiona hwemapfupa.
Nhasi, nekuda kwekuchembera kwese, nhamba yevarwere vanotambura nehosha dzemapfupa (zvakadai sekuputsika kwevakwegura, zvirwere zvejoint degenerative, uye osteoporosis) yakawedzera zvikuru (1, 2).Nokudaro, masangano ezvokurapa anonyanya kukosha pakuvhiyiwa kwemapfupa, kusanganisira orthopedic implants ye screws, ndiro, misumari uye majoini ekugadzira (3, 4).Nekudaro, zvechinyakare mapfupa emapfupa anonzi anogona kubatwa nebhakitiriya adhesion uye biofilm kuumbwa, izvo zvinogona kukonzera chirwere chekuvhiya nzvimbo (SSI) mushure mekuvhiyiwa (5, 6).Kana iyo biofilm ichinge yaumbwa pamusoro peiyo orthopedic implant, kubviswa kweiyo biofilm kunova kwakaoma zvakanyanya kunyangwe nekushandiswa kukuru kwemishonga inorwisa mabhakitiriya.Nokudaro, zvinowanzokonzera zvirwere zvakanyanya mushure mekushanda (7, 8).Pamusana pematambudziko ari pamusoro apa, kurapwa kwezviputi zvinotapukira kunofanira kusanganisira kushandiswazve, kusanganisira kubviswa kwezvose zvinomera uye zviputi zvakapoteredza;saka, murwere achatambura kurwadziwa kwakanyanya uye dzimwe njodzi (9, 10).
Kugadzirisa mamwe ematambudziko aya, zvinodhaka-eluting orthopedic implants zvakagadziridzwa kudzivirira hutachiona nekubvisa mabhakitiriya akabatanidzwa pamusoro (11, 12).Nekudaro, iyo zano ichiri kuratidza akati wandei miganhu.Zvave zvichinzi kuisirwa kwenguva refu kwezvinodhaka-eluting implants kwakakonzera kukuvadzwa kwematishu akatenderedza uye kwakakonzera kuzvimba, izvo zvinogona kutungamirira kune necrosis (13, 14).Pamusoro pezvo, izvo organic zvinonyungudutsa zvinogona kunge zviripo mushure mekugadzirwa kwezvinodhaka-eluting orthopedic implants, izvo zvinorambidzwa zvachose neUS Food and Drug Administration, zvinoda mamwe matanho ekunatsa kuti asangane nemiyero yayo (15).Mishonga-eluting implants inonetsa kuburitswa kwezvinodhaka, uye nekuda kwekuremerwa kwavo kwezvinodhaka, kushandiswa kwenguva refu kwemushonga hazvigoneke (16).
Imwe nzira yakajairika ndeyekupfekedza kuisirwa neantifouling polymer kudzivirira biological nyaya uye mabhakitiriya kuti asanamatira pamusoro (17).Semuyenzaniso, ma zwitterionic mapolima akwezva kutarisa nekuda kwekusanamira kwawo zvimiro kana asangana nemapuroteni eplasma, maseru, uye mabhakitiriya.Zvisinei, ine zvimwe zvipimo zvine chokuita nekugadzikana kwenguva refu uye kusimba kwemagetsi, izvo zvinodzivisa kushandiswa kwayo kunoshanda mumapfupa emapfupa, kunyanya nekuda kwekutsvaira kwemagetsi panguva yekuvhiya (18, 19).Pamusoro pezvo, nekuda kweiyo biocompatibility yakakwira, kushaikwa kwekuvhiyiwa kwekubvisa, uye nzvimbo yekuchenesa nzvimbo kuburikidza nekuora, mapfupa emapfupa akagadzirwa nezvinhu zvinogadziriswa zvakashandiswa (20, 21).Munguva yekuora, zvisungo zvekemikari pakati pepolymer matrix zvinoputswa uye kuvharwa kubva pamusoro, uye zvinonamatira zvinochenesa pamusoro.Nekudaro, anti-biological fouling nekucheneswa kwepamusoro kunoshanda munguva pfupi.Pamusoro pezvo, zvinhu zvakawanda zvinobatika zvinosanganisira poly(lactic acid-glycolic acid copolymer) (PLGA), polylactic acid (PLA) uye magnesium-based alloys ichaita kusaenzana biodegradation uye kukukurwa mumuviri, izvo zvinozokanganisa kugadzikana kwemichina.(makumi maviri nemaviri).Mukuwedzera, iyo biodegradable plate fragments inopa nzvimbo yebhakitiriya yekubatanidza, iyo inowedzera mukana wehutachiona munguva refu.Iyi njodzi yekushatiswa kwemechini uye hutachiona inodzikamisa kushandiswa kunoshanda kwekuvhiya kwepurasitiki (23).
Superhydrophobic (SHP) nzvimbo dzinotevedzera maitiro ehierarchical emashizha elotus ave mhinduro inogona kuitika kune anti-fouling nzvimbo (24, 25).Kana iyo SHP yakanyudzwa mumvura, mabhuru emhepo anobatwa, nekudaro achigadzira homwe dzemhepo uye kudzivirira kunamatira kwebhakitiriya (26).Zvisinei, zvidzidzo zvenguva pfupi zvakaratidza kuti SHP pamusoro ine zvipingamupinyi zvine chokuita ne mechanical durability uye kugadzikana kwenguva refu, iyo inodzivisa kushandiswa kwayo mumishonga yekurapa.Uyezve, homwe dzemhepo dzichanyungudika uye dzinorasikirwa ne-anti-fouling zvimiro, nekudaro zvichikonzera kunamatira kwebhakitiriya nekuda kwenzvimbo yakakura yeSHP (27, 28).Munguva ichangopfuura, Aizenberg nevamwe vaaishanda navo vakaunza nzira yekuvandudza ye-anti-biofouling pamusoro pekuputira nekugadzira nzvimbo yakatsetseka yakafemerwa neNepenthes pitcher chirimwa (29, 30).Iyo yakatsetseka pamusoro inoratidza kugadzikana kwenguva refu pasi pe hydraulic mamiriro, yakanyanya mvura inodzinga zvinwiwa zvipenyu, uye ine yekuzvigadzirisa.Nekudaro, hapana nzira yekuisa iyo yekupotera kune yakaoma-yakaita yekurapa implant, uye haina kuratidzwa kutsigira kupora kwematishu akakuvadzwa mushure mekusimwa.
Pano, isu tinosuma lubricated orthopedic implant surface (LOIS), micro/nano-structured orthopedic implant surface uye yakanyatsobatanidzwa neine lubricant layer yakaonda kuti irege kubatanidzwa nekuvhiyiwa kwepurasitiki Zvirwere zvebhakitiriya, zvakadai sekugadzirisa kuputsika.Nokuti iyo fluorine-inoshanda micro / nano-level chimiro chakanyatsogadzirisa lubricant pachigadziro, yakagadziridzwa LOIS inogona kudzoreredza zvizere kusungirirwa kwezvinwiwa zvakasiyana-siyana uye kuchengetedza anti-fouling kushanda kwenguva yakareba.LOIS machira anogona kuiswa kune zvigadzirwa zveakasiyana maumbirwo akagadzirirwa bhonzo synthesis.Iyo yakanakisa anti-biofouling zvimiro zveLOIS kurwisa mabhakitiriya ebiofilm [Pseudomonas aeruginosa uye methicillin-resistant Staphylococcus aureus (MRSA)] uye zvinhu zvipenyu (masero, mapuroteni uye calcium) zvakasimbiswa mu vitro.Muyero wekunamatira wekunamatira kwakawanda kune substrate iri pasi pe1%.Uye zvakare, kunyangwe mushure mekushushikana kwemagetsi senge kukwenya kwepamusoro kunoitika, kuzvirapa kunokonzerwa nemafuta anopinza kunobatsira kuchengetedza hunhu hwayo hwekudzivirira.Iyo mechanic durability bvunzo mhinduro inoratidza kuti kunyangwe mushure mekugadzirisa uye makemikari ekugadzirisa, iyo yakazara simba haizodzikisirwe zvakanyanya.Pamusoro pezvo, in vitro kuyedza inotevedzera kushushikana kwemagetsi munzvimbo yekuvhiya yakaitwa kuratidza kuti LOIS inokwanisa kumirisana nekushushikana kwakasiyana-siyana kunoitika panguva yekuvhiyiwa kwepurasitiki.Pakupedzisira, takashandisa tsuro-yakavakirwa mu vivo femoral fracture modhi, iyo yakaratidza kuti LOIS ine yepamusoro antibacterial zvimiro uye biocompatibility.Radiological uye histological mhedzisiro yakasimbisa kuti yakagadzika lubricant maitiro uye anti-biofouling zvivakwa mukati memavhiki e4 mushure mekusimwa inogona kuwana inoshanda yekurwisa hutachiona uye immune kupukunyuka kuita pasina kunonoka kupora kwemapfupa.
Mufananidzo 1A unoratidza dhiyabhorosi dhiyagiramu yeLOIS yakagadziridzwa, iyo yakasimwa ine micro/nano-scale zvimiro mune tsuro femoral fracture model kuti isimbise yayo yakanakisa anti-biological fouling uye anti-infection properties.Nzira yebiomimetic inoitwa kutevedzera pamusoro pechirimwa chehari yemvura, uye kudzivirira biofouling nekubatanidza lubricant layer mukati meiyo micro/nano chimiro chepamusoro.Nzvimbo yakabayiwa nemafuta inogona kuderedza kusangana pakati pezvinhu zvipenyu nepamusoro.Nokudaro, nekuda kwekuumbwa kwezvisungo zvemakemikari zvakagadzikana pamusoro pepamusoro, zvine maitiro akanaka ekurwisa uye kugadzikana kwenguva refu.Nekuda kweizvozvo, iyo anti-biofouling zvimiro zvenzvimbo yekuzora inobvumira akasiyana anoshanda mashandisirwo mubiomedical research.Nekudaro, tsvakiridzo yakadzama pamusoro pekuti iyi yakakosha nzvimbo inopindirana sei mumuviri haisati yapera.Nokuenzanisa LOIS neakashama substrates in vitro achishandisa albumin uye biofilm bacteria, iyo isina-adhesiveness yeLOIS inogona kusimbiswa (Mufananidzo 1B).Uye zvakare, nekukungurutsa madonhwe emvura pane yakatemerwa isina chinhu substrate uye LOIS substrate (Mufananidzo S1 uye Movie S1), iyo biological kusvibiswa kuita kunogona kuratidzwa.Sezvinoratidzwa mumufananidzo wefluorescence microscope, iyo yakafumurwa substrate yakaiswa mukumiswa kweprotein uye bhakitiriya yakaratidza huwandu hukuru hwezvinhu zvipenyu zvinonamatira pamusoro.Nekudaro, nekuda kweakanakisa anti-biofouling zvivakwa, LOIS kashoma kuratidza chero fluorescence.Kuti isimbise maitiro ayo e-anti-biofouling uye anti-infection, LOIS yakashandiswa pamusoro pekugadzirwa kwemapfupa emapfupa ekugadzirisa mapfupa (mahwendefa uye screws) uye akaiswa mumuenzaniso wekuputsika kwetsuro.Isati yasimwa, yakashama orthopaedic implant uye LOIS zvakaiswa mukumisikidzwa kwebhakitiriya kwemaawa gumi nemaviri.Iyo pre-incubation inova nechokwadi chekuti biofilm inoumbwa pamusoro peiyo yakazaruka implant yekuenzanisa.Mufananidzo 1C unoratidza mufananidzo wenzvimbo yekuputsika mavhiki e4 mushure mekusimwa.Kuruboshwe, tsuro ine mbesa isina kugadzirwa yemapfupa yakaratidza mwero wakakomba wekuzvimba nekuda kwekuumbwa kwebiofilm pamusoro peiyo inoiswa.Mhedzisiro yakapesana yakaonekwa mutsuro dzakasimwa neLOIS, kureva kuti, matishu akapoteredza eLOIS haana kuratidza zviratidzo zvehutachiona kana zviratidzo zvekuzvimba.Mukuwedzera, chifananidzo chemaziso chiri kuruboshwe chinoratidza nzvimbo yekuvhiya yetsuro ine kuiswa pachena, zvichiratidza kuti hapana mazamu akawanda aripo pamusoro pekuiswa pachena kwakawanikwa pamusoro peLOIS.Izvi zvinoratidza kuti LOIS ine kugadzikana kwenguva refu uye inokwanisa kuchengetedza iyo anti-biological fouling uye anti-adhesion properties.
(A) Schematic diagiramu yeLOIS nekusimwa kwayo mune yetsuro femoral fracture modhi.(B) Fluorescence microscopy mufananidzo weprotein uye bacterial biofilm panzvimbo isina chinhu uye LOIS substrate.4 mavhiki mushure mekusimwa, (C) mufananidzo wemifananidzo wenzvimbo yekuputsika uye (D) mufananidzo weX-ray (wakasimbiswa nerectangle tsvuku).Mufananidzo unofadza: Kyomin Chae, Yonsei University.
Tsuro dzakasiirwa, dzakaiswa pachena dzakasimwa zvisina kunaka dzakaratidza kupora kwemapfupa pasina zviratidzo zvekuzvimba kana utachiona.Nekune rimwe divi, SHP inodyara pre-yakaiswa mukumisikidzwa kwebhakitiriya inoratidza kuzvimba kwakabatana nehutachiona pamatishu akatenderedza.Izvi zvinogona kunge zvakakonzerwa nekusakwanisa kwayo kuvharidzira kunamatira kwebhakitiriya kwenguva yakareba (Mufananidzo S2).Kuti aratidze kuti LOIS haina kukanganisa maitiro ekuporesa, asi inhibits zvirwere zvinogona kuitika zvine chokuita nekusimwa, mifananidzo ye-X-ray yematrix akaonekwa uye LOIS panzvimbo yekuputsika yakaenzaniswa (Mufananidzo 1D).Mufananidzo weX-ray weiyo bare positive implant yakaratidza kuenderera mberi kwemitsara yeosteolysis, zvichiratidza kuti bhonzo harina kupora zvachose.Izvi zvinoratidza kuti hurongwa hwekudzoreredza pfupa hunogona kunonoka zvakanyanya nekuda kwekuzvimba kunoenderana nehutachiona.Mukupesana, yakaratidza kuti tsuro dzakasimwa neLOIS dzakanga dzapora uye hadzina kuratidza nzvimbo ipi neipi iri pachena yokuputsika.
Kuti ugadzire mishonga yekurapa nekugadzikana kwenguva refu uye kushanda (kusanganisira kuramba biofouling), kuedza kwakawanda kwakaitwa.Zvisinei, kuvapo kwezvinhu zvakasiyana-siyana zvebhayoloji uye simba rekunamatira kwematishu kunomisa kuvandudzwa kwemaitiro avo ekiriniki akavimbika.Kuti tikunde zvikanganiso izvi, isu takagadzira micro/nano layered chimiro uye chemikemikari yakagadziridzwa nzvimbo, iyo inogadziridzwa nekuda kwepamusoro capillary simba uye makemikari affinity kuchengetedza yakatsetseka lubricant kusvika pamwero mukuru.Mufananidzo 2A unoratidza iyo yakazara kugadzira maitiro eLOIS.Kutanga, gadzirira giredhi rekurapa Stainless simbi (SS) 304 substrate.Kechipiri, iyo micro/nano chimiro chinoumbwa paSS substrate nemakemikari etching uchishandisa hydrofluoric acid (HF) mhinduro.Kuti udzorere kushorwa kweSS, nitric acid (HNO3) mhinduro (31) inoshandiswa kugadzirisa iyo yakatemerwa substrate.Passivation inokwidziridza corrosion resistance yeSS substrate uye inoderedza zvakanyanya corrosion process iyo inogona kuderedza kuita kwese kweLOIS.Zvadaro, nekugadzira self-assembled monolayer (SAM) ine 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane (POTS), nzvimbo yacho inogadziriswa nemakemikari kuti ivandudze kuwirirana kwemakemikari pakati pepamusoro uye yakatsetseka lubricant Affinity.Iko kugadziridzwa kwepamusoro kunoderedza zvakanyanya simba repamusoro reiyo yakagadzirwa micro/nano-scale yakarongeka nzvimbo, iyo inofanana nepamusoro pesimba remafuta anotsvedzerera.Izvi zvinobvumira lubricant kuti inyatsonyorova, nokudaro ichigadzira yakagadzikana lubricant layer pamusoro.Iyo yakagadziridzwa pamusoro inoratidza yakawedzera hydrophobicity.Mhedzisiro yacho inoratidza kuti iyo inotsvedza lubricant inoratidza yakagadzikana maitiro paLOIS nekuda kweiyo yakakwira makemikari affinity uye capillary simba rinokonzerwa neiyo micro/nano chimiro (32, 33).Iko kuchinja kwemaziso pamusoro peSS mushure mekushandurwa kwepamusoro uye jekiseni remafuta zvakadzidzwa.Iyo micro/nano layered structure yakaumbwa pamusoro inogona kukonzera shanduko yekuona uye kusviba pamusoro.Chiitiko ichi chinoratidzwa nekuwedzeredzwa kwechiedza chekuparadzira mhedzisiro pane yakaomarara, iyo inowedzera kupararira kuratidzwa kunokonzerwa nechiedza chekuteya meshini (34).Mukuwedzera, mushure mokunge mafuta aiswa jekiseni, LOIS inova yakasviba.Iyo lubricating layer inokonzera kushomeka kwechiedza kuratidzwa kubva kune substrate, nekudaro kusviba iyo LOIS.Kuti ugone kugonesa iyo microstructure/nanostructure kuratidza diki diki inotsvedza angle (SA) kuti iwane anti-biofouling performance, scanning electron microscopy (SEM) uye atomic pairs dzakashandiswa kuita dzakasiyana HF etching nguva (0, 3)., 15 uye 60 maminitsi) Simba Maikorosikopu (AFM) (Mufananidzo 2B).SEM uye AFM mifananidzo inoratidza kuti mushure menguva pfupi ye etching (3 maminetsi e etching), iyo isina chinhu substrate yakagadzira kusaenzana nano-scale roughness.Kushata kwepamusoro kunoshanduka nenguva yekucheka (Mufananidzo S3).Iyo nguva-inosiyana-siyana curve inoratidza kuti kushata kwepamusoro kunoramba kuchiwedzera uye kunosvika pakakwirira pamaminetsi gumi nemashanu ekucheka, uyezve kuderera kushoma kwehushonga hunoonekwa pamaminetsi makumi matatu ekucheka.Panguva ino, iyo nano-level roughness inodzingirwa kure, nepo micro-level roughness inokura nesimba, zvichiita kuti roughness ichinje zvakanyanya kugadzikana.Mushure mekucheka kweanopfuura maminetsi makumi matatu, kumwe kuwedzera kwehukasha kunoonekwa, kunotsanangurwa zvakadzama sezvinotevera: SS inoumbwa nesimbi, yakasanganiswa nezvinhu zvinosanganisira iron, chromium, nickel, molybdenum nezvimwe zvinhu zvakawanda.Pakati pezvinhu izvi, iron, chromium uye molybdenum inoita basa rakakosha mukugadzira micron/nano-scale roughness paSS neHF etching.Mumatanho ekutanga ekuora, iron uye chromium zvinonyanya kuita ngura nekuti molybdenum ine yakakwirira corrosion resistance pane molybdenum.Sezvo iyo etching inofambira mberi, iyo etching solution inosvika pakuwedzeredza kwenzvimbo, ichigadzira fluorides uye oxides inokonzerwa ne etching.Fluoride uye oxide inonaya uye pakupedzisira inodzoreredza pamusoro, ichigadzira kushata kwepamusoro mu micron/nano renji (31).Iyi micro/nano-level roughness inoita basa rinokosha mukuzvipodza zvinhu zveLOIS.Iyo miviri miviri yepamusoro inoburitsa synergistic mhedzisiro, inowedzera zvakanyanya capillary simba.Chiitiko ichi chinobvumira lubricant kuti ipinde pamusoro zvakagadzikana uye inobatsira pakuzviporesa zvinhu (35).Kuumbwa kwe roughness kunoenderana ne etching nguva.Pasi pemaminetsi e10 etching, nzvimbo yacho inongova nano-scale roughness, iyo isina kukwana kubata mafuta akakwana kuti ave nebiofouling resistance (36).Kune rumwe rutivi, kana nguva yekucheka inopfuura maminitsi makumi matatu, iyo nano-scale roughness inoumbwa nekugadzirisazve iron uye chromium ichanyangarika, uye chete micro-scale roughness inoramba ichikonzerwa ne molybdenum.Iyo yakanyanyisa-etched surface inoshaya nano-scale roughness uye inorasikirwa ne synergistic effect ye-two-stage roughness, iyo inokanganisa maitiro ekuzvirapa kweLOIS.Zviyero zveSA zvakaitwa pamasubstrates ane nguva dzakasiyana dzekuisa kuratidza kurwisa-kukanganisa kuita.Mhando dzakasiyana dzezvinwiwa dzakasarudzwa zvichibva pane viscosity uye pamusoro pesimba, kusanganisira deionized (DI) mvura, ropa, ethylene glycol (EG), ethanol (EtOH) uye hexadecane (HD) (Figure S4).Iyo nguva-inosiyana-siyana etching pateni inoratidza kuti kune dzakasiyana zvinwiwa zvine akasiyana epamusoro masimba uye viscosities, iyo SA yeLOIS mushure memaminitsi gumi nemashanu ekucheka ndiyo yakaderera.Naizvozvo, LOIS yakagadziridzwa kuti iite maminetsi gumi nemashanu kuti igadzire micron uye nano-scale roughness, iyo inokodzera kuchengetedza kusimba kweiyo lubricant uye yakanakisa anti-fouling zvivakwa.
(A) Schematic dhizaini yechina-nhanho yekugadzira maitiro eLOIS.Iyo inset inoratidza iyo SAM yakaumbwa pane substrate.(B) SEM uye AFM mifananidzo, inoshandiswa kukwenenzvera iyo micro/nano chimiro cheiyo substrate pasi peakasiyana etching nguva.X-ray photoelectron spectroscopy (XPS) spectra ye (C) Cr2p uye (D) F1s mushure mekufamba kwepamusoro uye SAM coating.kana, arbitrary unit.(E) Inomiririra mifananidzo yemadonhwe emvura pane isina, etched, SHP uye LOIS substrates.(F) Iko kuonana kona (CA) uye SA kuyerwa kwezvinwiwa zvine kusawirirana kwepamusoro kwakasiyana paSHP neLOIS.Data inoratidzwa sezvinoreva ± SD.
Zvadaro, kuitira kusimbisa shanduko yemakemikari epamusoro, X-ray photoelectron spectroscopy (XPS) yakashandiswa kuongorora shanduko yekemikari yepasi pe substrate mushure mekuputira kwega kwega.Mufananidzo 2C inoratidza iyo XPS kuyerwa mhedzisiro yeHF yakamiswa pamusoro uye iyo HNO 3 yakarapwa pamusoro.Iwo maviri mahombe epamusoro ari pa587.3 uye 577.7 eV anogona kuverengerwa kune Cr-O bond iripo muchromium oxide layer, ndiwo mutsauko mukuru kubva kuHF etched surface.Izvi zvinonyanya kukonzerwa nekushandiswa kwesimbi uye chromium fluoride pamusoro neHNO3.Iyo HNO3-yakavakirwa etching inobvumira chromium kuumba passivating oxide layer pamusoro, izvo zvinoita kuti yakakamirwa SS irambe isingadzivirirwe nekuora.Mumufananidzo 2D, XPS spectra yakawanikwa kuti isimbise kuti fluorocarbon-based silane yakaumbwa pamusoro mushure meiyo SAM coating, iyo ine yakanyanya kukwirisa mvura yekudzinga kunyangwe EG, ropa uye EtOH.Iyo SAM coating inopedzwa nekuita silane inoshanda mapoka ane hydroxyl mapoka akaumbwa ne plasma kurapwa.Somugumisiro, kuwedzera kukuru kweCF2 neCF3 peaks kwakaonekwa.Simba rinosunga riri pakati pe286 ne296 eV rinoratidza kuti kugadziridzwa kwemakemikari kwakapedzwa zvinobudirira neSAM coating.SHP inoratidza yakakura CF2 (290.1 ​​​​eV) uye CF3 (293.3 eV) yepamusoro, iyo inokonzerwa nefluorocarbon-based silane yakaumbwa pamusoro.Mufananidzo 2E inoratidza anomiririra optical mifananidzo yekusangana kwekona (CA) kuyerwa kwemapoka akasiyana emvura yakasvibiswa inosangana neyakashama, etched, SHP, uye LOIS.Iyi mifananidzo inoratidza kuti iyo etched surface inova hydrophilic nekuda kweiyo micro / nano chimiro chakaumbwa nemakemikari etching kuitira kuti deionized mvura ipinde muchimiro.Zvisinei, kana iyo substrate yakaputirwa neSAM, iyo substrate inoratidza simba repellency yemvura, saka SHP yepamusoro inoumbwa uye nzvimbo yekubatana pakati pemvura nepamusoro idiki.Pakupedzisira, kuderera kweCA kwakaonekwa muLOIS, iyo inogona kuverengerwa nekupinda kwemafuta mukati meiyo microstructure, nekudaro ichiwedzera nzvimbo yekusangana.Kuti uratidze kuti nzvimbo yacho ine yakanakisa yekudzinga mvura uye isinganamire zvivakwa, LOIS yakaenzaniswa neSHP substrate nekuyera CA neSA uchishandisa zvinwiwa zvakasiyana (Mufananidzo 2F).Mhando dzakasiyana dzezvinwiwa dzakasarudzwa zvichibva pane viscosity uye pamusoro pesimba, kusanganisira deionized mvura, ropa, EG, EtOH uye HD (Mufananidzo S4).CA yekuyera mibairo inoratidza kuti kana CA ichienda kuHD, kukosha kwekudzikisa kweCA, uko CA ine simba rakaderera repamusoro.Mukuwedzera, iyo LOIS yeCA yose yakaderera.Nekudaro, kuyerwa kweSA kunoratidza chiitiko chakasiyana zvachose.Kunze kwemvura ine ionized, zvinwiwa zvese zvinonamatira kune SHP substrate pasina kutsvedza.Kune rumwe rutivi, LOIS inoratidza SA yakaderera zvikuru, apo mvura yose ikarereka pakona yakaderera kupfuura 10° kusvika 15°, mvura yose inokunguruka.Izvi zvinoratidza zvakasimba kuti kusanamatira kweLOIS kuri nani pane kweSHP pamusoro.Pamusoro pezvo, LOIS coatings inoiswawo kune akasiyana marudzi emidziyo, zvinosanganisira titanium (Ti), polyphenylsulfone (PPSU), polyoxymethylene (POM), polyether ether ketone (PEEK) uye bioabsorbable polymers (PLGA), Izvo zvinoiswa orthopedic zvinhu (Mufananidzo. S5)).Mifananidzo yakatevedzana yemadonhwe pazvinhu zvakabatwa neLOIS inoratidza kuti anti-biofouling zvimiro zveLOIS zvakafanana pane ese substrates.Mukuwedzera, mhedzisiro yekuyera yeCA neSA inoratidza kuti iyo isina-adhesive properties yeLOIS inogona kushandiswa kune zvimwe zvinhu.
Kuti usimbise maitiro e-anti-fouling eLOIS, marudzi akasiyana-siyana e-substrates (kusanganisira asina kupfeka, etched, SHP uye LOIS) akaiswa nePseudomonas aeruginosa uye MRSA.Aya mabhakitiriya maviri akasarudzwa seanomiririra mabhakitiriya echipatara, izvo zvinogona kutungamirira pakuumbwa kwebiofilms, zvichiita kuti SSI (37).Mufananidzo 3 (A uye B) unoratidza fluorescence microscope mifananidzo uye colony forming unit (CFU) mibairo yekuyera ye substrates incubated mubhakitiriya kuturika kwenguva pfupi (12 maawa) uye kwenguva refu (72 maawa), zvichiteerana.Munguva pfupi, mabhakitiriya anoumba masumbu uye anokura muhukuru, achizvifukidza nehutu-sezvinhu uye kudzivirira kubviswa kwavo.Zvisinei, panguva ye72-hour incubation, utachiona hunokura uye hunoita nyore kupararira kuti hugadzire mamwe makoroni kana mapoka.Nokudaro, zvinogona kufungidzirwa kuti 72-hour incubation ndeyenguva refu uye ndiyo nguva yakakodzera yekudzivirira kuumba biofilm yakasimba pamusoro pepamusoro (38).Munguva pfupi, nzvimbo yakavharidzirwa uye pamusoro peSHP yakaratidza kusungirirwa kwebhakitiriya, iyo yakaderedzwa ne25% kusvika ku50% kana ichienzaniswa neyakashama substrate.Zvisinei, nekuda kwekuita kwayo kwakanakisisa kwe-anti-biofouling uye kugadzikana, LOIS haina kuratidza bhakitiriya biofilm adhesion munguva pfupi uye kwenguva refu.The schematic diagram (Figure 3C) inotsanangura tsananguro ye-anti-biological fouling mechanism ye etching solution, SHP neLOIS.Iko fungidziro ndeyekuti iyo yakatemerwa substrate ine hydrophilic zvimiro ichava nenzvimbo yakakura kupfuura iyo isina chinhu substrate.Naizvozvo, kunamatira kwakawanda kwebhakitiriya kuchaitika pane yakamiswa substrate.Nekudaro, zvichienzaniswa neyakashama substrate, iyo yakamisirwa substrate ine yakanyanya kushoma biofilm yakaumbwa pamusoro.Izvi zvinodaro nekuti mamorekuru emvura anosunga zvakasimba kune hydrophilic pamusoro uye anoita semafuta emvura, nokudaro achikanganisa kunamatira kwebhakitiriya munguva pfupi (39).Nekudaro, iyo layer yemamorekuru emvura yakatetepa uye inonyungudika mukumiswa kwebhakitiriya.Nokudaro, mvura ye molecular layer inonyangarika kwenguva yakareba, zvichiita kuti iwedzere kunamatira kwebhakitiriya uye kupararira.Kune SHP, nekuda kweiyo pfupi-yenguva isiri-yekunyorova zvinhu, kunamatira kwebhakitiriya kunodzivirirwa.Iyo yakaderedzwa bhakitiriya adhesion inogona kuverengerwa homwe dzemhepo dzakavharirwa mune yakaturikidzana chimiro uye yakaderera pamusoro pesimba, nekudaro kuderedza kusangana pakati peiyo kumiswa kwebhakitiriya uye pamusoro.Nekudaro, kunamatira kwebhakitiriya kwakawanda kwakaonekwa muSHP nekuti yakarasikirwa neayo anti-fouling zvimiro kwenguva yakareba.Izvi zvinonyanya kukonzerwa nekunyangarika kwehomwe dzemhepo nekuda kwehydrostatic pressure uye kuparara kwemhepo mumvura.Izvi zvinonyanya kukonzerwa nekunyangarika kwehomwe dzemhepo nekuda kwekuparara uye yakaturikidzana chimiro chinopa yakakura nzvimbo yekunamatira (27, 40).Kusiyana neaya ma substrates maviri ane mhedzisiro yakakosha pakugadzikana kwenguva refu, iyo lubricant lubricant iri muLOIS inopinzwa muiyo micro / nano chimiro uye haizonyangarika kunyangwe munguva refu.Mafuta ekuzora akazadzwa nemicro/nano zvimiro akanyatsogadzikana uye anokwezvwa zvakasimba pamusoro pepamusoro nekuda kwehupamhi hwemakemikari affinity, nokudaro kudzivirira kunamatira kwebhakitiriya kwenguva yakareba.Mufananidzo S6 unoratidza mufananidzo wakakomberedzwa microscope mufananidzo wemafuta-akaiswa substrate yakanyudzwa muphosphate buffered saline (PBS).Mifananidzo inoenderera inoratidza kuti kunyange mushure memaawa 120 ekuzununguka zvishoma (120 rpm), iyo lubricant layer paLOIS inoramba isingachinji, ichiratidza kugadzikana kwenguva refu pasi pemamiriro ekuyerera.Izvi zvinokonzerwa nehupamhi hwemakemikari affinity pakati pe fluorine-based SAM coating uye perfluorocarbon-based lubricant, kuitira kuti chigadziko chakagadzikana chemafuta chigone kuumbwa.Naizvozvo, iyo anti-fouling performance inochengetedzwa.Mukuwedzera, iyo substrate yakaedzwa inopesana nemapuroteni anomiririra (albumin uye fibrinogen), ari muplasma, masero anonyatsoenderana nekushanda kwemuviri (macrophages uye fibroblasts), uye izvo zvine chokuita nekugadzirwa kwemapfupa.Izvo zviri mu calcium zvakanyanya.(Mufananidzo 3D, 1 uye 2, uye Mufananidzo S7) (41, 42).Mukuwedzera, iyo fluorescence microscope mifananidzo yeadhesion test ye fibrinogen, albumin uye calcium yakaratidza maitiro akasiyana ekunamatira eboka rimwe nerimwe re substrate (Mufananidzo S8).Panguva yekugadzirwa kwepfupa, mapfupa emapfupa uye calcium achangobva kuumbwa anogona kukomberedza kuisirwa kwemapfupa, izvo zvisingangoita kuti kubvisa kuoma, asi zvinogona kukonzera kukuvadza kusingatarisirwi kumurwere panguva yekubvisa.Naizvozvo, mazinga akaderera ecalcium anoisa pamapfupa mahwendefa uye screws anobatsira pakuvhiya kwemapfupa kunoda kubviswa kwekudyara.Zvichienderana nehuwandu hwenzvimbo yakasungirirwa zvichienderana nehukuru hwefluorescence uye kuverenga kwesero, takasimbisa kuti LOIS inoratidza yakanakisa anti-biofouling zvivakwa zvezvinhu zvese zvipenyu zvichienzaniswa nemamwe ma substrates.Zvinoenderana nemhedzisiro yezviyedzo zve in vitro, iyo anti-biological fouling LOIS inogona kuiswa kune orthopedic implants, iyo isingangodzivise hutachiona hunokonzerwa nebiofilm bacteria, asi zvakare kuderedza kuzvimba kunokonzerwa neanoshanda immune system.
(A) Fluorescence microscope mifananidzo yeboka rega rega (yakashama, yakamiswa, SHP neLOIS) yakavharirwa muPseudomonas aeruginosa uye MRSA kumiswa kwe12 uye 72 maawa.(B) Nhamba yeadherent CFU yePseudomonas aeruginosa neMRSA pamusoro peboka rimwe nerimwe.(C) Schematic diagram ye-anti-biological fouling mechanism yenguva pfupi uye yakareba-etching, SHP uye LOIS.(D) (1) Nhamba ye fibroblasts yakanamatira kune imwe neimwe substrate uye fluorescence microscope mifananidzo yemasero yakanamatira kune yakashama uye LOIS.(2) Adhesion test yemapuroteni ane chokuita nekudzivirira, albumin uye calcium inobatanidzwa muhutano hwemapfupa (* P <0.05, ** P <0.01, *** P <0.001 uye **** P <0.0001).ns, hazvina kukosha.
Panyaya yezvisingadzivisike zvakasimbiswa zvinonetsa, kusimba kwemagetsi kwagara kuri dambudziko guru rekushandiswa kweantifouling coatings.Traditional anti-sewage gel nzira dzakavakirwa pamapolima ane yakaderera mvura solubility uye fragility.Naizvozvo, ivo vanowanzo konzeresa kushushikana kwemagetsi mune biomedical application.Naizvozvo, mechanically durable antifouling coatings inoramba ichinetsa kune zvikumbiro senge orthopedic implants (43, 44).Mufananidzo 4A (1) unoratidza miviri mikuru yekushungurudzika inoshandiswa kune orthopedic implants, kusanganisira scratching (shear stress) uye kumanikidzika nemufananidzo wemaziso wezvakakanganiswa implant inogadzirwa nechisimba.Semuyenzaniso, kana sikuruu yakasimbiswa nescrewdriver, kana kuti chiremba anovhiya akabata bhonzo ndiro zvakasimba netwuma tweezers uye achishandisa simba rekumanikidza, ndiro yepurasitiki yebhonzo ichakuvadzwa nekukweshwa pazvikero zvemacro uye micro/nano (Mufananidzo 4A, 2) .Kuti uedze kana iyo inogadzirwa LOIS inogona kumira kukuvadza uku panguva yekuvhiyiwa kwepurasitiki, nanoindentation yakaitwa kuenzanisa kuoma kweiyo isina chinhu substrate uye LOIS pane micro / nano scale kuti idzidze mechanical properties ye micro / nano structure Impact (Mufananidzo. 4B).Iyo schematic dhizaini inoratidza akasiyana deformation maitiro eLOIS nekuda kwekuvapo kweiyo micro / nano zvimiro.Simba-kudzinga curve yakadhirowewa zvichienderana nemhedzisiro ye nanoindentation (Mufananidzo 4C).Mufananidzo webhuruu unomiririra iyo substrate isina chinhu, iyo inongoratidzira zvishoma deformation, sezvinoonekwa nehupamhi indentation kudzika kwe0.26-μm.Kune rumwe rutivi, kuwedzera zvishoma nezvishoma mu nanoindentation simba uye kutama kunoonekwa muLOIS (red curve) inogona kuratidza zviratidzo zvekuderedzwa kwema mechanical properties, zvichiita kuti nanoindentation yakadzika ye1.61μm.Izvi zvinodaro nekuti iyo micro / nano chimiro chiripo muLOIS chinopa yakadzama kufambira mberi nzvimbo yemuromo we nanoindenter, saka deformation yayo yakakura kupfuura iyo isina chinhu substrate.Konsta-Gdoutos et al.(45) anotenda kuti nekuda kwekuvapo kwezvimiro, nanoindentation uye micro/nano roughness zvinotungamira kune zvisina kujairika nanoindentation curves.Iyo nzvimbo ine shaded inoenderana neasina kujairika deformation curve inofungidzirwa kune iyo nanostructure, nepo iyo isina-shaded nzvimbo inonzi ine microstructure.Iyi deformation inogona kukuvadza microstructure / nanostructure yekubata lubricant uye inokanganisa kuita kwayo anti-fouling.Kuti udzidze kukanganisa kwekukuvara paLOIS, kukuvara kusingadzivisiki kune micro / nano zvimiro zvakadzokororwa mumuviri panguva yekuvhiyiwa kwepurasitiki.Nokushandisa ropa uye mapuroteni adhesion bvunzo, kugadzikana kweiyo anti-biofouling zvimiro zveLOIS mushure me in vitro inogona kutsanangurwa (Mufananidzo 4D).Mutsara wemifananidzo yemaziso inoratidza kukuvara kwakaitika pedyo nemakomba ega ega substrate.Muedzo wekunamatira ropa wakaitwa kuratidza mhedzisiro yekukuvara kwemagetsi pane anti-biofouling coating (Mufananidzo 4E).Kunge SHP, iyo anti-fouling zvivakwa zvakarasika nekuda kwekukuvara, uye LOIS inoratidza yakanakisa anti-fouling zvivakwa nekudzinga ropa.Izvi zvinodaro nekuti, nekuti simba repamusoro rinofambiswa necapillary chiito chinovhara nzvimbo yakakuvadzwa, kuyerera kweiyo microstructured lubricant lubricant inodzoreredza anti-fouling zvivakwa (35).Muitiro mumwe chete wakaonekwa muprotein adhesion test uchishandisa albumin.Munzvimbo yakakanganisika, kusungirirwa kweprotein pamusoro peSHP kunoonekwa zvakanyanya, uye nekuyera nzvimbo yayo yekuvhara, inogona kuverengwa sehafu yezinga rekunamatira re substrate isina chinhu.Kune rumwe rutivi, LOIS yakachengetedza maitiro ayo e-anti-biofouling pasina kukonzera kunamatira (Mufananidzo 4, F uye G).Pamusoro pezvo, pamusoro peiyo screw inowanzoiswa kune yakasimba mechanical stress, senge kuchera, saka takadzidza kugona kweLOIS coating kuti irambe yakasimba pa screw in vitro.Mufananidzo 4H unoratidza optical mifananidzo yeakasiyana screws, anosanganisira akashama, SHP uye LOIS.Rectangle dzvuku inomiririra nzvimbo inotarirwa iyo yakasimba mechanical stress inoitika panguva yekuiswa kwemapfupa.Kufanana neprotein adhesion test yeplate, fluorescence microscope inoshandiswa kufananidzira mapuroteni adhesion uye kuyera nzvimbo yekuvhara kuratidza kuvimbika kweLOIS coating, kunyange pasi pesimba rakasimba remagetsi (Mufananidzo 4, I uye J).Iyo LOIS-yakarapwa screws inoratidza yakanakisa anti-fouling kuita, uye inenge isina protein inonamatira pamusoro.Kune rimwe divi, kunamatira kweprotein kwaionekwa mumascrews asina chinhu uye SHP screws, uko kuvharika kwenzvimbo yeSHP screws yaive chikamu chimwe muzvitatu cheiyo yeare screws.Mukuwedzera, iyo orthopedic implant inoshandiswa pakugadzirisa inofanira kuva nemagetsi yakasimba kuti idzivirire kushushikana kunoshandiswa panzvimbo yekuputsika, sezvinoratidzwa mumufananidzo 4K.Naizvozvo, bvunzo yekukotama yakaitwa kuti ione mhedzisiro yekugadziriswa kwemakemikari pane zvemichina.Mukuwedzera, izvi zvinoitwa kuti uchengetedze kushushikana kwakagadziriswa kubva pakusimwa.Isa vertical mechanical force kusvika iyo implant yanyatsopetwa uye kushushikana-kurovera curve kunowanikwa (Mufananidzo 4L, 1).Zvivakwa zviviri zvinosanganisira Young's modulus uye flexural simba zvakaenzaniswa pakati pekushama uye LOIS substrates sezviratidzo zvesimba ravo remagetsi (Mufananidzo 4L, 2 uye 3).Young's modulus inoratidza kugona kwechinhu kutsungirira shanduko yemagetsi.Iyo Yechidiki's modulus yega yega substrate ndeye 41.48±1.01 uye 40.06±0.96 GPa, zvichiteerana;mutsauko wakaonekwa ungangoita 3.4%.Pamusoro pezvo, zvinoshumwa kuti simba rekukotama, iro rinotarisa kuoma kwechinhu, i102.34±1.51 GPa yeiyo isina chinhu substrate uye 96.99±0.86 GPa yeSHP.Iyo isina chinhu substrate inosvika 5.3% yakakwirira.Iko kuderera kudiki kwema mechanical properties kunogona kukonzerwa nenotch effect.Mune iyo notch mhedzisiro, iyo micro / nano roughness inogona kuita seti yeti notches, zvichitungamira kune yemuno kushushikana kunetseka uye kukanganisa ma mechanic zvimiro zvekuisa (46).Zvisinei, zvichibva pakuti kuoma kwe cortical bone yemunhu kunonzi kuri pakati pe7.4 ne31.6 GPa, uye yakayerwa LOIS modulus inodarika yemunhu cortical bone (47), LOIS inokwana kutsigira kuputsika uye iyo yose The mechanical properties inokanganiswa zvishoma nekugadziriswa kwepamusoro.
(A) Schematic diagram ye (1) the mechanical stress inoshandiswa kune orthopedic implant panguva yekushanda, uye (2) mufananidzo wemaziso weiyo yakakuvadzwa orthopedic implant.(B) Schematic diagram ye nano-mechanical properties kuyerwa ne nanoindentation uye LOIS panzvimbo isina chinhu.(C) Nanoindentation force-displacement curve yenzvimbo isina chinhu uye LOIS.(D) Mushure mekuedza in vitro, tevedzera mifananidzo yemaziso emhando dzakasiyana-siyana dzemapfupa emapfupa (nzvimbo yakakuvadzwa inoratidzirwa nerectangle tsvuku) kutevedzera kushushikana kwemagetsi kunokonzerwa panguva yekushanda.(E) Ropa adhesion test uye (F) protein adhesion test yeakakuvadzwa orthopedic plate group.(G) Edza kuvharwa kwenzvimbo yeprotein inonamatira kune ndiro.(H) Mifananidzo yeOptical yemhando dzakasiyana dze orthopedic screws mushure meiyo in vitro kuyedza.(I) Protein adhesion test yekudzidza kutendeseka kweakasiyana machira.(J) Edza nzvimbo yekuvhara kweprotein inonamatira kune screw.(K) Kufamba kwetsuro kunoitirwa kuunza kushushikana kwakasimba pabhonzo rakatyoka.(L) (1) Bend bvunzo mhinduro uye mifananidzo yemaziso pamberi uye mushure mekukotama.Musiyano mu (2) Mudiki's modulus uye (3) kukotama simba pakati pekuiswa isina chinhu uye SHP.Data inoratidzwa sezvinoreva ± SD (*P <0.05, **P <0.01, ***P <0.001 uye ****P <0.0001).Mufananidzo unofadza: Kyomin Chae, Yonsei University.
Mumakiriniki mamiriro, hutachiona hwakawanda hunosangana nezvinhu zvipenyu uye nzvimbo dzemaronda dzinobva kune yakakura, yakakura biofilms (48).Nokudaro, US Centers for Disease Control and Prevention inofungidzira kuti 65% yezvirwere zvose zvevanhu zvine chokuita nebiofilms (49).Muchiitiko ichi, zvinodikanwa kupa in vivo yekuyedza dhizaini inopa inowirirana biofilm kuumbwa pamusoro peiyo implant.Nokudaro, takagadzira tsuro femoral fracture model umo ma orthopedic implants akanga agara akaiswa mukusimudzirwa kwebhakitiriya uye akazosimwa mumakumbo etsuro kuti adzidze anti-fouling properties yeLOIS mu vivo.Nekuda kwezvinhu zvitatu zvinotevera zvakakosha, hutachiona hwehutachiona hunokonzerwa nekare-tsika panzvimbo pejekiseni rakananga rekumiswa kwebhakitiriya: (i) Chirwere chekudzivirira chetsuro chakasimba kupfuura chevanhu;saka, jekiseni yebhakitiriya suspensions uye planktonic bacteria inogoneka Izvo hazvina simba pakuumbwa kwebiofilms.(Ii) Planktonic mabhakitiriya anonyanya kubatwa nemishonga inorwisa mabhakitiriya, uye mishonga inorwisa mabhakitiriya inowanzoshandiswa mushure mekuvhiyiwa;pakupedzisira, (iii) iyo planktonic bacteria kuturika inogona kuderedzwa nemvura yemuviri wemhuka (50).Nekufanogadzirira kudyara mukumiswa kwebhakitiriya kusati kwaiswa, tinokwanisa kunyatsodzidza zvinokuvadza zvehutachiona hwehutachiona uye maitiro emuviri wekunze (FBR) pamapfupa ekuporesa maitiro.Tsuro dzakabayirwa mavhiki mana mushure mekusimwa, nekuti iyo osseointegration yakakosha pakupora kwemapfupa ichapedzwa mukati memavhiki mana.Zvadaro, ma implants akabviswa kubva kutsuro kuitira zvidzidzo zvezasi.Mufananidzo 5A unoratidza nzira yekuwedzera kwebhakitiriya.Iyo ine hutachiona hwemapfupa inoiswa mumuviri.Nekuda kwekutanga kuvharirwa mukusimwa kwebhakitiriya, tsuro nhanhatu panhanhatu dzakadyarwa dzakaiswa dzisina kupfeka dzakabatwa nehutachiona, asi hapana kana tsuro yakadyarwa neLOIS-yakagadziriswa implants yakabatwa.Utachiona hwehutachiona hunoenderera mberi mumatanho matatu, kusanganisira kukura, kukura uye kupararira (51).Kutanga, mabhakitiriya akabatanidzwa anoberekana uye anokura pamusoro, uyezve mabhakitiriya anoumba biofilm paanobudisa extracellular polymer (EPS), amyloid uye extracellular DNA.Biofilm haingopindiri chete nekupinda kwemishonga inorwisa mabhakitiriya, asi inokurudzirawo kuunganidza ma antibiotic-degrading enzymes (akadai se β-lactamase) (52).Pakupedzisira, iyo biofilm inoparadzira mabhakitiriya akakura mukati mematishu akatenderedza.Nokudaro, utachiona hunoitika.Mukuwedzera, kana mutumbi wekune imwe nyika ukapinda mumuviri, hutachiona hunogona kukonzera simba rekudzivirira zvirwere rinogona kukonzera kupisa kwakanyanya, kurwadziwa, uye kuderera kwekudzivirira.Mufananidzo 5B unopa mhedziso yeFBR inokonzerwa nekuiswa kweothopedic implant, panzvimbo yekuzvidzivirira kwezvirwere zvinokonzerwa nehutachiona hwehutachiona.Iwo immune system inoziva iyo yakaiswa semuviri wekune dzimwe nyika, uyezve inoita kuti masero nematishu aite kuti avhare muviri wekunze (53).Mumazuva ekutanga eFBR, matrix ekupa akaumbwa pamusoro peiyo orthopedic implants, izvo zvakaita kuti adsorption yefibrinogen.Iyo adsorbed fibrinogen inobva yaumba yakanyanyisa dense fibrin network, iyo inokurudzira kubatanidzwa kwe leukocytes (54).Kana iyo fibrin network ichinge yaumbwa, kuzvimba kwakanyanya kuchaitika nekuda kwekupinzwa kweneutrophils.Muchiito ichi, zvakasiyana-siyana zve cytokines zvakadai se tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) uye IL-β inosunungurwa, uye monocytes inotanga kupinda panzvimbo yekuisa uye kusiyanisa mumasero makuru.Phage (41, 55, 56).Kuderedza FBR kwagara kuri kunetsa nekuti yakawandisa FBR inogona kukonzera acute uye kusingaperi kuzvimba, izvo zvinogona kutungamirira kumatambudziko anouraya.Kuitira kuongorora kukanganisa kwehutachiona hwehutachiona mumisumbu yakatenderedza isina kuisirwa uye LOIS, hematoxylin uye eosin (H&E) uye Masson trichrome (MT) tsvina yakashandiswa.Kutsuro dzakasimwa dzine ma substrates akashama, hosha dzemabhakitiriya dzakanyanya kuenderera mberi, uye masiraidhi ematishu eH&E ainyatsoratidza maabscesses uye necrosis inokonzerwa nekuzvimba.Kune rumwe rutivi, iyo yakanyanya kusimba anti-biofouling pamusoro LOIS inhibits bhakitiriya kunamatira, saka hairatidzi zviratidzo zvehutachiona uye inoderedza kuzvimba (Mufananidzo 5C).Migumisiro yeMT staining yakaratidza maitiro akafanana.Zvisinei, MT staining yakaratidzawo edema mutsuro dzakasimwa neLOIS, zvichiratidza kuti kupora kwava kuda kuitika (Mufananidzo 5D).Kuti udzidze dhigirii rekudzivirira kwemuviri, immunohistochemical (IHC) tsvina yakaitwa uchishandisa cytokines TNF-α uye IL-6 yakabatana nemhinduro yekudzivirira.Chimiro chisina kupfeka chisina kupfeka chakanga chisina kuvharwa nebhakitiriya chakafananidzwa neLOIS yakaratidzwa kune mabhakitiriya asi isina utachiona kuti idzidze nzira yekuporesa mukushayikwa kwehutachiona hwehutachiona.Mufananidzo 5E unoratidza mufananidzo wakajeka weIHC siraidhi inoratidza TNF-α.Nzvimbo yebrown inomiririra mhinduro yezvirwere zvemuviri, zvichiratidza kuti immune system muLOIS yakaderedzwa zvishoma.Pamusoro pezvo, kutaura kweIL-6 muLOIS kwaive kwakaderera pane kutaura kwakashata kweasina kusimira asina kupfeka (Mufananidzo 5F).Kutaura kwe cytokine kwakaverengerwa nekuyera nzvimbo yeantibody staining inoenderana ne cytokine (Mufananidzo 5G).Zvichienzaniswa netsuro dzakafumurwa kune dzakasiiwa dzisina kunaka, mazinga ekutaura kwetsuro dzakasimwa neLOIS aive akaderera, achiratidza musiyano unoreva.Kuderera kwekutaura kwe cytokine kunoratidza kuti nguva yakareba, yakagadzikana yeAnti-fouling properties yeLOIS haina kungobatana chete nekudzivirira kwehutachiona hwehutachiona, asiwo nekuderera kweFBR, iyo inokonzerwa nemacrophages inonamatira kune substrate (53), 57 , 58).Naizvozvo, iyo yakaderedzwa immune system nekuda kweiyo immune evasion properties yeLOIS inogona kugadzirisa mhedzisiro mushure mekudyarwa, senge yakawandisa immune system mushure mekuvhiyiwa kwepurasitiki.
(A) Chimiro chechimiro chemaitiro ekugadzirwa kwebiofilm uye kupararira pamusoro pehutachiona hwehutachiona hwehutachiona.eDNA, extracellular DNA.(B) Schematic diagram ye immune response mushure mekuisa orthopedic implant.(C) H & E staining uye (D) MT kusvibiswa kwematishu akapoteredzwa emapfupa emapfupa akaiswa ane bare positive uye LOIS.IHC ye-immune-related cytokines (E) TNF-α uye (F) IL-6 mifananidzo yakasvibiswa yetsuro dzisina kupfeka-negative uye LOIS-dzakaiswa.(G) Quantification ye cytokine kutaura nenharaunda yekuvhara chiyero (** P <0.01).
Iyo biocompatibility yeLOIS nemhedzisiro yayo pamapfupa ekuporesa maitiro akaongororwa mu vivo uchishandisa diagnostic imaging [x-ray uye micro-computed tomography (CT)] uye osteoclast IHC.Mufananidzo 6A unoratidza maitiro ekuporesa kwepfupa anosanganisira matanho matatu akasiyana: kuzvimba, kugadzirisa, uye kugadzirisa.Kana kutyoka kuchiitika, masero ekuputika uye fibroblasts achapinda mukati mebhonzo rakatsemuka uye otanga kukura kuva vascular tissue.Munguva yekugadzirisa, ingrowth yevascular tissue inopararira pedyo nenzvimbo yekuputsika.Vascular tissue inopa chikafu chekugadzirwa kwepfupa idzva, iyo inonzi callus.Chikamu chekupedzisira chepfupa rekuporesa ndiyo danho rekugadzirisa, umo ukuru hwe callus hunoderedzwa kusvika kuhukuru hwemafupa akajairika nekubatsirwa kwekuwedzera kwehuwandu hweosteoclasts inoshandiswa (59).Matatu-dimensional (3D) kuvakwazve kwenzvimbo yakaputsika yakaitwa uchishandisa micro-CT scans kuti ione kusiyana kwehuwandu hwekuita callus muboka rimwe nerimwe.Chengetedza kuyambuka-chikamu chefemu kuti uone ukobvu hwe callus yakatenderedza bhonzo rakatyoka (Mufananidzo 6, B uye C).X-rays yakashandiswawo kuongorora nzvimbo dzakaputsika dzemapoka ose vhiki imwe neimwe kuti vaone maitiro akasiyana-siyana ekugadzirisa mapfupa muboka rimwe nerimwe (Mufananidzo S9).Callus uye mapfupa akakura anoratidzwa mubhuruu / girini nenyanga dzenzou, zvichiteerana.Mazhinji matishu akapfava anoseferwa kunze ane preset chikumbaridzo.Nude positive uye SHP yakasimbisa kuumbwa kwechidimbu chiduku che callus yakakomberedza nzvimbo yekuputsika.Kune rimwe divi, iyo yakafumurwa isina kunaka yeLOIS uye nzvimbo yekutsemuka yakakomberedzwa nekobvu callus.Micro-CT mifananidzo yakaratidza kuti kuumbwa kwe callus kwakadziviswa nehutachiona hwehutachiona uye hutachiwana hunokonzerwa nehutachiona.Izvi zvinodaro nekuti immune system inotungamira kuporeswa kwekukuvara kweseptic kunokonzerwa nekuzvimba kwakabatana nehutachiona, pane kupora kwemapfupa (60).IHC uye Tartrate-resistant Acid Phosphatase (TRAP) tsvina yakaitwa kuti ione osteoclast basa uye bone resorption (Mufananidzo 6D) (61).Mashoma chete akaiswa osteoclasts ane ruvara rwepepuru akawanikwa mune asina kupfeka zvakanaka uye SHP.Kune rumwe rutivi, akawanda akaiswa osteoclasts akaonekwa padyo neakashama akanaka uye akakura mapfupa eLOIS.Ichi chiitiko chinoratidza kuti pamberi pe osteoclasts, iyo callus yakapoteredza nzvimbo yekuputsika iri kutarisana nechisimba chekugadzirisa maitiro (62).Iyo bhonzo vhoriyamu uye osteoclast kutaura nzvimbo ye callus yakayerwa kuenzanisa nhanho yekuumbwa kwecallus yakatenderedza nzvimbo yekutyoka mumapoka ese, kuitira kuverengera iyo micro-CT scan uye IHC mhinduro (Mufananidzo 6E, 1 uye 2).Sezvinotarisirwa, zvisizvo zvisina kupfeka uye kuumbwa kwe callus muLOIS zvaive zvakanyanya kudarika mune mamwe mapoka, zvichiratidza kuti kugadziriswa kwepfupa kwakanaka kwakaitika (63).Mufananidzo S10 unoratidza mufananidzo wenzvimbo yekuvhiya, iyo MT inosvibisa mhedzisiro yetishu yakaunganidzwa padyo nescuru, uye TRAP inosvibisa mhedzisiro inoratidzira screw-bone interface.Mune substrate isina chinhu, callus yakasimba uye fibrosis formation yakaonekwa, nepo LOIS-yakagadziriswa implant yakaratidza nzvimbo isina kubatwa.Saizvozvowo, zvichienzaniswa nezvakaipa zvisina kupfeka, fibrosis yakaderera yakaonekwa mumatsuro akaiswa neLOIS, sezvinoratidzwa nemiseve yakachena.Uye zvakare, iyo yakasimba edema (yebhuruu museve) inogona kuverengerwa kune immune evasion zvivakwa zveLOIS, nekudaro kuderedza kuzvimba kwakanyanya.Iyo isiri-tsvimbo yakakomberedza kuisirwa uye yakaderedzwa fibrosis inoratidza kuti nzira yekubvisa iri nyore, izvo zvinowanzokonzera kumwe kutsemuka kana kuzvimba.Iyo bhonzo yekuporesa maitiro mushure mekubviswa screw yakaongororwa ne osteoclast chiitiko pane screw-bone interface.Ose ari maviri bhonzo rakashama uye iyo LOIS implant interface yakabata mazinga akafanana eosteoclasts kuti aenderere mberi nekuporesa kwemapfupa, zvichiratidza kuti iyo LOIS coating haina mhedzisiro yakaipa pakupora kwebhonzo kana immune system.Kuti asimbise kuti kugadziriswa kwepamusoro kunoitwa paLOIS hakuvhiringidzi nekugadzirisa kwepfupa, kuongororwa kwe-X-ray kwakashandiswa kuenzanisa kuporesa kwepfupa retsuro neoni dzakafumurwa dzisina kunaka uye mavhiki e6 ekuiswa kweLOIS (Mufananidzo 6F).Zvigumisiro zvakaratidza kuti zvichienzaniswa neboka risina utachiona risina hutachiona, LOIS yakaratidza chiyero chakafanana chekuporesa kwepfupa, uye pakanga pasina zviratidzo zvakajeka zvekuputsika (inopfuurira osteolysis line) mumapoka ose maviri.
(A) Schematic diagiramu yemapfupa ekuporesa maitiro mushure mekuputsika.(B) Musiyano muchiyero chekuumba callus yeboka rimwe nerimwe repamusoro uye (C) mufananidzo wepamuchinjikwa wenzvimbo yekuputsika.(D) TRAP tsvina yekuona osteoclast chiitiko uye bone resorption.Kubva pane basa reTRAP, kuumbwa kwekunze callus ye cortical bone yakaongororwa ne (E) (1) micro-CT uye (2) osteoclast basa.(F) Vhiki 6 mushure mekusimwa, mifananidzo ye-X-ray yepfupa rakaputsika rekuonekwa kusina kunaka (kusimbiswa nerectangle yakatsvuka yakatsvuka) uye LOIS (yakasimbiswa neblue dashed rectangle).Statistical analysis yakaitwa neone-way analysis of variance (ANOVA).* P <0.05.** P <0.01.
Muchidimbu, LOIS inopa mhando nyowani yeantibacterial utachiona zano uye immune yekupunyuka coating ye orthopedic implants.Zvakajairwa orthopedic implants neSHP functionalization inoratidza zvenguva pfupi-anti-biofouling zvivakwa, asi hazvigone kuchengetedza zvivakwa zvavo kwenguva yakareba.Iyo superhydrophobicity ye substrate inoteya mabhubhu emhepo pakati pebhakitiriya neiyo substrate, nekudaro kugadzira homwe dzemhepo, nekudaro kudzivirira kutapukira kwebhakitiriya.Zvisinei, nekuda kwekupararira kwemhepo, idzi homwe dzemhepo dzinobviswa nyore nyore.Kune rimwe divi, LOIS yakanyatso ratidza kugona kwayo kudzivirira hutachiona hunoenderana nebiofilm.Naizvozvo, nekuda kweanopesana-kurambwa maitiro eiyo lubricant layer yakaiswa mukati meiyo layered micro / nano chimiro chepamusoro, hutachiona hune hutachiona hunogona kudzivirirwa.Yakasiyana-siyana maitiro emhando anosanganisira SEM, AFM, XPS uye CA zviyero zvinoshandiswa kukwenenzvera LOIS kugadzira mamiriro.Uye zvakare, LOIS inogona zvakare kuiswa kune akasiyana biological zvinhu zvinowanzoshandiswa muothopedic kugadzirisa michina, sePLGA, Ti, PE, POM uye PPSU.Zvadaro, LOIS yakaedzwa mu vitro kuratidza yayo anti-biofouling zvimiro kurwisa mabhakitiriya uye zvinhu zvipenyu zvine chekuita nekudzivirira kwemuviri.Mhedzisiro yacho inoratidza kuti ine yakanakisa antibacterial uye anti-biofouling mhedzisiro kana ichienzaniswa neyakashama implant.Mukuwedzera, LOIS inoratidza simba remagetsi kunyange mushure mekushandisa mechanical stress, iyo isingadzivisiki mukuvhiya kwepurasitiki.Nekuda kwekuzvipodza kwemafuta ekuzora pamusoro peiyo micro / nano chimiro, LOIS yakabudirira kuchengetedza yayo anti-biological fouling properties.Kuti udzidze biocompatibility uye antibacterial properties yeLOIS in vivo, LOIS yakasimwa mukati metsuro femur kwemavhiki mana.Hapana hutachiona hwehutachiona hwakaonekwa mutsuro dzakasimwa neLOIS.Mukuwedzera, kushandiswa kweIHC kwakaratidza kuderedzwa kwehutano hwemunharaunda, zvichiratidza kuti LOIS haidziviriri kuporesa kwepfupa.LOIS inoratidza yakanakisa antibacterial uye immune evasion properties, uye yakaratidza kudzivirira zvinobudirira kuumbwa kwebiofilm isati yasvika uye panguva yekuvhiyiwa kwemapfupa, kunyanya kugadzirwa kwemapfupa.Nekushandisa tsuro bone marrow inflammatory femoral fracture model, mhedzisiro yehutachiona hunoenderana nebiofilm pamapfupa ekuporesa maitiro anokonzerwa ne pre-incubated implants akadzidzwa zvakanyanya.Sechidzidzo chemunguva yemberi, mutsva mu vivo modhi inodiwa kudzidza hutachiona hunogona kuitika mushure mekudyarwa kuti unzwisise zvizere uye kudzivirira hutachiona hunoenderana nebiofilm panguva yese yekupora.Mukuwedzera, osteoinduction ichiri dambudziko risina kugadziriswa mukubatanidzwa neLOIS.Kumwe kutsvagurudza kunodiwa kusanganisa kunamatira kwakasarudzika kwemaosteoinductive masero kana mushonga wekuvandudza neLOIS kukunda dambudziko.Pakazere, LOIS inomiririra inovimbisa orthopedic implant coating ine mechanic kusimba uye yakanakisa anti-biofouling zvivakwa, izvo zvinogona kuderedza SSI uye immune side mhedzisiro.
Geza iyo 15mm x 15mm x 1mm 304 SS substrate (Dong Kang M-Tech Co., Korea) muacetone, EtOH uye DI mvura kwemaminetsi gumi nemashanu kubvisa zvinosvibisa.Kuti ugadzire chimiro chemicro/nano-level pamusoro, iyo substrate yakacheneswa inonyudzwa mu48% kusvika 51% HF solution (DUKSAN Corp., South Korea) pa50°C.Nguva yekucheka inosiyana kubva ku0 kusvika ku60 maminetsi.Zvadaro, iyo etched substrate yakacheneswa nemvura yakasvibiswa uye yakaiswa mu 65% HNO3 (Korea DUKSAN Corp.) mhinduro pa 50 ° C kwemaminitsi makumi matatu kuti iite chromium oxide passivation layer pamusoro.Mushure mokunge passivation, substrate inoshambidzwa nemvura yakasvibiswa uye yakaomeswa kuti iwane substrate ine dhizaini yakagadzirwa.Zvadaro, substrate yakaratidzwa kune oxygen plasma (100 W, 3 maminetsi), uye pakarepo yakanyudzwa mumushonga we8.88 mM POTS (Sigma-Aldrich, Germany) mutoluene pakamuri tembiricha kwemaawa gumi nemaviri.Zvadaro, substrate yakavharwa nePOTS yakacheneswa neEtOH, uye yakanyungudutswa pa150 ° C kwemaawa maviri kuti iwane dense POTS SAM.Mushure mekuvhara kweSAM, chigadziko chemafuta chakaumbwa pa substrate nekushandisa perfluoropolyether lubricant (Krytox 101; DuPont, USA) ine vhoriyamu yekurodha ye20 μm / cm 2. Usati washandisa, sefa iyo lubricant kuburikidza ne 0.2 micron filter.Bvisa mafuta akawandisa nekurerekera pakona ye45° kwemaminitsi gumi nemashanu.Iyo imwecheteyo nzira yekugadzira yakashandiswa kune orthopedic implants yakagadzirwa ne304 SS (yekukiya ndiro uye cortical locking screw; Dong Kang M-Tech Co., Korea).Ese ma implants e orthopedic akagadzirirwa kuenderana ne geometry ye tsuro femur.
Iyo yepamusoro morphology ye substrate uye orthopedic implants yakaongororwa nemunda emission SEM (Ongorora F50, FEI, USA) uye AFM (XE-100, Park Systems, South Korea).Hushasha hwepamusoro (Ra, Rq) hunoyerwa nekuwanza nzvimbo ye20 μm ne20 μm (n=4).An XPS (PHI 5000 VersaProbe, ULVAC PHI, Japan) system yakashongedzerwa neAl Kα X-ray sosi ine saizi ye100μm2 yakashandiswa kuongorora chimiro chemakemikari epamusoro.Iyo CA yekuyera sisitimu yakashongedzerwa ine simba rekutora mufananidzo kamera (SmartDrop, FEMTOBIOMED, ​​​​South Korea) yakashandiswa kuyera mvura CA neSA.Pachiyero chega chega, 6 kusvika 10 μl yemadonhwe (mvura yakasvibiswa, ropa rebhiza, EG, 30% ethanol, uye HD) inoiswa pamusoro kuyera CA.Kana iyo inclination angle ye substrate inowedzera pakukurumidza kwe 2 ° / s (n = 4), SA inoyerwa kana donhwe rinodonha.
Pseudomonas aeruginosa [American Type Culture Collection (ATCC) 27853] uye MRSA (ATCC 25923) yakatengwa kubva kuATCC (Manassas, Virginia, USA), uye tsika yemasheya yakachengetwa pa -80 ° C.Isati yashandiswa, tsika yechando yaiiswa mumuto wesoya wetrypsin-thawed (Komed, Korea) pa37 ° C kwemaawa gumi nemasere uye wozotamiswa kaviri kuti uiite.Mushure mokuputira, tsika yacho yakasimbiswa pa10,000 rpm kwemaminitsi gumi pa4 ° C uye yakashambidzwa kaviri nePBS (pH 7.3) mhinduro.Iyo centrifuged tsika inobva yaiswa paropa agar plates (BAP).MRSA nePseudomonas aeruginosa dzakagadzirirwa husiku uye dzakagadzirwa muLuria-Bertani muto.Iko kusanganiswa kwePseudomonas aeruginosa uye MRSA muinoculum kwaive kwakatemwa neCFU yekumisikidzwa mu serial dilutions pane agar.Zvadaro, gadzirisa kusungirirwa kwebhakitiriya kune 0.5 McFarland standard, iyo yakaenzana ne108 CFU/ml.Wobva waderedza kushanda kwebhakitiriya kumiswa ka100 kusvika ku106 CFU/ml.Kuti uedze antibacterial adhesion properties, substrate yakasvibiswa pa 121 ° C kwemaminitsi gumi nemashanu isati yashandiswa.Iyo substrate yakabva yaendeswa kune 25 ml yebhakitiriya kuturika uye yakavharwa pa 37 ° C nekuzunungusa kwakasimba (200 rpm) kwe12 uye 72 maawa.Mushure mekuita, substrate imwe neimwe yakabviswa kubva muincubator uye yakashambidzwa katatu nePBS kubvisa chero mabhakitiriya anoyangarara pamusoro.Kuti uone iyo biofilm pa substrate, iyo biofilm yakagadziriswa ne methanol uye yakasvibiswa ne 1 ml yecrimidine orenji kwemaminitsi maviri.Zvadaro microscope yefluorescence (BX51TR, Olympus, Japan) yakashandiswa kutora mifananidzo yezvakasvibiswa biofilm.Kuti kuenzanisa iyo biofilm pa substrate, masero akabatanidzwa akaparadzaniswa kubva kune substrate nebead vortex nzira, iyo yaionekwa seyo nzira yakanakisisa yekubvisa mabhakitiriya akabatanidzwa (n = 4).Uchishandisa sterile forceps, bvisa substrate kubva pakukura wepakati uye tapa ndiro yetsime kuti ubvise mvura yakawandisa.Masero akasungirirwa akasununguka akabviswa nekugeza kaviri nePBS isina utachiona.Imwe neimwe substrate yakabva yaendeswa kuchubhu yekuyedza isina utachiona ine 9 ml ye0.1% protein ept saline (PSW) uye 2 g ye20 kusvika 25 isina girazi mabeads (0.4 kusvika 0.5 mm mudhayamita).Yakazovharwa kwemaminetsi matatu kuti ibvise maseru kubva kumuenzaniso.Mushure mekuputika, kumiswa kwacho kwakaderedzwa 10-yakapetwa ne 0.1% PSW, uyezve 0.1 ml yega yega dilution yakaiswa paBAP.Mushure memaawa makumi maviri nemana ekuvharirwa pa37 ° C, CFU yakaverengwa nemaoko.
Kune maseru, mbeva fibroblasts NIH/3T3 (CRL-1658; American ATCC) uye mbeva macrophages RAW 264.7 (TIB-71; American ATCC) yakashandiswa.Shandisa Dulbecco's modified Eagle medium (DMEM; LM001-05, Welgene, Korea) kune tsika mbeva fibroblasts uye kuwedzera ne 10% mhuru serum (S103-01, Welgene) uye 1% penicillin-streptomycin (PS ; LS202-02, Welgene (Welgene) Shandisa DMEM kutsika mbeva macrophages, yakawedzerwa ne10% fetal bovine serum (S001-01, Welgene) uye 1% PS Isa iyo substrate mumatanhatu-tsime tsika ndiro , Uye inoculate masero pa 105 masero / cm2. Masero akaiswa usiku hwose pa 37 ° C uye 5% CO2 Kusvibisa masero, masero akaiswa ne 4% paraformaldehyde kwemaminitsi makumi maviri uye akaiswa mu 0.5% Triton X Incubate kwemaminitsi mashanu mu -100 pa 37 ° C kwemaminitsi makumi matatu mushure mekugadzirisa, shandisa substrate ne 4',6-diamino-2-phenylindole (H -1200, Vector Laboratories, UK) VECTASHIELD fixation medium (n = 4 per cell). , fluorescein, fluorescein isothiocyanate-albumin (A9771, Sigma-Aldrich, Germany) uye plasma yevanhu The Alexa Fluor 488-conjugated fibrinogen (F13191, Invitrogen, USA) yakanyungudutswa muPBS (10 mM, pH 7.4).Izvo zvakasanganiswa zvealbumin uye fibrinogen zvaive 1 uye 150 μg / ml, zvichiteerana.Mushure me substrate Usati wanyura muprotein solution, sukurudza nePBS kuti udzorere mvura pamusoro.Zvadaro nyudza ma substrates ose mundiro yemvura mitanhatu ine mushonga weprotein uye incuite pa 37 ° C kwemaminitsi makumi matatu ne90.Mushure mekuvhara, Iyo substrate yakabva yabviswa kubva muprotein solution, yakashambidzwa zvinyoro nePBS 3 nguva, uye yakagadziriswa ne4% paraformaldehyde (n = 4 yeprotein imwe neimwe).Kune calcium, sodium chloride (0.21 M) uye potassium phosphate (3.77 mM) ) Yakanyungudutswa mumvura yakasvibiswa.Iyo pH yemhinduro yakagadziridzwa ku2.0 nekuwedzera hydrochloride mhinduro (1M).Ipapo calcium chloride (5.62 mM) yakanyungudika mune mhinduro.Nekuwedzera 1M tris(hydroxymethyl) -amino Methane inogadzirisa pH yemhinduro ku7.4.Nyura ma substrates ese mundiro ine matsime matanhatu akazadzwa ne 1.5× calcium phosphate solution uye bvisa kubva mumhinduro mushure memaminitsi makumi matatu.Nekusvibisa, 2 g Alizarin Red S (CI 58005) Sanganisa ne 100 ml yemvura yakasvibiswa.Zvadaro, shandisa 10% ammonium hydroxide kugadzirisa pH kusvika 4. Dhayisa substrate neAlizarin Red solution kwemaminitsi mashanu, uye wozozunungusa dhayi yakawandisa uye blot.Mushure mokuzunungusa, bvisa substrate.Nyaya yacho inopera mvura, yozonyudzwa muacetone kwemaminitsi mashanu, yozonyudzwa mune acetone-xylene (1: 1) mhinduro kwemaminitsi mashanu, uye pakupedzisira yakashambidzwa nexylene (n = 4).Fluorescence microscope (Axio Imager) ine × 10 uye × 20 chinangwa lenzi inoshandiswa..A2m, Zeiss, Germany) mifananidzo yese substrates.ImageJ/FIJI (https://imagej.nih.gov/ij/) yakashandiswa kuyera data rekunamatira rezvinhu zvipenyu paboka rega rega renzvimbo ina dzakasiyana dzekufungidzira.Shandura mifananidzo yese kuti ive yebhinari mifananidzo ine yakamisikidzwa zvikumbaridzo zve substrate kuenzanisa.
A Zeiss LSM 700 confocal maikorosikopu yakashandiswa kutarisa kugadzikana kweiyo lubricant layer muPBS mu reflection mode.Iyo fluorine-yakavakirwa SAM-yakavharwa girazi sampu ine jekiseni rekuzora rakanyudzwa mumhinduro yePBS, uye yakaedzwa ichishandisa orbital shaker (SHO-1D; Daihan Scientific, South Korea) pasi pemamiriro ekudedera akapfava (120 rpm).Wobva watora sampuli uye tarisa kurasikirwa kwemafuta nekuyera kurasikirwa kwechiedza chakaratidzwa.Kuti uwane mifananidzo yefluorescence mu reflection mode, sampuli inoratidzirwa kune 633 nm laser uye inounganidzwa, nokuti chiedza chicharatidzwa kumashure kubva kumuenzaniso.Iwo masampuli akayerwa panguva dzenguva dze0, 30, 60, uye 120 maawa.
Kuti uone kushanda kwemaitiro ekugadzirisa kwepamusoro pane nanomechanical properties ye orthopedic implants, nanoindenter (TI 950 TriboIndenter, Hysitron, USA) yakashongedzerwa nemativi matatu emapiramidhi akaita seBerkovich diamond tip yakashandiswa kuyera nanoindenedione.Mutoro wepamusoro ndeye 10 mN uye nzvimbo iri 100μmx 100μm.Pazviyero zvose, nguva yekutakura uye yekudzikisa ndeye 10 s, uye nguva yekubata pasi peak indentation load ndeye 2 s.Tora zviyero kubva kunzvimbo shanu dzakasiyana uye utore avhareji.Kuti uongorore mashandiro esimba remagetsi pasi pemutoro, bvunzo yekupeta-yetatu-yekupeta yakaitwa pachishandiswa muchina wekuyedza wepasirese (Instron 5966, Instron, USA).Iyo substrate inomanikidzwa pamwero wenguva dzose we10 N / s nekuwedzera mutoro.Iyo Bluehill Universal software chirongwa (n = 3) yakashandiswa kuverenga iyo inoshanduka modulus uye yakanyanya kudzvinyirira kushushikana.
Kuti uteedzere maitiro ekuvhiya uye ane hukama kukuvara kwemuchina wakakonzerwa panguva yekuvhiya, maitiro ekuvhiya akaitwa in vitro.Mafemu akaunganidzwa kubva kune akaurayiwa New Zealand tsuro chena.Iyo femur yakacheneswa uye yakagadziriswa mu4% paraformaldehyde kwevhiki 1.Sezvakatsanangurwa mumhuka yekuedza nzira, iyo yakagadziriswa femur yakavhiyiwa.Mushure mekuvhiyiwa, iyo orthopedic implant yakanyudzwa muropa (bhiza ropa, KISAN, Korea) kwe10 s kusimbisa kana kunamatira kweropa kwakaitika mushure mekukuvadzwa kwemagetsi (n = 3).
Tsuro chena makumi maviri neina dzeNew Zealand (huremu 3.0 kusvika 3.5kg, avhareji yemwedzi mitanhatu) dzakakamurwa zvisina tsarukano kuita mapoka mana: nude negative, nude positive, SHP neLOIS.Maitiro ese anosanganisira mhuka akaitwa zvinoenderana nehunhu hweInstitutional Animal Care uye Use Committee (IACUC yakabvumidzwa, KOREA-2017-0159).The orthopedic implant ine chivharo ndiro ine maburi mashanu (kureba 41 mm, hupamhi 7 mm uye ukobvu 2 mm) uye cortical locking screws (kureba 12 mm, dhayamita 2.7 mm) kugadzirisa pakatyoka.Kunze kweaya mahwendefa uye screws anoshandiswa mubare-negative boka, mahwendefa ese uye screws zvakaiswa muMRSA suspension (106 CFU/ml) kwemaawa gumi nemaviri.Boka rakashama-negative (n = 6) rakabatwa nemafuta asina kupfeka pamusoro pepamusoro pasina kutarisana nekusimudzirwa kwebhakitiriya, sekutonga kusina kunaka kwehutachiona.Boka rakashama rakanaka (n = 6) rakabatwa nechinhu chisina kuchena chepamusoro chakaiswa kune mabhakitiriya sekutonga kwakanaka kwehutachiona.Boka reSHP (n = 6) rakarapwa nehutachiona hwakafumurwa SHP implants.Pakupedzisira, boka reLOIS rakabatwa nebhakitiriya-yakafumurwa LOIS implants (n = 6).Mhuka dzose dzinochengetwa mukeji, uye zvokudya zvakawanda nemvura zvinopiwa.Tsuro dzisati dzavhiyiwa dzaitsanya kwemaawa gumi nemaviri.Mhuka dzacho dzakange dzave anesthetized ne intramuscular injection ye xylazine (5mg / kg) uye intravenous injection ye paclitaxel (3mg / kg) yekupinza.Mushure maizvozvi, shandisa 2% isoflurane uye 50% kusvika 70% yeokisijeni yezvokurapa (kuyerera kwe2 L / min) kuburikidza nehurongwa hwekufema kuchengetedza anesthesia.Inosimwa kuburikidza nenzira yakananga kune lateral femur.Mushure mekubviswa kwebvudzi uye povidone-iodine disinfection yeganda, kuchekwa kwe 6 cm kureba kwakaitwa kunze kwekuruboshwe pakati femur.Nekuvhura mukaha uripo pakati pemamhasuru anovhara femu, chikadzi chinobuda pachena.Isa ndiro pamberi pechikadzi shaft uye uigadzirise nemakumbo mana.Mushure mekugadzirisa, shandisa saha blade (1 mm gobvu) kugadzira zvakaputsika munzvimbo iri pakati pegomba rechipiri negomba rechina.Pakupera kwekuvhiyiwa, ronda raigezwa nesaline uye rakavharwa ne sutures.Tsuro imwe neimwe yaibayiwa pasi peganda neenrofloxacin (5 mg/kg) yakanyungudutswa chikamu chimwe muzvitatu mumunyu.Postoperative X-rays yefemur yakatorwa mumhuka dzose (0, 7, 14, 21, 28, uye 42 mazuva) kusimbisa osteotomy yefupa.Mushure mekudzika kweanesthesia, mhuka dzese dzakaurayiwa ne intravenous KCl (2 mmol/kg) pamazuva makumi maviri nemasere nemakumi mana nemaviri.Mushure mekuurayiwa, femur yakaongororwa ne micro-CT kuti ione uye ienzanise maitiro ekuporesa pfupa uye kuumbwa kwemapfupa matsva pakati pemapoka mana.
Mushure mekuurayiwa, zvinyoro zvinyoro zvaive zvakanangana nemapfupa emapfupa akaunganidzwa.Iyo matishu yakagadziriswa mu10% isina kwayakarerekera buffered formalin usiku humwe uye yakazopera mvura muEtOH.Mishonga yakasvibiswa yakaiswa muparafini uye yakakamurwa pahupamhi hwe 40 μm uchishandisa microtome (400CS; EXAKT, Germany).Kuti uone hutachiona, H & E staining uye MT staining zvakaitwa.Kuti utarise mhinduro yeanotambira, matishu akaiswa muchikamu akaiswa netsuro anti-TNF-α primary antibody (AB6671, Abcam, USA) uye tsuro anti-IL-6 (AB6672; Abcam, USA), uye ndokubatwa nehorseradish.Oxidase.Isa iyo avidin-biotin complex (ABC) staining system kune zvikamu maererano nemirairo yemugadziri.Kuti iite senge brown reaction chigadzirwa, 3,3-diaminobenzidine yakashandiswa muzvikamu zvese.Digital slide scanner (Pannoramic 250 Flash III, 3DHISTECH, Hungary) yakashandiswa kuona zvimedu zvese, uye ma substrates mana muboka rega rega akaongororwa nesoftware yeImageJ.
Mifananidzo ye-X-ray yakatorwa mumhuka dzose mushure mekuvhiyiwa uye vhiki imwe neimwe kutarisa kuporesa kuputsika (n = 6 neboka).Mushure mekuurayiwa, high-resolution micro-CT yakashandiswa kuverenga kuumbwa kwe callus yakakomberedza femur mushure mekuporesa.Iyo femur yakawanikwa yakacheneswa, yakagadziriswa mu4% paraformaldehyde kwemazuva matatu, uye yakasvibiswa mu75% ethanol.Mabhonzo akapera mvura akabva aongororwa nekushandisa micro-CT (SkyScan 1173, Brooke Micro-CT, Kandy, Belgium) kugadzira 3D voxel mifananidzo (2240 ​​× 2240 pixels) yebhonzo sample.Shandisa 1.0 mm Al sefa kuderedza ruzha rwechiratidzo uye shandisa kugadzirisa kwepamusoro kune ese scans (E = 133 kVp, I = 60 μA, nguva yekubatanidza = 500 ms).Nrecon software (version 1.6.9.8, Bruker microCT, Kontich, Belgium) yakashandiswa kugadzira vhoriyamu ye3D yemuenzaniso wakaongororwa kubva kune yakawanikwa 2D lateral projection.Kuti uongororwe, iyo 3D yakagadziridzwa mufananidzo yakakamurwa kuita 10mm × 10mm × 10mm cubes zvichienderana nenzvimbo yekutyoka.Verenga callus kunze kwe cortical bone.DataViewer (version 1.5.1.2; Bruker microCT, Kontich, Belgium) software yakashandiswa kudhijita redirect the scanned bone volume, uye CT-Analyzer (version 1.14.4.1; Bruker microCT, Kontich, Belgium) software yakashandiswa pakuongorora.Iyo yehukama x-ray absorption coefficients mune yakakura pfupa uye callus inosiyaniswa nekuwanda kwavo, uye ipapo vhoriyamu ye callus inoverengerwa (n = 4).Kuti usimbise kuti biocompatibility yeLOIS haina kunonoka kuporesa kwepfupa, kuwedzera X-ray uye micro-CT kuongororwa kwakaitwa mumatsuro maviri: asina kupfeka-negative uye LOIS mapoka.Mapoka maviri aya akaurayiwa muvhiki yechitanhatu.
Mafemu kubva kumhuka dzakabayirwa akaunganidzwa uye akaiswa mu4% paraformaldehyde kwemazuva matatu.The orthopedic implant inobva yanyatsobviswa kubva kufemu.Iyo femur yakagadziriswa kwemazuva makumi maviri nerimwe nekushandisa 0.5 M EDTA (EC-900, National Diagnostics Corporation).Ipapo iyo decalcified femur yakanyudzwa muEtOH kuita kuti ishaye mvura.Iyo femur isina mvura yakabviswa mu xylene uye yakaiswa muparafini.Ipapo sampuro yakachekwa neotomatiki rotary microtome (Leica RM2255, Leica Biosystems, Germany) ine ukobvu hwe3 μm.Kune TRAP staining (F6760, Sigma-Aldrich, Germany), zvikamu zvakasarudzwa zvakasvibiswa, zvakadzorerwa uye zvakaputirwa muTRAP reagent pa 37 ° C kweawa imwe.Mifananidzo yakatorwa pachishandiswa siraidhi scanner (Pannoramic 250 Flash III, 3DHISTECH, Hungary) uye yakaverengerwa nekuyera kuvharwa kwenzvimbo yenzvimbo yakasvibiswa.Mukuedza kwega kwega, angangoita mana substrates muboka rega rega akaongororwa neImageJ software.
Kuongorora kukosha kwehuwandu kwakaitwa nekushandisa GraphPad Prism (GraphPad Software Inc., USA).Unpaired t-test uye imwe-nzira yekuongorora yekusiyana (ANOVA) yakashandiswa kuyedza misiyano pakati pemapoka ekuongorora.Chiyero chekukosha chinoratidzwa mumufananidzo sezvinotevera: * P <0.05, ** P <0.01, *** P <0.001 uye ****P <0.0001;NS, hapana musiyano wakakosha.
Zvekuwedzera zvechinyorwa ichi, ndapota ona http://advances.sciencemag.org/cgi/content/full/6/44/eabb0025/DC1
Ichi chinyorwa chekuwana chakavhurika chakagoverwa pasi pemitemo yeCreative Commons Attribution-Non-Commercial License, iyo inobvumira kushandiswa, kugovera uye kubereka mune chero svikiro, chero kushandiswa kusiri kwekutsvaga kwekutengesa uye chirevo ndechekuti yekutanga. basa rakarurama.Reference.
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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
Iyo antibacterial uye immune immune coatings ye orthopedic implants inogona kuderedza hutachiona uye mhinduro dzekudzivirira zvirwere zvinokonzerwa nehutachiona.
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
Iyo antibacterial uye immune immune coatings ye orthopedic implants inogona kuderedza hutachiona uye mhinduro dzekudzivirira zvirwere zvinokonzerwa nehutachiona.
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