Kwa odwala omwe akuchitidwa opaleshoni ya mafupa a mafupa, matenda opatsirana ndi mabakiteriya ndi mayankho a chitetezo cha mthupi omwe amayambitsa matenda nthawi zonse akhala akupha moyo.Zamoyo zodziwika bwino zimatha kuipitsidwa ndi tizilombo toyambitsa matenda, zomwe zimapangitsa kuti mabakiteriya alowe pamalo ovulala ndikuyambitsa matenda pambuyo pa opaleshoni.Chifukwa chake, pakufunika kufunikira kokhala ndi anti-infection ndi zokutira zopulumukira za chitetezo chamthupi zoyika mafupa.Apa, tapanga ukadaulo wapamwamba wosintha malo oyika mafupa opangidwa ndi mafupa otchedwa Lubricated Orthopedic Implant Surface (LOIS), omwe amawuziridwa ndi malo osalala a mbiya zamitengo.LOIS ili ndi nthawi yayitali komanso yolimba yamadzimadzi yothamangitsa zakumwa ndi zinthu zachilengedwe (kuphatikiza ma cell, mapuloteni, calcium ndi mabakiteriya).Kuphatikiza apo, tidatsimikizira kulimba kwamakina motsutsana ndi zikwawu ndi mphamvu yokonza poyerekezera kuwonongeka kosapeweka pa opaleshoni ya in vitro.Mtundu wa fupa la fupa la kalulu wotupa wa fracture wa chikazi unagwiritsidwa ntchito pophunzira bwino za anti-biological scaling ndi anti-infection mphamvu ya LOIS.Tikuwona kuti LOIS, yomwe ili ndi anti-biofouling properties komanso kulimba kwa makina, ndi sitepe yopita patsogolo pa opaleshoni yopanda matenda ya mafupa.
Masiku ano, chifukwa cha ukalamba wonse, chiwerengero cha odwala omwe ali ndi matenda a mafupa (monga fractures okalamba, matenda osokonekera a mafupa, ndi mafupa osteoporosis) chawonjezeka kwambiri (1, 2).Chifukwa chake, mabungwe azachipatala amawona kuti opaleshoni ya mafupa ndi yofunika kwambiri, kuphatikiza zomangira za mafupa, zomangira, misomali ndi mfundo zopangira (3, 4).Komabe, ma implants achikhalidwe am'mafupa akuti amatha kulumikizidwa ndi mabakiteriya komanso mapangidwe a biofilm, omwe angayambitse matenda opangira opaleshoni (SSI) pambuyo pa opaleshoni (5, 6).Pamene biofilm imapangidwa pamwamba pa mafupa opangidwa ndi mafupa, kuchotsa biofilm kumakhala kovuta kwambiri ngakhale pogwiritsa ntchito mlingo waukulu wa maantibayotiki.Chifukwa chake, nthawi zambiri zimayambitsa matenda oopsa a postoperative (7, 8).Chifukwa cha mavuto omwe ali pamwambawa, chithandizo cha ma implants omwe ali ndi kachilombo chiyenera kuphatikizapo kubwezeretsanso, kuphatikizapo kuchotsa ma implants onse ndi minyewa yozungulira;motero, wodwalayo amamva kupweteka kwambiri komanso zoopsa zina (9, 10).
Kuti athetse ena mwamavutowa, ma implants a mafupa opangidwa ndi mankhwala apangidwa kuti ateteze matenda pochotsa mabakiteriya omwe amakhala pamwamba (11, 12).Komabe, njirayo ikuwonetsabe malire angapo.Zanenedwa kuti kuyika kwa nthawi yayitali kwa ma implants owonjezera mankhwala kwadzetsa kuwonongeka kwa minofu yozungulira ndikuyambitsa kutupa, komwe kungayambitse necrosis (13, 14).Kuphatikiza apo, zosungunulira za organic zomwe zitha kukhalapo pambuyo popanga zoyika za mafupa, zomwe ndizoletsedwa ndi US Food and Drug Administration, zimafunikira njira zina zoyeretsera kuti zikwaniritse miyezo yake (15).Ma implants otulutsa mankhwala osokoneza bongo amakhala ovuta kuti atulutse mankhwalawo moyenera, ndipo chifukwa cha kuchuluka kwawo kwamankhwala osokoneza bongo, kugwiritsa ntchito mankhwalawa kwanthawi yayitali sikutheka (16).
Njira ina yodziwika bwino ndikuyika choyikapocho ndi polima kuti ateteze zinthu zachilengedwe ndi mabakiteriya kuti asamamatire pamwamba (17).Mwachitsanzo, ma polima a zwitterionic akopa chidwi chifukwa cha kusamata kwawo akakumana ndi mapuloteni a plasma, ma cell, ndi mabakiteriya.Komabe, ili ndi malire okhudzana ndi kukhazikika kwanthawi yayitali komanso kulimba kwamakina, zomwe zimalepheretsa kugwiritsa ntchito kwake m'mitsempha ya mafupa, makamaka chifukwa cha kukwapula kwamakina panthawi ya opaleshoni (18, 19).Kuphatikiza apo, chifukwa cha kuchuluka kwa biocompatibility, kusowa kofunikira kwa opaleshoni yochotsa, komanso kuyeretsa pamwamba kudzera mu dzimbiri, ma implants opangidwa ndi mafupa opangidwa ndi zinthu zosawonongeka zagwiritsidwa ntchito (20, 21).Panthawi ya dzimbiri, zomangira za mankhwala pakati pa ma polima amathyoledwa ndikuchotsedwa pamwamba, ndipo zomangirazo zimayeretsa pamwamba.Komabe, kuipitsidwa kwa anti-biological pakuyeretsa pamwamba kumakhala kothandiza pakanthawi kochepa.Kuphatikiza apo, zinthu zambiri zomwe zimatha kuyamwa kuphatikiza poly(lactic acid-glycolic acid copolymer) (PLGA), polylactic acid (PLA) ndi ma aloyi opangidwa ndi magnesiamu zidzawonongeka komanso kukokoloka kwa thupi, zomwe zingasokoneze kukhazikika kwamakina.(makumi awiri ndimphambu ziwiri).Kuonjezera apo, zidutswa za mbale zowonongeka zimapereka malo oti mabakiteriya agwirizane, zomwe zimawonjezera mwayi wa matenda m'kupita kwanthawi.Chiwopsezo cha kuwonongeka kwamakina ndi matenda chimachepetsa kugwiritsa ntchito opaleshoni yapulasitiki (23).
Ma superhydrophobic (SHP) omwe amatsanzira mawonekedwe apamwamba a masamba a lotus akhala njira yothetsera malo odana ndi kuipitsa (24, 25).Malo a SHP akamizidwa mumadzimadzi, mpweya umatsekeka, motero kupanga matumba a mpweya ndikuletsa kumatira kwa mabakiteriya (26).Komabe, kafukufuku waposachedwapa wasonyeza kuti SHP pamwamba ili ndi zovuta zokhudzana ndi kukhazikika kwa makina ndi kukhazikika kwa nthawi yaitali, zomwe zimalepheretsa kugwiritsidwa ntchito kwake mu implants zachipatala.Kuphatikiza apo, matumba amlengalenga amatha kusungunuka ndikutaya katundu wawo wotsutsa, zomwe zimapangitsa kuti mabakiteriya azimatira chifukwa cha malo akulu a SHP (27, 28).Posachedwa, Aizenberg ndi anzake adayambitsa njira yatsopano yopangira anti-biofouling pamwamba pakupanga malo osalala owuziridwa ndi chomera cha Nepenthes pitcher (29, 30).Malo osalala amawonetsa kukhazikika kwanthawi yayitali pansi pamikhalidwe ya hydraulic, ndimadzimadzi kwambiri othamangitsa zamadzimadzi achilengedwe, ndipo amakhala ndi zodzikonza zokha.Komabe, palibe njira yogwiritsira ntchito zokutira ku choyikapo chachipatala chofanana ndi chovuta, komanso sichitsimikiziridwa kuthandizira kuchira kwa minofu yowonongeka pambuyo pa kuikidwa.
Pano, tikuyambitsa mafuta opangidwa ndi mafupa opangidwa ndi mafupa (LOIS), malo opangidwa ndi micro / nano-structured orthopedic implant pamwamba ndipo amaphatikizidwa mwamphamvu ndi mafuta ochepa kwambiri kuti asagwirizane ndi opaleshoni ya pulasitiki Matenda a bakiteriya, monga fracture fixation.Chifukwa chakuti mawonekedwe a fluorine-functionalized micro / nano-level amakonza bwino mafuta odzola pamapangidwewo, LOIS yotukuka imatha kuthamangitsa kumamatira kwa zakumwa zosiyanasiyana ndikusunga ntchito yotsutsa kwa nthawi yayitali.Zovala za LOIS zitha kugwiritsidwa ntchito kuzinthu zamitundu yosiyanasiyana zomwe zimapangidwira kupanga mafupa.Mphamvu zabwino kwambiri zolimbana ndi biofouling za LOIS motsutsana ndi mabakiteriya a biofilm [Pseudomonas aeruginosa ndi Staphylococcus aureus (MRSA) wolimbana ndi methicillin] ndi zinthu zachilengedwe (maselo, mapuloteni ndi calcium) zatsimikiziridwa mu vitro.Kumamatira kwamphamvu kwambiri ku gawo lapansi ndikochepera 1%.Kuonjezera apo, ngakhale kupsinjika kwamakina monga kukanda pamwamba kumachitika, kudzichiritsa komwe kumachitika chifukwa cha mafuta olowera kumathandizira kukhalabe ndi anti-fouling.Zotsatira zoyesa kulimba kwamakina zikuwonetsa kuti ngakhale zitasintha kapangidwe kake ndi mankhwala, mphamvu zonse sizidzachepetsedwa kwambiri.Kuphatikiza apo, kuyesa kwa in vitro komwe kumatengera kupsinjika kwamakina m'malo opangira opaleshoni kunachitika kutsimikizira kuti LOIS imatha kupirira zovuta zamakina zomwe zimachitika panthawi ya opaleshoni ya pulasitiki.Pomaliza, tidagwiritsa ntchito kalulu wozikidwa mu vivo femoral fracture model, yomwe idatsimikizira kuti LOIS ili ndi antibacterial properties komanso biocompatibility.Zotsatira za radiological ndi histological zatsimikizira kuti khalidwe lokhazikika la mafuta odzola komanso anti-biofouling katundu mkati mwa masabata a 4 mutatha kuikidwa akhoza kukwaniritsa bwino anti-infection ndi chitetezo chamthupi kuthawa ntchito popanda kuchedwa kuchiritsa fupa.
Chithunzi 1A chikuwonetsa chojambula chojambula cha LOIS yopangidwa, yomwe imayikidwa ndi micro/nano-scale structures mu kalulu femoral fracture model kuti atsimikizire bwino kwambiri anti-biological fouling ndi anti-infection properties.Njira ya biomimetic imachitika kuti ifananize pamwamba pa chomera champhika wamadzi, ndikuletsa biofouling mwa kuphatikiza mafuta osanjikiza mkati mwa micro/nano ya pamwamba.Pamwamba pa jekeseni wothira mafuta amatha kuchepetsa kukhudzana pakati pa zinthu zamoyo ndi pamwamba.Choncho, chifukwa cha mapangidwe okhazikika azitsulo zamagetsi pamtunda, ali ndi ntchito yabwino yotsutsa komanso kukhazikika kwa nthawi yaitali.Zotsatira zake, zotsutsana ndi biofouling pamalo opaka mafuta zimalola kugwiritsa ntchito zinthu zosiyanasiyana pakufufuza zamankhwala.Komabe, kafukufuku wochuluka wa momwe malo apaderawa amagwirira ntchito m'thupi sanamalizidwe.Poyerekeza LOIS ndi magawo amaliseche mu vitro pogwiritsa ntchito albumin ndi mabakiteriya a biofilm, kusamata kwa LOIS kungatsimikizidwe (Chithunzi 1B).Kuphatikiza apo, pakugudubuza madontho amadzi pagawo lopanda kanthu komanso gawo la LOIS (Chithunzi S1 ndi Movie S1), magwiridwe antchito achilengedwe amatha kuwonetsedwa.Monga momwe chithunzi cha maikulosikopu cha fluorescence, gawo lapansi lowonekera lomwe lidakulungidwa mu kuyimitsidwa kwa mapuloteni ndi mabakiteriya adawonetsa kuchuluka kwa zinthu zamoyo zomwe zimamatira pamwamba.Komabe, chifukwa cha zinthu zake zabwino kwambiri zothana ndi biofouling, LOIS simawonetsa fulorosenti iliyonse.Pofuna kutsimikizira kuti ali ndi anti-biofouling ndi anti-infection, LOIS inagwiritsidwa ntchito pamwamba pa mafupa a mafupa kuti apange mafupa (mbale ndi zomangira) ndikuyikidwa mu chitsanzo cha fracture ya kalulu.Asanakhazikitsidwe, implant wamaliseche wa mafupa ndi LOIS adayikidwa mu kuyimitsidwa kwa bakiteriya kwa maola 12.Pre-incubation imatsimikizira kuti biofilm imapangidwa pamwamba pa implant yowonekera kuti ifanane.Chithunzi 1C chikuwonetsa chithunzi cha malo ophwanyika masabata a 4 mutatha kuikidwa.Kumanzere, kalulu wokhala ndi mafupa opanda kanthu amawonetsa kutupa kwakukulu chifukwa cha mapangidwe a biofilm pamwamba pa implant.Chotsatira chake chinawonedwa mu akalulu oikidwa ndi LOIS, ndiko kuti, minofu yozungulira ya LOIS sinasonyeze zizindikiro za matenda kapena zizindikiro za kutupa.Kuonjezera apo, chithunzi cha kuwala kumanzere chimasonyeza malo opangira opaleshoni ya kalulu ndi choyikapo chowonekera, chosonyeza kuti palibe zomatira zambiri zomwe zilipo pamwamba pa kuyika kowonekera komwe kunapezeka pamwamba pa LOIS.Izi zikuwonetsa kuti LOIS ili ndi kukhazikika kwa nthawi yayitali ndipo ili ndi mphamvu yosunga zonyansa zotsutsana ndi biological and anti-adhesion properties.
(A) Chithunzi chojambula cha LOIS ndi kuikidwa kwake mu chitsanzo cha kuthyoka kwa chikazi cha kalulu.(B) Fluorescence microscopy chithunzi cha mapuloteni ndi bakiteriya biofilm pamalo opanda kanthu ndi LOIS gawo lapansi.Masabata a 4 atayikidwa, (C) chithunzi chazithunzi cha malo ophwanyika ndi (D) chithunzi cha X-ray (chowonetsedwa ndi rectangle yofiira).Chithunzi chovomerezeka: Kyomin Chae, Yonsei University.
Akalulu ovunditsidwa, omwe adayikidwa molakwika adawonetsa kuchira kwa mafupa popanda zizindikiro za kutupa kapena matenda.Kumbali inayi, SHP imayikidwa kale mu kuyimitsidwa kwa bakiteriya kumawonetsa kutupa kokhudzana ndi matenda pamitu yozungulira.Izi zitha kukhala chifukwa chakulephera kwake kuletsa kumamatira kwa mabakiteriya kwa nthawi yayitali (Chithunzi S2).Pofuna kutsimikizira kuti LOIS sichimakhudza machiritso, koma imalepheretsa matenda omwe angakhalepo okhudzana ndi kuikidwa, zithunzi za X-ray za matrix odziwika bwino ndi LOIS pa malo ophwanyika anafaniziridwa (Chithunzi 1D).Chithunzi cha X-ray cha implant chopanda kanthu chinasonyeza mizere yosalekeza ya osteolysis, kusonyeza kuti fupa silinachiritsidwe kwathunthu.Izi zikusonyeza kuti kuchira kwa mafupa kungachedwe kwambiri chifukwa cha kutupa kokhudzana ndi matenda.M’malo mwake, zinasonyeza kuti akalulu amene anaikidwa ndi LOIS anali atachira ndipo sanasonyeze malo ophwanyika oonekeratu.
Pofuna kupanga ma implants azachipatala okhala ndi kukhazikika kwanthawi yayitali komanso magwiridwe antchito (kuphatikiza kukana biofouling), zoyesayesa zambiri zapangidwa.Komabe, kukhalapo kwa zinthu zosiyanasiyana zamoyo ndi mphamvu zomata minofu kumachepetsa kukula kwa njira zawo zamankhwala odalirika.Kuti tithane ndi zofooka izi, tapanga mawonekedwe ang'onoang'ono / nano osanjikiza ndi malo osinthidwa ndi mankhwala, omwe amakometsedwa chifukwa cha mphamvu yayikulu ya capillary ndi kuyanjana kwamankhwala kuti tisunge mafuta osalala kwambiri.Chithunzi 2A chikuwonetsa njira yonse yopangira LOIS.Choyamba, konzani chitsulo chosapanga dzimbiri chamankhwala (SS) 304 gawo lapansi.Kachiwiri, mawonekedwe ang'onoang'ono / nano amapangidwa pagawo la SS ndi etching yamankhwala pogwiritsa ntchito yankho la hydrofluoric acid (HF).Pofuna kubwezeretsa kukana kwa dzimbiri kwa SS, njira ya nitric acid (HNO3) (31) imagwiritsidwa ntchito pokonza gawo lapansi lokhazikika.Passivation imakulitsa kukana kwa dzimbiri kwa gawo lapansi la SS ndipo imachepetsa kwambiri njira ya dzimbiri yomwe ingachepetse ntchito yonse ya LOIS.Kenaka, popanga monolayer wodzipangira yekha (SAM) ndi 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane (POTS), pamwamba pake amasinthidwa ndi mankhwala kuti apititse patsogolo mgwirizano wa mankhwala pakati pa pamwamba ndi mafuta osalala Affinity.Kusinthidwa kwapamwamba kumachepetsa kwambiri mphamvu ya pamwamba pa malo opangidwa ndi micro / nano-scale, omwe amafanana ndi mphamvu ya pamwamba pa mafuta osalala.Izi zimathandiza kuti mafutawo anyowedwe kotheratu, potero kupanga wosanjikiza wokhazikika wamafuta pamwamba.Malo osinthidwa amawonetsa kuwonjezereka kwa hydrophobicity.Zotsatira zikuwonetsa kuti mafuta oterera amawonetsa machitidwe okhazikika pa LOIS chifukwa cha kuyanjana kwamphamvu kwamankhwala ndi mphamvu ya capillary yomwe imayambitsidwa ndi kapangidwe ka micro/nano (32, 33).Kusintha kwa kuwala pamwamba pa SS pambuyo pa kusinthidwa kwapamwamba ndi jekeseni wa mafuta kunaphunziridwa.Mapangidwe ang'onoang'ono / nano omwe amapangidwa pamwamba amatha kusintha mawonekedwe ndikudetsa pamwamba.Chochitikachi chimachitika chifukwa cha kuwonjezereka kwa kuwala kwa kuwala pamtunda woyipa, komwe kumawonjezera kuwunikira komwe kumachitika chifukwa cha njira yotsekera kuwala (34).Kuphatikiza apo, mafuta akatha kubayidwa, LOIS imakhala yakuda.Kupaka mafuta kumapangitsa kuwala kocheperako kuwonekera kuchokera ku gawo lapansi, potero kudetsa LOIS.Pofuna kukhathamiritsa ma microstructure/nanostructure kuti awonetse kagawo kakang'ono kwambiri kotsetsereka (SA) kuti akwaniritse ntchito yolimbana ndi biofouling, scanning electron microscopy (SEM) ndi ma atomiki awiriawiri adagwiritsidwa ntchito popanga nthawi zosiyanasiyana za HF etching (0, 3)., Mphindi 15 ndi 60) Limbikitsani Maikulosikopu (AFM) (Chithunzi 2B).Zithunzi za SEM ndi AFM zikuwonetsa kuti patangopita nthawi yochepa (3 mphindi za etching), gawo lapansi lopanda kanthu lapanga roughness ya nano-scale yosiyana.Kuuma kwapamwamba kumasintha ndi nthawi yokhotakhota (Chithunzi S3).Kupindika kosiyanasiyana kwa nthawi kumawonetsa kuti kuuma kwapamwamba kumapitilirabe kukula ndikufikira pachimake pa mphindi 15 za etching, ndiyeno kutsika pang'ono kwa roughness kumawonedwa pakangodutsa mphindi 30.Panthawiyi, roughness ya nano-level imachotsedwa, pamene zovuta zazing'ono zimakula mwamphamvu, zomwe zimapangitsa kuti kusinthako kusinthe kukhala kokhazikika.Pambuyo etching kwa mphindi zoposa 30, kuwonjezereka kwina kwa roughness kumawonedwa, yomwe ikufotokozedwa mwatsatanetsatane motere: SS imapangidwa ndi chitsulo, chosakanikirana ndi zinthu monga chitsulo, chromium, faifi tambala, molybdenum ndi zinthu zina zambiri.Mwa zinthu izi, chitsulo, chromium ndi molybdenum zimagwira ntchito yofunika kwambiri popanga ma micron/nano-scale roughness pa SS ndi HF etching.Kumayambiriro kwa dzimbiri, chitsulo ndi chromium zimakhala ndi dzimbiri chifukwa molybdenum imakhala ndi mphamvu yolimbana ndi dzimbiri kuposa molybdenum.Pamene etching ikupita patsogolo, yankho la etching limafika pakuchulukirachulukira komweko, ndikupanga ma fluoride ndi ma oxide omwe amayamba chifukwa cha etching.Fluoride ndi okusayidi zimatsika ndipo pamapeto pake zimayikanso pamwamba, kupanga khwinya pamtunda wa micron/nano (31).Vutoli laling'ono / nano-level limakhala ndi gawo lofunikira pakudzichiritsa nokha kwa LOIS.Pamwamba wapawiri sikelo umatulutsa synergistic zotsatira, kwambiri kuonjezera capillary mphamvu.Izi zimalola kuti mafutawo alowe pansi mokhazikika komanso amathandizira kudzichiritsa okha (35).Mapangidwe a roughness zimadalira etching nthawi.Pansi pa mphindi 10 zowotchera, pamwamba pamakhala roughness ya nano-scale yokha, yomwe sikokwanira kuti ikhale ndi mafuta okwanira kuti isakanize biofouling (36).Komano, ngati etching nthawi kuposa mphindi 30, nano-mulingo roughness wopangidwa ndi redeposition wa chitsulo ndi chromium zidzatha, ndipo kokha yaying'ono roughness adzakhala chifukwa cha molybdenum.Kuchuluka kwapamwamba kulibe nano-scale roughness ndipo imataya mphamvu ya synergistic ya roughness ya magawo awiri, yomwe imakhudza kwambiri makhalidwe odzichiritsa okha a LOIS.Miyezo ya SA idachitidwa pazigawo zokhala ndi nthawi zosiyanasiyana zowonetsera kuti zitsimikizire kuti magwiridwe antchito a anti-fouling.Mitundu yosiyanasiyana yamadzimadzi idasankhidwa kutengera kukhuthala ndi mphamvu yapamtunda, kuphatikiza madzi a deionized (DI), magazi, ethylene glycol (EG), ethanol (EtOH) ndi hexadecane (HD) (Chithunzi S4).Njira yosinthira nthawi ikuwonetsa kuti pazamadzimadzi zosiyanasiyana zokhala ndi mphamvu ndi ma viscosity osiyanasiyana, SA ya LOIS pambuyo pa mphindi 15 zakukhazikika ndiyotsika kwambiri.Chifukwa chake, LOIS imakokedwa kuti ipangike kwa mphindi 15 kuti ipange roughness ya micron ndi nano-scale, yomwe ili yoyenera kusunga kulimba kwamafuta ndi zinthu zabwino zotsutsana ndi zonyansa.
(A) Chithunzi chojambula cha njira zinayi zopangira LOIS.Tsambali likuwonetsa SAM yopangidwa pagawo.(B) Zithunzi za SEM ndi AFM, zomwe zimagwiritsidwa ntchito kukhathamiritsa kapangidwe ka micro/nano kagawo kakang'ono ka nthawi zosiyanasiyana.Zithunzi za X-ray photoelectron spectroscopy (XPS) za (C) Cr2p ndi (D) F1s pambuyo podutsa pamwamba ndi zokutira za SAM.kapena, unit mokhazikika.(E) Zithunzi zoyimilira za madontho amadzi opanda kanthu, okhazikika, a SHP ndi LOIS.(F) Mulingo wa contact angle (CA) ndi SA muyeso wa zakumwa zokhala ndi zovuta zapamtunda zosiyanasiyana pa SHP ndi LOIS.Zambiri zimawonetsedwa ngati ± SD.
Kenaka, pofuna kutsimikizira kusintha kwa mankhwala a pamwamba, X-ray photoelectron spectroscopy (XPS) inagwiritsidwa ntchito pophunzira kusintha kwa mankhwala a gawo lapansi pambuyo popaka pamwamba pamtundu uliwonse.Chithunzi 2C chikuwonetsa zotsatira za muyeso wa XPS wa malo okhazikika a HF ndi malo otetezedwa a HNO 3.Mapiri akulu akulu awiri pa 587.3 ndi 577.7 eV atha kukhala chifukwa cha chomangira cha Cr-O chomwe chilipo mu chromium oxide wosanjikiza, womwe ndiye kusiyana kwakukulu kuchokera ku HF etched surface.Izi zimachitika makamaka chifukwa chomwa chitsulo ndi chromium fluoride pamtunda ndi HNO3.Etching yochokera ku HNO3 imalola chromium kuti ipange gawo losanjikiza la oxide pamwamba, zomwe zimapangitsa kuti SS yokhazikika ikhale yolimba ku dzimbiri.Mu Chithunzi 2D, mawonekedwe a XPS adapezedwa kuti atsimikizire kuti silane yochokera ku fluorocarbon idapangidwa pamtunda pambuyo pakupaka kwa SAM, komwe kumakhala ndi kuthamangitsa kwamadzi kwambiri ngakhale kwa EG, magazi ndi EtOH.Kupaka kwa SAM kumatsirizidwa pochita magulu ogwira ntchito a silane ndi magulu a hydroxyl opangidwa ndi mankhwala a plasma.Zotsatira zake, kuwonjezeka kwakukulu kwa nsonga za CF2 ndi CF3 kunawonedwa.Mphamvu zomangirira pakati pa 286 ndi 296 eV zikuwonetsa kuti kusintha kwamankhwala kwamalizidwa bwino ndi zokutira za SAM.SHP imasonyeza nsonga zazikulu za CF2 (290.1 ​​​​eV) ndi CF3 (293.3 eV), zomwe zimayambitsidwa ndi silane yopangidwa ndi fluorocarbon yomwe imapangidwa pamwamba.Chithunzi 2E chikuwonetsa zithunzi zoyimilira za miyeso yolumikizana (CA) yamagulu osiyanasiyana amadzi opangidwa ndi madzi osagwirizana ndi opanda, etched, SHP, ndi LOIS.Zithunzizi zikuwonetsa kuti malo okhazikika amakhala a hydrophilic chifukwa cha kachipangizo kakang'ono / nano komwe kamapangidwa ndi etching yamankhwala kuti madzi a deionized alowe mu kapangidwe kake.Komabe, gawo lapansili litaphimbidwa ndi SAM, gawo lapansi likuwonetsa kuthamangitsa madzi mwamphamvu, kotero SHP yapamtunda imapangidwa ndipo malo olumikizana pakati pa madzi ndi pamwamba ndi ochepa.Pomaliza, kuchepa kwa CA kunawonedwa ku LOIS, komwe kungabwere chifukwa cha kulowa kwa mafuta mu microstructure, potero kumawonjezera malo okhudzana.Pofuna kutsimikizira kuti pamwamba ili ndi katundu wabwino kwambiri wamadzimadzi komanso osamata, LOIS inafanizidwa ndi gawo lapansi la SHP poyesa CA ndi SA pogwiritsa ntchito zakumwa zosiyanasiyana (Chithunzi 2F).Mitundu yosiyanasiyana yamadzimadzi idasankhidwa potengera kukhuthala komanso mphamvu yapamtunda, kuphatikiza madzi osungunuka, magazi, EG, EtOH ndi HD (Chithunzi S4).Zotsatira za muyeso wa CA zimasonyeza kuti pamene CA imakonda HD, mtengo wochepetsera wa CA, kumene CA ili ndi mphamvu zochepa kwambiri padziko lapansi.Kuphatikiza apo, LOIS ya CA yonse ndiyotsika.Komabe, muyeso wa SA ukuwonetsa chodabwitsa chosiyana kwambiri.Kupatula madzi ionized, zakumwa zonse zimamatira ku gawo lapansi la SHP popanda kutsika.Kumbali ina, LOIS imawonetsa SA yotsika kwambiri, pomwe madzi onse akapendekeka pakona yotsika kuposa 10 ° mpaka 15 °, madzi onse amagudubuzika.Izi zikuwonetsa mwamphamvu kuti kusamata kwa LOIS kuli bwino kuposa SHP pamwamba.Kuphatikiza apo, zokutira za LOIS zimagwiritsidwanso ntchito ku mitundu yosiyanasiyana ya zida, kuphatikiza titaniyamu (Ti), polyphenylsulfone (PPSU), polyoxymethylene (POM), polyether ether ketone (PEEK) ndi ma polima a bioabsorbable (PLGA), Ndizinthu zopanga mafupa (Chithunzi. S5)).Zithunzi zotsatizana za madontho pazinthu zomwe zathandizidwa ndi LOIS zimasonyeza kuti anti-biofouling katundu wa LOIS ndi ofanana pa magawo onse.Kuonjezera apo, zotsatira za kuyeza kwa CA ndi SA zimasonyeza kuti zinthu zopanda zomatira za LOIS zingagwiritsidwe ntchito pazinthu zina.
Pofuna kutsimikizira zotsutsana ndi zowononga za LOIS, mitundu yosiyanasiyana ya magawo (kuphatikizapo opanda kanthu, etched, SHP ndi LOIS) adayikidwa ndi Pseudomonas aeruginosa ndi MRSA.Mabakiteriya awiriwa adasankhidwa ngati mabakiteriya oyimira chipatala, omwe angayambitse kupanga ma biofilms, omwe amatsogolera ku SSI (37).Chithunzi 3 (A ndi B) chikuwonetsa zithunzi za microscope ya fluorescence ndi zotsatira za kuyeza kwa colony forming unit (CFU) za magawo omwe amaphatikizidwa mu kuyimitsidwa kwa bakiteriya kwa nthawi yochepa (maola 12) ndi nthawi yayitali (maola 72), motsatira.M'kanthawi kochepa, mabakiteriya amapanga masango ndikukula kukula kwake, akudziphimba ndi zinthu zonga mamina ndikuletsa kuchotsedwa kwawo.Komabe, mkati mwa maola 72 ma incubation, mabakiteriya amakhwima ndi kukhala osavuta kumwazikana kuti apange magulu ambiri kapena magulu.Choncho, zikhoza kuganiziridwa kuti makulitsidwe a maola 72 ndi nthawi yayitali ndipo ndi nthawi yoyenera yopangira biofilm pamwamba (38).M'kanthawi kochepa, malo okhazikika komanso pamwamba pa SHP adawonetsa kumatira kwa bakiteriya, komwe kunachepetsedwa pafupifupi 25% mpaka 50% poyerekeza ndi gawo lapansi lopanda kanthu.Komabe, chifukwa cha ntchito yake yabwino kwambiri yotsutsa-biofouling ndi kukhazikika, LOIS sinasonyeze kumatira kwa mabakiteriya a biofilm mu nthawi yochepa komanso yayitali.Chithunzi chojambula (Chithunzi 3C) chikufotokozera kufotokozera kwa njira yowonongeka ya anti-biological ya etching solution, SHP ndi LOIS.Lingaliro ndiloti gawo lapansi lokhazikika lomwe lili ndi mphamvu za hydrophilic lidzakhala ndi malo okulirapo kuposa gawo lapansi lopanda kanthu.Chifukwa chake, kuphatikiza kwa bakiteriya kudzachitika pagawo lokhazikika.Komabe, poyerekeza ndi gawo lapansi lopanda kanthu, gawo lapansi lokhazikika limakhala ndi biofilm yochepa kwambiri yopangidwa pamwamba.Izi ndichifukwa choti mamolekyu amadzi amamangiriza mwamphamvu pamwamba pa hydrophilic ndikuchita ngati mafuta opangira madzi, motero amasokoneza kumamatira kwa mabakiteriya pakanthawi kochepa (39).Komabe, wosanjikiza wa mamolekyu madzi ndi woonda kwambiri ndi sungunuka mu suspensions bakiteriya.Choncho, madzi maselo wosanjikiza kutha kwa nthawi yaitali, kumabweretsa kwambiri mabakiteriya adhesion ndi kuchulukana.Kwa SHP, chifukwa cha nthawi yayitali yosanyowetsa, kumamatira kwa bakiteriya kumaletsedwa.Kutsika kwa bakiteriya kumamatira kumatha kukhala chifukwa cha matumba a mpweya omwe amatsekeredwa m'malo osanjikiza komanso mphamvu zotsika pansi, potero kuchepetsa kukhudzana pakati pa kuyimitsidwa kwa bakiteriya ndi pamwamba.Komabe, kumatira kwakukulu kwa bakiteriya kunawonedwa mu SHP chifukwa idataya mphamvu zake zotsutsa kwa nthawi yayitali.Izi zimachitika makamaka chifukwa cha kutha kwa matumba a mpweya chifukwa cha kuthamanga kwa hydrostatic komanso kusungunuka kwa mpweya m'madzi.Izi zimachitika makamaka chifukwa cha kutha kwa matumba a mpweya chifukwa cha kusungunuka ndi mawonekedwe osanjikiza omwe amapereka malo okulirapo omatira (27, 40).Mosiyana ndi magawo awiriwa omwe amakhudza kwambiri kukhazikika kwa nthawi yayitali, mafuta odzola omwe ali mu LOIS amalowetsedwa mu micro / nano dongosolo ndipo sadzatha ngakhale kwa nthawi yaitali.Mafuta odzaza ndi makina ang'onoang'ono / nano amakhala okhazikika kwambiri ndipo amakopeka kwambiri pamtunda chifukwa cha kugwirizana kwawo kwa mankhwala, potero amalepheretsa kumamatira kwa mabakiteriya kwa nthawi yaitali.Chithunzi cha S6 chikuwonetsa chithunzi chowoneka bwino cha maikulosikopu chamafuta opaka mafuta omizidwa mu mchere wa phosphate buffered saline (PBS).Zithunzi zopitirira zimasonyeza kuti ngakhale pambuyo pa maola a 120 akugwedezeka pang'ono (120 rpm), mafuta osanjikiza pa LOIS amakhalabe osasintha, kusonyeza kukhazikika kwa nthawi yaitali pansi pa mikhalidwe yothamanga.Izi ndichifukwa cha kuyanjana kwakukulu kwa mankhwala pakati pa zokutira za SAM zochokera ku fluorine ndi mafuta opangidwa ndi perfluorocarbon, kotero kuti wosanjikiza wokhazikika wamafuta apangidwe.Choncho, ntchito yotsutsa-fouling imasungidwa.Kuonjezera apo, gawo lapansili linayesedwa motsutsana ndi mapuloteni oimira (albumin ndi fibrinogen), omwe ali mu plasma, maselo ogwirizana kwambiri ndi chitetezo cha mthupi (macrophages ndi fibroblasts), ndi omwe amagwirizana ndi mapangidwe a mafupa.Zomwe zili mu calcium ndizochuluka kwambiri.(Chithunzi 3D, 1 ndi 2, ndi Chithunzi S7) (41, 42).Kuonjezera apo, zithunzi za microscope ya fluorescence zoyesa zomatira za fibrinogen, albumin ndi calcium zinasonyeza makhalidwe osiyanasiyana a gulu lirilonse la gawo lapansi (Chithunzi S8).Pakupanga fupa, fupa ndi kashiamu zomwe zangopangidwa kumene zimatha kuzungulira fupa la mafupa, zomwe sizimangopangitsa kuti kuchotsa kumakhala kovuta, komanso kungayambitse vuto losayembekezereka kwa wodwalayo panthawi yochotsa.Choncho, kuchepa kwa calcium m'mafupa ndi zomangira ndizopindulitsa pa opaleshoni ya mafupa yomwe imafuna kuchotsa implants.Kutengera kuchuluka kwa malo olumikizidwa kutengera mphamvu ya fluorescence komanso kuchuluka kwa maselo, tidatsimikizira kuti LOIS imawonetsa zinthu zabwino kwambiri zothana ndi biofouling pazachilengedwe zonse poyerekeza ndi magawo ena.Malinga ndi zotsatira za kuyesa kwa in vitro, anti-biological fouling LOIS ingagwiritsidwe ntchito pazitsulo za mafupa, zomwe sizingalepheretse matenda oyambitsidwa ndi mabakiteriya a biofilm, komanso kuchepetsa kutupa chifukwa cha chitetezo cha mthupi.
(A) Zithunzi za ma microscope za Fluorescence za gulu lililonse (amaliseche, zokhazikika, SHP ndi LOIS) zolumikizidwa mu Pseudomonas aeruginosa ndi kuyimitsidwa kwa MRSA kwa maola 12 ndi 72.(B) Chiwerengero cha CFU chotsatira cha Pseudomonas aeruginosa ndi MRSA pamwamba pa gulu lirilonse.(C) Chithunzi chojambula cha anti-biological fouling mechanism ya etching yaifupi komanso yayitali, SHP ndi LOIS.(D) (1) Chiwerengero cha fibroblasts anamamatira aliyense gawo lapansi ndi fluorescence microscope zithunzi za maselo amamatira anabala ndi LOIS.(2) Kuyesedwa kwa adhesion kwa mapuloteni okhudzana ndi chitetezo cha mthupi, albumin ndi calcium zomwe zimakhudzidwa ndi machiritso a mafupa (* P <0.05, ** P <0.01, *** P <0.001 ndi **** P <0.0001).ns, osafunikira.
Pankhani ya kupsinjika kosalephereka kokhazikika, kukhazikika kwamakina nthawi zonse kwakhala vuto lalikulu pakuyika zokutira zoletsa.Njira zachikhalidwe zotsutsana ndi zonyansa za gel osakaniza zimachokera pa ma polima okhala ndi madzi otsika komanso osalimba.Chifukwa chake, nthawi zambiri amakhudzidwa ndi kupsinjika kwamakina muzogwiritsira ntchito zamankhwala.Chifukwa chake, zokutira zolimba zamakina zimakhalabe zovuta pazogwiritsa ntchito monga ma implants a mafupa (43, 44).Chithunzi cha 4A (1) chikuwonetsa mitundu iwiri ikuluikulu ya kupsinjika komwe kumagwiritsidwa ntchito pazitsulo za mafupa, kuphatikizapo kukwapula (kumeta ubweya wa ubweya) ndi kuponderezedwa ndi chithunzi cha kuwala kwa implant yowonongeka yopangidwa ndi forceps.Mwachitsanzo, pamene wononga wononga ndi screwdriver, kapena pamene dotolo agwira fupa mbale zolimba ndi tweezers ndi compressive mphamvu, pulasitiki mafupa mbale adzakhala kuwonongeka ndi kukandidwa pa onse macro ndi micro/nano masikelo (Chithunzi 4A, 2) .Pofuna kuyesa ngati LOIS yopangidwa ikhoza kupirira zowonongeka izi panthawi ya opaleshoni ya pulasitiki, nanoindentation inkachitidwa kuti afanizire kuuma kwa gawo lapansi lopanda kanthu ndi LOIS pa micro / nano scale kuti aphunzire makina a micro / nano structure Impact (Chithunzi. 4b).Chithunzi chojambula chikuwonetsa machitidwe osiyanasiyana a LOIS chifukwa cha kupezeka kwa ma micro/nano.Mphepete mwa kusuntha kwa mphamvu inakokedwa kutengera zotsatira za nanoindentation (Chithunzi 4C).Chithunzi cha buluu chikuyimira gawo lapansi lopanda kanthu, lomwe limangowonetsa kupindika pang'ono, monga momwe zimawonekera ndikuya kwakuya kwa 0.26-μm.Kumbali ina, kuwonjezereka kwapang'onopang'ono kwa mphamvu ya nanoindentation ndi kusamuka komwe kumawonedwa mu LOIS (njira yofiyira) kungasonyeze zizindikiro za kuchepa kwa makina, zomwe zimapangitsa kuya kwa nanoindentation kwa 1.61μm.Izi ndichifukwa choti mawonekedwe ang'onoang'ono / nano omwe ali mu LOIS amapereka malo opita patsogolo kwambiri pansonga ya nanoindenter, kotero kusinthika kwake kumakhala kwakukulu kuposa gawo lapansi lopanda kanthu.Konsta-Gdoutos et al.(45) amakhulupirira kuti chifukwa cha kukhalapo kwa nanostructures, nanoindentation ndi micro / nano roughness kumabweretsa ma curve osagwirizana.Dera lokhala ndi mthunzi limagwirizana ndi njira yopindika yosasinthika yomwe imadziwika ndi mawonekedwe a nanostructure, pomwe malo osakhala ndi mthunzi amachokera ku microstructure.Kupindika kumeneku kumatha kuwononga microstructure/nanostructure of the hold lubricant ndikusokoneza magwiridwe ake odana ndi kuipitsa.Kuti muphunzire za kuwonongeka kwa LOIS, kuwonongeka kosalephereka kwazinthu zazing'ono / nano kunabwerezedwa m'thupi panthawi ya opaleshoni ya pulasitiki.Pogwiritsa ntchito kuyesa kwa magazi ndi mapuloteni, kukhazikika kwa anti-biofouling katundu wa LOIS pambuyo pa vitro kungadziwike (Chithunzi 4D).Zithunzi zingapo zowoneka bwino zikuwonetsa kuwonongeka komwe kunachitika pafupi ndi mabowo a gawo lililonse.Kuyesedwa kwa magazi kunachitidwa kuti awonetse zotsatira za kuwonongeka kwa makina pa anti-biofouling zokutira (Chithunzi 4E).Monga SHP, zotsutsana ndi zowonongeka zimatayika chifukwa cha kuwonongeka, ndipo LOIS imasonyeza zinthu zabwino kwambiri zotsutsana ndi kuipitsa pobweza magazi.Izi zili choncho chifukwa, chifukwa mphamvu yapamtunda imayendetsedwa ndi capillary yomwe imaphimba malo owonongeka, kutuluka kwa lubricant ya microstructured lubricant kumabwezeretsa anti-fouling properties (35).Mchitidwe womwewo udawonedwa pakuyezetsa kwa mapuloteni pogwiritsa ntchito albumin.M'dera lowonongeka, kumatira kwa mapuloteni pamwamba pa SHP kumawonedwa kwambiri, ndipo poyesa kufalikira kwa malo ake, kungathe kuwerengedwa ngati theka la mlingo womatira wa gawo lapansi lopanda kanthu.Kumbali inayi, LOIS idasungabe zinthu zake zotsutsana ndi biofouling popanda kuyambitsa kumamatira (Chithunzi 4, F ndi G).Kuphatikiza apo, pamwamba pa wononga nthawi zambiri amakhala ndi kupsinjika kwamphamvu kwamakina, monga kubowola, kotero tidaphunzira kuthekera kwa zokutira za LOIS kukhalabe pa screw mu vitro.Chithunzi 4H chikuwonetsa zithunzi zowoneka bwino za zomangira zosiyanasiyana, kuphatikiza zopanda kanthu, SHP ndi LOIS.Rectangle yofiyira imayimira malo omwe amayang'ana pomwe kupsinjika kwamphamvu kwamakina kumachitika panthawi yoyika mafupa.Mofanana ndi kuyesa kwa mapuloteni a mbale, microscope ya fluorescence imagwiritsidwa ntchito kufotokozera mapuloteni a mapuloteni ndikuyesa malo ophimba kuti atsimikizire kukhulupirika kwa nsalu ya LOIS, ngakhale pansi pa kupsinjika kwamphamvu kwa makina (Chithunzi 4, I ndi J).Zomangira zopangidwa ndi LOIS zimawonetsa magwiridwe antchito abwino kwambiri oletsa kuyipitsa, ndipo pafupifupi palibe mapuloteni omwe amamatira pamwamba.Kumbali ina, kumatira kwa mapuloteni kunkawoneka mu zomangira zopanda kanthu ndi zomangira za SHP, pomwe zomangira za SHP zinali gawo limodzi mwa magawo atatu a zomangira zopanda kanthu.Kuphatikiza apo, kuyika kwa mafupa komwe kumagwiritsidwa ntchito kukonzanso kuyenera kukhala kolimba mwamakina kuti athe kupirira kupsinjika komwe kumagwiritsidwa ntchito pamalo osweka, monga momwe tawonetsera pa Chithunzi 4K.Chifukwa chake, kuyezetsa kopindika kunachitika kuti adziwe momwe kusintha kwamankhwala kumakhudzira makina.Kuonjezera apo, izi zimachitidwa kuti mukhalebe ndi nkhawa yokhazikika kuchokera ku implant.Ikani mphamvu yoyimirira mpaka choyikapo chikulungidwa bwino ndipo piritsi la kupsinjika likupezeka (Chithunzi 4L, 1).Katundu awiri kuphatikiza Young modulus ndi flexural mphamvu anafaniziridwa pakati bare ndi LOIS gawo monga zizindikiro za mphamvu mawotchi awo (Chithunzi 4L, 2 ndi 3).Young's modulus ikuwonetsa kuthekera kwa zinthu kupirira kusintha kwamakina.Modulus ya Young pagawo lililonse ndi 41.48±1.01 ndi 40.06±0.96 GPa, motsatana;kusiyana komwe kumawonedwa ndi pafupifupi 3.4%.Kuphatikiza apo, akuti mphamvu yopindika, yomwe imatsimikizira kulimba kwa zinthuzo, ndi 102.34±1.51 GPa pagawo lopanda kanthu ndi 96.99±0.86 GPa ya SHP.Gawo lopanda kanthu ndi pafupifupi 5.3% kukwezeka.Kuchepa pang'ono kwa zinthu zamakina kumatha kuyambitsidwa ndi mawonekedwe a notch.M'mawonekedwe a notch, kuyabwa kwapang'ono / nano kumatha kukhala ngati ma notches, zomwe zimatsogolera kupsinjika kwanuko komanso kukhudza momwe makina amapangidwira (46).Komabe, potengera kuti kuuma kwa fupa la cortical fupa laumunthu kumanenedwa kuti kuli pakati pa 7.4 ndi 31.6 GPa, ndipo LOIS modulus yoyezera imaposa mafupa a cortical (47), LOIS ndi yokwanira kuthandizira kupasuka ndi zida zamakina zimakhudzidwa pang'ono ndi kusinthidwa kwapamwamba.
(A) Chithunzi chojambula cha (1) kupsyinjika kwa makina komwe kumagwiritsidwa ntchito pazitsulo za mafupa panthawi ya opaleshoni, ndi (2) chithunzi cha kuwala kwa mafupa owonongeka.(B) Chithunzi chojambula cha nano-mechanical properties muyeso ndi nanoindentation ndi LOIS pamtunda wopanda kanthu.(C) Nanoindentation mphamvu-kusuntha kokhota pamalo opanda kanthu ndi LOIS.(D) Pambuyo pakuyesa kwa in vitro, yerekezerani zithunzi zamitundu yosiyanasiyana ya mafupa a mafupa (malo owonongeka amawonetsedwa ndi rectangle yofiira) kuti muyese kupsinjika kwamakina komwe kumachitika panthawi ya opareshoni.(E) Kuyesedwa kwa magazi ndi (F) kuyesa kwa mapuloteni a gulu lowonongeka la mafupa a mafupa.(G) Yezerani kuchuluka kwa mapuloteni omwe akumatira ku mbale.(H) Zithunzi zowoneka bwino zamitundu yosiyanasiyana ya zomangira za mafupa pambuyo pa kuyesa kwa in vitro.(I) Kuyesedwa kwa mapuloteni kuti muphunzire kukhulupirika kwa zokutira zosiyanasiyana.(J) Yezerani kuchuluka kwa puloteni yomwe imamatira ku screw.(K) Kuyenda kwa kalulu kumapangidwira kuti pakhale kupanikizika kosasunthika pafupa losweka.(L) (1) Pindani zotsatira zoyesa ndi zithunzi zowoneka bwino musanapindike komanso mutatha.Kusiyana kwa (2) Young's modulus ndi (3) mphamvu yopindika pakati pa implant yopanda kanthu ndi SHP.Deta imawonetsedwa ngati ± SD (*P <0.05, **P <0.01, ***P <0.001 ndi ****P <0.0001).Chithunzi chovomerezeka: Kyomin Chae, Yonsei University.
M'malo azachipatala, mabakiteriya ambiri amalumikizana ndi zida zamoyo ndi malo ovulala amachokera ku ma biofilms okhwima, okhwima (48).Choncho, US Centers for Disease Control and Prevention akuti 65% ya matenda onse a anthu ndi okhudzana ndi biofilms (49).Pankhaniyi, ndikofunikira kupereka mawonekedwe oyesera mu vivo omwe amapereka mawonekedwe ofanana a biofilm pamwamba pa implant.Choncho, tinapanga chitsanzo cha fracture cha kalulu cha chikazi cha kalulu chomwe ma implants a mafupa adayikidwa kale mu kuyimitsidwa kwa bakiteriya ndiyeno amaikidwa m'matumbo a kalulu kuti aphunzire za anti-fouling za LOIS mu vivo.Chifukwa cha mfundo zitatu zofunikazi, matenda a bakiteriya amayamba chifukwa cha chikhalidwe chisanachitike osati jekeseni mwachindunji ya kuyimitsidwa kwa mabakiteriya: (i) Chitetezo cha mthupi cha akalulu ndi champhamvu mwachibadwa kuposa cha anthu;Choncho, jekeseni wa suspensions bakiteriya ndi mabakiteriya planktonic n'zotheka Zilibe mphamvu pa mapangidwe biofilms.(Ii) Mabakiteriya a planktonic amatha kutenga maantibayotiki, ndipo mankhwala opha tizilombo amagwiritsidwa ntchito pambuyo pa opaleshoni;Pomaliza, (iii) kuyimitsidwa kwa mabakiteriya a planktonic kumatha kuchepetsedwa ndi madzi amthupi la nyama (50).Mwa kukulitsa kukhazikitsidwa kwa bakiteriya mu kuyimitsidwa kwa bakiteriya tisanakhazikitsidwe, titha kuphunzira mozama zotsatira zoyipa za matenda a bakiteriya ndi machitidwe a thupi lakunja (FBR) pa machiritso a mafupa.Akalulu anaperekedwa nsembe kwa masabata a 4 atayikidwa, chifukwa osseointegration yofunikira kuti mafupa achiritsidwe adzatsirizidwa mkati mwa masabata a 4.Kenako, ma implants anachotsedwa kwa akalulu kuti akaphunzire kumunsi kwa mtsinje.Chithunzi 5A chikuwonetsa momwe mabakiteriya amachulukira.Kuyika kwa mafupa omwe ali ndi kachilombo kumalowetsedwa m'thupi.Chifukwa cha makulitsidwe chisanadze mu kuyimitsidwa kwa bakiteriya, akalulu asanu ndi limodzi mwa asanu ndi limodzi oikidwa ndi implants wamaliseche anali ndi kachilomboka, pamene akalulu omwe anaikidwa ndi ma implants opangidwa ndi LOIS analibe matenda.Matenda a mabakiteriya amapitilira munjira zitatu, kuphatikiza kukula, kukhwima ndi kubalalitsidwa (51).Choyamba, mabakiteriya ophatikizidwa amaberekana ndikukula pamwamba, ndiyeno mabakiteriya amapanga biofilm pamene amatulutsa polymer extracellular (EPS), amyloid ndi extracellular DNA.Biofilm sikuti imangosokoneza kulowa kwa maantibayotiki, komanso imalimbikitsa kudziunjikira kwa ma enzyme owononga ma antibiotic (monga β-lactamase) (52).Pomaliza, biofilm imafalitsa mabakiteriya okhwima m'magulu ozungulira.Choncho, matenda amapezeka.Kuonjezera apo, pamene thupi lachilendo limalowa m'thupi, matenda omwe angayambitse chitetezo champhamvu cha mthupi angayambitse kutupa kwakukulu, kupweteka, ndi kuchepa kwa chitetezo cha mthupi.Chithunzi 5B chimapereka chithunzithunzi cha FBR yoyambitsidwa ndi kuyika kwa mafupa a mafupa, osati kuyankha kwa chitetezo cha mthupi chifukwa cha matenda a bakiteriya.Chitetezo cha mthupi chimazindikira choyikacho ngati thupi lachilendo, kenako chimapangitsa kuti ma cell ndi minyewa igwirizane kuti itseke thupi lakunja (53).M'masiku oyambilira a FBR, matrix operekera adapangidwa pamwamba pa ma implants a mafupa, zomwe zidapangitsa kuti fibrinogen adsorption.The adsorbed fibrinogen ndiye imapanga netiweki wandiweyani kwambiri wa fibrin, womwe umalimbikitsa kulumikizidwa kwa leukocyte (54).Pamene maukonde a fibrin apangidwa, kutupa kwakukulu kudzachitika chifukwa cha kulowetsedwa kwa neutrophils.Pa sitepe iyi, ma cytokines osiyanasiyana monga tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) ndi IL-β amamasulidwa, ndipo ma monocyte amayamba kulowetsa malo oyikapo ndikusiyanitsa m'maselo akuluakulu.Phazi (41, 55, 56).Kuchepetsa FBR kwakhala kovuta nthawi zonse chifukwa FBR yochulukirapo imatha kuyambitsa kutupa koopsa komanso kosatha, komwe kungayambitse zovuta zakupha.Pofuna kuwunika momwe matenda a bakiteriya amakhudzira minyewa yozungulira choyikacho chopanda kanthu ndi LOIS, madontho a hematoxylin ndi eosin (H&E) ndi Masson trichrome (MT) adagwiritsidwa ntchito.Kwa akalulu oikidwa ndi magawo opanda kanthu, matenda owopsa a bakiteriya amakula, ndipo zithunzi zamtundu wa H & E zimasonyeza bwino zilonda ndi necrosis chifukwa cha kutupa.Kumbali inayi, malo amphamvu kwambiri odana ndi biofouling LOIS amalepheretsa kumamatira kwa bakiteriya, kotero samawonetsa zizindikiro za matenda ndipo amachepetsa kutupa (Chithunzi 5C).Zotsatira za MT staining zidawonetsa zomwezo.Komabe, madontho a MT adawonetsanso edema mu akalulu omwe adayikidwa ndi LOIS, kuwonetsa kuti kuchira kwatsala pang'ono kuchitika (Chithunzi 5D).Pofuna kuphunzira kuchuluka kwa chitetezo cha mthupi, kutsekemera kwa immunohistochemical (IHC) kunkagwiritsidwa ntchito pogwiritsa ntchito cytokines TNF-α ndi IL-6 zokhudzana ndi chitetezo cha mthupi.Kuyika kwamaliseche kwamaliseche komwe kunalibe mabakiteriya kunafanizidwa ndi LOIS yomwe idawonetsedwa ndi mabakiteriya koma osayambukiridwa kuti iphunzire kuchiritsa popanda matenda a bakiteriya.Chithunzi 5E chikuwonetsa chithunzi chazithunzi cha IHC chomwe chikuwonetsa TNF-α.Malo a bulauni amaimira chitetezo cha mthupi, kusonyeza kuti chitetezo cha mthupi mu LOIS chimachepetsedwa pang'ono.Kuphatikiza apo, mawu a IL-6 mu LOIS anali ocheperako poyerekeza ndi mawu olakwika amaliseche osabala (Chithunzi 5F).Mafotokozedwe a cytokine adawerengedwa poyezera dera la antibody lolingana ndi cytokine (Chithunzi 5G).Poyerekeza ndi akalulu omwe amawonekera ku ma implants olakwika, milingo ya mawu a akalulu oikidwa ndi LOIS anali otsika, kusonyeza kusiyana kwakukulu.Kuchepa kwa mawu a cytokine kukuwonetsa kuti kukhazikika kwa nthawi yayitali, kokhazikika kotsutsana ndi kuipitsidwa kwa LOIS sikungokhudzana ndi kuletsa kwa matenda a bakiteriya, komanso kuchepa kwa FBR, komwe kumayambitsidwa ndi macrophages kumamatira ku gawo lapansi (53, 57 ndime 58).Chifukwa chake, kuchepa kwa chitetezo chamthupi chifukwa cha chitetezo cha chitetezo cha mthupi cha LOIS kumatha kuthana ndi zotsatirapo pambuyo pa kuikidwa, monga kuyankha mochuluka kwa chitetezo chamthupi pambuyo pa opaleshoni ya pulasitiki.
(A) Chithunzi chojambula cha njira yopangira biofilm ndikufalikira pamwamba pa choyikapo cha mafupa omwe ali ndi kachilombo.eDNA, extracellular DNA.(B) Chithunzi chojambula chachitetezo cha chitetezo chamthupi pambuyo pa kuyika kwa mafupa.(C) Kudetsa kwa H & E ndi (D) Kudetsa kwa MT kwa minyewa yozungulira ya ma implants a mafupa okhala ndi zabwino komanso LOIS.IHC ya ma cytokines okhudzana ndi chitetezo chamthupi (E) TNF-α ndi (F) IL-6 ndi zithunzi zojambulidwa za akalulu opanda maliseche ndi LOIS-implanted.(G) Kuchulukitsa kwa mawu a cytokine ndi kuyeza kwa dera (** P <0.01).
The biocompatibility ya LOIS ndi zotsatira zake pa mafupa machiritso anayesedwa mu vivo pogwiritsa ntchito chithunzithunzi cha matenda [x-ray ndi micro-computed tomography (CT)] ndi osteoclast IHC.Chithunzi 6A chikuwonetsa njira yochiritsira mafupa yomwe imaphatikizapo magawo atatu osiyana: kutupa, kukonza, ndi kukonzanso.Pamene fracture ichitika, maselo otupa ndi fibroblasts adzalowa mu fupa losweka ndikuyamba kukula mu minofu ya mitsempha.Panthawi yokonzanso, ingrowth ya minofu ya mitsempha imafalikira pafupi ndi malo ophwanyika.Minofu ya mitsempha imapereka zakudya zopangira mafupa atsopano, omwe amatchedwa callus.Gawo lomaliza la machiritso a mafupa ndilo gawo lokonzanso, momwe kukula kwa callus kumachepetsedwa mpaka kukula kwa fupa lachibadwa mothandizidwa ndi kuwonjezeka kwa osteoclasts (59).Kumanganso katatu (3D) kwa malo ophwanyidwa kunkagwiritsidwa ntchito pogwiritsa ntchito ma micro-CT scans kuti muwone kusiyana kwa msinkhu wa callus mapangidwe mu gulu lirilonse.Yang'anani mbali ya fupa la femur kuti muwone makulidwe a callus yozungulira fupa losweka (Chithunzi 6, B ndi C).Ma X-ray ankagwiritsidwanso ntchito kufufuza malo ophwanyidwa a magulu onse mlungu uliwonse kuti ayang'ane njira zosiyanasiyana zotsitsimutsa mafupa mu gulu lirilonse (Chithunzi S9).Callus ndi mafupa okhwima akuwonetsedwa mu buluu / wobiriwira ndi minyanga ya njovu, motsatira.Minofu yofewa yambiri imasefedwa ndi malo oikidwiratu.Wamaliseche wabwino ndi SHP adatsimikizira kupangidwa kwa callus yaying'ono kuzungulira malo ophwanyika.Kumbali inayi, zowoneka bwino za LOIS ndi malo ophwanyika akuzunguliridwa ndi callus wandiweyani.Zithunzi za Micro-CT zimasonyeza kuti mapangidwe a callus analepheretsedwa ndi matenda a bakiteriya komanso kutupa kwa matenda.Izi ndichifukwa choti chitetezo chamthupi chimayika patsogolo machiritso a kuvulala kwa septic komwe kumachitika chifukwa cha kutupa kokhudzana ndi matenda, m'malo mochira mafupa (60).IHC ndi Tartrate-resistant Acid Phosphatase (TRAP) madontho adachitidwa kuti ayang'ane zochitika za osteoclast ndi resorption ya fupa (Chithunzi 6D) (61).Ndi ma osteoclasts ochepa okha opangidwa ndi utoto wofiirira omwe adapezeka muzamaliseche ndi SHP.Kumbali inayi, ma osteoclast ambiri omwe adatsegulidwa adawonedwa pafupi ndi mafupa amaliseche abwino komanso okhwima a LOIS.Chodabwitsa ichi chikuwonetsa kuti pamaso pa osteoclasts, callus yozungulira malo ophwanyidwa akukumana ndi kukonzanso kwachiwawa (62).Kuchuluka kwa fupa ndi mawonekedwe a osteoclast a callus adayesedwa kuti afananize kuchuluka kwa mapangidwe a callus kuzungulira malo ophwanyika m'magulu onse, kuti athe kuwerengera zotsatira za micro-CT scan ndi IHC (Chithunzi 6E, 1 ndi 2).Monga momwe zikuyembekezeredwa, zoipa zamaliseche ndi mapangidwe a callus ku LOIS zinali zapamwamba kwambiri kuposa magulu ena, kusonyeza kuti kukonzanso bwino kwa mafupa kunachitika (63).Chithunzi S10 chikuwonetsa chithunzi chowonekera cha malo opangira opaleshoni, chotsatira chodetsa cha MT cha minofu yomwe yasonkhanitsidwa pafupi ndi screw, ndi chotsatira chodetsa cha TRAP chowunikira mawonekedwe a screw-bone.Mu gawo lapansi lopanda kanthu, ma callus amphamvu ndi mapangidwe a fibrosis adawonedwa, pomwe choyikapo chopangidwa ndi LOIS chinawonetsa malo osasunthika.Mofananamo, poyerekeza ndi zoipa zamaliseche, kutsika kwa fibrosis kunawonedwa mu akalulu oikidwa ndi LOIS, monga momwe mivi yoyera imasonyezera.Kuphatikiza apo, edema yolimba (muvi wabuluu) imatha kukhala chifukwa cha chitetezo cha mthupi cha LOIS, potero kuchepetsa kutupa kwakukulu.Kupanda ndodo pafupi ndi implant ndi kuchepetsa fibrosis kumasonyeza kuti kuchotsako kumakhala kosavuta, zomwe nthawi zambiri zimabweretsa fractures kapena kutupa.Njira yochiritsira mafupa pambuyo pochotsa zowononga idawunikidwa ndi ntchito ya osteoclast pa mawonekedwe a screw-bone.Onse fupa lopanda kanthu ndi mawonekedwe a LOIS implant amatengera magawo ofanana a osteoclasts kuti apititse patsogolo machiritso a mafupa, zomwe zikuwonetsa kuti zokutira za LOIS zilibe vuto lililonse pamachiritso a mafupa kapena kuyankha kwa chitetezo chamthupi.Pofuna kutsimikizira kuti kusinthidwa kwapamwamba komwe kumachitidwa pa LOIS sikumasokoneza njira yochiritsira mafupa, kufufuza kwa X-ray kunagwiritsidwa ntchito poyerekeza machiritso a mafupa a akalulu omwe ali ndi ma ion owoneka bwino komanso masabata a 6 a LOIS implantation (Chithunzi 6F).Zotsatirazo zinasonyeza kuti poyerekeza ndi gulu losakhudzidwa lamaliseche losakhudzidwa, LOIS inasonyeza mlingo womwewo wa machiritso a mafupa, ndipo panalibe zizindikiro zoonekeratu za kupasuka (mzere wopitirira wa osteolysis) m'magulu onsewa.
(A) Chithunzi chojambula cha machiritso a mafupa pambuyo pa kusweka.(B) Kusiyana kwa mlingo wa mapangidwe a callus a gulu lililonse lapamwamba ndi (C) chithunzi cha mtanda wa malo ophwanyika.(D) MTENGO wodetsa kuti muwone mwatsatanetsatane zochitika za osteoclast ndi kupuma kwa mafupa.Kutengera ntchito ya TRAP, kupangidwa kwa callus yakunja kwa fupa la cortical kunayesedwa mochulukira ndi (E) (1) micro-CT ndi (2) ntchito ya osteoclast.(F) Masabata a 6 atayikidwa, zithunzi za X-ray za fupa losweka la fupa lowonekera (zowonetsedwa ndi rectangle yofiira) ndi LOIS (zosonyezedwa ndi rectangle ya blue dashed).Kusanthula kwachiwerengero kunachitidwa ndi njira imodzi yowunikira kusiyana (ANOVA).* P <0.05.** P <0.01.
Mwachidule, LOIS imapereka njira yatsopano yothanirana ndi mabakiteriya komanso zokutira zachitetezo cha chitetezo chamthupi pakuyika kwa mafupa.Ma implants ochiritsira ochiritsira mafupa okhala ndi SHP functionalization amawonetsa nthawi yayitali anti-biofouling katundu, koma sangathe kusunga katundu wawo kwa nthawi yayitali.The superhydrophobicity wa gawo lapansi misampha thovu mpweya pakati mabakiteriya ndi gawo lapansi, potero kupanga matumba mpweya, potero kupewa matenda bakiteriya.Komabe, chifukwa cha kufalikira kwa mpweya, matumba a mpweyawa amachotsedwa mosavuta.Kumbali inayi, LOIS yatsimikizira kuti imatha kuteteza matenda okhudzana ndi biofilm.Choncho, chifukwa cha zotsutsana ndi kukana kwa mafuta osanjikiza omwe amalowetsedwa mu mawonekedwe a micro / nano, kutupa kokhudzana ndi matenda kungapewedwe.Njira zosiyanasiyana zamakhalidwe kuphatikiza miyeso ya SEM, AFM, XPS ndi CA imagwiritsidwa ntchito kukhathamiritsa mikhalidwe yopangira LOIS.Kuphatikiza apo, LOIS itha kugwiritsidwanso ntchito kuzinthu zosiyanasiyana zachilengedwe zomwe zimagwiritsidwa ntchito kwambiri pazida zowongolera mafupa, monga PLGA, Ti, PE, POM ndi PPSU.Kenako, LOIS idayesedwa mu vitro kuti itsimikizire kuti ili ndi anti-biofouling motsutsana ndi mabakiteriya ndi zinthu zachilengedwe zokhudzana ndi kuyankha kwa chitetezo chamthupi.Zotsatira zikuwonetsa kuti ili ndi antibacterial komanso anti-biofouling zotsatira poyerekeza ndi implant yopanda kanthu.Kuphatikiza apo, LOIS ikuwonetsa mphamvu zamakina ngakhale mutagwiritsa ntchito kupsinjika kwamakina, zomwe sizingalephereke mu opaleshoni ya pulasitiki.Chifukwa chodzichiritsa chokha chamafuta omwe ali pamwamba pa micro/nano structure, LOIS idasunga bwino zinthu zake zotsutsana ndi biological.Kuti muphunzire za biocompatibility ndi antibacterial properties za LOIS mu vivo, LOIS inayikidwa mu kalulu femur kwa masabata 4.Palibe matenda a bakiteriya omwe adawonedwa mu akalulu oikidwa ndi LOIS.Kuphatikiza apo, kugwiritsa ntchito IHC kunawonetsa kuchepa kwa chitetezo cham'deralo, kuwonetsa kuti LOIS sikulepheretsa kuchiritsa kwa mafupa.LOIS imawonetsa kwambiri antibacterial komanso chitetezo chamthupi, ndipo zatsimikiziridwa kuti zimalepheretsa mapangidwe a biofilm asanayambe komanso panthawi ya opaleshoni ya mafupa, makamaka pakuphatikizika kwa mafupa.Pogwiritsa ntchito fupa la fupa la kalulu lomwe limayambitsa kuphulika kwa chikazi, zotsatira za matenda okhudzana ndi biofilm pa machiritso a mafupa opangidwa ndi implants pre-incubated implants anaphunzira mozama.Monga kafukufuku wamtsogolo, mtundu watsopano wa vivo ukufunika kuti uphunzire matenda omwe angatheke atayikidwa kuti amvetse bwino ndikupewa matenda okhudzana ndi biofilm panthawi yonse ya machiritso.Kuphatikiza apo, osteoinduction akadali vuto losathetsedwa pakuphatikizana ndi LOIS.Kufufuza kwina kumafunika kuphatikiza kuphatikiza kosankha kwa maselo osteoinductive kapena mankhwala obwezeretsanso ndi LOIS kuti athane ndi vutoli.Ponseponse, LOIS imayimira zokutira zolimbitsa thupi zolimbitsa thupi zokhala ndi mphamvu zamakina komanso anti-biofouling katundu, zomwe zimatha kuchepetsa SSI ndi zotsatira za chitetezo chamthupi.
Sambani gawo la 15mm x 15mm x 1mm 304 SS (Dong Kang M-Tech Co., Korea) mu acetone, EtOH ndi madzi a DI kwa mphindi 15 kuti muchotse zonyansa.Kuti apange mawonekedwe ang'onoang'ono / nano-level pamwamba, gawo lapansi loyeretsedwa limamizidwa mu 48% mpaka 51% HF solution (DUKSAN Corp., South Korea) pa 50 ° C.Nthawi yophika imasiyanasiyana kuchokera ku 0 mpaka 60 mphindi.Kenaka, gawo lapansi lokhazikika linatsukidwa ndi madzi osungunuka ndikuyikidwa mu 65% HNO3 (Korea DUKSAN Corp.) yankho pa 50 ° C kwa mphindi 30 kuti apange chromium oxide passivation wosanjikiza pamwamba.Pambuyo pa passivation, gawo lapansi limatsukidwa ndi madzi a deionized ndikuwumitsa kuti lipeze gawo lapansi ndi mawonekedwe osanjikiza.Kenako, gawo lapansili lidawonetsedwa ndi plasma ya okosijeni (100 W, 3 mphindi), ndipo nthawi yomweyo adamizidwa mu njira ya 8.88 mm POTS (Sigma-Aldrich, Germany) mu toluene kutentha kwa maola 12.Kenako, gawo lapansi lophimbidwa ndi POTS lidatsukidwa ndi EtOH, ndikulowetsedwa pa 150 ° C kwa maola awiri kuti mupeze POTS SAM wandiweyani.Pambuyo popaka SAM, mafuta opangira mafuta anapangidwa pa gawo lapansi pogwiritsa ntchito mafuta a perfluoropolyether (Krytox 101; DuPont, USA) ndi kuchuluka kwa 20 μm / cm 2. Musanagwiritse ntchito, sungani mafutawo kudzera mu fyuluta ya micron 0.2.Chotsani mafuta ochulukirapo popendekera pakona ya 45 ° kwa mphindi 15.Njira yopangira yomweyi idagwiritsidwanso ntchito pakuyika kwa mafupa opangidwa ndi 304 SS (mbale yotsekera ndi screw locking cortical; Dong Kang M-Tech Co., Korea).Ma implants onse a mafupa amapangidwa kuti agwirizane ndi geometry ya kalulu femur.
Maonekedwe a pamwamba a gawo lapansi ndi zoikamo za mafupa adawunikidwa ndi SEM emission ya m'munda (Yendetsani F50, FEI, USA) ndi AFM (XE-100, Park Systems, South Korea).Kukula kwapamtunda (Ra, Rq) kumayesedwa ndikuchulukitsa dera la 20 μm ndi 20 μm (n=4).Dongosolo la XPS (PHI 5000 VersaProbe, ULVAC PHI, Japan) lomwe lili ndi gwero la Al Kα X-ray lokhala ndi malo a 100μm2 linagwiritsidwa ntchito kusanthula kapangidwe kake ka mankhwala.Makina oyezera a CA okhala ndi kamera yojambula zithunzi (SmartDrop, FEMTOBIOMED, ​​​​South Korea) adagwiritsidwa ntchito kuyeza madzi a CA ndi SA.Pa muyeso uliwonse, 6 mpaka 10 μl ya madontho (madzi opangidwa ndi deionized, magazi a akavalo, EG, 30% ethanol, ndi HD) amayikidwa pamwamba kuti ayeze CA.Pamene mbali yokhotakhota ya gawo lapansi ikukwera pa liwiro la 2 ° / s (n = 4), SA imayesedwa pamene dontho likugwa.
Pseudomonas aeruginosa [American Type Culture Collection (ATCC) 27853] ndi MRSA (ATCC 25923) anagulidwa kuchokera ku ATCC (Manassas, Virginia, USA), ndipo chikhalidwe cha katundu chinasungidwa pa -80 ° C .Musanagwiritse ntchito, chikhalidwe chozizira chimakulungidwa mu msuzi wa soya wa trypsin-thawed (Komed, Korea) pa 37 ° C kwa maola 18 ndikusamutsidwa kawiri kuti ayambitse.Pambuyo pa makulitsidwe, chikhalidwecho chinali centrifuged pa 10,000 rpm kwa mphindi 10 pa 4 ° C ndikutsuka kawiri ndi yankho la PBS (pH 7.3).Chikhalidwe cha centrifuged chimasinthidwa pama mbale a agar amagazi (BAP).MRSA ndi Pseudomonas aeruginosa adakonzedwa usiku wonse ndikutukulidwa mu msuzi wa Luria-Bertani.Kuchuluka kwa Pseudomonas aeruginosa ndi MRSA mu inoculum kunatsimikiziridwa mochulukira ndi CFU ya kuyimitsidwa kwa serial dilutions pa agar.Kenako, sinthani kuchuluka kwa mabakiteriya kukhala 0,5 McFarland muyezo, womwe ndi wofanana ndi 108 CFU/ml.Ndiye kuchepetsa ntchito bakiteriya kuyimitsidwa 100 nthawi 106 CFU/ml.Kuyesa antibacterial adhesion properties, gawo lapansili lidatsekedwa pa 121 ° C kwa mphindi 15 musanagwiritse ntchito.Gawolo linasamutsidwa ku 25 ml ya kuyimitsidwa kwa bakiteriya ndikumangirira pa 37 ° C ndi kugwedeza kwakukulu (200 rpm) kwa maola 12 ndi 72.Pambuyo pa makulitsidwe, gawo lililonse limachotsedwa mu chofungatira ndikutsuka katatu ndi PBS kuchotsa mabakiteriya oyandama pamtunda.Kuti muwone biofilm pa gawo lapansi, biofilm idakhazikitsidwa ndi methanol ndikuyipitsidwa ndi 1 ml ya crimidine lalanje kwa mphindi ziwiri.Kenako microscope ya fluorescence (BX51TR, Olympus, Japan) idagwiritsidwa ntchito kujambula zithunzi za biofilm yodetsedwa.Pofuna kuwerengera biofilm pa gawo lapansi, maselo ophatikizidwa adalekanitsidwa ndi gawo lapansi ndi njira ya bead vortex, yomwe inkaonedwa kuti ndiyo njira yabwino kwambiri yochotsera mabakiteriya ophatikizidwa (n = 4).Pogwiritsa ntchito steril forceps, chotsani gawo lapansi pamalo okulirapo ndikudina chitsimecho kuti muchotse madzi ochulukirapo.Maselo omangika momasuka adachotsedwa ndikutsuka kawiri ndi PBS wosabala.Gawo lirilonse linasamutsidwa ku chubu choyesera chosabala chomwe chili ndi 9 ml ya 0.1% protein ept saline (PSW) ndi 2 g ya 20 mpaka 25 mikanda yagalasi yosabala (0.4 mpaka 0.5 mm m'mimba mwake).Kenako idasinthidwa kwa mphindi za 3 kuti ichotse ma cell kuchokera ku zitsanzo.Pambuyo pa vortexing, kuyimitsidwa kunachepetsedwa kawiri kawiri ndi 0.1% PSW, ndiyeno 0.1 ml ya dilution iliyonse idayikidwa pa BAP.Pambuyo pa maola 24 a incubation pa 37 ° C, CFU inawerengedwa pamanja.
Kwa maselo, mbewa fibroblasts NIH/3T3 (CRL-1658; American ATCC) ndi mouse macrophages RAW 264.7 (TIB-71; American ATCC) anagwiritsidwa ntchito.Gwiritsani ntchito Dulbecco's modified Eagle medium (DMEM; LM001-05, Welgene, Korea) ku chikhalidwe cha mbewa fibroblasts ndikuwonjezera ndi 10% seramu ya ng'ombe (S103-01, Welgene) ndi 1% penicillin-streptomycin (PS ; LS202-02, Welgene (Welgene) Gwiritsani ntchito DMEM ku chikhalidwe cha mbewa macrophages, chowonjezera 10% fetal bovine serum (S001-01, Welgene) ndi 1% PS Ikani gawo lapansi mu mbale ya chikhalidwe cha maselo asanu ndi limodzi , Ndipo ikani maselo pa 105 maselo / cm2. Maselowo adayikidwa usiku wonse pa 37 ° C ndi 5% CO2 Kudetsa ma cell, ma cell adayikidwa ndi 4% paraformaldehyde kwa mphindi 20 ndikuyika mu 0.5% Triton X Incubate kwa mphindi 5 mu -100 pa 37 ° C kwa mphindi 30 Pambuyo pa kuyika, gwiritsani ntchito gawo lapansi ndi 4',6-diamino-2-phenylindole (H -1200, Vector Laboratories, UK) VECTASHIELD fixation medium (n = 4 pa selo). , fluorescein, fluorescein isothiocyanate-albumin (A9771, Sigma-Aldrich, Germany) ndi madzi a m'magazi a anthu The Alexa Fluor 488-conjugated fibrinogen (F13191, Invitrogen, USA) inasungunuka mu PBS (10 mM, pH 7.4).Kuchuluka kwa albumin ndi fibrinogen kunali 1 ndi 150 μg/ml, motero.Pambuyo pa gawo lapansi Musanamize mu njira ya mapuloteni, muzimutsuka ndi PBS kuti mubwezeretse madzi pamwamba.Kenaka mizani zigawo zonse mu mbale ya zitsime zisanu ndi chimodzi zomwe zili ndi madzi a puloteni ndipo muyike pa 37 ° C kwa mphindi 30 ndi 90.Pambuyo pa kukulitsidwa, Gawoli linachotsedwa ku njira ya mapuloteni, kutsukidwa mofatsa ndi PBS 3 nthawi, ndikuyika ndi 4% paraformaldehyde (n = 4 pa mapuloteni aliwonse).Kwa calcium, sodium chloride (0.21 M) ndi potaziyamu phosphate (3.77 mM) ) Anasungunuka m'madzi opanda madzi.PH ya yankho idasinthidwa kukhala 2.0 powonjezera yankho la hydrochloride (1M).Kenako calcium chloride (5.62 mmM) idasungunuka mu yankho.Powonjezera 1M tris(hydroxymethyl) -amino Methane imasintha pH ya yankho kukhala 7.4.Miwiritsani magawo onse mu mbale ya zitsime zisanu ndi imodzi yodzazidwa ndi 1.5× calcium phosphate solution ndikuchotsa mu njirayo pakatha mphindi 30.Popaka utoto, 2 g Alizarin Red S (CI 58005) Sakanizani ndi 100 ml ya madzi osakaniza.Kenaka, gwiritsani ntchito 10% ammonium hydroxide kuti musinthe pH mpaka 4. Dyani gawo lapansi ndi njira ya Alizarin Red kwa mphindi 5, kenaka gwedezani utoto wochuluka ndikupukuta.Pambuyo pogwedeza, chotsani gawo lapansi.Zinthuzo zimachotsedwa madzi m'thupi, kenako zimamizidwa mu acetone kwa mphindi 5, kenako zimamizidwa mu njira ya acetone-xylene (1: 1) kwa mphindi 5, ndikutsukidwa ndi xylene (n = 4).Ma microscope a fluorescence (Axio Imager) okhala ndi magalasi a × 10 ndi × 20 amagwiritsidwa ntchito..A2m, Zeiss, Germany) zithunzi zonse zapansi.ImageJ/FIJI (https://imagej.nih.gov/ij/) idagwiritsidwa ntchito kuwerengera deta yomatira ya zinthu zamoyo pagulu lililonse la madera anayi osiyanasiyana ojambulira.Sinthani zithunzi zonse kukhala zithunzi zamabizinesi okhala ndi malire okhazikika kuti mufananize ndi gawo lapansi.
Zeiss LSM 700 confocal microscope idagwiritsidwa ntchito kuyang'anira kukhazikika kwa mafuta osanjikiza mu PBS mumayendedwe owunikira.Magalasi opangidwa ndi fluorine opangidwa ndi SAM okhala ndi wosanjikiza wothira mafuta adamizidwa mu yankho la PBS, ndikuyesedwa pogwiritsa ntchito orbital shaker (SHO-1D; Daihan Scientific, South Korea) pansi pamikhalidwe yogwedezeka pang'ono (120 rpm).Ndiye kutenga chitsanzo ndi kuwunika imfa ya lubricant ndi kuyeza imfa ya kunanyezimira kuwala.Kuti mupeze zithunzi za fluorescence mumawonekedwe owonetsera, chitsanzocho chimawonekera ku 633 nm laser ndiyeno nkusonkhanitsidwa, chifukwa kuwala kudzawonekeranso kuchokera ku chitsanzo.Zitsanzozo zinayesedwa panthawi ya 0, 30, 60, ndi maola 120.
Pofuna kudziwa momwe kusintha kwamadzi kumakhudzira mawonekedwe a nanomechanical a implants of orthopedic implants, nanoindenter (TI 950 TriboIndenter, Hysitron, USA) yokhala ndi nsonga ya diamondi ya Berkovich yokhala ndi mbali zitatu ya piramidi inagwiritsidwa ntchito kuyeza nanoindenedione.Kulemera kwakukulu ndi 10 mN ndipo dera ndi 100μmx 100μm.Pamiyezo yonse, nthawi yotsitsa ndi kutsitsa ndi 10 s, ndipo nthawi yogwira pansi pamtundu wapamwamba kwambiri ndi 2 s.Tengani miyeso kuchokera m'malo asanu ndikutenga avareji.Pofuna kuyesa mphamvu yamakina yomwe ili pansi pa katundu, kuyesa kwapakati pa mfundo zitatu kunachitika pogwiritsa ntchito makina oyesera padziko lonse lapansi (Instron 5966, Instron, USA).Gawo lapansi limapanikizidwa pafupipafupi 10 N / s ndikuwonjezera katundu.Pulogalamu yamapulogalamu ya Bluehill Universal (n = 3) idagwiritsidwa ntchito kuwerengera ma flexural modulus komanso kupsinjika kwakukulu.
Pofuna kutsanzira ndondomeko ya opaleshoniyo komanso kuwonongeka kwa makina komwe kumachitika panthawi ya opaleshoniyo, ntchitoyo inkachitika mu vitro.Nyamazi zinasonkhanitsidwa kuchokera ku akalulu oyera omwe anaphedwa ku New Zealand.Femur idatsukidwa ndikuyikidwa mu 4% paraformaldehyde kwa sabata imodzi.Monga tafotokozera mu njira yoyesera nyama, femur yokhazikika idachitidwa opaleshoni.Opaleshoniyo itatha, implantation ya mafupa idamizidwa m'magazi (magazi a akavalo, KISAN, Korea) kwa 10 s kuti atsimikizire ngati kuphatikizika kwa magazi kunachitika pambuyo povulala kwamakina (n = 3).
Akalulu oyera aamuna a 24 a ku New Zealand (kulemera kwa 3.0 mpaka 3.5kg, zaka zapakati pa miyezi 6) adagawidwa mwachisawawa m'magulu anayi: maliseche amaliseche, amaliseche, SHP ndi LOIS.Njira zonse zokhudzana ndi zinyama zinkachitidwa motsatira mfundo za Komiti ya Institutional Animal Care and Use Committee (IACUC yovomerezeka, KOREA-2017-0159).Kuyika kwa mafupa kumapangidwa ndi mbale yokhoma yokhala ndi mabowo asanu (utali wa 41 mm, m'lifupi 7 mm ndi makulidwe 2 mm) ndi zomangira zokhotakhota (kutalika kwa 12 mm, m'mimba mwake 2.7 mm) kuti ziwonongeke.Kupatula mbale ndi zomangira zomwe zimagwiritsidwa ntchito mu gulu lopanda kanthu, mbale zonse ndi zomangira zidakulungidwa mu kuyimitsidwa kwa MRSA (106 CFU/ml) kwa maola 12.Gulu lopanda maliseche (n = 6) linachitidwa ndi ma implants amaliseche amaliseche popanda kukhudzana ndi kuyimitsidwa kwa bakiteriya, monga kulamulira koipa kwa matenda.Gulu lopanda kanthu (n = 6) lidathandizidwa ndi choyikapo chopanda kanthu chowululidwa ndi mabakiteriya ngati njira yowongolera matenda.Gulu la SHP (n = 6) linathandizidwa ndi ma implant a SHP omwe ali ndi bakiteriya.Potsirizira pake, gulu la LOIS linathandizidwa ndi ma implants a LOIS opangidwa ndi mabakiteriya (n = 6).Zinyama zonse zimasungidwa mu khola, ndipo chakudya ndi madzi ambiri amaperekedwa.Opaleshoni isanachitike, akalulu adasala kudya kwa maola 12.Zinyamazo zinagwidwa ndi jekeseni wa intramuscular wa xylazine (5mg / kg) ndi jekeseni wa paclitaxel (3mg / kg) kuti apangidwe.Pambuyo pake, perekani 2% isoflurane ndi 50% mpaka 70% ya okosijeni yachipatala (kuthamanga kwa 2 L / min) kupyolera mu dongosolo la kupuma kuti mukhale ndi anesthesia.Imayikidwa kudzera mu njira yolunjika ku lateral femur.Pambuyo pochotsa tsitsi ndikuchotsa povidone-iodine pakhungu, kudula pafupifupi masentimita 6 kunapangidwa kunja kwa chikazi chapakati chakumanzere.Potsegula mpata pakati pa minofu yomwe imaphimba chikazi, femur imawonekera bwino.Ikani mbale kutsogolo kwa shaft yachikazi ndikuyikonza ndi zomangira zinayi.Pambuyo pokonza, gwiritsani ntchito tsamba la macheka (1 mm wandiweyani) kuti mupange chophukacho pakati pa dzenje lachiwiri ndi dzenje lachinayi.Kumapeto kwa opaleshoniyo, chilondacho chinatsukidwa ndi saline ndikutsekedwa ndi sutures.Kalulu aliyense anabayidwa jekeseni pansi pa khungu ndi enrofloxacin (5 mg/kg) kuchepetsedwa gawo limodzi mwa magawo atatu a saline.Ma X-ray a postoperative a femur adatengedwa mu nyama zonse (0, 7, 14, 21, 28, ndi masiku 42) kuti atsimikizire osteotomy ya fupa.Pambuyo pa opaleshoni yakuya, nyama zonse zidaphedwa ndi mtsempha wa KCl (2 mmol/kg) pamasiku 28 ndi 42.Pambuyo pa kuphedwa, femur inafufuzidwa ndi micro-CT kuti ione ndikufanizira machiritso a mafupa ndi mapangidwe atsopano a mafupa pakati pa magulu anayi.
Pambuyo pa kuphedwa, minyewa yofewa yomwe imalumikizana mwachindunji ndi mafupa a mafupa anasonkhanitsidwa.Minofuyo idakhazikitsidwa mu 10% yosalowerera ndale ya formalin usiku umodzi kenako imachotsedwa madzi mu EtOH.Minofu yopanda madzi inayikidwa mu parafini ndikuyika pa makulidwe a 40 μm pogwiritsa ntchito microtome (400CS; EXAKT, Germany).Kuti muwonetsetse matendawa, madontho a H&E ndi madontho a MT adachitidwa.Pofuna kuyang'ana yankho la wolandirayo, minofu yomwe ili ndi zigawozo inayikidwa ndi kalulu anti-TNF-α primary antibody (AB6671, Abcam, USA) ndi kalulu anti-IL-6 (AB6672; Abcam, USA), ndiyeno amachiritsidwa ndi horseradish.Oxidase.Ikani makina oyipitsa a avidin-biotin complex (ABC) m'magawo molingana ndi malangizo a wopanga.Kuti awoneke ngati chinthu chopangidwa ndi bulauni, 3,3-diaminobenzidine idagwiritsidwa ntchito mbali zonse.Sikina yojambulira ya digito (Pannoramic 250 Flash III, 3DHISTECH, Hungary) idagwiritsidwa ntchito powonera magawo onse, ndipo magawo osachepera anayi pagulu lililonse adawunikidwa ndi pulogalamu ya ImageJ.
Zithunzi za X-ray zinatengedwa mu zinyama zonse pambuyo pa opaleshoni ndi sabata iliyonse kuti ziwone machiritso a fracture (n = 6 pa gulu).Pambuyo pa kuphedwa, mkulu-resolution micro-CT inagwiritsidwa ntchito kuwerengera mapangidwe a callus mozungulira femur pambuyo pochiritsa.Femur yomwe idapezedwa idatsukidwa, kuyikidwa mu 4% paraformaldehyde kwa masiku atatu, ndikuthiridwa madzi mu 75% ethanol.Mafupa opanda madziwa adafufuzidwa pogwiritsa ntchito micro-CT (SkyScan 1173, Brooke Micro-CT, Kandy, Belgium) kuti apange zithunzi za 3D voxel (2240 ​​× 2240 pixels) za chitsanzo cha fupa.Gwiritsani ntchito fyuluta ya 1.0 mm Al kuti muchepetse phokoso lazizindikiro ndikugwiritsa ntchito kusanja kwakukulu pamasikidwe onse (E = 133 kVp, I = 60 μA, nthawi yophatikiza = 500 ms).Mapulogalamu a Nrecon (mtundu wa 1.6.9.8, Bruker microCT, Kontich, Belgium) adagwiritsidwa ntchito kupanga voliyumu ya 3D ya zitsanzo zojambulidwa kuchokera ku 2D lateral projection yomwe inapezedwa.Kuti muwunike, chithunzi chomangidwanso cha 3D chimagawidwa mu cubes 10mm × 10mm × 10mm malinga ndi malo ophwanyika.Werengani callus kunja kwa fupa la kortical.Mapulogalamu a DataViewer (mtundu wa 1.5.1.2; Bruker microCT, Kontich, Belgium) adagwiritsidwa ntchito kuti alowetsenso voliyumu ya fupa yojambulidwa, ndipo mapulogalamu a CT-Analyzer (mtundu 1.14.4.1; Bruker microCT, Kontich, Belgium) anagwiritsidwa ntchito pofufuza.Ma coefficients amtundu wa x-ray mu mafupa okhwima ndi callus amasiyanitsidwa ndi kachulukidwe kake, ndiyeno kuchuluka kwa callus kumawerengedwa (n = 4).Pofuna kutsimikizira kuti biocompatibility ya LOIS sikuchedwa kuchiritsa mafupa, kusanthula kowonjezera kwa X-ray ndi micro-CT kunachitika mu akalulu awiri: magulu amaliseche ndi LOIS.Magulu onsewa anaphedwa pa sabata lachisanu ndi chimodzi.
Nyama zamphongo zochokera ku nyama zoperekedwa nsembe zinasonkhanitsidwa ndikuyikidwa mu 4% paraformaldehyde kwa masiku atatu.Kuyika kwa mafupa kumachotsedwa mosamala kuchokera ku chikazi.Femur idasinthidwa kwa masiku 21 pogwiritsa ntchito 0.5 M EDTA (EC-900, National Diagnostics Corporation).Kenako femur yochotsedwa idamizidwa mu EtOH kuti ikhale yopanda madzi.Chikazi chopanda madzi chinachotsedwa mu xylene ndikuyikidwa mu parafini.Kenako chitsanzocho chidadulidwa ndi ma rotary microtome (Leica RM2255, Leica Biosystems, Germany) yokhala ndi makulidwe a 3 μm.Kwa madontho a TRAP (F6760, Sigma-Aldrich, Germany), zitsanzo zogawanikazo zinachotsedwa, kubwezeretsedwa ndi kulowetsedwa mu TRAP reagent pa 37 ° C kwa ola la 1.Zithunzi zinapezedwa pogwiritsa ntchito sikisini ya silaidi (Pannoramic 250 Flash III, 3DHISTECH, Hungary) ndi kuyesedwa poyesa kufalikira kwa dera lothimbirira.Pakuyesa kulikonse, magawo osachepera anayi pagulu lililonse adawunikidwa ndi pulogalamu ya ImageJ.
Kusanthula kufunikira kwachiwerengero kudachitika pogwiritsa ntchito GraphPad Prism (GraphPad Software Inc., USA).Mayeso osayembekezeka a t ndi kusanthula kwa njira imodzi (ANOVA) adagwiritsidwa ntchito kuyesa kusiyana pakati pa magulu owunika.Mulingo wofunikira ukuwonetsedwa pachithunzichi motere: *P<0.05, **P<0.01, ***P<0.001 ndi ****P<0.0001;NS, palibe kusiyana kwakukulu.
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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
Zovala za antibacterial ndi chitetezo cha mthupi zomwe zimapangidwira mafupa zimatha kuchepetsa matenda ndi mayankho a chitetezo chamthupi omwe amayamba chifukwa cha matenda.
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
Zovala za antibacterial ndi chitetezo cha mthupi zomwe zimapangidwira mafupa zimatha kuchepetsa matenda ndi mayankho a chitetezo chamthupi omwe amayamba chifukwa cha matenda.
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