Maka ndị ọrịa na-arụ ọrụ ịwa ahụ ọkpụkpụ ọkpụkpụ, ọrịa nje bacteria na nzaghachi mgbochi ọrịa na-ebute bụ mgbe niile ihe egwu na-eyi ndụ egwu.Ihe ndị na-eme ihe ndị dị ndụ na-enwekarị mmerụ ahụ, nke na-eme ka nje bacteria na-abanye n'ógbè ahụ merụrụ ahụ ma na-ebute ọrịa mgbe ịwachara ahụ.Ya mere, ọ dị mkpa ngwa ngwa ịmepụta mgbochi ọrịa na mgbochi mgbapụ mkpuchi maka ọkpụkpụ orthopedic.N'ebe a, anyị emepụtala nkà na ụzụ mgbanwe elu elu maka ọkpụkpụ ọkpụkpụ nke a na-akpọ Lubricated Orthopedic Implant Surface (LOIS), bụ nke sitere n'ike mmụọ nsọ dị nro nke ndị na-akụ olulu mmiri.LOIS nwere mmiri mmiri na-adịte aka na nke siri ike maka ụdị mmiri dị iche iche na ihe ndị dị ndụ (gụnyere mkpụrụ ndụ, protein, calcium na bacteria).Na mgbakwunye, anyị kwadoro ịdịte aka n'ibu megide ncha na idozi ike site n'ime ka mmebi ahụ na-agaghị ekwe omume n'oge ịwa ahụ in vitro.A na-eji ụdị mgbawa femoral na-egbuke egbuke nke oke bekee mee ihe iji mụọ nke ọma mgbochi nje na-egbochi ọrịa nke LOIS.Anyị na-eche na LOIS, nke nwere ihe na-egbochi biofouling na ịdịte aka n'ibu, bụ nzọụkwụ na-aga n'ihu na ịwa ahụ ọkpụkpụ na-enweghị ọrịa.
Taa, n'ihi ịka nká n'ozuzu, ọnụ ọgụgụ ndị ọrịa na-arịa ọrịa orthopedic (dị ka mgbaji ndị agadi, ọrịa nkwonkwo degenerative, na ọkpụkpụ ọkpụkpụ) abawanyela nke ukwuu (1, 2).Ya mere, ụlọ ọrụ ahụike na-etinye aka dị ukwuu na ịwa ahụ ọkpụkpụ, gụnyere ọkpụkpụ ọkpụkpụ nke skru, efere, mbọ na nkwonkwo artificial (3, 4).Otú ọ dị, a na-akọ na ọkpụkpụ ọkpụkpụ omenala na-enwe ike ịnweta adhesion bacteria na nhazi biofilm, nke nwere ike ịkpata ọrịa ebe ịwa ahụ (SSI) mgbe a wachara ya (5, 6).Ozugbo a na-emepụta biofilm n'elu ihe ọkpụkpụ nke ọkpụkpụ, mwepụ nke biofilm na-esiwanye ike ọbụna na iji nnukwu ọgwụ nje mee ihe.Ya mere, ọ na-ebutekarị ọrịa na-efe efe mgbe ịwachara ahụ (7, 8).N'ihi nsogbu ndị a dị n'elu, ọgwụgwọ nke ihe ndị na-ebute ọrịa kwesịrị ịgụnye ịmaliteghachi ọrụ, gụnyere iwepụ ihe niile etinyere na anụ ahụ gbara ya gburugburu;ya mere, onye ọrịa ahụ ga-enwe nnukwu mgbu na ụfọdụ ihe ize ndụ (9, 10).
Iji dozie ụfọdụ n'ime nsogbu ndị a, a na-emepụta ọgwụ orthopedic na-emepụta ọgwụ iji gbochie ọrịa site n'iwepụ nje bacteria ndị dị n'elu (11, 12).Agbanyeghị, atụmatụ a ka na-egosi oke oke.A kọwawo na ntinye ogologo oge nke ihe ndị na-emepụta ọgwụ na-emepụta ihe na-eme ka ọkpụkpụ dị gburugburu na-eme ka ọ bụrụ ihe na-egbuke egbuke, nke nwere ike iduga necrosis (13, 14).Na mgbakwunye, ihe mgbaze organic nwere ike ịdị mgbe emechara usoro nrụpụta orthopedic nke ọgwụ na-eme ka ọ bụrụ nke US Food and Drug Administration machibidoro, chọrọ usoro nchacha ọzọ iji mezuo ụkpụrụ ya (15).Ihe ntinye ọgwụ na-eme ka ọ bụrụ ihe ịma aka maka ntọhapụ nke ọgwụ na-achịkwa, na n'ihi oke ọgwụ ha na-ebu, ngwa ngwa nke ọgwụ agaghị ekwe omume (16).
Usoro ọzọ a na-ahụkarị bụ iji ihe na-eme ihe na-eme ihe na-eme ka ọ ghara ịkụnye ihe n'osisi ahụ iji gbochie ihe ndị dị ndụ na nje bacteria ịrapara n'elu (17).Dịka ọmụmaatụ, ndị polymers zwitterionic adọtala nlebara anya n'ihi ihe ndị na-adịghị ejikọta ha mgbe ha na protein, sel, na nje bacteria na-ejikọta na plasma.Otú ọ dị, ọ nwere ụfọdụ njedebe metụtara nkwụsi ike na-adịte aka na nkwụsi ike nke ígwè ọrụ, nke na-egbochi ngwa ya bara uru na orthopedic implants, karịsịa n'ihi nchacha ígwè n'oge usoro ịwa ahụ (18, 19).Tụkwasị na nke ahụ, n'ihi ịdị elu ya dị elu, enweghị mkpa maka ịwa ahụ na-ewepu, na ihe ndị na-ehicha ihe dị n'elu site na corrosion, a na-eji ihe ndị na-emepụta ihe na-emepụta orthopedic mee ihe (20, 21).N'oge corrosion, kemịkalụ kemịkalụ dị n'etiti polymer matrix na-agbajikwa ma kewapụ ya n'elu, na ndị na-akwado ya na-ehicha elu.Otú ọ dị, mgbochi nje nje site na ihicha elu dị irè n'ime obere oge.Na mgbakwunye, ọtụtụ ihe ndị na-anabata ya gụnyere poly (lactic acid-glycolic acid copolymer) (PLGA), polylactic acid (PLA) na alloys dabeere na magnesium ga-enweta mbibi na-enweghị isi na mbibi na ahụ, nke ga-emetụta nkwụsi ike n'ibu.(iri abụọ na abụọ).Tụkwasị na nke ahụ, iberibe efere ndị dị ndụ na-enye ebe nje bacteria na-ejikọta, nke na-eme ka ohere nke ibute ọrịa na-adịte aka.Ihe ize ndụ a nke mmebi akụrụngwa na ọrịa na-egbochi ngwa bara uru nke ịwa ahụ plastik (23).
Elu Superhydrophobic (SHP) nke na-eṅomi usoro nhazi nke akwụkwọ lotus aghọwo ihe ngwọta maka ebe ndị na-adịghị mma (24, 25).Mgbe elu SHP na-emikpu n'ime mmiri mmiri, ikuku ikuku ga-abụ ọnyà, si otú ahụ na-akpụ akpa ikuku na igbochi nje bacteria (26).Otú ọ dị, nnyocha ndị e mere n'oge na-adịbeghị anya egosiwo na elu SHP nwere ihe ndị na-adịghị mma metụtara ịdịte aka n'ibu na nkwụsi ike ogologo oge, nke na-egbochi ngwa ya na ntinye ọgwụ.Ọzọkwa, akpa ikuku ga-agbaze ma tụfuo ihe ndị na-emebi emebi, si otú a na-eme ka adhesion nje gbasaa karịa n'ihi nnukwu elu nke SHP (27, 28).N'oge na-adịbeghị anya, Aizenberg na ndị ọrụ ibe webatara usoro ọhụrụ nke mgbochi biofouling n'elu mkpuchi site na ịmepụta elu dị nro sitere n'ike mmụọ nsọ nke Nepenthes pitcher osisi (29, 30).Elu dị larịị na-egosi nkwụsi ike ogologo oge n'okpuru ọnọdụ hydraulic, bụ oke mmiri na-egbochi mmiri mmiri ndụ, ma nwee ihe na-edozi onwe ya.Otú ọ dị, ọ dịghị ụzọ a ga-esi tinye mkpuchi na ntinye ọgwụ na-edozi ahụ, ma ọ bụ na-egosi na ọ kwadoro usoro ọgwụgwọ nke anụ ahụ mebiri emebi mgbe etinyere ya.
N'ebe a, anyị na-ewebata lubricated orthopedic implant surface (LOIS), micro/nano-structured orthopedic implant surface and tightly jikọtara ya na obere mmanu mmanu oyi akwa iji gbochie ya njikọ na plastic ịwa ahụ nje nje nje, dị ka mgbaji na-edozi.N'ihi na Fluorine-functionalized micro/nano-level Ọdịdị na-edozi mmanu na nhazi ahụ, LOIS nke mepere emepe nwere ike ịkwụsị nchikota nke mmiri dị iche iche ma nọgide na-arụ ọrụ mgbochi ruo ogologo oge.Enwere ike itinye mkpuchi LOIS na ihe dị iche iche ezubere maka njikọ ọkpụkpụ.Ihe kacha mma na-egbochi biofouling nke LOIS megide nje bacteria biofilm [Pseudomonas aeruginosa na methicillin na-eguzogide Staphylococcus aureus (MRSA)] na ihe ndị dị ndụ (sel, protein na calcium) ka akwadoro na vitro.Ọnụọgụ adhesion nke nnukwu nnabata na mkpụrụ bụ ihe na-erughị 1%.Na mgbakwunye, ọbụlagodi mgbe nrụgide n'ibu dị ka ịcha elu na-apụta, ọgwụgwọ onwe onye nke mmanụ na-abanye n'ime ya na-enyere aka ịnọgide na-enwe ihe ndị na-emebi emebi.Nsonaazụ ule ịdịte aka n'ibu na-egosi na ọ bụrụgodị na emezigharị nhazi na kemịkal, ike agaghị ebelata nke ukwuu.Tụkwasị na nke ahụ, a na-eme nnwale in vitro nke na-eme ka nrụgide ọrụ na gburugburu ebe ịwa ahụ pụta iji gosi na LOIS nwere ike iguzogide nrụgide ọrụ dị iche iche nke na-eme n'oge ịwa ahụ plastik.N'ikpeazụ, anyị na-eji oke bekee dabere na vivo femoral fracture model, nke gosipụtara na LOIS nwere ihe ndị na-egbu egbu na-egbu egbu na biocompatibility.Nsonaazụ redio na akụkọ ihe mere eme gosipụtara na omume mmanu mmanu kwụsiri ike na ihe mgbochi biofouling n'ime izu 4 ka etinyechara ya nwere ike nweta ọrụ mgbochi ọrịa dị mma yana mgbanarị mgbapụ na-egbu oge na usoro ọgwụgwọ ọkpụkpụ.
Ọgụgụ 1A na-egosi eserese schematic nke LOIS mepere emepe, bụ nke etinyere ya na ihe owuwu micro/nano na ụdị mgbawa femoral nke oke bekee iji kwado ọmarịcha ihe nhụsianya nke ndu na ihe mgbochi ọrịa.A na-eme usoro biomimetic iji mee ka elu nke osisi ite mmiri, na igbochi biofouling site n'itinye akwa mmanu mmanu n'ime micro/nano nhazi nke elu.N'elu ịgbanye mmanụ mmanu nwere ike ibelata kọntaktị dị n'etiti ihe ndị dị ndụ na elu.Ya mere, n'ihi nguzobe nke ike kemịkalụ agbụ n'elu, ọ nwere magburu onwe antifouling arụmọrụ na ogologo oge kwụsie ike.N'ihi ya, ihe mgbochi biofouling nke elu mmanu mmanu na-enye ohere dị iche iche dị irè na nyocha biomedical.Agbanyeghị, nyocha sara mbara banyere etu ihu elu a pụrụ iche si emekọrịta ihe n'ime ahụ emezubeghị.Site n'ịtụle LOIS na ihe ndị gba ọtọ na vitro na-eji albumin na nje bacteria biofilm, enwere ike ikwenye na anaghị ejikọta LOIS (Njirimara 1B).Na mgbakwunye, site n'ịkwapụ ụmụ irighiri mmiri na mkpụrụ osisi na-enweghị isi na mkpụrụ osisi LOIS (Figure S1 na Movie S1), enwere ike igosipụta arụmọrụ mmetọ nke ndu.Dị ka egosiri na foto microscope nke fluorescence, mkpụrụ ekpughere ekpughere na nkwusioru nke protein na nje bacteria gosipụtara nnukwu ihe dị ndụ na-arapara n'elu.Agbanyeghị, n'ihi ọmarịcha ihe mgbochi biofouling ya, LOIS anaghị egosipụta ọ bụla fluorescence.Iji kwado ihe mgbochi biofouling na mgbochi ọrịa, a na-etinye LOIS n'elu ihe ọkpụkpụ nke ọkpụkpụ maka ọkpụkpụ ọkpụkpụ (efere na kposara) ma tinye ya n'ụdị mgbawa oke bekee.Tupu etinye ya, a na-etinye ọkpụkpụ orthopedic gba ọtọ na LOIS na nkwụsị nke nje maka awa 12.Mgbochi nke mbụ na-eme ka a hụ na a na-emepụta biofilm n'elu ihe a na-ekpuchi anya maka ntụnyere.Ọgụgụ 1C na-egosi foto nke saịtị mgbawa 4 izu mgbe etinyere ya.N'aka ekpe, oke bekee nke nwere orthopedic na-enweghị ihe ọ bụla gosipụtara oke ọkụ n'ihi nhazi nke biofilm n'elu ihe a na-etinye.Achọpụtara ihe dị iche na oke bekee etinyere na LOIS, ya bụ, anụ ahụ gbara gburugburu nke LOIS egosighi akara nke ọrịa ma ọ bụ ihe mgbaàmà nke mbufụt.Tụkwasị na nke ahụ, ihe oyiyi anya nke dị n'aka ekpe na-egosi ebe ịwa ahụ nke oke bekee na-ekpuchi ya, na-egosi na ọ dịghị ọtụtụ ihe nrịbama dị n'elu ebe a na-ahụ anya n'elu LOIS.Nke a na-egosi na LOIS nwere nkwụsi ike na-adịte aka ma nwee ike ịnọgide na-enwe ihe ndị na-emebi ihe na-emebi ihe na ihe mgbochi.
(A) Eserese eserese nke LOIS na ntinye ya na ụdị mgbawa femoral oke bekee.(B) Ihe onyonyo nke fluorescence microscopy nke protein na biofilm nje na-efe efe n'elu ala na mkpụrụ osisi LOIS.Izu 4 ka etinyechara ya, (C) onyonyo onyonyo nke saịtị mgbaji na (D) onyonyo X-ray (nke akụkụ anọ na-acha uhie uhie gosipụtara).Foto site n'ikike: Kyomin Chae, Mahadum Yonsei.
The igba ogwu, kpughere na-adịghị mma kụnyere oke bekee gosiri a nkịtị ọkpụkpụ ọgwụgwọ usoro na-enweghị ihe ọ bụla ihe ịrịba ama nke mbufụt ma ọ bụ ọrịa.N'aka nke ọzọ, SHP implants tupu etinye ya na nkwusioru nje na-egosipụta mbufụt metụtara ọrịa na anụ ahụ gbara ya gburugburu.Enwere ike ịsị na nke a bụ enweghị ike igbochi nje bacteria ogologo oge (Figure S2).Iji gosi na LOIS adịghị emetụta usoro ọgwụgwọ ahụ, ma na-egbochi ọrịa ndị nwere ike ime ka ọ bụrụ ihe ọkụkụ, ihe oyiyi X-ray nke matrix dị mma ekpughere na LOIS na ebe mgbawa ka atụnyere (Njirimara 1D).Onyonyo X-ray nke etinyere nke ọma na-egosi na ahịrị osteolysis na-aga n'ihu, na-egosi na ọkpụkpụ ahụ agwọbeghị kpamkpam.Nke a na-egosi na usoro mgbake ọkpụkpụ nwere ike ịdị ogologo oge n'ihi mbufụt metụtara ọrịa.N'ụzọ megidere nke ahụ, o gosiri na oke bekee etinyere na LOIS agbakeela ma egosighi ebe mgbawa doro anya.
Iji zụlite ihe ntinye ọgwụ na nkwụsi ike na ọrụ ogologo oge (gụnyere iguzogide biofouling), emeela ọtụtụ mgbalị.Otú ọ dị, ọnụnọ nke ihe ndị dị ndụ dị iche iche na ihe dị iche iche nke adhesion anụ ahụ na-egbochi mmepe nke ụzọ ha nwere ike ịdabere na ụlọ ọgwụ.Iji merie adịghị ike ndị a, anyị emepụtala ihe owuwu micro/nano na-agbanwe agbanwe na elu nke kemịkalụ, bụ nke kachasị mma n'ihi ike capillary dị elu na njikọ kemịkalụ iji mee ka mmanu na-egbuke egbuke ruo n'ókè kasị ukwuu.Ọgụgụ 2A na-egosi usoro nrụpụta n'ozuzu nke LOIS.Mbụ, kwadebe ọgwụ igwe anaghị agba nchara (SS) 304 mkpụrụ.Nke abuo, a na-etolite ihe owuwu micro/nano na SS substrate site na etching kemịkalụ site na iji ngwọta hydrofluoric acid (HF).Iji weghachi nguzogide corrosion nke SS, a na-eji ihe ngwọta nitric acid (HNO3) (31) mee ihe iji hazie mkpụrụ osisi etched.Passivation na-akwalite nguzogide corrosion nke SS substrate ma na-ebelata usoro corrosion nke nwere ike ibelata arụmọrụ LOIS n'ozuzu ya.Mgbe ahụ, site n'ịmepụta monolayer nke onwe ya (SAM) na 1H, 1H, 2H, 2H-perfluorooctyltriethoxysilane (POTS), a na-emezi elu elu nke kemịkalụ iji meziwanye mmekọrịta kemịkalụ n'etiti elu na ezigbo mmanu mmanu Affinity.Mgbanwe nke elu na-ebelata ike elu nke elu a haziri ahazi nke micro/nano, nke dabara na ike elu nke mmanu dị nro.Nke a na-enye ohere ka mmiri na-agba mmiri kpamkpam, si otú ahụ na-eme ka oyi akwa siri ike n'elu.N'elu gbanwetụrụ na-egosipụta ike hydrophobicity.Nsonaazụ na-egosi na mmanye na-amị amị na-egosipụta omume kwụsiri ike na LOIS n'ihi njikọ kemịkalụ dị elu na ike capillary nke micro/nano Ọdịdị kpatara (32, 33).Mgbanwe ngwa anya dị n'elu SS mgbe emezigharị elu na ntụtụ mmanu mmanu amụrụ.Ọdịdị micro/nano oyi akwa nke emebere n'elu nwere ike ime mgbanwe anya ma mee ka elu gbaa ọchịchịrị.A na-ekwu na ihe omume a bụ n'ihi mmụba ọkụ na-agbasawanye n'elu ala ahụ siri ike, nke na-eme ka nlegharị anya na-agbasa site na usoro ọnyà ọkụ (34).Na mgbakwunye, mgbe agbachara mmanụ mmanu ahụ, LOIS na-agba ọchịchịrị.Okpokoro mmanu mmanu na-eme ka obere ìhè pụta ìhè site na mkpụrụ, si otú ahụ na-agbaji LOIS.Iji bulite microstructure/nanostructure iji gosi obere sliding angle (SA) iji nweta arụmọrụ mgbochi biofouling, a na-eji nyocha eletrọnịkị microscopy (SEM) na atomiki abụọ mee oge HF etching dị iche iche (0, 3)., 15 na 60 nkeji) Force Microscope (AFM) (Foto 2B).Onyonyo SEM na AFM na-egosi na mgbe obere oge etching (nkeji 3 nke etching), mkpụrụ osisi ahụ na-enweghị isi etolitela nano na-enweghị isi.Ọkpụkpụ elu na-agbanwe site na oge etching (Figure S3).Ogologo oge na-agbanwe agbanwe na-egosi na nhụsianya elu na-aga n'ihu na-abawanye ma rute ọnụ ọgụgụ kasị elu na nkeji iri na ise nke etching, na mgbe ahụ, ọ bụ naanị ntakịrị mbelata na uru adịghị ike ka a na-ahụ na nkeji 30 nke etching.N'oge a, nano-larịị adịghị ike na-apụ apụ, ebe ọkpụkpụ micro-level na-etolite nke ọma, na-eme ka ọkpụkpụ na-agbanwe agbanwe.Mgbe etching maka ihe karịrị nkeji 30, a na-ahụ mmụba ọzọ na ike siri ike, nke a kọwara n'ụzọ zuru ezu dị ka ndị a: SS bụ ígwè, nke nwere ihe ndị gụnyere ígwè, chromium, nickel, molybdenum na ọtụtụ ihe ndị ọzọ.N'ime ihe ndị a, ígwè, chromium na molybdenum na-ekere òkè dị mkpa n'ịmepụta micron/nano-scale roughness na SS site na HF etching.Na mmalite nke corrosion, ígwè na chromium na-abụkarị corrosion n'ihi na molybdenum nwere nguzogide corrosion dị elu karịa molybdenum.Ka etching ahụ na-aga n'ihu, ngwọta etching na-erute oke mmiri ozuzo na mpaghara, na-emepụta fluorides na oxides nke etching kpatara.Fluoride na oxide na-ebuli ma mechaa depụtaghachi n'elu, na-akpụ akpụ n'elu na oke micron/nano (31).Nke a micro/nano-level roughness na-arụ ọrụ dị mkpa na njirimara ọgwụgwọ onwe nke LOIS.Igwe elu okpukpu abụọ na-emepụta mmetụta synergistic, na-abawanye ike capillary nke ukwuu.Ihe omume a na-eme ka mmanu mmanu banye n'elu nke ọma ma na-enye aka na-agwọ onwe ya (35).Nhazi nke roughness na-adabere na oge etching.N'okpuru nkeji 10 nke etching, elu nwere naanị nano-scale roughness, nke na-ezughị iji jide mmanu zuru ezu iji nwee nguzogide biofouling (36).N'aka nke ọzọ, ọ bụrụ na oge etching gafere 30 nkeji, nano-scale roughness kpụrụ site redeposition nke ígwè na chromium ga-apụ n'anya, na ọ bụ naanị micro-scale roughness ga-anọgide n'ihi molybdenum.N'elu-etched elu enweghị nano-ọnụ ọgụgụ roughness ma na-efunahụ synergistic mmetụta nke abụọ-ogbo roughness, nke na-adịghị emetụta ndị na-agwọ onwe àgwà nke LOIS.A na-eme nha SA na ihe ndị nwere oge etching dị iche iche iji gosipụta arụmọrụ mgbochi.A na-ahọrọ ụdị mmiri dị iche iche dabere na viscosity na ike elu, gụnyere mmiri deionized (DI), ọbara, ethylene glycol (EG), ethanol (EtOH) na hexadecane (HD) (Figure S4).Usoro etching nke na-agbanwe oge na-egosi na maka mmiri mmiri dị iche iche nwere ike dị iche iche na viscosities dị iche iche, SA nke LOIS mgbe nkeji iri na ise nke etching gasịrị bụ nke kasị ala.Ya mere, a na-ahazi LOIS ka ọ na-etch maka nkeji 15 iji mepụta micron na nano-scale roughness, nke kwesịrị ekwesị maka ịnọgide na-adịte aka nke mmanu mmanu na ihe ndị na-adịghị mma.
(A) Eserese eserese nke usoro nrụpụta nzọụkwụ anọ nke LOIS.Ntinye ahụ na-egosi SAM emebere na mkpụrụ.(B) Ihe onyonyo SEM na AFM, nke a na-eji na-ebuli usoro micro/nano nke mkpụrụ n'okpuru oge etching dị iche iche.X-ray photoelectron spectroscopy (XPS) ụdịdị (C) Cr2p na (D) F1s mgbe elu passivation na SAM mkpuchi.au, ngalaba aka ike.(E) Onyonyo nnochite anya nke ụmụ irighiri mmiri n'elu ihe efu, etched, SHP na LOIS substrates.(F) Igwe kọntaktị (CA) na nha SA nke mmiri mmiri nwere esemokwu dị iche iche na SHP na LOIS.A na-akọwapụta data dị ka ọ pụtara ± SD.
Mgbe ahụ, iji gosi na mgbanwe nke chemical Njirimara nke elu, X-ray photoelectron spectroscopy (XPS) na-eji na-amụ mgbanwe na chemical mejupụtara nke substrate elu mgbe ọ bụla mkpuchi mkpuchi.Ọgụgụ 2C na-egosi nsonaazụ nha XPS nke elu HF etched na elu HNO 3 mesoro.Enwere ike ịpụta ọnụ ọnụ abụọ kachasị elu na 587.3 na 577.7 eV na njikọ Cr-O dị na oyi akwa chromium oxide, nke bụ isi ihe dị iche na elu HF etched.Nke a na-abụkarị n'ihi oriri nke ígwè na chromium fluoride n'elu site na HNO3.Etching dabeere na HNO3 na-enye ohere ka chromium mepụta oyi akwa oxide na-agafe agafe n'elu, nke na-eme ka SS etched ọzọ na-eguzogide corrosion.Na eserese 2D, enwetara spectra XPS iji gosi na e guzobere silane nke sitere na fluorocarbon n'elu mgbe mkpuchi SAM gasịrị, nke nwere oke mmiri na-asọ oyi ọbụlagodi EG, ọbara na EtOH.A na-emecha mkpuchi SAM site na ịmeghachi ìgwè ndị na-arụ ọrụ silane na ìgwè hydroxyl nke e guzobere site na ọgwụgwọ plasma.N'ihi ya, a hụrụ mmụba dị ukwuu na ọnụ ọgụgụ CF2 na CF3.Ike njikọ dị n'etiti 286 na 296 eV na-egosi na emechala mgbanwe kemịkalụ nke ọma site na mkpuchi SAM.SHP na-egosi nnukwu CF2 (290.1 ​​eV) na CF3 (293.3 eV), nke sitere na silane nke dabeere na fluorocarbon kpụrụ n'elu.Ọgụgụ 2E na-egosi nha anya ihe nleba anya nke akụkụ kọntaktị (CA) maka otu dị iche iche nke mmiri deionized na kọntaktị na ihe efu, etched, SHP, na LOIS.Onyonyo ndị a na-egosi na elu etched na-aghọ hydrophilic n'ihi nhazi micro/nano nke etching kemịkalụ na-eme ka mmiri na-amịpụta na-abanye n'ime ihe owuwu ahụ.Otú ọ dị, mgbe a na-ekpuchi ihe nkedo ahụ na SAM, ihe ntinye ahụ na-egosiputa mmiri siri ike, ya mere a na-emepụta SHP elu na mpaghara kọntaktị n'etiti mmiri na elu dị ntakịrị.N'ikpeazụ, a hụrụ mbelata CA na LOIS, nke nwere ike ịsị na ntinye nke mmanu mmanu n'ime microstructure, si otú a na-amụba mpaghara kọntaktị.Iji gosi na elu ahụ nwere ezigbo mmiri na-agbapụta mmiri na ihe ndị na-adịghị mma, e jiri ya tụnyere LOIS na SHP substrate site n'iji CA na SA na-eji mmiri dị iche iche (Njirimara 2F).A na-ahọrọ ụdị mmiri dị iche iche dabere na viscosity na ike elu, gụnyere mmiri deionized, ọbara, EG, EtOH na HD (Figure S4).Nsonaazụ nha CA na-egosi na mgbe CA na-eche HD, uru mbelata nke CA, ebe CA nwere ike elu kacha ala.Na mgbakwunye, LOIS nke mkpokọta CA dị ala.Agbanyeghị, nha SA na-egosi ihe dị iche kpamkpam.Ewezuga mmiri ionized, mmiri niile na-arapara na mkpụrụ SHP na-enweghị isi.N'aka nke ọzọ, LOIS na-egosi SA dị ala, ebe mgbe mmiri mmiri niile na-atụgharị n'akụkụ dị ala karịa 10 ° ruo 15 °, mmiri mmiri niile ga-apụ.Nke a na-egosi nke ọma na enweghị nrapado nke LOIS dị mma karịa nke elu SHP.Tụkwasị na nke ahụ, a na-etinyekwa mkpuchi LOIS n'ụdị ihe dị iche iche, gụnyere titanium (Ti), polyphenylsulfone (PPSU), polyoxymethylene (POM), polyether ether ketone (PEEK) na polymers bioabsorbable (PLGA), Ha bụ ihe ọkpụkpụ ọkpụkpụ (Figure). S5)).Onyonyo dị n'usoro nke ụmụ irighiri mmiri na ihe ndị LOIS na-emeso na-egosi na ihe mgbochi biofouling nke LOIS bụ otu na ihe niile.Tụkwasị na nke ahụ, nsonaazụ nha nke CA na SA na-egosi na a pụrụ itinye ihe ndị na-adịghị mma nke LOIS na ihe ndị ọzọ.
Iji kwado njirimara mgbochi mmebi nke LOIS, ụdị mkpụrụ dị iche iche (gụnyere bare, etched, SHP na LOIS) ka etinyere na Pseudomonas aeruginosa na MRSA.A họọrọ nje bacteria abụọ a dị ka nje bacteria na-anọchite anya ụlọ ọgwụ, nke nwere ike iduga n'ichepụta biofilms, na-eduga na SSI (37).Onyonyo 3 (A na B) na-egosi onyonyo microscope fluorescence yana nsonaazụ nrụpụta nke okpuru ọchịchị agbakwunyere na nkwusioru nje maka obere oge (awa 12) na ogologo oge (awa 72), n'otu n'otu.N'ime oge dị mkpirikpi, nje bacteria ga-etolite ụyọkọ ma na-eto eto, na-ekpuchi onwe ha na ihe ndị yiri imi ma gbochie iwepụ ha.Otú ọ dị, n'ime awa iri asaa na abụọ, nje bacteria ahụ ga-eto eto ma dị mfe ịgbasa ka ha wee nwee ọtụtụ ógbè ma ọ bụ ụyọkọ.Ya mere, enwere ike iche na ntinye awa 72 na-adị ogologo oge ma bụrụ oge ntinye nke kwesịrị ekwesị iji mepụta biofilm siri ike n'elu (38).N'ime oge dị mkpirikpi, elu etched na elu SHP gosipụtara adhesion nje bacteria, bụ nke belatara ihe dịka 25% ruo 50% ma e jiri ya tụnyere ihe na-adịghị ahụkebe.Otú ọ dị, n'ihi arụmọrụ na nkwụsi ike ya dị mma, LOIS egosighi adhesion biofilm nke nje na obere oge na ogologo oge.Eserese schematic (Onyonyo 3C) na-akọwa nkọwa nke usoro mmebi ihe mgbochi nke ihe ngwọta etching, SHP na LOIS.Echiche a bụ na mkpụrụ osisi etched nwere ihe ndị nwere hydrophilic ga-enwe ebe dị elu karịa mkpụrụ osisi efu.Ya mere, adhesion ọzọ nke nje ga-eme na mkpụrụ osisi etched.Otú ọ dị, e jiri ya tụnyere mkpụrụ osisi na-adịghị ahụkebe, mkpụrụ osisi a kpụrụ akpụ enwechaghị obere biofilm emebere n'elu.Nke a bụ n'ihi na ụmụ irighiri mmiri na-ejikọta nke ọma na elu hydrophilic ma na-eme ihe dị ka mmanụ mmanu maka mmiri, si otú a na-egbochi njikọ nke nje bacteria na obere oge (39).Otú ọ dị, oyi akwa nke ụmụ irighiri mmiri dị ezigbo mkpa ma na-agbaze na nkwụsị nke nje.Ya mere, oyi akwa molekụla mmiri na-apụ n'anya ruo ogologo oge, na-eduga n'ọtụtụ nje bacteria na mmụba.Maka SHP, n'ihi njirimara ya na-adịghị adịte aka, a na-egbochi nje bacteria.Enwere ike ịsị na ntachi obi nke nje bacteria na-ebelata site na akpa ikuku tọrọ n'ime usoro dị larịị na ike ala dị ala, si otú a na-ebelata kọntaktị n'etiti nkwusioru nje na elu.Otú ọ dị, a hụrụ adhesion nke nje bacteria na SHP n'ihi na ọ tụfuru ihe ndị na-eme ka ọ ghara imebi ya ruo ogologo oge.Nke a bụ isi n'ihi nkwụsị nke akpa ikuku n'ihi nrụgide hydrostatic na mgbasa nke ikuku na mmiri.Nke a na-abụkarị n'ihi mwepu nke akpa ikuku n'ihi mgbasa na nhazi ọkwa nke na-enye ebe dị elu maka adhesion (27, 40).N'adịghị ka ihe abụọ a na-enwe mmetụta dị mkpa na nkwụsi ike ogologo oge, a na-agbanye mmanụ na-ekpo ọkụ dị na LOIS n'ime micro / nano Ọdịdị na ọ gaghị efu ọbụna n'ime ogologo oge.Ihe mmanu mmanu juputara na ihe owuwu micro/nano kwụsiri ike ma na-adọrọ adọrọ n'elu ya n'ihi njikọ kemịkalụ ha dị elu, si otú a na-egbochi nnabata nje ruo ogologo oge.Ọgụgụ S6 na-egosi nlegharị anya ihe onyonyo confocal confocal nke mkpụrụ etinyere mmanụ mmanu na-emikpu na saline buffered phosphate (PBS).Onyonyo na-aga n'ihu na-egosi na ọbụlagodi mgbe awa 120 nke ntakịrị ịma jijiji (120 rpm), akwa mmanu mmanu na LOIS na-anọgide na-agbanweghị, na-egosi nkwụsi ike ogologo oge n'okpuru ọnọdụ mmiri.Nke a bụ n'ihi oke kemịkalụ dị n'etiti mkpuchi SAM nke fluorine na mmanụ mmanu nke perfluorocarbon, nke mere na enwere ike ịmepụta oyi akwa siri ike.Ya mere, a na-ejigide arụ ọrụ mgbochi.Tụkwasị na nke ahụ, a nwalere mkpụrụ osisi ahụ megide protein ndị nnọchiteanya (albumin na fibrinogen), nke dị na plasma, sel ndị nwere njikọ chiri anya na ọrụ mgbochi (macrophages na fibroblasts), na ndị metụtara ọkpụkpụ ọkpụkpụ.Ọdịnaya nke calcium dị oke elu.(Foto 3D, 1 na 2, na foto S7) (41, 42).Tụkwasị na nke ahụ, ihe oyiyi microscope nke fluorescence nke ule adhesion maka fibrinogen, albumin na calcium gosipụtara àgwà adhesion dị iche iche nke otu mkpụrụ nke ọ bụla (Figure S8).N'oge ọkpụkpụ ọkpụkpụ, ọkpụkpụ ọkpụkpụ na calcium nke e kpụrụ ọhụrụ nwere ike gbaa gburugburu ọkpụkpụ orthopedic, nke ọ bụghị nanị na-eme ka iwepụ ihe siri ike, ma ọ pụkwara ịkpata onye ọrịa ahụ na-atụghị anya ya n'oge usoro mwepụ.Ya mere, obere ego nke calcium na efere ọkpụkpụ na skru na-aba uru maka ịwa ahụ ọkpụkpụ nke na-achọ iwepụ ihe ọkụkụ.Dabere na ọnụọgụ nke mpaghara agbakwunyere dabere na ike fluorescence na ọnụ ọgụgụ cell, anyị kwadoro na LOIS na-egosi ezigbo ihe mgbochi biofouling maka ihe niile dị ndụ ma e jiri ya tụnyere ihe ndị ọzọ.Dị ka nsonaazụ nke nnwale in vitro si kwuo, a pụrụ itinye LOIS na-emebi ihe ndị na-egbochi ihe ndị dị ndụ na ọkpụkpụ orthopedic, nke nwere ike ọ bụghị nanị igbochi ọrịa nje bacteria na-akpata, kamakwa ibelata mbufụt nke usoro ahụ ji alụso ọrịa ọgụ na-akpata.
(A) Onyonyo microscope fluorescence nke otu ọ bụla (ọtọ, etched, SHP na LOIS) etinyere na nkwusioru Pseudomonas aeruginosa na MRSA maka awa 12 na 72.(B) Ọnụ ọgụgụ nke ndị na-akwado CFU nke Pseudomonas aeruginosa na MRSA n'elu otu ọ bụla.(C) Eserese atụmatụ nke usoro mmebi ihe mgbochi nke ndụ nke etching dị mkpụmkpụ na ogologo oge, SHP na LOIS.(D) (1) Ọnụ ọgụgụ nke fibroblasts rapaara na nke ọ bụla mkpụrụ na fluorescence ihe oyiyi microscope nke mkpụrụ ndụ na-arapara n'ahụ na LOIS.(2) Nnwale adhesion nke protein ndị metụtara mgbochi, albumin na calcium na-etinye aka na usoro ọgwụgwọ ọkpụkpụ (* P <0.05, ** P <0.01, *** P <0.001 na **** P <0.0001).ns, ọ dịghị mkpa.
N'ihe banyere nrụgide gbadoro ụkwụ na-enweghị atụ, ịdịte aka n'ibu bụ mgbe niile bụ isi ihe ịma aka maka ntinye nke mkpuchi mgbochi.Ụzọ gel na-egbochi nsị mmiri na-emekarị na-adabere na polymers nwere obere mmiri solubility na fragility.Ya mere, ha na-enwekarị ike ịnweta nrụgide n'ibu na ngwa biomedical.Ya mere, ihe mkpuchi mgbochi na-adịgide adịgide na-adịgide adịgide na-anọgide na-abụ ihe ịma aka maka ngwa ndị dị ka ọkpụkpụ ọkpụkpụ (43, 44).Ọnụọgụ 4A (1) na-egosiputa ụdị nrụgide abụọ a na-etinye na ntinye nke orthopedic, gụnyere ịcha (nrụgide nrụgide) na mkpakọ na ihe oyiyi anya nke ihe nkwụnye mebiri emebi na-emepụta site na forceps.Dịka ọmụmaatụ, mgbe ejiri screwdriver mechie ihe nkedo ahụ, ma ọ bụ mgbe dọkịta na-awa ahụ na-ejide efere ọkpụkpụ nke ọma na tweezers ma tinye ike mkpakọ, efere ọkpụkpụ plastik ga-emebi ma kpoo ya na macro na micro / nano scales (Nyocha 4A, 2) .Iji nwalee ma LOIS arụpụtara ọ nwere ike idi mmebi ndị a n'oge ịwa ahụ rọba, a na-eme nanoindentation iji tụnyere ike nke mkpụrụ osisi efu na LOIS na nha micro / nano iji mụọ ihe eji arụ ọrụ nke micro / nano Ọdịdị Mmetụta (Ọnọdụ). 4B).Eserese eserese a na-egosi omume nrụrụ dị iche iche nke LOIS n'ihi ọnụnọ nke micro/nano owuwu.Edepụtara usoro nchụpụ ike na-adabere na nsonaazụ nanoindentation (Njirimara 4C).Onyonyo na-acha anụnụ anụnụ na-anọchi anya mkpụrụ osisi na-enweghị ihe ọ bụla, nke na-egosi naanị nrụrụ dị ntakịrị, dịka a na-ahụ ya site na omimi ntinye kachasị nke 0.26-μm.N'aka nke ọzọ, mmụba nke nwayọọ nwayọọ na ike nanoindentation na nchụpụ a na-ahụ na LOIS (uhie uhie) nwere ike igosi ihe ịrịba ama nke ihe ndị na-emepụta ihe na-ebelata, na-ebute nanoindentation omimi nke 1.61μm.Nke a bụ n'ihi na ihe owuwu micro/nano dị na LOIS na-enye ohere ọganihu miri emi maka ọnụ nanoindenter, ya mere nrụrụ ya dị ukwuu karịa nke mkpụrụ osisi efu.Konsta-Gdoutos et al.(45) kwenyere na n'ihi ọnụnọ nke nanostructures, nanoindentation na micro / nano roughness na-eduga na ntụgharị nanoindentation na-adịghị mma.Mpaghara shaded kwekọrọ na ngbanwe na-adịghị mma nke a na-akpọ nanostructure, ebe ebe a na-enweghị ndò na-ekwu na microstructure.Nrụrụ a nwere ike mebie microstructure/nanostructure nke mmanu na-ejide ma na-emetụta arụmọrụ ya na-adịghị mma.Iji mụọ mmetụta nke mmebi na LOIS, mmebi a na-apụghị izere ezere nke micro/nano ka e megharịrị n'ime ahụ n'oge ịwa ahụ rọba.Site n'iji nyocha ọbara na protein adhesion, nkwụsi ike nke ihe mgbochi biofouling nke LOIS mgbe in vitro nwere ike ikpebi (Njirimara 4D).Usoro onyonyo anya na-egosi mmebi nke mere n'akụkụ oghere nke mkpụrụ nke ọ bụla.A na-eme nyocha adhesion ọbara iji gosipụta mmetụta nke mmebi n'ibu na mkpuchi mgbochi biofouling (Figure 4E).Dị ka SHP, ihe ndị na-eme ka ọ ghara imebi ihe na-efunahụ ya n'ihi mmebi, na LOIS na-egosipụtakwa ihe ndị na-adịghị mma nke ọma site n'iweghachi ọbara.Nke a bụ n'ihi na, n'ihi na ike elu na-akpali site na capillary edinam na-ekpuchi ebe mebiri emebi, na-erugharị na mmanu mmanu microstructured na-eweghachi ihe ndị na-adịghị mma (35).A hụrụ otu ihe a na-eme na nnwale nrapanye protein site na iji albumin.N'ebe ahụ mebiri emebi, a na-ahụta adhesion nke protein n'elu SHP, na site n'ịtụle mkpuchi mpaghara ya, enwere ike ịgụta ya dị ka ọkara nke ọkwa adhesion nke mkpụrụ osisi efu.N'aka nke ọzọ, LOIS nọgidere na-enwe ihe ndị na-egbochi biofouling n'emeghị ka adhesion (Njirimara 4, F na G).Tụkwasị na nke ahụ, a na-enwekarị nrụgide ọrụ siri ike n'elu ihe nkedo ahụ, dị ka mkpọpu mmiri, n'ihi ya, anyị mụrụ ikike nke mkpuchi LOIS ka ọ ghara ịdịgidere na screw in vitro.Ọgụgụ 4H na-egosi onyonyo anya nke skru dị iche iche, gụnyere iferi, SHP na LOIS.Rektangulu uhie na-anọchi anya ebe a na-achọsi ike bụ ebe nrụgide ọrụ siri ike na-eme n'oge ịkụnye ọkpụkpụ.Dị ka ule adhesion protein nke efere ahụ, a na-eji microscope fluorescence na-ese onyinyo protein adhesion ma tụọ ebe mkpuchi iji gosipụta iguzosi ike n'ezi ihe nke mkpuchi LOIS, ọbụlagodi n'okpuru nrụgide siri ike (Nyocha 4, I na J).Ihe nkedo nke LOIS na-emeso skru na-egosipụta ọmarịcha arụmọrụ na-adịghị mma, ọ fọrọ nke nta ka ọ bụrụ na ọ dịghị protein na-arapara n'elu.N'aka nke ọzọ, a na-ahụ adhesion protein na skru efu na SHP skru, ebe ebe mkpuchi nke SHP skru bụ otu ụzọ n'ụzọ atọ nke skru efu.Tụkwasị na nke ahụ, ọkpụkpụ ọkpụkpụ nke a na-eji emezi ihe kwesịrị ịdị ike nke ọma iji nagide nrụgide a na-etinye na saịtị mgbawa, dị ka egosiri na Figure 4K.Ya mere, a na-eme ule na-ehulata iji chọpụta mmetụta nke mgbanwe kemịkal na ihe ndị na-emepụta ihe.Tụkwasị na nke a, a na-eme nke a iji nọgide na-enwe nrụgide siri ike site na ntinye.Tinye ike igwe kwụ ọtọ ruo mgbe etinyere ya nke ọma wee nweta usoro nrụgide nrụgide (Foto 4L, 1).Atụnyere akụrụngwa abụọ gụnyere modulus nke Young na ike mgbanwe n'etiti ihe efu na LOIS dị ka ihe na-egosi ike ọrụ ha (Njirimara 4L, 2 na 3).Modules nke ntorobịa na-egosi ikike nke ihe iji guzogide mgbanwe igwe.Modules nke Young nke mkpụrụ nke ọ bụla bụ 41.48± 1.01 na 40.06±0.96 GPA, n'otu n'otu;ihe dị iche a hụrụ bụ ihe dịka 3.4%.Tụkwasị na nke ahụ, a na-akọ na ike na-ehulata, nke na-ekpebi ike nke ihe ahụ, bụ 102.34 ± 1.51 GPa maka ihe efu na 96.99 ± 0.86 GPa maka SHP.Mkpụrụ osisi efu dị ihe dịka 5.3% elu.Enwere ike ịkpata ntakịrị mbelata na akụrụngwa akụrụngwa site na mmetụta notch.Na mmetụta notch, micro/nano roughness nwere ike na-eme dị ka set nke notches, na-eduga na mgbakasị uche uche na mpaghara na-emetụta n'ibu Njirimara nke implant (46).Otú ọ dị, dabere n'eziokwu ahụ bụ na a na-akọ na isi ike nke ọkpụkpụ cortical mmadụ dị n'etiti 7.4 na 31.6 GPa, na LOIS modulus a tụrụ karịa nke ọkpụkpụ cortical mmadụ (47), LOIS zuru ezu iji kwado mgbaji ahụ na n'ozuzu ya. Mechanical Njirimara na-obere emetụta elu mgbanwe.
(A) Eserese atụmatụ nke (1) nrụsi ọrụ ike etinyere n'ọkpụkpụ ọkpụkpụ n'oge a na-arụ ọrụ, na (2) ihe onyonyo anya nke ihe ọkpụkpụ ọkpụkpụ mebiri emebi.(B) Eserese eserese nke nano-mechanical Properties nke nanoindentation na LOIS n'elu elu efu.(C) Nanoindentation ike-mgbanwe akụkụ nke elu efu na LOIS.(D) Mgbe nnwale in vitro gachara, mee ka onyonyo anya nke ụdị dị iche iche nke efere orthopedic dị iche iche (a na-egosipụta mpaghara mebiri emebi na rectangle na-acha uhie uhie) iji mee ka nrụgide igwe kpatara n'oge ọrụ ahụ.(E) Nnwale nnabata ọbara na (F) nnwale nnabata protein nke otu efere ọkpụkpụ ahụ mebiri emebi.(G) Tulee ebe mkpuchi protein na-arapara na efere ahụ.(H) Onyonyo anya nke ụdị skru orthopedic dị iche iche mgbe nnwale in vitro gasịrị.(I) Nnwale adhesion protein iji mụọ iguzosi ike n'ezi ihe nke mkpuchi dị iche iche.(J) Tulee mkpuchi mpaghara nke protein na-arapara na skru.(K) Ntugharị nke oke bekee bu n'obi na-ebute nrụgide na-adịgide adịgide na ọkpụkpụ gbajiri agbaji.(L) (1) Kwuo nsonaazụ ule na onyonyo anya tupu na mgbe agbachara.Ọdịiche dị na (2) modulus nke ntorobịa na (3) ike na-ehulata n'etiti ntinye efu na SHP.Ekwuputara data dị ka ± SD (* P <0.05, ** P <0.01, *** P <0.001 na **** P <0.0001).Foto site n'ikike: Kyomin Chae, Mahadum Yonsei.
N'ọnọdụ ụlọ ọgwụ, ọtụtụ kọntaktị nje bacteria na ihe ndị dị ndụ na ebe ọnya na-abịa site na tozuru okè, tozuru okè biofilms (48).Ya mere, Centlọ Ọrụ Maka Nchịkwa na Mgbochi Ọrịa US na-eme atụmatụ na 65% nke ọrịa mmadụ niile metụtara biofilms (49).N'okwu a, ọ dị mkpa ịnye ihe nyocha in vivo nke na-enye nhazi biofilm na-agbanwe agbanwe n'elu ihe ntinye.Ya mere, anyị mepụtara ụdị mgbawa femoral nke oke bekee nke ọkpụkpụ orthopedic na-ebute ụzọ na nkwụsị nke nje bacteria wee tinye ya na femurs oke bekee iji mụọ ihe ndị na-adịghị mma nke LOIS na vivo.N'ihi eziokwu atọ ndị a dị mkpa, nje na-efe efe na-ebute site na tupu omenala kama ịgba ọgwụ mgbochi nje bacteria kpọmkwem: (i) Usoro ahụ ji alụso ọrịa ọgụ nke oke bekee siri ike karịa nke mmadụ;ya mere, ịgba ọgwụ mgbochi nje na nje bacteria planktonic ga-ekwe omume Ọ nweghị mmetụta ọ bụla na nhazi nke biofilms.(Ii) Nje bacteria Planktonic na-enwekarị ike ịnweta ọgwụ nje, a na-ejikarị ọgwụ nje eme ihe mgbe a wachara ya ahụ;N'ikpeazụ, (iii) mmiri ahụ anụ ahụ nwere ike ịgbanye nkwụsị nke nje bacteria planktonic (50).Site n'ịmepụta ihe a na-etinye na nkwusioru nje tupu etinye ya, anyị nwere ike nyochaa nke ọma mmetụta ọjọọ nke ọrịa nje na mmeghachi omume anụ ahụ nke mba ọzọ (FBR) na usoro ọgwụgwọ ọkpụkpụ.A na-achụrụ oke bekee 4 izu mgbe etinyere ya, n'ihi na osseointegration dị mkpa maka usoro ọgwụgwọ ọkpụkpụ ga-agwụ n'ime izu 4.Mgbe ahụ, a na-ewepụ ihe ndị a na-etinye na rabbits maka ọmụmụ ihe ala.Ọgụgụ 5A na-egosi usoro mgbasa nke nje bacteria.A na-ewebata ihe ọkpụkpụ ọkpụkpụ nke butere ya n'ime ahụ.N'ihi mmalite mmalite na nkwusioru nje bacteria, isii n'ime oke bekee isii nke etinyere na ihe mgbochi gba ọtọ bu ọrịa, ebe ọ dịghị nke ọ bụla n'ime oke bekee nke etinyere na LOIS na-agwọ ọrịa.Ọrịa nje bacteria na-aga n'ihu na usoro atọ, gụnyere uto, maturation na mgbasa (51).Nke mbụ, nje bacteria na-ejikọta ya na-amụba ma na-eto n'elu, mgbe ahụ nje bacteria na-emepụta biofilm mgbe ha na-ewepụ polymer extracellular (EPS), amyloid na extracellular DNA.Biofilm ọ bụghị nanị na-egbochi ntinye nke ọgwụ nje, kamakwa na-akwalite nchịkọta nke enzymes na-emerụ emerụ (dị ka β-lactamase) (52).N'ikpeazụ, biofilm na-agbasa nje bacteria ndị tozuru okè n'ime anụ ahụ gbara ya gburugburu.Ya mere, ọrịa na-apụta.Tụkwasị na nke ahụ, mgbe ahụ mba ọzọ na-abanye n'ime ahụ, ọrịa nke nwere ike ime ka ọ bụrụ ihe mgbochi siri ike nwere ike ime ka ọzịza siri ike, mgbu, na mbelata mgbochi.Ọgụgụ 5B na-enye nkọwapụta nke FBR kpatara site na ntinye nke orthopedic implant, kama nzaghachi mgbochi nke ọrịa nje kpatara.Usoro ahụ ji alụso ọrịa ọgụ na-amata ntinye etinyere dị ka onye mba ọzọ, wee mee ka mkpụrụ ndụ na anụ ahụ meghachi omume iji kpuchie ahụ mba ọzọ (53).N'oge mmalite nke FBR, a na-emepụta matrix ọkọnọ n'elu ọkpụkpụ ọkpụkpụ, nke mere ka ọ bụrụ mgbasa ozi nke fibrinogen.Fibrinogen adsorbed wee mepụta netwọk fibrin dị oke egwu, nke na-akwalite ntinye nke leukocytes (54).Ozugbo e guzobere netwọk fibrin, nnukwu mbufụt ga-eme n'ihi ntinye nke neutrophils.N'ime usoro a, a na-ahapụ ụdị cytokines dị iche iche dị ka tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) na IL-β, na monocytes na-amalite ịbanye n'ime ebe a na-etinye ihe ma na-ekewa n'ime nnukwu sel.Nkeji (41, 55, 56).Mbelata FBR bụ ihe ịma aka mgbe niile n'ihi na oke FBR nwere ike ibute nnukwu ọnya na-adịghị ala ala, nke nwere ike ibute nsogbu na-egbu egbu.Iji chọpụta mmetụta nke nje na-efe efe na anụ ahụ gbara ebe a na-etinye ihe na-enweghị ihe ọkụkụ na LOIS, a na-eji hematoxylin na eosin (H&E) na Masson trichrome (MT).Maka oke bekee etinyere na ihe efu, ọrịa nje siri ike na-aga n'ihu, na slide anụ ahụ H&E gosipụtara n'ụzọ doro anya abscesses na necrosis nke mbufụt kpatara.N'aka nke ọzọ, oke mgbochi mgbochi biofouling elu LOIS na-egbochi nje bacteria, n'ihi ya, ọ dịghị ihe ịrịba ama nke ọrịa na-ebelata mbufụt (Figure 5C).Nsonaazụ nke ntụpọ MT gosipụtara otu omume ahụ.Agbanyeghị, ntụpọ MT gosikwara edema na oke bekee etinyere na LOIS, na-egosi na mgbake na-aga ime (Njirimara 5D).Iji mụọ ogo nzaghachi mgbochi ọrịa, a na-eme ntụpọ immunohistochemical (IHC) site na iji cytokines TNF-a na IL-6 metụtara nzaghachi mgbochi.A na-atụnye ihe na-adịghị mma nke gba ọtọ nke na-adịghị ekpughe na nje bacteria na LOIS nke ekpughere na nje bacteria ma ọ bụghị nje iji mụọ usoro ọgwụgwọ na-enweghị ọrịa nje.Ọgụgụ 5E na-egosi onyonyo anya nke ihe mmịfe IHC nke na-egosipụta TNF-α.Mpaghara aja aja na-anọchite anya nzaghachi mgbochi ahụ, na-egosi na nzaghachi mgbochi na LOIS dị ntakịrị belata.Tụkwasị na nke ahụ, okwu IL-6 na LOIS dị ntakịrị karịa okwu ọjọọ nke ịgba ọtọ na-adịghị mma (Njirimara 5F).A kọwapụtara okwu nke cytokine site n'ịtụ mpaghara ntụpọ mgbochi nke dabara na cytokine (Figure 5G).E jiri ya tụnyere oke bekee ekpughere na ntinye na-adịghị mma, ọkwa ngosipụta nke oke bekee etinyere na LOIS dị ala, na-egosi ọdịiche bara uru.Mbelata nke okwu cytokine na-egosi na ogologo oge, ihe na-emebi ihe na-adịghị mma nke LOIS abụghị nanị ihe jikọrọ ya na mgbochi nke ọrịa nje, kamakwa na mbelata nke FBR, nke na-ebute site na macrophages na-agbaso na mkpụrụ (53, 57, 58).Ya mere, mmeghachi omume mgbochi nke mbelata n'ihi njirimara mgbapụ nke LOIS nwere ike dozie mmetụta ndị dị na ya mgbe etinyere ya, dị ka nzaghachi mgbochi oke mgbe ịwa ahụ plastik gasịrị.
(A) Eserese eserese nke usoro nhazi biofilm wee gbasaa n'elu ihe ọkpụkpụ ọkpụkpụ nwere ọrịa.eDNA, DNA extracellular.(B) Eserese nhazi nke nzaghachi mgbochi mgbe etinyere orthopedic.(C) H&E ntanye na (D) MT ntanye nke anụ ahụ gbara ya gburugburu nke ọkpụkpụ ọkpụkpụ nwere ezigbo ihe na LOIS.IHC nke cytokines ndị na-alụso ọrịa ọgụ (E) TNF-a na (F) IL-6 bụ ihe onyonyo nwere ntụpọ nke oke bekee gba ọtọ na-adịghị mma na LOIS.(G) Ọnụ okwu cytokine site na nha mkpuchi mpaghara (** P <0.01).
A na-enyocha biocompatibility nke LOIS na mmetụta ya na usoro ọgwụgwọ ọkpụkpụ na vivo site na iji nchọpụta nchọpụta [x-ray na micro-computed tomography (CT)] na osteoclast IHC.Ọnụ ọgụgụ 6A na-egosi usoro ọgwụgwọ ọkpụkpụ gụnyere usoro atọ dị iche iche: mbufụt, nrụzi, na nhazigharị.Mgbe mgbawa na-eme, mkpụrụ ndụ na-egbuke egbuke na fibroblasts ga-abanye n'ime ọkpụkpụ gbajiri agbaji wee malite ịmalite n'ime anụ ahụ vaskụla.N'oge a na-arụzi ọrụ, ntinye nke anụ ahụ vaskụla na-agbasa n'akụkụ ebe mgbawa ahụ.Anụ ahụ akwara na-enye nri maka ịmepụta ọkpụkpụ ọhụrụ, nke a na-akpọ callus.Oge ikpeazụ nke usoro ọgwụgwọ ọkpụkpụ bụ nhazigharị nhazi, nke a na-ebelata nha nke callus na ọkpụkpụ nkịtị site n'enyemaka nke mmụba na ọkwa nke osteoclasts na-arụ ọrụ (59).A na-emezigharị akụkụ atọ (3D) nke ebe mgbawa site na iji nyocha micro-CT iji hụ ọdịiche dị na ọkwa nke nhazi callus na otu ọ bụla.Leba anya n'ofe akụkụ nke femur ka ị hụ oke nke callus gburugburu ọkpụkpụ gbajiri agbaji (Foto 6, B na C).A na-ejikwa ụzarị X na-enyocha ebe mgbawa nke otu niile kwa izu iji hụ usoro nhazi ọkpụkpụ dị iche iche na otu ọ bụla (Nyocha S9).A na-egosi callus na ọkpụkpụ ndị tozuru okè na-acha anụnụ anụnụ / akwụkwọ ndụ akwụkwọ ndụ na ọdụm, n'otu n'otu.A na-ehichapụ ọtụtụ anụ ahụ dị nro site n'ọnụ ụzọ edobere ya.Ịgba ọtọ ziri ezi na SHP kwadoro nhazi nke obere callus gburugburu ebe mgbawa ahụ.N'aka nke ọzọ, ekpughere adịghị mma nke LOIS na ebe mgbaji ahụ bụ nnukwu callus gbara ya gburugburu.Ihe onyonyo Micro-CT gosiri na a na-egbochi nguzobe callus site na nje nje na mbufụt metụtara ọrịa.Nke a bụ n'ihi na usoro ahụ ji alụso ọrịa ọgụ na-ebute ụzọ ọgwụgwọ nke mmerụ ahụ septic kpatara site na mbufụt metụtara ọrịa, kama mgbake ọkpụkpụ (60).A na-eme ntụpọ IHC na Tartrate Acid Phosphatase (TRAP) iji hụ ọrụ osteoclast na resorption ọkpụkpụ (Nyocha 6D) (61).Naanị ole na ole osteoclasts na-arụ ọrụ na-acha odo odo ka ahụrụ na ihe gba ọtọ na SHP.N'aka nke ọzọ, a hụrụ ọtụtụ osteoclasts na-arụ ọrụ n'akụkụ ọkpụkpụ gba ọtọ na nke tozuru okè nke LOIS.Ihe omume a na-egosi na n'ihu osteoclasts, callus gburugburu ebe mgbawa ahụ na-eme mgbanwe na-eme ihe ike (62).A na-atụle ụda ọkpụkpụ na mpaghara osteoclast nke callus iji tụnyere ọkwa nke nhazi callus gburugburu ebe mgbawa ahụ n'ìgwè niile, iji chọpụta micro-CT scan na nsonaazụ IHC (Figure 6E, 1 na 2).Dị ka a tụrụ anya ya, ihe ndị na-adịghị mma na nhazi callus dị na LOIS dị elu karịa na ndị ọzọ, na-egosi na ọkpụkpụ ọkpụkpụ dị mma mere (63).Ọnụọgụ S10 na-egosi onyonyo anya nke saịtị ịwa ahụ, nsonaazụ MT na-enweta nke anụ ahụ anakọtara n'akụkụ ihe nkedo ahụ, yana nsonaazụ ntụpọ TRAP na-eme ka ọ pụta ìhè ngbanwe ọkpụkpụ-ọkpụkpụ.N'ime mkpụrụ osisi na-adịghị ahụkebe, a hụrụ nguzobe callus na fibrosis siri ike, ebe ihe a na-etinye na LOIS na-egosi elu na-adịghị ejikọta ya.N'otu aka ahụ, ma e jiri ya tụnyere ihe na-adịghị mma, a na-ahụ fibrosis dị ala na oke bekee etinyere na LOIS, dị ka akụ na-acha ọcha na-egosi.Tụkwasị na nke a, enwere ike ịsị na edema siri ike (akụ na-acha anụnụ anụnụ) nwere ike ịpụta na njirimara mgbapụ nke LOIS, si otú a na-ebelata mbufụt siri ike.Ebe a na-adịghị ahụkebe na gburugburu ebe a na-etinye ya na fibrosis na-ebelata na-egosi na usoro iwepụ ya dị mfe, nke na-ebutekarị mgbawa ma ọ bụ mbufụt ndị ọzọ.A na-enyocha usoro ọgwụgwọ ọkpụkpụ mgbe ewepụsịrị screw site na ọrụ osteoclast na screw-bone interface.Ma ọkpụkpụ na-adịghị ahụkebe na LOIS implant interface na-etinye uche na ọkwa osteoclasts yiri nke ahụ iji nwetakwuo ọgwụgwọ ọkpụkpụ, na-egosi na mkpuchi LOIS enweghị mmetụta ọjọọ na ọgwụgwọ ọkpụkpụ ma ọ bụ nzaghachi mgbochi.Iji gosi na ngbanwe elu nke a na-eme na LOIS adịghị egbochi usoro ọgwụgwọ ọkpụkpụ, a na-eji nyocha X-ray tụnyere ọgwụgwọ ọkpụkpụ nke rabbits na ions na-adịghị mma na 6 izu nke ntinye LOIS (Nyocha 6F).Nsonaazụ gosiri na ma e jiri ya tụnyere otu ndị na-adịghị ahụkebe ịgba ọtọ, LOIS gosipụtara otu ogo nke ọgwụgwọ ọkpụkpụ, ọ dịghịkwa ihe ịrịba ama doro anya nke mgbaji (akara osteolysis na-aga n'ihu) na otu abụọ ahụ.
(A) Eserese eserese nke usoro ọgwụgwọ ọkpụkpụ mgbe agbajichara.(B) Ọdịiche dị n'ogo nke nhazi callus nke otu elu nke ọ bụla na (C) ihe oyiyi cross-section nke saịtị mgbawa.(D) Nchacha TRAP iji jiri anya nke uche hụ ọrụ osteoclast na ọkpụkpụ ọkpụkpụ.Dabere na ọrụ TRAP, a na-enyocha nhazi nke callus mpụga nke ọkpụkpụ cortical site na (E) (1) micro-CT na (2) ọrụ osteoclast.(F) 6 izu mgbe etinyechara ya, onyonyo X-ray nke ọkpụkpụ gbajiri agbaji nke ihe na-adịghị mma nke ekpughere (nke rectangle na-acha uhie uhie na-egosipụta) na LOIS (nke akụkụ anọ na-acha anụnụ anụnụ gosipụtara).Emere nyocha ọnụ ọgụgụ site na nyocha otu ụzọ nke iche iche (ANOVA).P <0.05.** P <0.01.
Na nkenke, LOIS na-enye ụdị ọhụrụ nke atụmatụ nje nje bacteria na mkpuchi mgbanarị maka mgbaba ọkpụkpụ.Nkwụnye orthopedic nke ọdịnala nwere ọrụ SHP na-egosipụta ihe mgbochi biofouling dị mkpụmkpụ, mana enweghị ike idobe akụrụngwa ha ogologo oge.Ọnyà superhydrophobicity nke mkpụrụ ahụ na-ejide afụ ikuku n'etiti nje bacteria na mkpụrụ, si otú ahụ na-akpụ akpa ikuku, si otú ahụ gbochie ọrịa nje.Otú ọ dị, n'ihi mgbasa nke ikuku, a na-ewepụ akpa ikuku ndị a ngwa ngwa.N'aka nke ọzọ, LOIS egosila nke ọma ikike ya igbochi ọrịa ndị metụtara biofilm.Ya mere, n'ihi mgbochi mgbochi nke oyi akwa mmanu nke a na-agbanye n'ime elu micro/nano , nwere ike igbochi mbufụt metụtara ọrịa.A na-eji ụzọ njirimara dị iche iche gụnyere nha SEM, AFM, XPS na CA iji kwalite ọnọdụ nrụpụta LOIS.Tụkwasị na nke a, a pụkwara itinye LOIS n'ihe dị iche iche nke ihe ndị dị ndụ nke a na-ejikarị eme ihe n'ịgwọ orthopedic, dị ka PLGA, Ti, PE, POM na PPSU.Mgbe ahụ, a nwalere LOIS na vitro iji gosipụta ihe mgbochi biofouling ya megide nje bacteria na ihe ndị dị ndụ metụtara nzaghachi mgbochi.Nsonaazụ na-egosi na ọ nwere ezigbo antibacterial na anti-biofouling mmetụta ma e jiri ya tụnyere nke a na-etinye n'ime ihe efu.Tụkwasị na nke ahụ, LOIS na-egosi ike arụ ọrụ ọbụna mgbe etinyechara nrụgide n'ibu, bụ nke a na-apụghị izere ezere na ịwa ahụ plastik.N'ihi ihe na-agwọ onwe ya nke mmanu mmanu n'elu ihe owuwu micro/nano, LOIS jisiri ike na-eme ihe na-emebi ihe na-emebi ihe.Iji mụọ biocompatibility na antibacterial Njirimara nke LOIS in vivo, LOIS ka etinyere n'ime oke bekee femur maka izu anọ.A hụghị ọrịa nje na oke bekee etinyere na LOIS.Tụkwasị na nke ahụ, iji IHC gosipụtara mbelata nke nzaghachi mgbochi nke mpaghara, na-egosi na LOIS anaghị egbochi usoro ọgwụgwọ ọkpụkpụ.LOIS na-egosiputa ihe ndị dị mma na-egbochi nje bacteria na nke na-adịghị alụso ọrịa ọgụ, ma gosi na ọ na-egbochi ịmepụta biofilm nke ọma tupu na n'oge ịwa ahụ ọkpụkpụ, karịsịa maka njikọ ọkpụkpụ.Site n'iji ụmị ọkpụkpụ oke bekee na-egbuke egbuke ụdị mgbawa femoral, mmetụta nke ọrịa ndị metụtara biofilm na usoro ọgwụgwọ ọkpụkpụ na-ebute site n'ịkụnye ihe ndị a na-ebute ụzọ.Dị ka ọmụmụ ihe n'ọdịnihu, a chọrọ ụdị ọhụrụ in vivo iji mụọ ọrịa nwere ike ime mgbe etinyere ya ka ọ ghọta nke ọma ma gbochie ọrịa ndị metụtara biofilm n'oge usoro ọgwụgwọ dum.Na mgbakwunye, osteoinduction ka bụ ihe ịma aka a na-edozighị na njikọta na LOIS.Ọ dị mkpa nyocha ọzọ iji jikọta nchikota nhọrọ nke mkpụrụ ndụ osteoinductive ma ọ bụ ọgwụ na-emegharị ahụ na LOIS iji merie ihe ịma aka ahụ.N'ozuzu, LOIS na-anọchite anya mkpuchi mkpuchi orthopedic na-ekwe nkwa nke nwere ike ịrụ ọrụ na ihe ndị dị mma na-egbochi biofouling, nke nwere ike ibelata SSI na mmetụta mgbochi.
Sachaa mkpụrụ 15mm x 15mm x 1mm 304 SS (Dong Kang M-Tech Co., Korea) na acetone, EtOH na mmiri DI maka nkeji iri na ise iji wepụ ihe mmetọ.Iji mepụta usoro micro/nano-larịị n'elu, a na-emikpu mkpụrụ a kpochara na 48% ruo 51% HF solution (DUKSAN Corp., South Korea) na 50°C.Oge etching dị iche site na 0 ruo 60 nkeji.Mgbe ahụ, a na-eji mmiri deionized sachaa ihe etched ma tinye ya na 65% HNO3 (Korea DUKSAN Corp.) ngwọta na 50 ° C maka nkeji 30 iji mepụta oyi akwa chromium oxide passivation n'elu.Mgbe ngafe gachara, a na-asacha mkpụrụ ahụ na mmiri deionized wee kpoo ya ka ọ nweta mkpụrụ nwere ihe dị larịị.Na-esote, ekpughere mkpụrụ osisi ahụ na plasma oxygen (100 W, 3 nkeji), ma tinye ya ozugbo na ngwọta nke 8.88 mM POTS (Sigma-Aldrich, Germany) na toluene na ụlọ okpomọkụ maka awa 12.Mgbe ahụ, a na-eji EtOH sachaa mkpụrụ ahụ ekpuchiri POTS, wee kpochie ya na 150 Celsius maka awa 2 iji nweta nnukwu POTS SAM.Mgbe mkpuchi SAM gasịrị, a na-emepụta oyi akwa na-egbuke egbuke na mkpụrụ site n'itinye mmanụ aṅụ perfluoropolyether (Krytox 101; DuPont, USA) nke na-ebu ibu nke 20 μm / cm 2. Tupu ojiji, nyochaa mmanụ mmanu site na nzacha 0.2 micron.Wepu mmanu ngafe site na itughari n'akuku 45° maka nkeji iri na ise.A na-eji otu usoro mmepụta ihe maka orthopedic implants mere nke 304 SS (efere mkpọchi na cortical locking screw; Dong Kang M-Tech Co., Korea).Emebere ihe nkwụnye ọkpụkpụ niile ka ọ dabara na geometry nke femur oke bekee.
A na-enyocha morphology elu nke mkpụrụ na orthopedic emission SEM (Inspection F50, FEI, USA) na AFM (XE-100, Park Systems, South Korea).A na-atụle ịdị nro nke elu (Ra, Rq) site n'ịba ụba mpaghara nke 20 μm site na 20 μm (n=4).Ejiri usoro XPS (PHI 5000 VersaProbe, ULVAC PHI, Japan) nwere isi iyi X-ray nke Al Kα nwere nha nha 100μm2 iji nyochaa kemịkalụ elu.A na-eji sistemu nha CA nke nwere igwefoto na-ese foto dị ike (SmartDrop, FEMTOBIOMED, ​​South Korea) iji tụọ mmiri mmiri CA na SA.Maka nha ọ bụla, a na-etinye 6 ruo 10 μl nke ụmụ irighiri mmiri (mmiri deionized, ọbara ịnyịnya, EG, 30% ethanol, na HD) n'elu iji tụọ CA.Mgbe akụkụ ọchịchọ nke mkpụrụ ahụ na-abawanye na ọsọ nke 2°/s (n = 4), SA na-atụ mgbe mmiri mmiri dara.
Pseudomonas aeruginosa [American Type Culture Collection (ATCC) 27853] na MRSA (ATCC 25923) zụtara n'aka ATCC (Manassas, Virginia, USA), na-echekwa omenala ngwaahịa na -80 ° C.Tupu eji ya, a na-etinye omenala oyi kpọnwụrụ na broth soybean trypsin-thawed (Komed, Korea) na 37 Celsius maka awa 18 wee bufee ya ugboro abụọ ka ọ rụọ ọrụ.Mgbe emechara ya, a na-etinye omenala ahụ na 10,000 rpm maka nkeji 10 na 4 Celsius C wee saa ya ugboro abụọ na ngwọta PBS (pH 7.3).A na-emezi omenala centrifuged na efere agar ọbara (BAP).A kwadebere MRSA na Pseudomonas aeruginosa n'abali wee zụlite ya na broth Luria-Bertani.Ntinye uche nke Pseudomonas aeruginosa na MRSA n'ime inoculum bụ nke CFU kpebisiri ike nke nkwusioru na dilutions serial na agar.Mgbe ahụ, dozie ntinye nke nje bacteria na ọkọlọtọ 0.5 McFarland, nke ya na 108 CFU / ml.Mgbe ahụ, gbanye nkwusioru nje na-arụ ọrụ ugboro 100 na 106 CFU / ml.Iji nwalee njirimara nrapado nke nje bacteria, a na-agbazi mkpụrụ ahụ na 121 Celsius maka nkeji 15 tupu ejiri ya.A na-ebufe mkpụrụ ahụ na 25 ml nke nkwusioru nje ma tinye ya na 37 Celsius C na ịma jijiji siri ike (200 rpm) maka awa 12 na 72.Mgbe emechara ya, a na-ewepụ mkpụrụ ọ bụla na incubator wee saa ya ugboro atọ na PBS iji wepụ nje bacteria na-ese n'elu elu.Iji lelee biofilm na mkpụrụ, a na-edozi biofilm na methanol ma jiri 1 ml nke oroma crimidine mee ya maka nkeji 2.Mgbe ahụ, a na-eji microscope fluorescence (BX51TR, Olympus, Japan) see foto nke biofilm nwere ntụpọ.Iji mee ka ọnụọgụ biofilm dị na mkpụrụ osisi ahụ, a na-ekewa mkpụrụ ndụ ndị a na-ejikọta na mkpụrụ site na usoro bead vortex, bụ nke a na-ewere dị ka ụzọ kachasị mma iji wepụ nje bacteria na-ejikọta (n = 4).N'iji ike na-adịghị mma, wepụ mkpụrụ ahụ na ihe na-eto eto ma kpatụ efere olulu mmiri iji wepụ oke mmiri mmiri.E wepụrụ sel ndị etinyere nke ọma site na iji PBS na-adịghị ọcha saa ya ugboro abụọ.A bufere mkpụrụ nke ọ bụla na tube ule na-adịghị ọcha nke nwere 9 ml nke 0.1% protein ept saline (PSW) na 2 g nke 20 ruo 25 beads enyo enyo (0.4 ruo 0.5 mm na dayameta).A na-atụgharị ya maka nkeji 3 iji wepụ mkpụrụ ndụ n'ime ihe nlele ahụ.Mgbe vortexing gasịrị, a na-ejiri 0.1% PSW gbarie nkwusioru ahụ ugboro 10, wee tinye 0.1 ml nke mgbaze ọ bụla na BAP.Mgbe awa 24 nke incubation gasịrị na 37°C, ejiri aka gụọ CFU.
Maka mkpụrụ ndụ, ejiri fibroblasts mouse NIH / 3T3 (CRL-1658; American ATCC) na macrophages RAW 264.7 (TIB-71; American ATCC).Jiri Dulbecco's modified Eagle medium (DMEM; LM001-05, Welgene, Korea) na omenala fibroblasts òké wee gbakwunye na 10% nwa ehi serum (S103-01, Welgene) na 1% penicillin-streptomycin (PS; LS202-02, Welgene (Welgene) ). A na-etinye mkpụrụ ndụ n'otu ntabi anya na 37 ° C na 5% CO2 maka nsị cell, a na-edozi mkpụrụ ndụ na 4% paraformaldehyde maka nkeji 20 ma tinye ya na 0.5% Triton X Incubate maka nkeji 5 na -100 na 37 ° C maka nkeji 30 mgbe usoro mmechi ahụ gasịrị, jiri mkpụrụ osisi na 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) na plasma mmadụ The Alexa Fluor 488-conjugated fibrinogen (F13191, Invitrogen, USA) gbazere na PBS (10 mM, pH 7.4).Ọnụ ọgụgụ nke albumin na fibrinogen bụ 1 na 150 μg / ml, n'otu n'otu.Mgbe mkpụrụ ahụ gachara Tupu imikpu n'ime ihe ngwọta protein, sachaa ha na PBS iji mee ka mmiri dị n'elu.Mgbe ahụ, gbanye ihe niile dị n'ime efere isii nke nwere ngwọta protein ma tinye ya na 37 ° C maka nkeji 30 na 90.Mgbe emechara ya, a na-ewepụ mkpụrụ ahụ na ngwọta protein, jiri nwayọọ na-asa ya na PBS ugboro 3, ma dozie ya na 4% paraformaldehyde (n = 4 maka protein ọ bụla).Maka calcium, sodium chloride (0.21 M) na potassium phosphate (3.77 mM) na-agbaze na mmiri deionized.A na-edozi pH nke ngwọta na 2.0 site na ịgbakwunye ngwọta hydrochloride (1M).Mgbe ahụ, calcium chloride (5.62 mM) gbazere na ngwọta.Site na ịgbakwunye 1M tris (hydroxymethyl) -amino Methane na-edozi pH nke ngwọta na 7.4.Na-emikpu ihe niile n'ime efere isii nke ọma jupụtara na ngwọta calcium phosphate 1.5 × ma wepụ ya na ngwọta mgbe minit 30 gasịrị.Maka ntụpọ, 2 g Alizarin Red S (CI 58005) Gwakọta na 100 ml nke mmiri deionized.Mgbe ahụ, jiri 10% ammonium hydroxide dozie pH ka ọ bụrụ 4. Tinye ihe ngwọta Alizarin na-acha uhie uhie maka nkeji 5, wee kpochapụ ihe na-esi ísì ụtọ ma kpochaa.Mgbe usoro ịma jijiji ahụ gasịrị, wepụ mkpụrụ ahụ.A na-ekpochapụ ihe ahụ, wee gbanye ya na acetone maka nkeji 5, wee gbanye ya na acetone-xylene (1: 1) ngwọta maka nkeji 5, n'ikpeazụkwa na-asa ya na xylene (n = 4).A na-eji microscope fluorescence (Axio Imager) nwere oghere ebumnuche ×10 na ×20..A2m, Zeiss, Germany) na-ese onyinyo ihe niile.A na-eji ImageJ/FIJI (https://imagej.nih.gov/ij/) kọwaa data adhesion nke ihe ndị dị ndụ na otu ọ bụla nke mpaghara ihe onyonyo anọ dị iche iche.Tugharia onyonyo niile ka ọ bụrụ onyonyo ọnụọgụ abụọ nwere ọnụ ụzọ kwụ ọtọ maka ntụnyere mkpụrụ osisi.
Ejiri microscope confocal Zeiss LSM 700 iji nyochaa nkwụsi ike nke oyi akwa mmanu na PBS na ọnọdụ ntụgharị uche.A na-emikpu ihe nlele SAM mkpuchi fluorine nke nwere oyi akwa mmanye agbanyere n'ime ngwọta PBS, wee nwalee site na iji orbital shaker (SHO-1D; Daihan Scientific, South Korea) n'okpuru ọnọdụ ịma jijiji dị nro (120 rpm).Wee were nlele ahụ wee nyochaa ọnwụ nke mmanu mmanu site n'ịtụ ụda ọkụ na-egosipụta.Iji nweta ihe oyiyi fluorescence na ọnọdụ ntụgharị, a na-ekpughere ihe nlele ahụ na laser 633 nm wee kpokọta ya, n'ihi na ọkụ ga-egosipụta azụ site na nlele ahụ.A tụrụ ihe nlele ahụ n'oge oge nke 0, 30, 60, na 120 awa.
Iji chọpụta mmetụta nke usoro mgbanwe n'elu na njirimara nanomechanical nke orthopedic implants, a nanoindenter (TI 950 TriboIndenter, Hysitron, USA) nke nwere ọnụ ọnụ diamond Berkovich nwere akụkụ pyramid nwere akụkụ atọ dị n'akụkụ ọnụ iji tụọ nanoindenedione.Ọnụ kacha elu bụ 10mN na mpaghara ahụ bụ 100μmx 100μm.Maka nha niile, oge nbudata na nbudata bụ 10 s, na oge njide n'okpuru ibu ntinye kacha elu bụ 2 s.Were nha site na ebe dị iche iche ise wee were nkezi.Iji nyochaa arụmọrụ ike n'ibu n'okpuru ibu, a na-eji igwe nyocha nke ụwa (Instron 5966, Instron, USA).A na-ejikọta mkpụrụ ahụ na ọnụego 10 N / s mgbe niile na-ebuwanye ibu.A na-eji mmemme ngwanrọ Bluehill Universal (n = 3) gbakọọ modulus mgbanwe yana nrụgide mkpakọ kacha.
Iji mee ka usoro ọrụ ahụ na-emebi emebi na-arụ ọrụ na ihe ndị metụtara ya mere n'oge ọrụ ahụ, a na-arụ ọrụ ahụ na vitro.A nakọtara femurs ahụ n'aka oke bekee ọcha New Zealand gburu.A na-ehicha femur ma dozie ya na 4% paraformaldehyde maka izu 1.Dị ka akọwara n'usoro nnwale anụmanụ, a na-awa ahụ femur a kapịrị ọnụ.Mgbe arụ ọrụ ahụ gasịrị, a na-agbanye ọkpụkpụ orthopedic n'ime ọbara (ọbara ịnyịnya, KISAN, Korea) maka 10 s iji gosi ma ọ bụrụ na adhesions ọbara mere mgbe a na-etinye mmerụ ahụ (n = 3).
Ngụkọta 24 nwoke New Zealand ọcha oke bekee (arọ 3.0 ruo 3.5kg, nkezi afọ 6 ọnwa) kewara n'ụzọ anọ: ịgba ọtọ adịghị mma, ịgba ọtọ, SHP na LOIS.Emere usoro niile metụtara ụmụ anụmanụ dịka ụkpụrụ ụkpụrụ omume nke Kọmitii Nlekọta na ojiji nke Institutional Animal Care and Use Committee (IACUC kwadoro, KOREA-2017-0159).Ihe nkedo nke orthopedic na-agụnye efere mkpọchi nke nwere oghere ise (ogologo 41 mm, obosara 7 mm na ọkpụrụkpụ 2 mm) na mkpuchi mkpuchi cortical (ogologo 12 mm, dayameta 2.7 mm) maka nkwụsị mgbawa.Ewezuga efere na skru ndị ahụ ejiri n'ime otu adịghị mma, efere niile na skru ka etinyere na nkwusioru MRSA (106 CFU/ml) maka awa iri na abụọ.A na-emeso otu ndị na-adịghị mma (n=6) site na ntinye elu nke gba ọtọ na-enweghị ikpughe na nkwusioru nje, dị ka njikwa na-adịghị mma maka ọrịa.A na-emeso ndị ahụ na-adịghị mma (n = 6) site na ntinye n'elu ala nke ekpughere na nje bacteria dị ka njikwa dị mma maka ọrịa.A na-emeso òtù SHP (n = 6).N'ikpeazụ, a na-emeso òtù LOIS ahụ site na ntinye LOIS nke na-ekpughe nje bacteria (n = 6).A na-edobe anụmanụ niile n'ime ọnụ ụlọ, a na-enyekwa ọtụtụ nri na mmiri.Tupu ọrụ ahụ, a na-ebu ọnụ rabbits maka awa 12.A na-eme ka ụmụ anụmanụ na-agba ume site na intramuscular injection of xylazine (5mg/kg) na intravenous injection of paclitaxel (3mg/kg) maka ntinye.Mgbe nke ahụ gasịrị, nyefee 2% isoflurane na 50% ruo 70% oxygen ahụike (ọnụọgụ 2 L / min) site na usoro iku ume iji nọgide na-enwe anesthesia.A na-akụnye ya site na ịbịaru nso n'akụkụ femur.Mgbe ewepụchara ntutu isi na nsị nke povidone-iodine nke akpụkpọ ahụ, a na-eme mbepụ ihe dịka 6 cm ogologo n'èzí nke etiti femur ekpe.Site na imepe oghere dị n'etiti mọzụlụ na-ekpuchi femur, femur na-ekpughe nke ọma.Debe efere ahụ n'ihu ogwe femoral ma dozie ya na nkedo anọ.Mgbe emezichara, jiri eriri nkwọ (1mm gbara ọkpụrụkpụ) mee ka mgbaji dị n'etiti oghere nke abụọ na oghere nke anọ.Na njedebe nke ọrụ ahụ, a na-asacha ọnyá ahụ na saline ma mechie ya na sutures.A gbanyere oke bekee ọ bụla n'okpuru akpụkpọ ahụ na enrofloxacin (5 mg / kg) gbanyere otu ụzọ n'ụzọ atọ na saline.A na-ewere ụzarị X-ray nke femur mgbe arụchara ọrụ na anụmanụ niile (0, 7, 14, 21, 28, na 42 ụbọchị) iji kwado osteotomy nke ọkpụkpụ.Mgbe mgbawa miri emi gasịrị, KCl (2 mmol / kg) na-egbu anụ niile gburu anụmanụ n'ime ụbọchị 28 na 42.Mgbe e gbuchara ya, micro-CT na-enyocha femur ka ọ hụ ma tụnyere usoro ọgwụgwọ ọkpụkpụ na ọkpụkpụ ọhụrụ n'etiti otu anọ.
Mgbe e gbuchara ya, a na-achịkọta anụ ahụ dị nro nke na-ejikọta kpọmkwem na ọkpụkpụ orthopedic.Edobere anụ ahụ ahụ na 10% na-anọpụ iche nke formalin n'abali wee kpoo ya na EtOH.A na-etinye anụ ahụ akpọnwụ akpọnwụ na paraffin wee kewaa ya na ọkpụrụkpụ nke 40 μm site na iji microtome (400CS; EXAKT, Germany).Iji hụ ọrịa ahụ anya, a na-eme ntụpọ H&E na ntụpọ MT.Iji lelee nzaghachi onye ọbịa, a na-etinye anụ ahụ nke akụkụ ya na oke mgbochi TNF-α bụ isi (AB6671, Abcam, USA) na rabbit anti-IL-6 (AB6672; Abcam, USA), wee mesoo ya na horseradish.Oxidase.Tinye usoro ntụpọ avidin-biotin (ABC) na ngalaba dịka ntuziaka onye nrụpụta si dị.Ka ọ pụta ìhè dị ka ngwaahịa mmeghachi omume agba aja aja, 3,3-diaminobenzidine ejiri mee ihe n'akụkụ niile.A na-eji ihe nlebanya slide dijitalụ (Pannoramic 250 Flash III, 3DHISTECH, Hungary) were were were were anya nke uche nke mpekere niile, na opekata mpe ihe anọ dị n'ime otu ọ bụla ka ejiri ngwa ImageJ nyochara ya.
A na-ese foto X-ray n'ime anụmanụ niile mgbe a wachara ya ahụ yana kwa izu iji nyochaa ọgwụgwọ mgbaji (n=6 kwa otu).Mgbe e gbuchara ya, a na-eji micro-CT dị elu mee ihe iji gbakọọ nhazi nke callus gburugburu femur mgbe ọgwụgwọ gasịrị.A na-ehicha femur ahụ enwetara, dozie ya na 4% paraformaldehyde maka ụbọchị 3, wee kpoo ya na 75% ethanol.A na-enyocha ọkpụkpụ ndị ahụ akpọnwụwo site na iji micro-CT (SkyScan 1173, Brooke Micro-CT, Kandy, Belgium) iji mepụta ihe oyiyi voxel 3D (2240 ​​× 2240 pixels) nke ọkpụkpụ ọkpụkpụ.Jiri 1.0 mm Al filter iji belata mkpọtụ mgbaàmà wee tinye mkpebi dị elu na nyocha niile (E = 133 kVp, I = 60 μA, oge ntinye = 500 ms).Ejiri ngwanrọ Nrecon (ụdị 1.6.9.8, Bruker microCT, Kontich, Belgium) wepụta olu 3D nke nlele nyocha site na ntule akụkụ 2D enwetara.Maka nyocha, a na-ekewa onyonyo 3D arụgharịrị n'ime cubes 10mm × 10mm × 10mm dịka saịtị mgbawa siri dị.Gbakọọ callus n'èzí ọkpụkpụ cortical.DataViewer (ụdị 1.5.1.2; Bruker microCT, Kontich, Belgium) software e ji digitally redirectly nke ọkpụkpụ scanned olu, na CT-Analyzer (ụdị 1.14.4.1; Bruker microCT, Kontich, Belgium) software e ji mee nyocha.A na-amata ọnụọgụ mmịpụta x-ray dị n'ọkpụkpụ tozuru oke na callus site na njupụta ha, mgbe ahụ a na-atụle olu callus (n = 4).Iji gosi na bioocompatibility nke LOIS adịghị egbu oge usoro ọgwụgwọ ọkpụkpụ, a na-eme nyocha X-ray na micro-CT ọzọ na rabbits abụọ: ndị gba ọtọ-na-adịghị mma na LOIS.E gburu otu abụọ ahụ n'izu nke isii.
A na-anakọta femurs sitere na anụmanụ ndị a chụrụ n'àjà ma dozie ya na 4% paraformaldehyde maka ụbọchị 3.A na-ewepụzi ihe nkedo orthopedic nke ọma na femur.E mebiri femur maka ụbọchị 21 site na iji 0.5 M EDTA (EC-900, National Diagnostics Corporation).Mgbe ahụ, femur ahụ mebiri emebi na-emikpu na EtOH iji mee ka mmiri gwụ ya.E wepụrụ femur mmiri na-ekpo ọkụ na xylene ma tinye ya na paraffin.Mgbe ahụ, a na-eji microtome rotary na-akpaghị aka (Leica RM2255, Leica Biosystems, Germany) kechie ihe nlele ahụ na ọkpụrụkpụ nke 3 μm.Maka ntụpọ TRAP (F6760, Sigma-Aldrich, Germany), a na-ekpochapụ ihe nlele ndị ahụ, rehydrated ma tinye ya na TRAP reagent na 37 ° C maka awa 1.Enwetara onyonyo site na iji ihe nyocha slide (Pannoramic 250 Flash III, 3DHISTECH, Hungary) wee wepụta ya site n'ịtụ mpaghara mkpuchi nke ebe ntụpọ ahụ.N'ime nnwale ọ bụla, opekata mpe mkpụrụ anọ n'ime otu ọ bụla ka ejiri ngwa ImageJ nyochara.
Emere nyocha ọnụ ọgụgụ dị mkpa site na iji GraphPad Prism (GraphPad Software Inc., USA).A na-eji t-ule na-ejikọtaghị ọnụ na nyocha nke otu ụzọ (ANOVA) iji nwalee ọdịiche dị n'etiti otu nyocha.E gosipụtara ọkwa dị mkpa na ọnụ ọgụgụ a: * P <0.05, ** P<0.01, *** P<0.001 na ****P <0.0001;NS, enweghị nnukwu ọdịiche.
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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
Ihe mkpuchi nje bacteria na mgbapụ nke mgbochi nke ọkpụkpụ orthopedic nwere ike ibelata ọrịa na nzaghachi mgbochi nke ọrịa na-ebute.
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
Ihe mkpuchi nje bacteria na mgbapụ nke mgbochi nke ọkpụkpụ orthopedic nwere ike ibelata ọrịa na nzaghachi mgbochi nke ọrịa na-ebute.
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