本文關(guān)鍵詞：銀鍶摻雜納米羥基磷灰石的合成及骨修復(fù)基礎(chǔ)研究　出處：《河北大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 骨修復(fù) 納米羥基磷灰石 感染 炎癥 礦化

【摘要】：當(dāng)骨由于損傷或疾病而發(fā)生骨缺損后,引入骨修復(fù)材料以用于恢復(fù)骨骼完整性、填充空隙和增強(qiáng)骨修復(fù)能力就成為了迫切的需求。納米羥基磷灰石(HA)的組成和結(jié)構(gòu)與骨的礦物質(zhì)成分相似,具有生物相容性高、骨誘導(dǎo)性強(qiáng)、比表面積大、成骨性好、生物降解性強(qiáng)、骨整合性好等優(yōu)點(diǎn),應(yīng)用在骨缺損修復(fù)方面具有天然的優(yōu)勢(shì)。骨修復(fù)過程中可能會(huì)出現(xiàn)感染和炎癥,在HA中摻入銀和鍶可以防止感染以及促進(jìn)HA的礦化能力。本文采用水熱法以EDTA二鈉為模板合成了系列納米HA、銀摻雜納米HA、鍶摻雜納米HA和銀鍶共摻雜HA。使用TEM、SEM、XRD、FT-IR和EDS表征不同HA的形貌、結(jié)構(gòu)和組成。以大腸桿菌(E.Coli)和金黃色葡萄球菌(S.aureus)作為模型,評(píng)價(jià)了不同HA的抗菌性;以巨噬細(xì)胞(RAW264.7)和成骨細(xì)胞(MC3T3-E1)作為模型,評(píng)價(jià)了不同HA對(duì)細(xì)胞活力、成骨細(xì)胞ALP的表達(dá)及礦化結(jié)節(jié)能力和巨噬細(xì)胞的炎癥因子(TNF-α與IL-6)的表達(dá)的影響。本文具體的研究?jī)?nèi)容及結(jié)果主要包括以下三個(gè)方面:(1)合成了納米HA和銀摻雜納米HA,實(shí)驗(yàn)結(jié)果表明:合成的材料為納米級(jí)實(shí)心棒狀,平均直徑為60.1±4.8 nm,平均長(zhǎng)度為173.7±8.5 nm。不同質(zhì)量分?jǐn)?shù)的銀摻雜納米HA都對(duì)E.Coli和S.aureus具有抗菌作用,且銀摻雜質(zhì)量分?jǐn)?shù)越高,抗菌效果越好。銀摻雜納米HA對(duì)巨噬細(xì)胞和成骨細(xì)胞毒性較小,納米HA和Ag_(5.0)HA相對(duì)于對(duì)照組都促進(jìn)了ALP的表達(dá),表現(xiàn)出促成骨細(xì)胞分化的能力,HA相對(duì)于OS對(duì)照組的ALP表達(dá)量提高了449.8%,Ag_(5.0)HA相對(duì)于OS對(duì)照組的ALP表達(dá)量提高了432.1%,Ag_(5.0)HA的促成骨細(xì)胞分化的能力稍弱于HA。HA和Ag_(5.0)HA都促進(jìn)了礦化結(jié)節(jié)的生成,HA相對(duì)于OS對(duì)照組的礦化量提高了63.3%,Ag_(5.0)HA相對(duì)于OS對(duì)照組的礦化量提高了58.8%。HA和Ag_(5.0)HA都對(duì)RAW264.7細(xì)胞的細(xì)胞因子TNF-α的分泌有抑制作用,抑制率分別為18.7%和29.1%,Ag_(5.0)HA的抑制程度更顯著。HA對(duì)RAW264.7細(xì)胞的細(xì)胞因子IL-6的分泌沒有影響,而Ag_(5.0)HA明顯抑制了IL-6的表達(dá),抑制率為13.1%。(2)合成了納米HA和鍶摻雜納米HA,實(shí)驗(yàn)結(jié)果表明:合成的材料為納米級(jí)實(shí)心棒狀,鍶摻雜納米HA的平均直徑為38.6±2.2 nm,平均長(zhǎng)度為135.3±27.4 nm,均低于納米HA。鍶摻雜納米HA生物相容性好,納米HA和Sr_(5.0)HA相對(duì)于對(duì)照組都促進(jìn)了ALP的表達(dá),表現(xiàn)出促成骨細(xì)胞分化的能力,HA相對(duì)于OS對(duì)照組的ALP表達(dá)量提高了449.8%,Sr_(5.0)HA相對(duì)于OS對(duì)照組的ALP表達(dá)量提高了630.8%。Sr_(5.0)HA的促成骨細(xì)胞分化的能力強(qiáng)于HA,ALP表達(dá)量提高了181.0%,說明鍶摻雜能顯著提高成骨細(xì)胞成骨分化的能力。HA和Sr_(5.0)HA具有明顯的促礦化能力,HA相對(duì)于OS對(duì)照組的礦化量提高了63.3%,Sr_(5.0)HA相對(duì)于OS對(duì)照組的礦化量提高了136.2%。而Sr_(5.0)HA又比HA的礦化能力提高了72.9%,因此鍶摻雜能提高HA的礦化能力。HA和Sr_(5.0)HA都對(duì)RAW264.7細(xì)胞的細(xì)胞因子TNF-α的分泌產(chǎn)生了抑制作用,抑制率分別為18.7%和19.3%。HA對(duì)IL-6的分泌沒有影響,沒有顯著性差異,而Sr_(5.0)HA對(duì)IL-6的分泌有抑制作用,抑制率為11.3%。(3)合成了納米HA和銀鍶共摻雜納米HA,實(shí)驗(yàn)結(jié)果表明:合成的材料都是納米級(jí)實(shí)心棒狀,銀鍶共摻雜納米HA的平均直徑和平均長(zhǎng)度均低于納米HA,銀鍶共摻雜納米HA的平均直徑為37.0±4.8 nm,平均長(zhǎng)度為109.1±20.0 nm。不同鍶摻雜比的銀鍶共摻雜納米HA都對(duì)E.Coli和S.aureus有抗菌作用,對(duì)E.Coli的抑菌圈半徑在約6.6 mm到約6.9 mm之間,大于Ag_(5.0)HA對(duì)E.Coli的抑菌圈半徑;對(duì)S.aureus的抑菌圈半徑在約6.2 mm到約6.5 mm之間,大于Ag_(5.0)HA對(duì)S.aureus的抑菌圈半徑。銀鍶共摻雜HA毒性小,納米HA和Ag_(5.0)Sr_(5.0)HA相對(duì)于對(duì)照組都促進(jìn)了ALP的表達(dá),表現(xiàn)出促成骨細(xì)胞分化的能力,HA相對(duì)于OS對(duì)照組的ALP表達(dá)量提高了297.1%,Ag_(5.0)Sr_(5.0)HA相對(duì)于OS對(duì)照組的ALP表達(dá)量提高了451.1%。Ag_(5.0)Sr_(5.0)HA的促成骨細(xì)胞分化的能力強(qiáng)于HA,ALP表達(dá)量提高了154.0%,說明銀鍶摻共摻雜能顯著提高成骨細(xì)胞成骨分化的能力。HA和Ag_(5.0)Sr_(5.0)HA具有明顯的促礦化能力,HA相對(duì)于OS對(duì)照組的礦化量提高了63.3%,Ag_(5.0)Sr_(5.0)HA相對(duì)于OS對(duì)照組的礦化量提高了89.9%。而Ag_(5.0)Sr_(5.0)HA又比HA的礦化能力提高了26.6%,因此銀鍶共摻雜能提高HA的礦化能力。HA和Ag_(5.0)Sr_(5.0)HA都對(duì)RAW264.7細(xì)胞的細(xì)胞因子TNF-α的分泌產(chǎn)生了抑制作用,抑制率分別為18.7%和36.0%。HA對(duì)IL-6的分泌沒有影響,沒有顯著性差異,而Ag_(5.0)Sr_(5.0)HA對(duì)IL-6的分泌有抑制作用,抑制率為10.2%。銀鍶共摻雜賦予了納米HA抗菌、抗炎和促成骨分化礦化的能力。
[Abstract]:When the bone and bone defect due to injury or disease, the introduction of bone repair materials for restoration of bone integrity, filling the gap and enhance bone repair ability has become an urgent demand. Nano hydroxyapatite (HA) mineral composition and structure of bone and similar, with high biocompatibility, osteoinductivity. A large surface area, good osteogenic, biodegradable, has the advantages of good osseointegration, the application has a natural advantage in the repair of bone defects. There may be infection and inflammation in the bone repair process in HA doped with silver and strontium can prevent infection and promote the mineralization of HA. This paper uses the hydrothermal synthesis of HA nano template with two sodium EDTA, silver doped nano HA, nano HA and silver doped strontium strontium doped HA. using TEM, SEM, XRD, FT-IR and EDS characterization of different HA morphology, structure and composition. In Escherichia coli (E.Coli) and golden Aureus (S.aureus) as a model, the evaluation of different HA antibacterial; macrophages (RAW264.7) and osteoblasts (MC3T3-E1) as a model, the evaluation of different HA on cell viability, inflammatory cytokines ALP expression of osteoblasts and mineralized nodule ability and macrophages (TNF- alpha and IL-6) effects expression. Research contents and results of this paper include the following three aspects: (1) HA and nano silver doped nano HA was synthesized, the experimental results show that the synthesis of materials for nano solid rod, the average diameter is 60.1 + 4.8 nm, the average length of silver doped HA 173.7 + 8.5 different mass fractions of nm. has antibacterial effect on E.Coli and S.aureus, and the silver doping concentration is high, the better. The antibacterial effect of silver doped nano HA on macrophages and osteoblasts less toxicity of nano HA and Ag_ (5) HA for the control group are promoting ALP table As shown, the ability to achieve differentiation of bone cells, HA compared with OS control group the expression of ALP increased by 449.8%, Ag_ (5) HA compared with OS control group the expression of ALP increased by 432.1%, Ag_ (5) HA induced osteoblast differentiation ability weaker than that of HA.HA and Ag_ (5) HA Pro the formation of mineralized nodules, compared to the control group OS HA mineralization rate increased by 63.3%, Ag_ (5) HA relative to the amount of mineralization of OS group increased by 58.8%.HA and Ag_ (5) HA on RAW264.7 cells TNF- cytokine secretion inhibition, the inhibition rate were 18.7% and 29.1%, Ag_ (5) the degree of inhibition of HA more significantly IL-6 cytokines.HA on RAW264.7 cells have no effect on the secretion of Ag_, and (5) IL-6 significantly inhibited the expression of HA, the inhibition rate was 13.1%. (2) nano HA and strontium doped nano HA was synthesized, the experimental results show that the synthesis of nano solid material strontium doped nano rod. 綾矵A鐨勫鉤鍧囩洿寰勪負(fù)38.6鹵2.2 nm,騫沖潎闀垮害涓，

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