银锶掺杂纳米羟基磷灰石的合成及骨修复基础研究

发布时间：2018-01-13 17:44

  本文关键词：银锶掺杂纳米羟基磷灰石的合成及骨修复基础研究　出处：《河北大学》2017年硕士论文　论文类型：学位论文

  更多相关文章： 骨修复 纳米羟基磷灰石 感染 炎症 矿化

【摘要】：当骨由于损伤或疾病而发生骨缺损后,引入骨修复材料以用于恢复骨骼完整性、填充空隙和增强骨修复能力就成为了迫切的需求。纳米羟基磷灰石(HA)的组成和结构与骨的矿物质成分相似,具有生物相容性高、骨诱导性强、比表面积大、成骨性好、生物降解性强、骨整合性好等优点,应用在骨缺损修复方面具有天然的优势。骨修复过程中可能会出现感染和炎症,在HA中掺入银和锶可以防止感染以及促进HA的矿化能力。本文采用水热法以EDTA二钠为模板合成了系列纳米HA、银掺杂纳米HA、锶掺杂纳米HA和银锶共掺杂HA。使用TEM、SEM、XRD、FT-IR和EDS表征不同HA的形貌、结构和组成。以大肠杆菌(E.Coli)和金黄色葡萄球菌(S.aureus)作为模型,评价了不同HA的抗菌性;以巨噬细胞(RAW264.7)和成骨细胞(MC3T3-E1)作为模型,评价了不同HA对细胞活力、成骨细胞ALP的表达及矿化结节能力和巨噬细胞的炎症因子(TNF-α与IL-6)的表达的影响。本文具体的研究内容及结果主要包括以下三个方面:(1)合成了纳米HA和银掺杂纳米HA,实验结果表明:合成的材料为纳米级实心棒状,平均直径为60.1±4.8 nm,平均长度为173.7±8.5 nm。不同质量分数的银掺杂纳米HA都对E.Coli和S.aureus具有抗菌作用,且银掺杂质量分数越高,抗菌效果越好。银掺杂纳米HA对巨噬细胞和成骨细胞毒性较小,纳米HA和Ag_(5.0)HA相对于对照组都促进了ALP的表达,表现出促成骨细胞分化的能力,HA相对于OS对照组的ALP表达量提高了449.8%,Ag_(5.0)HA相对于OS对照组的ALP表达量提高了432.1%,Ag_(5.0)HA的促成骨细胞分化的能力稍弱于HA。HA和Ag_(5.0)HA都促进了矿化结节的生成,HA相对于OS对照组的矿化量提高了63.3%,Ag_(5.0)HA相对于OS对照组的矿化量提高了58.8%。HA和Ag_(5.0)HA都对RAW264.7细胞的细胞因子TNF-α的分泌有抑制作用,抑制率分别为18.7%和29.1%,Ag_(5.0)HA的抑制程度更显著。HA对RAW264.7细胞的细胞因子IL-6的分泌没有影响,而Ag_(5.0)HA明显抑制了IL-6的表达,抑制率为13.1%。(2)合成了纳米HA和锶掺杂纳米HA,实验结果表明:合成的材料为纳米级实心棒状,锶掺杂纳米HA的平均直径为38.6±2.2 nm,平均长度为135.3±27.4 nm,均低于纳米HA。锶掺杂纳米HA生物相容性好,纳米HA和Sr_(5.0)HA相对于对照组都促进了ALP的表达,表现出促成骨细胞分化的能力,HA相对于OS对照组的ALP表达量提高了449.8%,Sr_(5.0)HA相对于OS对照组的ALP表达量提高了630.8%。Sr_(5.0)HA的促成骨细胞分化的能力强于HA,ALP表达量提高了181.0%,说明锶掺杂能显著提高成骨细胞成骨分化的能力。HA和Sr_(5.0)HA具有明显的促矿化能力,HA相对于OS对照组的矿化量提高了63.3%,Sr_(5.0)HA相对于OS对照组的矿化量提高了136.2%。而Sr_(5.0)HA又比HA的矿化能力提高了72.9%,因此锶掺杂能提高HA的矿化能力。HA和Sr_(5.0)HA都对RAW264.7细胞的细胞因子TNF-α的分泌产生了抑制作用,抑制率分别为18.7%和19.3%。HA对IL-6的分泌没有影响,没有显著性差异,而Sr_(5.0)HA对IL-6的分泌有抑制作用,抑制率为11.3%。(3)合成了纳米HA和银锶共掺杂纳米HA,实验结果表明:合成的材料都是纳米级实心棒状,银锶共掺杂纳米HA的平均直径和平均长度均低于纳米HA,银锶共掺杂纳米HA的平均直径为37.0±4.8 nm,平均长度为109.1±20.0 nm。不同锶掺杂比的银锶共掺杂纳米HA都对E.Coli和S.aureus有抗菌作用,对E.Coli的抑菌圈半径在约6.6 mm到约6.9 mm之间,大于Ag_(5.0)HA对E.Coli的抑菌圈半径;对S.aureus的抑菌圈半径在约6.2 mm到约6.5 mm之间,大于Ag_(5.0)HA对S.aureus的抑菌圈半径。银锶共掺杂HA毒性小,纳米HA和Ag_(5.0)Sr_(5.0)HA相对于对照组都促进了ALP的表达,表现出促成骨细胞分化的能力,HA相对于OS对照组的ALP表达量提高了297.1%,Ag_(5.0)Sr_(5.0)HA相对于OS对照组的ALP表达量提高了451.1%。Ag_(5.0)Sr_(5.0)HA的促成骨细胞分化的能力强于HA,ALP表达量提高了154.0%,说明银锶掺共掺杂能显著提高成骨细胞成骨分化的能力。HA和Ag_(5.0)Sr_(5.0)HA具有明显的促矿化能力,HA相对于OS对照组的矿化量提高了63.3%,Ag_(5.0)Sr_(5.0)HA相对于OS对照组的矿化量提高了89.9%。而Ag_(5.0)Sr_(5.0)HA又比HA的矿化能力提高了26.6%,因此银锶共掺杂能提高HA的矿化能力。HA和Ag_(5.0)Sr_(5.0)HA都对RAW264.7细胞的细胞因子TNF-α的分泌产生了抑制作用,抑制率分别为18.7%和36.0%。HA对IL-6的分泌没有影响,没有显著性差异,而Ag_(5.0)Sr_(5.0)HA对IL-6的分泌有抑制作用,抑制率为10.2%。银锶共掺杂赋予了纳米HA抗菌、抗炎和促成骨分化矿化的能力。
[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. 绫矵A鐨勫钩鍧囩洿寰勪负38.6卤2.2 nm,骞冲潎闀垮害涓，

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