本文選題：殼聚糖 + 鈦��； 參考：《第二軍醫(yī)大學(xué)》2013年博士論文

【摘要】：背景 內(nèi)植物材料已在骨科得到廣泛應(yīng)用，內(nèi)植物周圍感染問題也日益突出。骨科內(nèi)植物表面宿主免疫低下、磨損顆粒損害周圍組織、細(xì)菌耐藥、生物膜形成等導(dǎo)致這類感染極難治療，也導(dǎo)致住院時(shí)間延長，增加患者痛苦，增加并發(fā)癥和死亡風(fēng)險(xiǎn)，也造成醫(yī)療資源的巨大支出。許多研究致力于內(nèi)植物表面改性，但往往存在抑菌能力欠佳、導(dǎo)致細(xì)菌耐藥、妨礙骨整合等缺陷。如能研制出新型抗菌內(nèi)植物表面，使其具備良好的抗菌能力與生物相容性，且可有效預(yù)防細(xì)菌粘附與生物膜形成，甚至可促進(jìn)骨整合，將有一定的研究與應(yīng)用前景。 第一部分 目的 合成不同取代度的氯乙酰硫脲羧甲基殼聚糖衍生物（CCMC），研究其抑菌性能、細(xì)胞毒性以及對(duì)骨髓間充質(zhì)干細(xì)胞（MSCs）增殖與分化的影響。 方法 1、以抑菌圈試驗(yàn)及抑菌率試驗(yàn)測試其對(duì)金黃色葡萄球菌、表皮葡萄球菌的體外抗菌性能； 2、以MTT比色法研究其對(duì)MC3T3-E1細(xì)胞的細(xì)胞毒性； 3、以MTT比色法測定其對(duì)MSCs增殖的影響； 4、以堿性磷酸酶活性和骨鈣素含量測定其對(duì)MSCs分化的影響。 結(jié)果 1、隨著CCMC取代度的增高，其抑菌活性也逐漸增強(qiáng)； 2、0.88取代度的CCMC具有體外細(xì)胞毒性，另兩種取代度的CCMC沒有體外細(xì)胞毒性； 3、0.24與0.53取代度的CCMC對(duì)MSCs的增殖和分化均有一定促進(jìn)作用，0.88取代度的CCMC對(duì)MSCs的增殖和分化均有一定抑制作用。 第二部分 目的 測定不同取代度的CCMC的最小抑菌濃度及最小生物膜清除濃度，以及特定濃度下對(duì)金黃色葡萄球菌以及表皮葡萄球菌生物膜抑制率及對(duì)生物膜活性的影響。 方法 1、以細(xì)菌定量法測定不同取代度的CCMC的最小抑菌濃度，以比色法測定各取代度CCMC的最小生物膜清除濃度； 2、以比色法測定各取代度CCMC在特定濃度下對(duì)三種生物膜的抑制率； 3、以熒光染色比較各取代度CCMC對(duì)表皮葡萄球菌生物膜活性的影響。 結(jié)果 1、隨著CCMC取代度的增高，最小抑菌濃度及最小生物膜清除濃度也逐漸減��； 2、相同濃度下，取代度越高，CCMC對(duì)三種生物膜的抑制率越高，0.88與0.53取代度的CCMC在較小濃度下即可有效抑制生物膜生長； 3、相同濃度下，取代度越高，CCMC對(duì)表皮葡萄球菌生物膜活性的抑制能力越強(qiáng)；隨著濃度的增高，各取代度CCMC對(duì)表皮葡萄球菌生物膜活性的抑制能力越強(qiáng)。 第三部分 目的 鈦合金表面共價(jià)結(jié)合0.53取代度的CCMC（Ti-CCMC），通過與鈦合金表面（Ti）的比較，研究這種鈦表面抑制生物膜的能力、對(duì)成骨細(xì)胞粘附與增殖的影響，，建立新西蘭大白兔感染模型，研究Ti-CCMC對(duì)感染局部骨組織的影響與全身性影響。 方法 1、以熒光染色法比較Ti-CCMC與Ti在體外對(duì)生物膜活性的影響； 2、以細(xì)菌定量法比較Ti-CCMC與Ti在體外對(duì)表皮葡萄球菌粘附的影響； 3、比較Ti-CCMC與Ti對(duì)MC3T3-E1細(xì)胞粘附與增殖的影響； 4、以細(xì)菌培養(yǎng)定量檢測法測定鈦棒周圍骨組織內(nèi)細(xì)菌含量； 5、比較兩組兔的體重變化，行血細(xì)菌培養(yǎng)、X線檢查，研究兩組兔全身性情況的差異； 6、對(duì)兩組兔股骨中段髓腔行革蘭氏染色，比較兩組細(xì)菌局部遷移的差異。 結(jié)果 1、Ti-CCMC體外對(duì)生物膜活性有明顯抑制作用； 2、Ti-CCMCC由一定的抑制表皮葡萄球菌粘附的能力； 3、Ti-CCMC可促進(jìn)MC3T3-E1細(xì)胞粘附與增殖； 4、Ti組骨組織內(nèi)見細(xì)菌生長，Ti-CCMC組未見細(xì)菌生長； 5、兩組兔體重比較無明顯差異，Ti組術(shù)后X線檢查發(fā)現(xiàn)股骨骨質(zhì)破壞，Ti-CCMC組未見明顯異常，未見骨溶解與透亮帶； 6、Ti組兔股骨中段髓腔行革蘭氏染色，可見革蘭氏藍(lán)染的細(xì)菌散在，Ti組未見細(xì)菌。 結(jié)論 殼聚糖�；螂逖苌锞哂辛己玫目咕芰Α⑸锵嗳菪耘c骨整合能力，可作為骨科內(nèi)植物表面抗菌的新型材料，具有一定的研究與應(yīng)用前景。
[Abstract]:background
Internal plant materials have been widely used in the Department of orthopedics, and the problems of the infection around plants are also becoming increasingly prominent. The low immunity of the plant surface host in the Department of orthopedics, the damage to the surrounding tissues, the resistance of the bacteria, the formation of biofilm, etc., lead to the extremely difficult treatment of this type of infection, and the prolonged hospitalization, the pain of the patients, the increase of the risk of complications and death. Many studies have contributed to the enormous expenditure of medical resources. Many studies have been devoted to the surface modification of plants, but they often have poor bacteriostasis, causing resistance to bacteria and impeding bone integration, such as developing a new type of antibacterial internal plant surface, making it good for antibacterial and biocompatibility, and effectively preventing bacterial adhesion and biofilm. It can even promote bone integration. It will have some research and application prospects.
Part one
objective
The synthesis of chloracetyl thiourea carboxymethyl chitosan derivative (CCMC) with different degrees of substitution was synthesized, and its antibacterial properties, cytotoxicity and the effect on the proliferation and differentiation of bone marrow mesenchymal stem cells (MSCs) were studied.
Method
1, the antibacterial activity of Staphylococcus aureus and Staphylococcus epidermidis was tested by inhibition zone test and inhibition rate test.
2, the cytotoxicity of MC3T3-E1 cells was studied by MTT colorimetry.
3, the effect of MTT on the proliferation of MSCs was determined by the colorimetric method.
4, the effects of alkaline phosphatase activity and osteocalcin content on MSCs differentiation were measured.
Result
1, with the increase of CCMC substitution degree, its antibacterial activity also increased.
CCMC with 2,0.88 substitution degree had cytotoxicity in vitro, and the other two CCMC of substitution degree had no cytotoxicity in vitro.
3,0.24 and 0.53 degree of substitution CCMC promoted the proliferation and differentiation of MSCs, and the 0.88 degree of substitution CCMC inhibited the proliferation and differentiation of MSCs.
The second part
objective
The minimum inhibitory concentration and minimum biofilm clearance concentration of CCMC with different degree of substitution, as well as the inhibitory rate of Staphylococcus aureus and Staphylococcus epidermidis in specific concentration and the effect on biofilm activity under specific concentration were measured.
Method
1, the minimum inhibitory concentration of CCMC with different degree of substitution was determined by bacterial quantitative method, and the minimum biofilm clearance concentration of each degree of substitution CCMC was determined by colorimetry.
2, the inhibition rates of three CCMC on specific biomembrane at different concentrations were determined by colorimetry.
3, the effects of substitution degree CCMC on the activity of Staphylococcus epidermidis biofilm were compared by fluorescence staining.
Result
1, with the increase of CCMC substitution degree, the minimum inhibitory concentration and minimum biofilm clearance concentration also decreased.
2, under the same concentration, the higher the degree of substitution, the higher the inhibition rate of CCMC to the three biofilms, and the inhibition of the biofilm growth under the smaller concentration of 0.88 and the 0.53 substitution degree.
3, under the same concentration, the higher the degree of substitution, the stronger the inhibition ability of CCMC to the activity of Staphylococcus epidermidis biofilm, and with the increase of concentration, the stronger the inhibition ability of each substitution degree CCMC to the activity of Staphylococcus epidermidis biofilm.
The third part
objective
The titanium alloy surface covalently combined with the 0.53 degree of substitution of CCMC (Ti-CCMC). By comparing with the titanium alloy surface (Ti), the effect of the titanium surface on the inhibition of the biofilm was studied. The effect on the adhesion and proliferation of osteoblasts was studied. The infection model of New Zealand white rabbits was established, and the effect of Ti-CCMC on the local bone tissue was studied and the effect of the whole body was studied.
Method
1, the effects of Ti-CCMC and Ti on biofilm activity in vitro were compared by fluorescence staining.
2, the influence of Ti-CCMC and Ti on the adhesion of Staphylococcus epidermidis in vitro was compared by bacterial quantitative method.
3, the effects of Ti-CCMC and Ti on adhesion and proliferation of MC3T3-E1 cells were compared.
4, bacterial culture was used to determine the bacterial content in bone tissue around titanium bar.
5, the weight changes of two groups of rabbits were compared, and blood bacterial culture and X-ray examination were performed to study the difference of the whole body condition between the two groups.
6, two groups of rabbits were treated with Gram staining in the middle femoral medullary cavity, and the difference of the local migration between the two groups was compared.
Result
1, Ti-CCMC significantly inhibited biofilm activity in vitro.
2, Ti-CCMCC inhibited the adhesion of Staphylococcus epidermidis to some extent.
3, Ti-CCMC can promote the adhesion and proliferation of MC3T3-E1 cells.
4, bacterial growth was observed in bone tissue of group Ti, and no bacterial growth was found in group Ti-CCMC.
5, there was no significant difference in body weight between the two groups. In group Ti, X-ray examination revealed femoral bone destruction, Ti-CCMC group showed no obvious abnormalities, no osteolysis and translucent zone.
6, in group Ti, the middle femoral medullary cavity of rabbits was stained with Gram stain. The bacteria of gram blue stain were scattered and no bacteria were found in group Ti.
conclusion
Chitosan acyl thiourea derivatives have good antibacterial ability, biocompatibility and bone integration ability, which can be used as new materials for antibacterial surface of plants in the Department of orthopedics, and has a certain research and application prospects.

【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R318.08

【共引文獻(xiàn)】

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2 遲菲;鄧叢靜;周建斌;;稻稈醋液成分分析及抑菌·殺菌性能研究[J];安徽農(nóng)業(yè)科學(xué);2010年16期

3 周先漢;張安;曾慶梅;程麗梅;宋俊驊;李琪玲;;超臨界CO_2殺滅芽孢工藝條件的優(yōu)化[J];安徽農(nóng)業(yè)科學(xué);2010年17期

4 楊麗萍;;醫(yī)院床墊細(xì)菌污染的調(diào)查分析[J];安徽衛(wèi)生職業(yè)技術(shù)學(xué)院學(xué)報(bào);2009年05期

5 周婭琴;楊廣嵐;吳磊;袁華玲;楊雪峰;;安徽省2004年部分醫(yī)院內(nèi)鏡消毒現(xiàn)狀調(diào)查[J];安徽預(yù)防醫(yī)學(xué)雜志;2006年04期

6 李國蘭;;一種消毒濕巾殺菌效果試驗(yàn)研究[J];安徽醫(yī)藥;2006年10期

7 姜桂芳;陳國田;崔銀娟;;采血場所消毒滅菌效果監(jiān)測與管理對(duì)策[J];蚌埠醫(yī)學(xué)院學(xué)報(bào);2011年06期

8 王云芳,任麗娟,秦萍;新生兒醫(yī)院感染的調(diào)查與分析[J];青島大學(xué)醫(yī)學(xué)院學(xué)報(bào);2000年03期

9 王云芳,劉沂,孔玉敏,秦萍;口腔門診HBsAg污染的檢測與評(píng)估[J];青島大學(xué)醫(yī)學(xué)院學(xué)報(bào);2001年03期

10 呂銳;蘇冬梅;孟林;鄭麗莎;;羅布麻纖維的抗菌性能研究[J];青島大學(xué)醫(yī)學(xué)院學(xué)報(bào);2006年01期

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4 張玉桃;;2009年龍門縣醫(yī)療保健機(jī)構(gòu)消毒質(zhì)量監(jiān)測結(jié)果分析[A];2010廣東省預(yù)防醫(yī)學(xué)會(huì)學(xué)術(shù)年會(huì)資料匯編[C];2010年

5 王奎濤;康志娟;趙海棟;吳海霞;魯楠;孫亞偉;邢士輝;;金屬在二氧化氯消毒體系下的腐蝕[A];二氧化氯研究與應(yīng)用--2010二氧化氯與水處理技術(shù)研討會(huì)論文集[C];2010年

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8 畢東杰;劉桂華;;手部衛(wèi)生管理環(huán)節(jié)的必要性[A];第一屆航天醫(yī)院管理論壇論文匯編[C];2007年

9 楊自芳;何其莊;施陽;吳四華;瞿思一;;殼寡糖銅的合成表征及生物活性研究[A];上海市化學(xué)化工學(xué)會(huì)2009年度學(xué)術(shù)年會(huì)論文集[C];2009年

10 溫憲芹;孟蔚;陳璐;孫啟華;李子堯;崔樹玉;;2005年山東省部分醫(yī)院牙科手機(jī)、車針消毒滅菌方式及效果調(diào)查[A];山東預(yù)防醫(yī)學(xué)會(huì)第二屆學(xué)術(shù)年會(huì)論文匯編[C];2006年

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7 任宇;山羊脂肪間充質(zhì)干細(xì)胞的體外分離培養(yǎng)與定向分化[D];內(nèi)蒙古大學(xué);2011年

8 張?jiān)?三種含氯消毒劑滅活飲用水中病毒的效果及其表現(xiàn)動(dòng)力學(xué)研究[D];復(fù)旦大學(xué);2011年

9 潘麗;HSV-1感染大鼠骨髓間充質(zhì)干細(xì)胞及形成潛伏感染的初步研究[D];天津醫(yī)科大學(xué);2008年

10 劉傳江;骨髓間充質(zhì)干細(xì)胞在治療大鼠慢性胰腺炎中的作用[D];山東大學(xué);2011年

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