本文選題：重組人骨形態(tài)發(fā)生蛋白-2　切入點：殼聚糖　出處：《蘭州大學(xué)》2015年碩士論文

【摘要】：目的：本研究殼聚糖通過乳化交聯(lián)形成rhBMP-2殼聚糖微球,復(fù)合在可降解的明膠海綿支架上,制備可降解骨形態(tài)發(fā)生蛋白復(fù)合貼膜,提高BMP有效誘導(dǎo)成骨性。研究方法：1、選用H2SO4作為乳化劑,通過乳化交聯(lián)法制備NaOH/rhBMP-2/殼聚糖微球,觀察殼聚糖微球的形態(tài)特征、細胞相容性以及其緩釋作用。2、制備rhBMP-2/殼聚糖微球/明膠海綿復(fù)合貼膜,將其與rhBMP-2/殼聚糖微球及明膠海綿對照研究,通過體外釋放實驗,觀察并分析復(fù)合膜的體外釋放情況。3、通過動物實驗骨缺損模型,觀察可降解骨形態(tài)發(fā)生蛋白復(fù)合貼膜緩釋能力及誘導(dǎo)成骨作用。將可降解骨形態(tài)發(fā)生蛋白復(fù)合貼膜植入新西蘭兔的橈骨缺損區(qū)域,在預(yù)定時間點,通過影像、組織學(xué)檢查,以及通過測定缺損區(qū)域骨樣本骨量、鈣量及堿性磷酸酶。觀察rhBMP-2/殼聚糖微球/明膠海綿誘導(dǎo)成骨的情況。應(yīng)用IBM SPSS19.0統(tǒng)計學(xué)軟件數(shù)據(jù)包對數(shù)據(jù)進行分析,組間數(shù)據(jù)采用均數(shù)±標(biāo)準(zhǔn)差(x±s)描述,組間數(shù)據(jù)的比較,采用單因素方差分析,檢測水準(zhǔn)a=0.05,p0.05具有統(tǒng)計學(xué)意義。結(jié)果：以殼聚糖為原料,通過乳化交聯(lián)負載rhBMP-2成功制備出NaOH-殼聚糖微球,其形態(tài)規(guī)整,分散均勻,微球的成球性良好,與單純可吸收明膠海綿復(fù)合支架比較,殼聚糖微球復(fù)合明膠海綿貼膜前期突釋小,中后期可持續(xù)緩慢釋放。通過MTT法檢查殼聚糖微球細胞毒性為1級,細胞相容性良好,以兔橈處建造骨缺損模型,通過大體觀察、影像學(xué)檢查,測試骨量、骨密度、鈣含量及堿性磷酸酶活性檢測,統(tǒng)計學(xué)分析結(jié)果提示：可降解骨形態(tài)發(fā)生蛋白-2復(fù)合貼膜可有效促進骨缺損修復(fù),較單純的明膠海綿及殼聚糖微球控釋作用佳。結(jié)論：1、以H2SO4為乳化劑,通過乳化交聯(lián)法制備的NaOH-殼聚糖微球具有良好的生物相容性和蛋白高吸附性。此法工藝簡便易行,殼聚糖微球穩(wěn)定性良好；2、以殼聚糖為原料,復(fù)合明膠海綿所制備的可降解重人組骨形態(tài)發(fā)生蛋白-2復(fù)合貼膜,經(jīng)體外釋放實驗、植入兔橈骨缺損模型觀察檢測分析,載rhBMP-2的NaOH-殼聚糖微球與帶正電的明膠海綿復(fù)合通過穩(wěn)定的靜電作用可有效提高復(fù)合膜的控釋作用,較單純的殼聚糖微球及明膠海綿有更好的誘導(dǎo)成骨性。
[Abstract]:Aim: in this study, rhBMP-2 chitosan microspheres were prepared by emulsification and crosslinking of chitosan onto degradable gelatin sponge scaffolds to prepare degradable bone morphogenetic protein composite film, so as to improve the osteogenicity induced by BMP.Methods H2SO4 was used as emulsifier to prepare NaOH/rhBMP-2/ chitosan microspheres by emulsification and crosslinking method. The morphology, cytocompatibility and slow-release of chitosan microspheres were observed. The rhBMP-2/ chitosan microspheres / gelatin sponge composite film was prepared.Compared with rhBMP-2/ chitosan microspheres and gelatin sponge, the in vitro release of the composite membrane was observed and analyzed, and the bone defect model was established by animal experiment.To observe the ability of biodegradable bone morphogenetic protein (BMP) composite membrane to release and to induce osteogenesis.The degradable bone morphogenetic protein composite patch was implanted into the radius defect area of New Zealand rabbits. The bone mass, calcium content and alkaline phosphatase in the defect area were measured by imaging, histological examination and bone samples at a predetermined time point.The osteogenesis of rhBMP-2/ chitosan microspheres / gelatin sponge was observed.IBM SPSS19.0 data package was used to analyze the data. The data between groups were described by mean 鹵standard deviation (x 鹵s). The comparison of data among groups and single factor analysis of variance (ANOVA) showed statistical significance.Results: NaOH- chitosan microspheres were successfully prepared by emulsifying and crosslinking loaded rhBMP-2 with chitosan as raw material. The microspheres were characterized by regular morphology, uniform dispersion and good spheroidization, compared with simple absorbable gelatin sponge composite scaffolds.Chitosan microspheres combined with gelatin sponge had little sudden release in the early stage and sustained and slow release in the middle and late stage.The cytotoxicity of chitosan microspheres was grade 1 by MTT assay. The rabbit model of bone defect was constructed at the radial of rabbit. Bone mass, bone mineral density, calcium content and alkaline phosphatase activity were measured by gross observation and imaging examination.The results showed that the biodegradable bone morphogenetic protein-2 composite film could effectively promote the repair of bone defect, which was better than that of simple gelatin sponge and chitosan microspheres.Conclusion the NaOH-chitosan microspheres prepared by emulsification and crosslinking with H2SO4 as emulsifier have good biocompatibility and high protein adsorption.The method is simple and convenient, and chitosan microspheres have good stability. The biodegradable heavy human group bone morphogenetic protein-2 composite membrane was prepared from chitosan as raw material and gelatin sponge, and was released in vitro.The observation and analysis of rabbit radial defect model showed that the combination of rhBMP-2 loaded NaOH-chitosan microspheres and positively charged gelatin sponge could effectively improve the controlled release effect of the composite membrane through stable electrostatic interaction.Compared with simple chitosan microspheres and gelatin sponges, the osteogenesis was better than that of chitosan microspheres and gelatin sponges.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R318.08

【參考文獻】

相關(guān)期刊論文 前1條

1 陳發(fā)明,吳織芬,金巖,杜巖,王國芳;重組人骨形態(tài)發(fā)生蛋白-2緩釋微球?qū)Τ晒羌毎淖饔肹J];中華實驗外科雜志;2005年02期

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本文編號：1715214