本文選題：骨髓間充質(zhì)干細胞 + 微囊泡�。� 參考：《山西醫(yī)科大學》2017年碩士論文

【摘要】：研究目的:構(gòu)建大鼠膝關(guān)節(jié)軟骨缺損模型,關(guān)節(jié)腔內(nèi)分別注射生理鹽水、BMSCs-MVs和BMSCs,通過檢測修復軟骨組織中的相關(guān)因子含量變化,比較各組軟骨損傷的修復效果,探討B(tài)MSCs-MVs對軟骨組織損傷修復的影響。研究方法:1、全骨髓貼壁法分離、培養(yǎng)SD大鼠的BMSCs并鑒定;2、用超速離心法從上述BMSCs培養(yǎng)液中提取微囊泡(microvesicles,MVs)并用電鏡觀察其形態(tài);3、構(gòu)建SD大鼠膝關(guān)節(jié)軟骨缺損模型,隨機分為六個大組(每組6只):假手術(shù)組、生理鹽水組、BMSCs組、BMSCs-MVs組(高、中、低3個劑量組)。每組雙膝關(guān)節(jié)腔內(nèi)注射分別注射不同實驗試劑,每周注射1次,連續(xù)4、8次,每組分別于注射4周、8周后各處死3只,收集標本備用;4、修復組織的大體觀察;組織切片染色(HE、糖原染色、甲苯胺藍染色);免疫組化觀察修復區(qū)Ⅱ型膠原、Ⅹ型膠原、MMP-13含量;修復組織Wakitani組織學評分;RT-PCR修復軟骨組織Ⅱ型膠原、Ⅹ型膠原、MMP-13含量測定。5、統(tǒng)計學分析:本研究全部數(shù)據(jù)用SPSS18.0統(tǒng)計軟件處理,計量資料用均數(shù)±標準差表示,用單因素方差分析行多組間比較,卡方檢驗行兩組間比較,P0.05為差異有顯著性意義。結(jié)果:1.大體觀,BMSCs組、低劑量組缺損填充較完整、有光澤、邊緣整齊,與周圍組織界限難以區(qū)分;2.低劑量組Ⅱ型膠原表達為陽性,糖胺多糖含量較假手術(shù)組減少,較生理鹽水組增多,Ⅹ型膠原、MMP-13含量較間充質(zhì)干細胞組、生理鹽水組減少;3.低劑量組改良Wakitani評分術(shù)后6周、10周低于生理鹽水組,且差異有統(tǒng)計學意義(P0.05),高于間充質(zhì)干細胞組,差異無統(tǒng)計學意義(P0.05);術(shù)后6周與術(shù)后10周相比,生理鹽水組、低劑量組、間充質(zhì)干細胞組Wakitani評分均降低,差異有統(tǒng)計學意義(P0.05);4.低劑量組術(shù)后6周、10周COL-II mRNA相對表達量與間充質(zhì)干細胞組相比差異無統(tǒng)計學差異(P0.05)。5.低劑量組術(shù)后6周、10周改良Wakitani評分、免疫組化、RT-PCR均較高劑量組、中劑量組表現(xiàn)更優(yōu),差異有統(tǒng)計學意義(P0.05);結(jié)論:骨髓間充質(zhì)干細胞源性微囊泡可以促進軟骨缺損的修復。骨髓間充質(zhì)干細胞可以通過旁分泌途徑促進軟骨修復。
[Abstract]:Objective: to establish the cartilage defect model of knee joint in rats, and to compare the repair effect of cartilage injury in each group by detecting the changes of relative factors in cartilage tissue by injecting normal saline into the articular cavity of BMSCs-MVs and BMSCs respectively. To investigate the effect of BMSCs-MVs on cartilage injury and repair. Methods BMSCs of SD rats was isolated and identified by the whole bone marrow adherent method. Microvesiclesus microvesicus MVss were extracted from the BMSCs medium by ultracentrifugation and the morphology of MVs3 was observed by electron microscope to establish the cartilage defect model of the knee joint of SD rats. They were randomly divided into six groups (6 rats in each group: sham operation group, normal saline group, BMSCs group, BMSCs-MVs group) (high, middle and low dose groups). Different experimental reagents were injected into the knee joint of each group once a week for 4 consecutive times. 3 rats in each group were killed after 4 weeks and 8 weeks of injection respectively. Tissue sections were stained with HEH, glycogen, toluidine blue staining; immunohistochemistry was used to observe the content of collagen 鈪，

本文編號：1825655