本文選題：三維細(xì)胞遷移 + 免疫反應(yīng)�。� 參考：《浙江大學(xué)》2017年碩士論文

【摘要】：細(xì)胞遷移作為細(xì)胞的基本功能之一,與人類及動(dòng)物的生物學(xué)和生理學(xué)行為息息相關(guān)。其中,細(xì)胞在三維環(huán)境下的遷移行為可以更好地描述細(xì)胞在體內(nèi)的真實(shí)生理過(guò)程,是組織工程與再生醫(yī)學(xué)領(lǐng)域中的重點(diǎn)和難點(diǎn)。本文構(gòu)建了全新的模型,借用Transwell小室研究不同種類細(xì)胞基于免疫反應(yīng)向傳統(tǒng)支架材料和刺激響應(yīng)智能水凝膠材料中的遷移行為。本文首先采用50ng/mL干擾素IFN-γ和不同濃度的脂多糖LPS(0-300ng/mL)對(duì)原代巨噬細(xì)胞(M0)進(jìn)行極化,分泌細(xì)胞因子(TNF-α和VEGF)誘導(dǎo)內(nèi)皮細(xì)胞向膠原-殼聚糖材料中進(jìn)行遷移。通過(guò)激光共聚焦顯微鏡表征發(fā)現(xiàn),隨著LPS濃度的提高,內(nèi)皮細(xì)胞可以向-10 ℃冷凍干燥支架材料中更深的區(qū)域遷移,且該變化趨勢(shì)與TNF-α的濃度變化趨勢(shì)相一致。另外,在50ng/mL IFN-γ和300ng/mL LPS條件下,刺激巨噬細(xì)胞來(lái)誘導(dǎo)內(nèi)皮細(xì)胞向不同孔結(jié)構(gòu)膠原-殼聚糖支架材料中遷移試驗(yàn)中發(fā)現(xiàn):內(nèi)皮細(xì)胞在-10 ℃支架(186.7±15.0μm)比在-20 ℃支架(108.1±13.0μm)中更容易遷移至更深的區(qū)域,證明了材料的孔徑對(duì)于細(xì)胞三維遷移有著重要的影響。為了研究pH響應(yīng)性智能材料在外界環(huán)境pH改變的情況下對(duì)不同種類細(xì)胞三維遷移的影響,制備了碳碳雙鍵和席夫堿鍵交聯(lián)的雙網(wǎng)絡(luò)水凝膠,并將該水凝膠置于不同的pH緩沖液(pH 7.4和pH 5)處理。結(jié)果發(fā)現(xiàn),pH 5處理后的水凝膠其溶脹率和網(wǎng)格大小有所提高,但是機(jī)械性能卻明顯降低。利用激光共聚焦顯微鏡研究不同種類細(xì)胞基于免疫反應(yīng)向不同pH緩沖液處理的材料中三維遷移行為,結(jié)果發(fā)現(xiàn)內(nèi)皮細(xì)胞(ECs)在pH5處理后的水凝膠中比在pH 7.4處理后的水凝膠中更容易遷移至更深的區(qū)域,而骨髓間充質(zhì)干細(xì)胞(BMSCs)細(xì)胞則在pH 7.4緩沖液處理后的水凝膠中遷移更深的距離。除了 pH響應(yīng)性材料外,光響應(yīng)材料也是智能響應(yīng)材料中應(yīng)用比較廣泛的一種。本文制備了碳碳雙鍵和香豆素基團(tuán)光二聚交聯(lián)的雙網(wǎng)絡(luò)水凝膠,并將該水凝膠在波長(zhǎng)為λ=254 nm的光下處理。結(jié)果發(fā)現(xiàn),光處理20min后的水凝膠其溶脹率和孔徑明顯提高,而機(jī)械性能明顯降低。通過(guò)激光共聚焦顯微鏡表征內(nèi)皮細(xì)胞在波長(zhǎng)為λ=254 nm的光處理前后的材料中三維遷移情況,結(jié)果發(fā)現(xiàn)內(nèi)皮細(xì)胞在光處理后的水凝膠中比在光處理前的水凝膠中更容易遷移至更深的區(qū)域,再一次驗(yàn)證了內(nèi)皮細(xì)胞更容易在模量較小的三維材料中遷移。本文借用該全新的實(shí)驗(yàn)?zāi)Ｐ吞骄苛擞绊懠?xì)胞三維遷移的因素(材料孔結(jié)構(gòu)、模量以及生物信號(hào)因子),為受損組織修復(fù)與再生提供了一定的實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Cell migration, as one of the basic functions of cells, is closely related to the biological and physiological behaviors of human beings and animals. Among them, the migration behavior of cells in three-dimensional environment can better describe the true physiological process of cells in vivo, and it is an important and difficult point in the field of tissue engineering and regenerative medicine. In this paper, a new model was constructed to study the migration behavior of different types of cells into traditional scaffold materials and stimulus-responsive smart hydrogel materials based on immunoreaction by using Transwell chamber. In this paper, 50 ng / mL interferon IFN- 緯 and different concentrations of lipopolysaccharide (LPS0-300 ng / mL) were used to polarize the primary macrophages (M0) and secrete cytokines TNF- 偽 and VEGF to induce the migration of endothelial cells into collagen-chitosan materials. The results of laser confocal microscopy showed that with the increase of LPS concentration, endothelial cells could migrate to the deeper region in the freeze-drying scaffold material at -10 鈩，

本文編號(hào)：2031507