【摘要】：骨組織工程研究為三維支架的植入提供了一種潛在的方法。然而,在三維支架植入骨再生方面仍然存在一些挑戰(zhàn),包括創(chuàng)建結(jié)構(gòu)和力學(xué)性能與自然骨相匹配的再生骨組織,以及成功實(shí)現(xiàn)支架與宿主骨的整合。其中,新骨形成和血管形成的程度被認(rèn)為是骨組織工程臨床應(yīng)用的關(guān)鍵因素。水凝膠的含水量豐富,與人體組織極為相似,可用作組織工程支架材料。然而,水凝膠的親水性表面不利于蛋白質(zhì)的吸附,不利于粘附和細(xì)胞的生長(zhǎng),從而不利于新骨和血管化的形成,因此利用胺基與醛基之間的希夫堿反應(yīng)將RGD接枝在多糖上對(duì)其表面親水性進(jìn)行改性。本研究使用RGD接枝氧化海藻酸鈉/N-琥珀酰殼聚糖(RGD-OSA/NSC)復(fù)合水凝膠為基體材料,以低強(qiáng)度脈沖超聲波(LIPUS)作為機(jī)械刺激的方法以期實(shí)現(xiàn)高水平的血管形成和新骨形成。為了評(píng)價(jià)LIPUS作用下復(fù)合水凝膠材料的血管化能力和成骨的性能,我們進(jìn)行了較系統(tǒng)的體外成骨誘導(dǎo)和內(nèi)皮誘導(dǎo)實(shí)驗(yàn)研究。結(jié)果顯示RGD-OSA/NSC復(fù)合水凝膠在細(xì)胞粘附,增殖以及分化等方面表現(xiàn)出良好的生物學(xué)性能。MTT細(xì)胞活性分析表明,LIPUS與RGD共同作用實(shí)驗(yàn)組細(xì)胞的總數(shù)量最多,成骨誘導(dǎo)中ALP活性／染色和礦化結(jié)節(jié)以及內(nèi)皮誘導(dǎo)中免疫染色檢測(cè)實(shí)驗(yàn)中,比較LIPUS-RGD-OSA/NCS實(shí)驗(yàn)組與Control-RGD-OSA/NCS實(shí)驗(yàn)組及LIPUS-OSA/NCS實(shí)驗(yàn)組,發(fā)現(xiàn)前者的陽(yáng)性率均高于后兩者之和,表明LIPUS和RGD在促進(jìn)hMSCs的增殖和分化過(guò)程中有協(xié)同作用。這些結(jié)果表明同時(shí)使用RGD修飾和LIPUS可能為骨組織工程支架植入過(guò)程中新骨形成和血管形成中提供一種有效地方法。
[Abstract]:Bone tissue engineering provides a potential method for the implantation of three-dimensional scaffolds. However, there are still some challenges in three-dimensional scaffold bone regeneration, including the creation of regenerated bone tissue with structural and mechanical properties matching with natural bone, and the successful integration of scaffold and host bone. Among them, the degree of new bone formation and angiogenesis is considered to be a key factor in clinical application of bone tissue engineering. Hydrogels are rich in water content and very similar to human tissues and can be used as scaffolds for tissue engineering. However, the hydrophilic surface of hydrogels is not conducive to protein adsorption, adhesion and cell growth, which is not conducive to the formation of new bone and vascularization. Therefore, the surface hydrophilicity of RGD was modified by the Schiff base reaction between amino group and aldehyde group. In this study, RGD grafted sodium alginate / sodium succinyl chitosan (RGD-OSA/NSC) composite hydrogel was used as matrix material, and low intensity pulsed ultrasonic (LIPUS) was used as mechanical stimulation method to achieve high level of angiogenesis and new bone formation. In order to evaluate the vascularization ability and osteogenic properties of the composite hydrogels under the action of LIPUS, we conducted a systematic in vitro osteogenesis and endothelial induction experiments. The results showed that RGD-OSA/NSC composite hydrogels exhibited good biological properties in cell adhesion, proliferation and differentiation. The positive rates of ALP activity / staining and mineralized nodules in osteogenic induction and the immunostaining in endothelial induction were compared between LIPUS-RGD-OSA/NCS group, Control-RGD-OSA/NCS experimental group and LIPUS-OSA/NCS experimental group, and the positive rates of the former group were higher than that of the latter two groups. These results suggest that LIPUS and RGD have synergistic effects in promoting the proliferation and differentiation of hMSCs. These results suggest that both RGD modification and LIPUS may provide an effective method for new bone formation and angiogenesis during bone tissue engineering stent implantation.
【學(xué)位授予單位】：西南交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R318.08

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