發(fā)布時(shí)間：2018-05-03 14:14

  本文選題：組織工程 + 堿性成纖維細(xì)胞生長(zhǎng)因子��； 參考：《第三軍醫(yī)大學(xué)》2014年碩士論文

【摘要】：先天性疾病、炎癥、感染、腫瘤以及創(chuàng)傷等可導(dǎo)致膀胱結(jié)構(gòu)和功能的缺失與損害，而傳統(tǒng)的腸道代膀胱會(huì)引起多種并發(fā)癥，如酸堿代謝紊亂、感染、結(jié)石甚至惡變等。組織工程技術(shù)的發(fā)展為膀胱重建帶來(lái)了全新的方法。目前，構(gòu)建的膀胱組織植入體內(nèi)后的快速血管化是組織工程膀胱面臨的主要問(wèn)題，利用在支架材料上復(fù)合生長(zhǎng)因子為組織工程膀胱體內(nèi)充分快速血管化提供了一種選擇。 堿性成纖維細(xì)胞生長(zhǎng)因子（basic fibroblast growth factor，bFGF）對(duì)創(chuàng)傷愈合、組織修復(fù)、血管形成起著重要促進(jìn)作用，但將bFGF簡(jiǎn)單吸附到材料上再植入體內(nèi)，體液的浸潤(rùn)和流動(dòng)造成因子擴(kuò)散，導(dǎo)致修復(fù)效果有限。因此，我們?cè)赽FGF的N端偶聯(lián)膠原結(jié)合區(qū)（Collagen binding domain，CBD），使其具有結(jié)合膠原能力，在體內(nèi)可以定點(diǎn)、持續(xù)釋放再生信號(hào)分子，同時(shí)外源的膠原支架材料也為細(xì)胞增殖和組織再生提供了三維空間。 本課題以大動(dòng)物比格犬進(jìn)行膀胱重建實(shí)驗(yàn)，，研究?jī)?nèi)容包括以下兩個(gè)部分： 第一部分：比格犬膀胱損傷模型制作及組織工程膀胱的重建。 將由中科院遺傳與發(fā)育研究所提供的CBD-bFGF和膠原支架材料，制備成CBD-bFGF/Collagen材料及PBS/Collagen材料。13只雄性成年比格犬分為假手術(shù)組（Sham operated，n=3）、單純膠原組（PBS/Collagen, n=5）和CBD-bFGF復(fù)合膠原材料組（CBD-bFGF/Collagen, n=5），取下腹部旁正中切口暴露膀胱，切除50%～60%膀胱，分別用CBD-bFGF/Collagen材料及PBS/Collagen材料進(jìn)行膀胱重建。術(shù)后1月內(nèi)單純膠原組及CBD-bFGF復(fù)合膠原材料組各有一條比格犬死亡，尸檢后提示原因?yàn)榘螂讎?yán)重感染導(dǎo)致膀胱破裂，進(jìn)一步引起腹腔感染而死亡。各組剩下比格犬全部存活，無(wú)尿漏、感染發(fā)生。 第二部分：兩種修復(fù)材料對(duì)比格犬膀胱重建修復(fù)作用評(píng)價(jià)。 術(shù)后6月，我們對(duì)所有比格犬的膀胱進(jìn)行了形態(tài)學(xué)、組織學(xué)及功能學(xué)檢測(cè)，評(píng)價(jià)兩種修復(fù)材料對(duì)比格犬膀胱重建修復(fù)作用。尿動(dòng)力學(xué)分析結(jié)果：膀胱最大容量和順應(yīng)性在PBS/Collagen組、CBD-bFGF/Collagen組、Sham operated組依次增加，3組間比較差異有統(tǒng)計(jì)學(xué)意義（P 0.05），而CBD-bFGF/Collagen組和Sham operated組逼尿肌最大壓力比較差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。靜脈腎盂造影及逆行膀胱造影結(jié)果：腎臟、輸尿管正常顯影，膀胱輪廓清晰，形態(tài)正常。大體標(biāo)本及組織學(xué)觀察結(jié)果：PBS/Collagen組膀胱有憩室及小梁形成，上皮層正常，黏膜下層組織增生，肌層再生不良；而CBD-bFGF/Collagen組僅有少量小梁形成，顯示了與對(duì)照組相似的正常3層組織結(jié)構(gòu)。 綜上所述，在比格犬膀胱重建修復(fù)實(shí)驗(yàn)中，CBD-bFGF/Collagen修復(fù)材料能夠更好的促進(jìn)膀胱組織再生和功能恢復(fù)，采用CBD-bFGF結(jié)合膠原支架材料構(gòu)建的組織工程膀胱是理想的膀胱替代修復(fù)材料。
[Abstract]:Congenital diseases, inflammation, infection, tumor and trauma can lead to the loss and damage of bladder structure and function. Traditional intestinal replacement of bladder can cause many complications, such as acid-base metabolic disorder, infection, stone and even malignant change. The development of tissue engineering technology has brought a new method for bladder reconstruction. At present, the rapid vascularization of the constructed bladder tissue after implantation is the main problem facing the tissue engineering bladder. Using the composite growth factor on the scaffold material provides a choice for the full rapid vascularization in the tissue engineering bladder. Basic fibroblast growth factor (bFGF) plays an important role in wound healing, tissue repair and angiogenesis. However, bFGF is simply adsorbed on the material and implanted into the body. The infiltration and flow of body fluids lead to factor diffusion. Resulting in limited repair effect. Therefore, we have the ability of binding collagen in the N-terminally coupled collagen binding region of bFGF, Collagen binding domain, so that we can continuously release regenerative signaling molecules at specific sites in the body. At the same time, exogenous collagen scaffolds also provide three-dimensional space for cell proliferation and tissue regeneration. In this study, the bladder reconstruction experiment was carried out with a large animal Beagle dog. The research includes the following two parts: The first part: the model of bladder injury in Beagle dog and the reconstruction of tissue engineering bladder. CBD-bFGF and collagen scaffolds to be provided by the Institute of Genetics and Development of the Chinese Academy of Sciences, A total of 13 male adult Beagle dogs were prepared into CBD-bFGF/Collagen and PBS/Collagen materials. They were divided into sham operated group (sham operated), collagen group (CBD-bFGF/Collagen / Collagen5) and CBD-bFGF composite glue raw material group (CBD-bFGF- / Collagen, nnbFGF / Collagen). The central incision of lower abdomen was taken to expose bladder, and 50 60% of bladder was removed. CBD-bFGF/Collagen and PBS/Collagen materials were used for bladder reconstruction. One beagle died in the collagen group and one in the CBD-bFGF raw material group within one month after the operation. The postmortem revealed that severe infection of the bladder resulted in rupture of the bladder and further death in the abdominal cavity. All the remaining Beagle dogs in each group survived, had no urine leakage and infection. Part two: evaluation of the effect of two kinds of restoration materials on bladder reconstruction in Beagle dog. Six months after operation, we examined the bladder morphology, histology and function of all Beagle dogs, and evaluated the effects of two kinds of restoration materials on bladder reconstruction in Beagle dogs. The results of urodynamic analysis showed that the maximal bladder volume and compliance were significantly increased in PBS/Collagen group (CBD-bFGF / Collagen group), but there was no significant difference in maximal detrusor pressure between CBD-bFGF/Collagen group and Sham operated group (P 0.05). The results of intravenous pyelography and retrograde cystography: normal development of kidney and ureter, clear outline of bladder and normal shape. The results of gross specimen and histological observation showed that the bladder had diverticulum and trabecular formation in the group of w: PBS / Collagen, normal epithelium, hyperplasia of submucous tissue and poor regeneration of myometrium, while in CBD-bFGF/Collagen group, only a few trabeculae were formed. A normal 3-layer structure similar to that of the control group was shown. In conclusion, CBD-bFGFr / Collagen can promote bladder tissue regeneration and function recovery better in the bladder reconstruction experiment of Beagle dog. The tissue engineering bladder constructed with CBD-bFGF combined with collagen scaffold is an ideal bladder replacement material.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R699.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 SHI Qin;GAO Wei;HAN XingLong;ZHU XueSong;SUN Jie;XIE Fang;HOU XiangLin;YANG HuiLin;DAI JianWu;CHEN Liang;;Collagen scaffolds modified with collagen-binding bFGF promotes the neural regeneration in a rat hemisected spinal cord injury model[J];Science China(Life Sciences);2014年02期

本文編號(hào)：1838832