本文選題：膠原支架　切入點(diǎn)：堿性成纖維生長因子　出處：《南京大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景：在臨床上,感染或?qū)m腔操作均可引起子宮內(nèi)膜損傷,造成內(nèi)膜再生障礙,引起宮腔粘連、閉經(jīng)等,并成為繼發(fā)性不孕重要原因之一。然而現(xiàn)階段的治療方法只能部分改善粘連,對內(nèi)膜的重建再生并沒有有效的手段。隨著組織工程的研究深入,最新的組織工程材料為修復(fù)受損內(nèi)膜提供了新的治療前景。本課題組利用膠原支架復(fù)合bFGF在大鼠子宮全層損傷修復(fù)中得到令人興奮的結(jié)果。本研究將著重于建立小型香豬子宮內(nèi)膜損傷模型以及利用組織工程方法對損傷后內(nèi)膜進(jìn)行修復(fù)。第一章膠原支架復(fù)合骨髓間充質(zhì)干細(xì)胞和膠原復(fù)合堿性成纖維生長因子對小型香豬子宮壁全層損傷后新鮮創(chuàng)面修復(fù)作用目的：研究空白膠原支架,復(fù)合bFGF膠原支架,復(fù)合香豬BMSC膠原支架對小型香豬子宮全層2cm×1cm機(jī)械性損傷創(chuàng)面的修復(fù)作用。方法：本研究包括小型香豬39只,共分為四組：自然修復(fù)組(子宮損傷后不進(jìn)行干預(yù)修復(fù)),膠原支架/PBS組(子宮損傷后用空白膠原膜修復(fù)),膠原支架/CBD-bFGF組(子宮損傷后用復(fù)合CBD-bFGF的膠原支架修復(fù)),膠原支架/BMSC組,(子宮損傷后用復(fù)合小型香豬骨髓間充質(zhì)干細(xì)胞的膠原支架修復(fù)),每組再分為切片組和生育組(單側(cè)損傷和雙側(cè)損傷)。術(shù)后90天開腹觀察子宮修復(fù)狀況,有無外翻,腹腔粘連程度,宮腔是否通暢等。將無外翻、創(chuàng)面修復(fù)完全的子宮取出,進(jìn)行組織學(xué)檢測,包括HE染色,免疫組化染色等。結(jié)果：(1)各組子宮均出現(xiàn)不同程度內(nèi)膜外翻,以自然修復(fù)組(90.3%)尤為嚴(yán)重,其余各組分別為空白膠原膜組(55%),膠原支架/CBD-bFGF組(63%),膠原支架／BMSC組(61%),各膠原支架修復(fù)組創(chuàng)面愈合率間無顯著性差異；(2)對比單側(cè)損傷和雙側(cè)損傷,除自然修復(fù)組外(創(chuàng)面愈合率分別為9.7%和0),其余各組均為單側(cè)修復(fù)率高于雙側(cè),空白膠原膜(66.7%和25%),膠原支架/CBD-bFGF組(36.7%和16.7%),膠原支架/BMSC組(44.4%和37.5%)；(3)HE和免疫組化顯示：膠原支架/PBS、膠原支架/CBD-bFGF組和膠原支架/BMSC組的肌層厚度優(yōu)于自然修復(fù)組,但三組兩者之間沒有明顯差異。子宮內(nèi)膜厚度四組之間沒有顯著性差異。結(jié)論：當(dāng)前采用的香豬子宮全層損傷大小及手術(shù)方式不適合進(jìn)一步的干預(yù)研究,需進(jìn)一步探究減輕手術(shù)部位炎癥反應(yīng)的方法；也需要探索香豬子宮全層損傷合適的大小。以確定能夠反應(yīng)子宮損傷干預(yù)效果的香豬模型。第二章建立穩(wěn)定有效的小型香豬子宮內(nèi)膜損傷模型目的：建立合適的大動(dòng)物子宮內(nèi)膜損傷模型為進(jìn)一步探究臨床組織工程修復(fù)子宮內(nèi)膜奠定基礎(chǔ)。方法：性成熟小型香豬4只麻醉后,暴露雙側(cè)子宮,剪開三面創(chuàng)口,創(chuàng)面大小分組設(shè)置為：長為2cm、3cm、4cm,寬為約管徑1/2-2/3。根據(jù)子宮長度,每側(cè)設(shè)置2-3個(gè)創(chuàng)面。分離漿膜與內(nèi)膜,剪去內(nèi)膜并將漿膜縫合于創(chuàng)緣。術(shù)后8周和12周分別開腹觀察子宮與周圍組織粘連情況,創(chuàng)面修復(fù)效果,管腔是否塌陷(通暢度)。將創(chuàng)面愈合的子宮取出并進(jìn)行組織學(xué)檢測,包括HE染色和免疫組化染色。結(jié)果：(1)改進(jìn)后的模型相對于子宮全層損傷模型,腹腔黏連嚴(yán)重程度明顯減輕,即使有少量創(chuàng)面粘連,也多為易于分離的疏松膜性粘連；克服了創(chuàng)面內(nèi)膜外翻,不愈合等情況。所有術(shù)后的子宮管腔均通暢。(2)術(shù)后8周,2cm、3cm的創(chuàng)面修復(fù)已有新生內(nèi)膜組織包括：有一定內(nèi)膜厚度,有血管和腺體形成,還有少量環(huán)形肌纖維長入。(3)術(shù)后12周,2cm創(chuàng)面修復(fù)已近于正常組織；3cm創(chuàng)面內(nèi)膜厚度、血管數(shù)量低于2cm創(chuàng)面,腺體結(jié)構(gòu)不成熟；4cm創(chuàng)面攣縮,管腔塌陷,僅有單層上皮細(xì)胞形成,肌層結(jié)構(gòu)紊亂,纖維增生顯著,瘢痕化明顯。結(jié)論：通過改進(jìn)后的手術(shù)方法能夠建立較為穩(wěn)定成熟的小型香豬子宮內(nèi)膜損傷模型。
[Abstract]:Background: in clinical infection, or intrauterine operation can cause endometrial damage, causing intimal aplasia caused by intrauterine adhesions, amenorrhea, and become one of the important causes of secondary infertility. However only treatment at this stage of the improved adhesion, reconstruction and regeneration of endometrium. With no effective means of tissue engineering provides in-depth, promising new organization of the new engineering materials for repair of damaged endometrium. The research group using collagen scaffolds bFGF get exciting results in rat uterus full-thickness injury repair. This study will focus on the establishment of miniature pig endometrial damage model and using tissue engineering methods of damage after the endometrial repair. The first chapter collagen scaffold combined with bone marrow mesenchymal stem cells and collagen combined with basic fibroblast growth factor on uterine wall injury in minipigs After the repair effect of fresh tissue Objective: To study the blank composite collagen scaffold, bFGF scaffold, BMSC scaffold composite repair effect of pig of miniature pig uterus total layer 2cm * 1cm mechanical injury wound. Methods: This study included 39 miniature pigs were divided into four groups: natural repair group (uterine injury is not intervention), repair of collagen scaffold /PBS group (uterine injury with blank collagen membrane repair, collagen scaffold (/CBD-bFGF group) after uterine injury with collagen scaffold to repair composite CBD-bFGF), group /BMSC (composite collagen scaffold, miniswines bone marrow mesenchymal stem cells with collagen scaffold to repair the uterus after injury, n = 16) as the slice group and the control group (unilateral and bilateral injury injury). After 90 days of open observation of uterine repair status, there is no valgus, the degree of adhesion, intrauterine patency. No valgus, wound repair complete uterus take Out for histological examination, including HE staining, immunohistochemical staining. Results: (1) there are different degrees of uterus intimal eversion, the natural repair group (90.3%) is particularly serious, the remaining groups were divided into blank collagen membrane group (55%), /CBD-bFGF group (63%) collagen scaffolds, collagen scaffold / group BMSC (61%), the collagen scaffold to repair the wound healing group had no significant difference between the rate; (2) comparison of unilateral injury and bilateral injury, in addition to the natural repair group (wound healing rates were 9.7% and 0), the other groups were unilateral repair rate is higher than the double side blank, collagen membrane (66.7% and 25% /CBD-bFGF group), collagen scaffold (36.7% and 16.7%), group /BMSC collagen scaffold (44.4% and 37.5%); (3) HE and immunohistochemistry showed that: /PBS collagen scaffold, muscle layer thickness is better than that of /CBD-bFGF group and collagen scaffold collagen scaffold /BMSC group natural recovery group, but no significant differences between the three groups. Both the uterus The endometrial thickness had no significant difference between the four groups. Conclusion: the pig uterus full-thickness injury size and operation mode is not suitable for further intervention studies, further explore methods to reduce surgical site inflammation; also need to explore the size of full-thickness damage suitable pig uterus. To determine the effect of pig model can uterine injury intervention the reaction. The second chapter is to establish a stable and effective miniswines endometrial injury model objective: to establish an animal model of endometrial lesion suitable for further clinical study of tissue engineering to repair endometrium foundation. Methods: 4 anesthetized mature miniature pigs after exposure of bilateral uterine, cut three wound, wound size packet set as long as 2cm: 3cm, 4cm, width of about 1/2-2/3. diameter according to the length of uterus, each side set up 2-3 wounds. Separating the serosa and intima, cut in film and The suture to the wound edge. Serous after 8 weeks and 12 weeks were performed to observe the uterine adhesion with surrounding tissue, the effect of wound repair, lumen collapse (patency). The wound healing of uterus and was removed for histological examination, including HE staining and immunohistochemical staining. Results: (1) improved the model is compared with the whole layer of the uterus damage model, adhesion severity significantly reduced, even if there is a small wound adhesion, mostly for loose membrane adhesion easy separation; overcome the wound healing without intimal eversion. All postoperative uterine lumen was unobstructed. (2) 8 weeks after surgery, 2cm 3cm has wound repair of neointimal tissue including: certain endometrial thickness with blood vessels and glands, as well as a small amount of circular muscle fiber ingrowth. (3) after 12 weeks, 2cm has been close to the normal tissue wound repair; wound 3cm endometrial thickness, the number of blood vessels below 2cm wound, Glandular structure is not mature; 4cm wound contracture, lumen collapse, only a monolayer of epithelial cells, muscle layer structure disorder, fiber hyperplasia, scarring significantly. Conclusion: the modified surgical approach to establish minipig endometrial damage model is more stable and mature.

【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R711.74

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