本文選題：關(guān)節(jié)軟骨 + 軟骨組織工程��； 參考：《浙江大學(xué)》2015年博士論文

【摘要】：肌肉骨骼和關(guān)節(jié)疾病影響著數(shù)千萬人的健康。關(guān)節(jié)軟骨是一種透明光滑的組織,它覆蓋在骨端關(guān)節(jié)頭的表面,起到減少相鄰兩骨間的摩擦等作用。但它是一種無血管無神經(jīng)的組織,所以一旦缺損,很難自發(fā)修復(fù)。關(guān)節(jié)軟骨損傷是臨床上十分常見的病癥,也是骨科和運動醫(yī)學(xué)中公認(rèn)的治療難題。由于軟骨無血管無神經(jīng),營養(yǎng)供應(yīng)不佳,傳統(tǒng)觀點認(rèn)為軟骨損傷無法自發(fā)修復(fù),如果不采取外加手段干預(yù)治療,會導(dǎo)致進一步的軟骨磨損,甚至引發(fā)骨關(guān)節(jié)炎；另一方面,盡管骨軟骨損傷有一定的自發(fā)修復(fù)能力,但修復(fù)組織常出現(xiàn)軟骨終末分化的現(xiàn)象,對其結(jié)構(gòu)和功能造成不可逆的損害,嚴(yán)重影響患者的生活工作質(zhì)量。針對目前軟骨損傷修復(fù)過程中碰到的難題,本研究分別構(gòu)建生物活性材料以開發(fā)針對性治療方案。 首先通過對新西蘭大白兔體內(nèi)造模,我們評估了部分軟骨損傷、全層軟骨損傷和骨軟骨損傷的自發(fā)修復(fù)能力。結(jié)果顯示部分軟骨損傷無自發(fā)修復(fù)能力；全層軟骨損傷有一定自發(fā)修復(fù)能力,且觀察到間質(zhì)干細(xì)胞(MSCs)從缺損邊緣向缺損中心流動；骨軟骨損傷有一定自發(fā)修復(fù)能力,但修復(fù)組織出現(xiàn)軟骨終末分化現(xiàn)象。 進而針對部分軟骨損傷修復(fù),通過使用軟骨損傷離體模型和生物材料模擬體內(nèi)微環(huán)境,評估了MSCs在部分和全層軟骨損傷表面的粘附性和細(xì)胞形態(tài),并進一步利用Transwell實驗評估外源性與內(nèi)源性SDF.1對體外培養(yǎng)細(xì)胞的誘導(dǎo)遷移作用。離體模型與體外實驗結(jié)果顯示,與軟骨微環(huán)境和二型膠原(col Ⅱ)支架相比,軟骨下骨微環(huán)境和一型膠原(col I)支架更有利于MSCs粘附,且SDF-1能夠顯著提高MSCs的遷移。在此基礎(chǔ)上構(gòu)建的col I復(fù)合SDF.1生物活性支架為部分軟骨缺損創(chuàng)造了合適的微環(huán)境,動物實驗驗證顯著提高了兔部分軟骨損傷自發(fā)修復(fù)能力。 最后針對骨軟骨損傷修復(fù),體外實驗結(jié)果顯示通過抑制經(jīng)典Wnt/β-catenin通路活性,PTHrP能夠顯著降低茜素紅染色和終末分化相關(guān)基因的表達(dá)。對新西蘭大白兔體內(nèi)骨軟骨損傷造模,缺損處移植膠原.蠶絲雙層支架,并在三個不同時間點關(guān)節(jié)腔注射PTHrP(術(shù)后4.6,7.9和10.12周)。術(shù)后4.6周PTHrP治療組的修復(fù)再生效果(軟骨和軟骨下骨的重建)要優(yōu)于其他時間點治療組,軟骨終末分化現(xiàn)象減少(鈣化、骨化、基質(zhì)降解),BMSCs成軟骨能力提高(細(xì)胞形態(tài)、col II和糖胺聚糖合成)。此外,PTHrP的應(yīng)用時間影響PTHrP受體的表達(dá),進而影響PTHrP的治療效果。 綜上,對于部分軟骨損傷,我們應(yīng)用col Ⅰ生物膜材料復(fù)合基質(zhì)細(xì)胞衍生因子SDF-1構(gòu)建生物活性材料促進C-MSCs和SM-MSCs的遷移粘附,進而開啟軟骨自發(fā)修復(fù)；對于骨軟骨損傷,我們應(yīng)用膠原.蠶絲雙層支架復(fù)合甲狀旁腺激素相關(guān)蛋白PTHrP抑制軟骨終末分化,促進成軟骨過程,進而提高軟骨自發(fā)修復(fù)質(zhì)量。該研究結(jié)果無論對于解釋軟骨損傷治療的生物學(xué)本質(zhì),還是最終臨床轉(zhuǎn)化治療軟骨損傷都有至關(guān)重要的意義。
[Abstract]:Musculoskeletal and joint diseases affect the health of millions of people. Articular cartilage is a transparent, smooth tissue that covers the surface of the end of the end of the bone and plays a role in reducing the friction between the two adjacent bones. But it is a kind of no vascular and non nerve tissue, so it is difficult to repair spontaneously once the defect is damaged. The articular cartilage injury is ten in clinical. The common disease is also a recognized treatment problem in the Department of orthopedics and sports medicine. Because the cartilage is no blood vessel and no nerve, the nutritional supply is poor, the traditional view that the cartilage damage can not be repaired spontaneously, if the treatment is not taken, it will lead to further cartilage wear and even osteoarthritis; on the other hand, despite the bone cartilage The injury has a certain ability to repair spontaneously, but the repair tissue often appears cartilage end differentiation, causing irreversible damage to its structure and function, which seriously affects the quality of life and work of the patients. In view of the problems encountered in the process of repair of cartilage injury, this study develops bioactive materials to develop targeted treatment schemes respectively.
First of all, we evaluated the ability of partial cartilage injury, total cartilage injury and spontaneous repair of osteochondral damage through the modeling of New Zealand white rabbits. Results showed that partial cartilage injury had no spontaneous repair ability, and there was a certain spontaneous repair ability of the whole layer cartilage injury, and the interstitial stem cells (MSCs) were observed from the defect edge to the defect. There is a spontaneous repair ability of osteochondral injury, but there is a phenomenon of cartilage terminal differentiation in repair tissue.
In view of the repair of partial cartilage damage, the adhesion and cell morphology of MSCs on the surface of partial and total cartilage damage were evaluated by using the model of cartilage damage in vitro and biomaterial to simulate the microenvironment in vivo, and the induced migration of exogenous and endogenous SDF.1 on the cultured cells was evaluated by Transwell experiment. The body model and in vitro experimental results show that the subchondral bone microenvironment and the type of collagen (Col I) scaffold are more conducive to MSCs adhesion compared with the cartilage microenvironment and the type two collagen (Col II) scaffold, and SDF-1 can significantly improve the migration of MSCs. On this basis, the col I compound SDF.1 bioactive scaffold has created a suitable part of the cartilage defect. The microenvironment of animal experiments proved that the ability of spontaneous repair of partial cartilage injury in rabbits was significantly improved.
At last, in response to the repair of osteochondral damage, in vitro experiments showed that by inhibiting the activity of the classic Wnt/ beta -catenin pathway, PTHrP could significantly reduce the expression of alizarin red staining and terminal differentiation related genes. PTHrP (4.6,7.9 and 10.12 weeks after operation). The regenerative effect of the PTHrP treatment group (cartilage and subchondral bone reconstruction) at 4.6 weeks after the operation was better than the other time point treatment group. The cartilage end differentiation was reduced (calcification, ossification, matrix degradation), and the energy of BMSCs was improved (cell morphology, col II and glycosaminoglycan synthesis). In addition, PTHrP The application time affects the expression of PTHrP receptor, thereby affecting the therapeutic effect of PTHrP.
To sum up, for partial cartilage damage, we use Col I biofilm material complex matrix cell derived factor SDF-1 to construct bioactive materials to promote the migration and adhesion of C-MSCs and SM-MSCs, and then open the cartilage to repair spontaneously; for osteochondral damage, we apply collagen. Silk double layer scaffold composite parathyroid hormone related protein PTHrP It is of great significance to explain the biological nature of cartilage damage treatment and to the final clinical transformation of cartilage damage.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R687.4;R318.08

【共引文獻】

相關(guān)期刊論文 前10條

1 張慧;程愛琳;;電紡納米纖維構(gòu)建組織工程支架研究新進展[J];材料導(dǎo)報;2009年09期

2 唐，

本文編號：2057469