本文選題：骨髓間充質(zhì)干細(xì)胞 + 異種�。� 參考：《遼寧醫(yī)學(xué)院》2011年碩士論文

【摘要】：目的 探討異種來(lái)源的骨髓間充質(zhì)干細(xì)胞(BMSCs)復(fù)合聚乳酸/羥基乙酸共聚物(PLGA)生物支架,加入轉(zhuǎn)化生長(zhǎng)因子-β1(TGF-β1)和軟骨形態(tài)發(fā)生蛋白(CDMP1)聯(lián)合誘導(dǎo)后修復(fù)甲狀軟骨缺損的能力,評(píng)價(jià)修復(fù)效果,為應(yīng)用軟骨組織工程技術(shù)修復(fù)軟骨缺損提供新的思路和方法。 方法 分別取20周人胚胎及剛出生乳兔的四肢骨骨髓,以密度梯度離心法分離出BMSCs,體外培養(yǎng)擴(kuò)增至第三代。倒置顯微鏡觀察BMSCs生長(zhǎng)狀態(tài),MTT檢測(cè)細(xì)胞活率。分別取第三代的BMSCs,制成細(xì)胞懸液,以5×108/ml的密度接種到PLGA生物支架上,將細(xì)胞支架復(fù)合物加入CDMP1和TGF-β1聯(lián)合誘導(dǎo)21天。取24只3月齡新西蘭白兔,體重3-4公斤,雌雄不限,隨機(jī)分為三組。全麻下制作甲狀軟骨缺損的模型,異種實(shí)驗(yàn)組植入人胚胎BMSCs支架復(fù)合物,同種實(shí)驗(yàn)組植入兔BMSCs支架復(fù)合物,空白對(duì)照組植入單純支架。術(shù)后4周,8周進(jìn)行大體觀察,HE染色,阿利新藍(lán)染色,免疫組化觀察缺損修復(fù)的效果。 結(jié)果 兩種BMSCs在生長(zhǎng)速度和形態(tài)上無(wú)明顯差異。細(xì)胞支架復(fù)合物植入缺損區(qū)后均有軟骨細(xì)胞生成,缺損處表面光滑平整,阿利新藍(lán)染色和免疫組化見(jiàn)兩組的新生細(xì)胞分泌軟骨細(xì)胞基質(zhì)糖氨聚糖(GAG)和Ⅱ型膠原量無(wú)明顯差異。單純支架組缺損處只有少量纖維組織修復(fù),表面凹陷粗糙,阿利新藍(lán)染色和免疫組化檢測(cè)幾乎無(wú)軟骨細(xì)胞基質(zhì)GAG和Ⅱ型膠原分泌。統(tǒng)計(jì)學(xué)顯示兩種BMSCs支架復(fù)合物修復(fù)喉軟骨缺損的效果差異無(wú)統(tǒng)計(jì)學(xué)意義(P 0.05),且均好于單純支架組,差異有統(tǒng)計(jì)學(xué)意義(P 0.05)。 結(jié)論 異種來(lái)源的BMSCs復(fù)合PLGA在TGF-β1和CDMP1聯(lián)合誘導(dǎo)下所得的組織工程化軟骨與同種來(lái)源的BMSCs所獲得的組織工程化軟骨修復(fù)喉軟骨缺損具有可比性。
[Abstract]:Purpose To explore the ability of bone marrow mesenchymal stem cells (BMSCs) combined with poly (lactic acid) / glycolic acid (PLGA) scaffold to repair thyroid cartilage defect induced by TGF- 尾 1 (transforming growth factor- 尾 1) and CDMP1 (cartilage morphogenetic protein). To evaluate the effect of cartilage repair and provide new ideas and methods for repairing cartilage defects by using cartilage tissue engineering technology. Method Bone marrow of limbs of 20 week human embryo and newborn rabbit were isolated by density gradient centrifugation. BMSCs were cultured and amplified to the third generation in vitro. The growth state of BMSCs was observed by inverted microscope. Cell suspensions were prepared from the third generation BMSCs and inoculated into PLGA scaffolds at the density of 5 脳 108/ml. The scaffold complexes were added to CDMP1 and TGF- 尾 1 for 21 days. 24 3-month old New Zealand white rabbits weighing 3-4 kg were randomly divided into three groups. The model of thyroid cartilage defect was made under general anesthesia. Human embryonic BMSCs scaffold complex was implanted in xenogeneic experimental group, rabbit BMSCs scaffold complex was implanted in allogenic experimental group, and pure scaffold was implanted in blank control group. At 4 weeks and 8 weeks after operation, HE staining, Alixin blue staining and immunohistochemistry were used to observe the effect of repairing defects. Result There was no significant difference in growth rate and morphology between the two kinds of BMSCs. Chondrocytes were formed after the scaffold complex was implanted into the defect area, and the surface of the defect was smooth and smooth. There was no significant difference between the two groups in the secretion of matrix glycosaminoglycan (GAG) and type 鈪，

本文編號(hào)：1907151