本文選題：肋軟骨 + 自體移植�。� 參考：《復旦大學》2010年碩士論文

【摘要】： 耳廓再造術(shù)中,肋軟骨以其取材方便,便于雕刻成型而被廣泛采用。自體軟骨移植雖無排斥反應,但仍存在再造耳廓變形、攣縮,與正常耳廓外觀不對稱等不良手術(shù)后果。胰島素樣生長因子-Ⅰ(IGF-Ⅰ)是調(diào)節(jié)軟骨細胞合成的重要生長因子之一,國內(nèi)外學者離體實驗證實它能刺激細胞的分裂增殖,增強細胞蛋白多糖及Ⅱ型膠原的合成。但針對IGF-Ⅰ對體內(nèi)自體移植的肋軟骨起何種作用,尚未見有文獻報道。 研究目的：評估IGF-Ⅰ凝膠對自體移植帶軟骨膜肋軟骨增殖及代謝的作用。 材料和方法 實驗動物及分組：健康新西蘭白兔24只,幼年及成年各12只,雌雄不限,幼年兔為1月齡,體重lkg左右,成年兔為6月齡,體重2.5kg左右。按年齡分為幼年組及成年組,各組中再隨機分為手術(shù)對照組、10ng/ml實驗組及50ng/ml實驗組,每組各4只。手術(shù)對照組直接將自體肋軟骨取出,移植于背部皮下；10ng/ml實驗組將自體肋軟骨取出后,與0.5ml的10ng/ml IGF-Ⅰ凝膠混合為軟骨-生長因子復合物后移植于背部皮下；50ng/ml實驗組將自體肋軟骨取出后,與0.5ml的50ng/mlIGF-Ⅰ凝膠混合為軟骨-生長因子復合物后移植于背部皮下。在手術(shù)后8周,再取術(shù)側(cè)對側(cè)生理情況下生長的肋軟骨作為空白對照組共同進行檢測。 凝膠制備及手術(shù)方法：纖維蛋白原(200mg/ml)、凝血酶(10u/ml)、IGF-Ⅰ(10、50ng/ml)混合制備成IGF-Ⅰ纖維蛋白凝膠,現(xiàn)用現(xiàn)配。實驗兔麻醉、消毒后取出第8、9肋軟骨,截為3段,按不同組別直接移植于背部皮下或混合不同濃度凝膠后移植于背部皮下。 標本采集及結(jié)果評估：移植8周后,動物處死,取出移植的肋軟骨,與空白對照組共同進行檢測。檢測項目包括：大體觀察、HE染色、透射電鏡檢查及軟骨羥脯氨酸分析。 統(tǒng)計學方法：使用stata 7.0統(tǒng)計軟件進行數(shù)據(jù)統(tǒng)計處理。對于各組所得到的軟骨羥脯氨酸含量的結(jié)果,運用完全隨機設(shè)計的單因素方差分析評價其4組間有無差異,各組間差異的評價通過t檢驗來實現(xiàn)。 結(jié)果 1.大體觀察：術(shù)后8周,幼年兔中,50ng/ml實驗組移植軟骨與周圍組織粘連較牢固,不易分離；手術(shù)對照組與10ng/ml實驗組移植軟骨與周圍組織無明顯粘連,較易分離。手術(shù)對照組與50ng/ml實驗組移植軟骨呈乳白色,半透明,略有彈性,與空白對照組軟骨相近；10ng/ml實驗組移植軟骨色澤偏黃,透明感及彈性均較差。成年兔中,各組差別不明顯,但軟骨彈性均較幼年軟骨差。 2.HE染色：幼年兔中,50ng/ml實驗組軟骨膜完整,骨原細胞較多,軟骨細胞增生活躍；手術(shù)對照組及10ng/ml實驗組軟骨膜結(jié)構(gòu)紊亂,細胞結(jié)構(gòu)少,軟骨細胞發(fā)生退行性變。成年兔中,50ng/ml實驗組軟骨膜細胞結(jié)構(gòu)較其余各組多,但4組軟骨均存在空泡形成及基質(zhì)分離現(xiàn)象。 3.透射電鏡檢查：幼年兔軟骨透射電鏡表現(xiàn)：在空白對照組及50ng/ml實驗組中,可見健康的軟骨細胞,軟骨與周圍組織交界處成纖維細胞少；手術(shù)對照組中,可見到壞死的軟骨細胞,與周圍組織交界處成纖維細胞活躍；10ng/ml實驗組中,可見到散在的鈣化灶,未發(fā)現(xiàn)正常的軟骨細胞。成年兔軟骨透射電鏡表現(xiàn)：空白對照組中,可見軟骨與周圍組織交界處有成熟的成纖維細胞；手術(shù)對照組中,可見軟骨與周圍組織交界處有壞死的軟骨細胞；10ng/ml實驗組中,也可見到軟骨細胞壞死,但軟骨膜細胞正常；50ng/ml實驗組中,可見健康的軟骨細胞,同時軟骨基質(zhì)增多明顯。 4.軟骨羥脯氨酸分析：幼年兔：空白對照組與50ng/ml實驗組間差別無統(tǒng)計學意義(P0.05),且均大于手術(shù)對照組和10ng/ml實驗組,差別有統(tǒng)計學意義(P0.05),手術(shù)對照組與10ng/ml實驗組之間差異無統(tǒng)計學意義(P0.05)。成年兔：各組間差異無統(tǒng)計學意義(P0.05)。 結(jié)論 1.在幼年兔中,50ng/ml濃度的IGF-Ⅰ凝膠可以保持其正常生理狀況下的增殖,和軟骨細胞合成細胞外基質(zhì)的能力,維持移植后的軟骨及軟骨膜細胞的正常表型。 2. 10ng/ml濃度的IGF-Ⅰ凝膠對幼年兔移植軟骨及軟骨膜細胞的生長沒有促進作用,而凝膠可能影響軟骨及軟骨膜細胞的正常增殖與代謝。 3.成年兔中,在大體觀察、光鏡、電鏡及生化檢查等各個方面,各組之間均無明顯差別,提示隨年齡增長,軟骨中的細胞成分減少,且處于靜息狀態(tài),軟骨及軟骨膜細胞對IGF-Ⅰ的敏感性降低。 4.幼年期行自體肋軟骨移植的效果優(yōu)于成年。
[Abstract]:During auricle reconstruction, costal cartilage is widely used for its convenience and convenience for carving. Although autologous cartilage transplantation has no rejection, there are still some adverse effects on the reconstruction of auricle deformation, contracture, and abnormal appearance of the auricle. Insulin-like growth factor - I (IGF- I) is an important growth factor for regulating the synthesis of cartilage cells. First, in vitro, domestic and foreign scholars have proved that it can stimulate cell division and proliferation and enhance the synthesis of cell proteoglycan and type II collagen. However, it has not been reported that the effect of IGF- I on autogenous autogenous costal cartilage has not been reported.
Objective: To evaluate the effect of IGF- I gel on the proliferation and metabolism of autogenous transplantation of costal cartilage.
Materials and methods
Experimental animals and groups: 24 healthy New Zealand white rabbits, young and adult 12, male and female, and male and female, young rabbit 1 month old, weight LKG, adult rabbit 6 month old, weight 2.5kg. They are divided into young group and adult group according to age. Each group is randomly divided into operation control group, 10ng/ml experiment group and 50ng/ml experimental group, 4 rats in each group. The operation is 4. The autogenous costal cartilage was removed and transplanted subcutaneously in the back. After taking out the autogenous costal cartilage in the 10ng/ml experimental group, the 10ng/ml IGF- I gel was mixed with the 0.5ml 10ng/ml IGF- I gel as the cartilage growth factor complex and transplanted subcutaneously into the back. The 50ng/ml experimental group removed the autogenous costal cartilage and mixed with the 0.5ml 50ng/mlIGF- I gel into cartilage growth. After 8 weeks of operation, the costal cartilage growing on the contralateral side was taken as a blank control group.
Gelatin preparation and operation methods: fibrinogen (200mg/ml), thrombin (10u/ml), IGF- I (10,50ng/ml) mixed with IGF- I fibrin gel, now used. Experimental rabbit anaesthesia, after disinfection of the 8,9 rib cartilage, cut into 3 segments, according to different groups directly transplanted in the back subcutaneous or mixed with different concentration gels after transplantation to the back subcutaneous.
Sample collection and results evaluation: after 8 weeks of transplantation, the animals were killed, the costal cartilage was removed and the control group was tested together with the blank control group. The test items included gross observation, HE staining, transmission electron microscopy and cartilage hydroxyproline analysis.
Statistical method: Using Stata 7 statistical software to carry out data statistical processing. For the results of the content of cartilage hydroxyproline in each group, there is no difference between the 4 groups by the single factor variance analysis of the complete random design. The evaluation of the differences between each group is realized by the t test.
Result
1. general observation: 8 weeks after operation, in the young rabbits, the graft cartilage of 50ng/ml experimental group was firmly adhered to the surrounding tissue, and it was not easy to separate. There was no obvious adhesion between the graft cartilage and the surrounding tissue in the operation control group and the 10ng/ml experimental group. The transplantation cartilage of the operation control group and the 50ng/ml experimental group were white, translucent and slightly elastic. In 10ng/ml experimental group, the color of the graft was yellow, and the transparency and elasticity were poor. In adult rabbits, the differences were not obvious, but the cartilage elasticity was worse than that of young ones.
2.HE staining: in young rabbits, the cartilage membrane of the 50ng/ml experimental group was complete, the osteogenic cells were more and the chondrocytes proliferated active. The cartilage membrane structure was disorganized in the operation control group and the 10ng/ml experimental group, the cell structure was less, the chondrocytes were degenerative. In the adult rabbit, the cartilage membrane cell structure of the 50ng/ml experimental group was more than the other groups, but the cartilage of the 4 groups existed in all the other groups. The formation of vacuoles and the separation of matrix.
3. transmission electron microscope examination: the transmission electron microscopy of young rabbit cartilage: in the blank control group and the 50ng/ml experimental group, the healthy chondrocytes and the fibroblasts at the junction of the surrounding tissue were found less. The necrotic chondrocytes could be seen in the operation control group, and the fibroblasts were active at the intersection of the surrounding tissues; in the 10ng/ml experimental group, In the blank control group, there were mature fibroblasts at the junction of the cartilage and surrounding tissue; in the operation control group, there was necrotic cartilage cells at the junction of the cartilage and surrounding tissue; in the 10ng/ml experimental group, the soft tissue could also be seen in the soft tissue. Osteocytes were necrotic, but the chondrocytes were normal. In the 50ng/ml experimental group, healthy chondrocytes were observed, and cartilage matrix increased significantly.
4. cartilage hydroxyproline analysis: young rabbits: there was no statistical difference between the blank control group and the 50ng/ml experimental group (P0.05), and all were larger than the operation control group and the 10ng/ml experimental group, the difference was statistically significant (P0.05), the difference between the control group and the 10ng/ml experimental group was not statistically significant (P0.05). Learning meaning (P0.05).
conclusion
1. in young rabbits, the 50ng/ml concentration of IGF- I gel can maintain its normal physiological condition, and the ability of chondrocytes to synthesize the extracellular matrix, and maintain the normal phenotype of the cartilage and the cartilage membrane cells after transplantation.
2. 10ng/ml concentration of IGF- I gel has no effect on the growth of transplanted cartilage and chondrocytes in young rabbits, but the gel may affect the normal proliferation and metabolism of cartilage and cartilage membrane cells.
In 3. adult rabbits, there were no significant differences in the general observation, light microscopy, electron microscopy and biochemical examination. It was suggested that the cell components in cartilage decreased with age and were resting, and the sensitivity of cartilage and cartilage membrane cells to IGF- I was reduced.
4. the effect of autogenous costal cartilage transplantation in childhood is better than that in adults.

【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R764.9

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