【摘要】：目的:周圍神經(jīng)損傷是臨床上較常見的致殘性疾病,直接損害了勞動力資源,給社會和家庭生活造成沉重的負(fù)擔(dān)。周圍神經(jīng)損傷修復(fù)后再生受多種因素的影響,吻合口處的微環(huán)境及局部的癜痕形成嚴(yán)重影響了神經(jīng)損傷后修復(fù)效果,為此,改善局部微環(huán)境及減少局部瘢痕有利于周圍神經(jīng)再生促進(jìn)神經(jīng)功能恢復(fù)。羊膜具有半透性,保證營養(yǎng)物質(zhì)順利透過,促進(jìn)雪旺細(xì)胞增殖,加速神經(jīng)軸索生長,抑制周圍成纖維細(xì)胞的生物活性,限制神經(jīng)周圍組織侵入神經(jīng)斷端,抑制周圍瘢痕形成,防止神經(jīng)粘連及神經(jīng)壓迫等。本實驗構(gòu)建大鼠坐骨神經(jīng)損傷模型,在顯微鏡下行神經(jīng)吻接術(shù),吻接處給予人新鮮羊膜包裹,通過大體觀察、光鏡觀察、透射電鏡、免疫組化、神經(jīng)電生理檢測、坐骨神經(jīng)功能指數(shù)等指標(biāo)分析,評價新鮮羊膜促進(jìn)周圍神經(jīng)損傷后修復(fù)再生的效果。方法:選用成年、健康SD大鼠60只,雌雄不限,體重220克~250克。取一側(cè)腿制備坐骨神經(jīng)急性損傷模型。隨機(jī)分A組(修復(fù)神經(jīng)后用人新鮮羊膜包裹吻合口),B組(修復(fù)神經(jīng)后用生物膜包裹吻合口),C組(修復(fù)神經(jīng)后不做任何處理)三組,每組20只。術(shù)后2、4、8和12周各行大體觀察、光鏡觀察、免疫組織化學(xué)檢測(s-100蛋白在神經(jīng)組織中的表達(dá)情況);分別于術(shù)后4、8、12周各行透射電鏡觀察、軸突成像分析(計數(shù)有髓神經(jīng)纖維數(shù)目、軸突直徑及髓鞘厚度)、復(fù)合肌肉動作電位檢測(測量并記錄潛伏期、波幅和神經(jīng)傳導(dǎo)速度)、計算坐骨神經(jīng)功能指數(shù)及其恢復(fù)率。所有數(shù)據(jù)均進(jìn)行統(tǒng)計學(xué)處理。結(jié)果:大體觀察:三組大鼠術(shù)后手術(shù)切口都未出現(xiàn)感染,7天后切口拆線,切口一期愈合;三組大鼠術(shù)后2周大部出現(xiàn)下肢稍腫脹,活動明顯減少且跛行。2周后腫脹、精神狀態(tài)及飲食逐漸恢復(fù),A、B組大鼠跛行時間較C組短,活動量恢復(fù)更早。A、B組和C組相比足底和足趾皮膚潰瘍大鼠數(shù)量少且潰瘍恢復(fù)時間短。術(shù)后2周羊膜、生物膜局部稍有吸收,4周大部吸收,8周完全吸收。A組、B組神經(jīng)和周圍組織稍有粘連且粘連較疏松,活動度可。C組神經(jīng)和周圍組織廣泛緊密粘連,鈍性不易分離,活動度差。光鏡觀察:術(shù)后2、4、8、12周A組、B組神經(jīng)恢復(fù)情況明顯優(yōu)于C組。電鏡觀察:術(shù)后4周A、B、C三組神經(jīng)纖維再生均不明顯,板層結(jié)構(gòu)均不清晰。術(shù)后8周、12周A、B組比C組神經(jīng)纖維再生數(shù)量更多、髓鞘厚度厚、板層結(jié)構(gòu)更清晰,軸突直徑更大。免疫組織化學(xué)檢測:A、B組中S-100蛋白表達(dá)及分布均優(yōu)于同時期C組。軸突圖像分析:A、B組神經(jīng)吻合口遠(yuǎn)端有髓神經(jīng)纖維直徑、髓鞘厚度及橫截面有髓神經(jīng)纖維數(shù)目均優(yōu)于同時期C組,結(jié)果有統(tǒng)計學(xué)意義(P0.05)。神經(jīng)電生理檢測:術(shù)后術(shù)側(cè)神經(jīng)的誘發(fā)電位潛伏期、波幅及傳導(dǎo)速度檢測示:A、B組比C組潛伏期短、波幅高及神經(jīng)傳導(dǎo)速度快,結(jié)果有統(tǒng)計學(xué)意義(P0.05)。坐骨神經(jīng)功能指數(shù):A組、B組坐骨神經(jīng)功能指數(shù)均明顯高于同時期C組,統(tǒng)計學(xué)分析有顯著性差異(P0.05)。結(jié)論:人羊膜作為一種生物材料修復(fù)周圍神經(jīng)損傷有比較好的效果�？梢詼p輕受損神經(jīng)與周圍組織的粘連和減少神經(jīng)吻合處的瘢痕形成;可促進(jìn)神經(jīng)纖維再生、軸突直徑增大、髓鞘增厚;可以減輕神經(jīng)切口處炎性反應(yīng)、免疫反應(yīng)。
[Abstract]:Objective: peripheral nerve injury is a common disabling disease in the clinic, which directly damages the labor resources and causes a heavy burden on social and family life. The regeneration of peripheral nerve injury is affected by many factors, and the microenvironment and the local scar at the anastomotic stoma seriously affect the repair effect after the nerve injury. For this reason, The improvement of local microenvironment and reduction of local scar is beneficial to the regeneration of peripheral nerve and the recovery of nerve function. The amniotic membrane has semimodialysis, ensures the smooth transmission of nutrients, promotes the proliferation of Schwann cells, accelerates the growth of the nerve axons, inhibits the biological activity of the peripheral fibroblasts, limits the surrounding tissue to invade the nerve broken ends and inhibits the surrounding tissue. In this experiment, the rat sciatic nerve injury model was constructed. The rat model of sciatic nerve injury was constructed, the nerve kissing was carried out under the microscope, and the fresh amniotic membrane was wrapped in the kissing place. The indexes of the fresh sheep were evaluated through gross observation, light microscopy, transmission electron microscopy, immunohistochemistry, neurophysiological test, and the sciatic nerve function index. The effect of membrane on regeneration of peripheral nerve injury. Methods: 60 healthy SD rats, male and female, 220 gram of body weight, 220 gram of body weight were used to prepare the acute injury model of the sciatic nerve in one side of the leg. The group of A (repaired the nerve after the nerve was repaired with the fresh amniotic membrane), and the group B (repair the nerve after the nerve membrane was wrapped with the anastomosis), and the C group (repair) The three groups, 20 rats in each group, were observed in 2,4,8 and 12 weeks after operation, with light microscopy and immunohistochemical staining (the expression of S-100 protein in the nerve tissue); the transmission electron microscopy was performed at the 4,8,12 weeks after the operation, and the axon imaging analysis (counting the number of myelinated nerve fibers, axon diameter and the thickness of myelin sheath) were observed, respectively. Combined muscle action potential detection (measuring and recording latency, amplitude and nerve conduction velocity) and calculating the sciatic nerve function index and its recovery rate. All data were statistically processed. Results: gross observation: three groups of rats were not infected after operation, 7 days after the incision was cut, the incision was healed, and the three groups of rats were 2 weeks after operation. There was a slight swelling of the lower extremities, decreased activity obviously and the limp.2 weeks after the swelling, the mental state and diet gradually resumed. The limping time of the group of A and B rats was shorter than that of the C group, and the amount of activity recovered earlier.A. The number of foot ulcers in the foot and toe of the B group and the C group was less and the recovery time was shorter. The amniotic membrane, the biofilm partially absorbed, and the large part of the biofilm after 2 weeks after the operation. Absorption, 8 weeks completely absorbed group.A, group B nerve and surrounding tissue slightly adhesions and adhesion more loose, activity can be.C group nerve and surrounding tissue widely close adhesion, blunt not easy to separate, poor activity. After 2,4,8,12 week A group, B group nerve recovery is obviously better than the C group. Electron microscope observation: 4 weeks A, B, C three groups of nerve fibers after the operation The regeneration was not obvious, and the lamellar structure was not clear. 8 weeks after the operation, 12 weeks A, the B group had more nerve regeneration than the C group, the thickness of the myelin sheath was thicker, the lamellar structure was clearer and the diameter of the axon was greater. The expression and distribution of S-100 protein in the group of B were better than that of the simultaneous C group. The axonal image analysis: A, B group of distal nerve anastomoses had medullary God. The diameter of the fibers, the thickness of myelin sheath and the number of myelinated nerve fibers in the cross section were better than those in the C group, and the results were statistically significant (P0.05). The neurophysiological tests showed that the latency of evoked potential, amplitude and conduction velocity of the postoperative lateral nerve were detected: A, the latency of the group B was shorter, the amplitude of wave was higher and the nerve conduction velocity was faster, and the results were statistically significant. (P0.05) the sciatic nerve function index: the function index of the sciatic nerve in group A and group B was significantly higher than that of the group C at the same time. There was a significant difference (P0.05). Conclusion: the amniotic membrane as a biological material has a better effect on the repair of peripheral nerve injury. It can reduce the adhesion of the damaged nerve to the surrounding tissue and reduce the nerve anastomosis. Scar formation; can promote nerve fiber regeneration, axon diameter increased, myelin thickening; can reduce the inflammation of nerve incision, immune response.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R651.3

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