构建Ⅰ型胶原蛋白神经导管及其在前臂正中神经损伤重建中的作用机制
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摘要
背景:Ⅰ型胶原蛋白是一种高分子材料,生物相容性、细胞亲和力强,能在特定的条件下发生降解。同时,经过交联处理、干预后能获得良好的机械性能,但是在前臂正中神经损伤中的重建效果缺乏研究。目的:探讨Ⅰ型胶原蛋白神经导管制备方法及在前臂正中神经损伤重建中的作用机制。方法:选择由贵州医科大学附属医院医学动物实验中心提供的SD大鼠40只,随机取10只大鼠设为假手术组,剩余30只SD大鼠采用激光诱导光化学反应建立大鼠前臂正中神经损伤动物模型,建模成功后随机分为阳性对照组10只、Ⅰ型胶原蛋白组10只和自体神经组10只。假手术组常规喂养,不参与建模;阳性对照组建模成功后不采取特殊处理,自行修复;Ⅰ型胶原蛋白组采用Ⅰ型胶原蛋白神经导管进行桥接,自体神经组采用自体神经进行桥接,比较各组修复效果。结果与结论:(1)倒置显微镜下可见交联前Ⅰ型胶原蛋白排列松散,呈蜂窝状不规则的孔隙及孔径,孔径大小为10-100μm,孔隙20-200μm,孔隙间质相对较薄,交联后Ⅰ型胶原蛋白排列致密,胶原纤维之间能形成相对规则的孔径、孔隙,大小为50-100μm,孔隙为20-200μm,孔隙间质增厚,空间结构发生明显的变化;(2)Ⅰ型胶原蛋白组与自体神经组修复后4,8,12周翻转及放置明尼苏达手灵巧度评定方法评分,均低于阳性对照组(P <0.05),高于假手术组(P <0.05);(3)Ⅰ型胶原蛋白组、自体神经组修复后12周波幅、潜伏期差异无显著性意义(P> 0.05),但均高于阳性对照组(P <0.05);(4)修复12周后,阳性对照组可见神经损伤部位明显,周围可见坏死组织;自体神经组未见损伤部位,周围坏死面积减少,恢复良好;Ⅰ型胶原蛋白组甲苯胺蓝染色下未见损伤部位,恢复良好;(5)结果提示,通过自制模具能成功制备Ⅰ型胶原蛋白神经导管,将其用于大鼠前臂正中神经损伤重建中能获得良好的修复效果,有助于促进神经损伤修复。
BACKGROUND: Type I collagen is a polymer material that has good biocompatibility and good cell affinity, and can degrade under certain conditions. It can also develop good mechanical properties after cross-linking, but it is less reported in the reconstruction of the injured median nerve of the forearm. OBJECTIVE: To explore the preparation method of type I collagen nerve conduit and its mechanism in the reconstruction following median nerve injury in the forearm. METHODS: A total of 40 Sprague-Dawley rats were selected from the Medical Animal Experimental Center, the Affiliated Hospital of Guizhou Medical University, 10 of which were randomly selected as sham surgery group. The remaining 30 rats were used to establish a rat model of median nerve injury in the forearm by laser-induced photochemical reaction. After successful modeling, the model rats were randomly divided into positive control group(n=10), type I collagen group (n=10) and autologous nerve group (n=10). The sham surgery group was routinely fed and did not participate in the modeling; the positive control group did not take special treatment after the successful modeling; the type I collagen group was subjected to bridging with type I collagen nerve conduit; and the autologous nerve group was subjected to bridging with autologous nerve. The repair effects were compared among groups. RESULTS AND CONCLUSION:(1) Under the inverted microscope, the type I collagen was loosely arranged before cross-linking, and it had honeycomb-shaped irregular pores with the pore size of 10-100 μm and the porosity of 20-200 μm, and the pore interstitial was relatively thin. After cross-linking, the type I collagen was densely arranged, the collagen fibers could form relatively regular pores with the pore size of 50-100 μm and the porosity of 20-200 μm, the interstitial mass was thickened, and the spatial structure changed significantly.(2) After 4, 8 and 12 weeks of repair, the scores on the Minnesota Manual Dexterity Test in the type I collagen and autologous nerve groups were significantly lower than those in the positive control group(P < 0.05) and higher than those in the sham surgery group (P < 0.05).(3) At 12 weeks after repair, there was no significant difference in amplitude and latency between the type I collagen group and the autologous nerve group (P > 0.05), but the amplitude and latency in both groups were significantly higher than those in the positive control group (P < 0.05).(4) At 12 weeks after repair, the nerve injury site surrounded by necrotic tissues was visible in the positive control group; no injury was found in the autologous nerve group, and the surrounding necrotic area decreased, indicating good recovery; no injury was shown by toluidine blue staining in the type II collagen group, indicating good recovery. Overall, the type I collagen nerve conduit can be successfully prepared by the self-made mold, and it can be used for the reconstruction following median nerve injury in the rat forearm, helping nerve repair.
BACKGROUND: Type I collagen is a polymer material that has good biocompatibility and good cell affinity, and can degrade under certain conditions. It can also develop good mechanical properties after cross-linking, but it is less reported in the reconstruction of the injured median nerve of the forearm. OBJECTIVE: To explore the preparation method of type I collagen nerve conduit and its mechanism in the reconstruction following median nerve injury in the forearm. METHODS: A total of 40 Sprague-Dawley rats were selected from the Medical Animal Experimental Center, the Affiliated Hospital of Guizhou Medical University, 10 of which were randomly selected as sham surgery group. The remaining 30 rats were used to establish a rat model of median nerve injury in the forearm by laser-induced photochemical reaction. After successful modeling, the model rats were randomly divided into positive control group(n=10), type I collagen group (n=10) and autologous nerve group (n=10). The sham surgery group was routinely fed and did not participate in the modeling; the positive control group did not take special treatment after the successful modeling; the type I collagen group was subjected to bridging with type I collagen nerve conduit; and the autologous nerve group was subjected to bridging with autologous nerve. The repair effects were compared among groups. RESULTS AND CONCLUSION:(1) Under the inverted microscope, the type I collagen was loosely arranged before cross-linking, and it had honeycomb-shaped irregular pores with the pore size of 10-100 μm and the porosity of 20-200 μm, and the pore interstitial was relatively thin. After cross-linking, the type I collagen was densely arranged, the collagen fibers could form relatively regular pores with the pore size of 50-100 μm and the porosity of 20-200 μm, the interstitial mass was thickened, and the spatial structure changed significantly.(2) After 4, 8 and 12 weeks of repair, the scores on the Minnesota Manual Dexterity Test in the type I collagen and autologous nerve groups were significantly lower than those in the positive control group(P < 0.05) and higher than those in the sham surgery group (P < 0.05).(3) At 12 weeks after repair, there was no significant difference in amplitude and latency between the type I collagen group and the autologous nerve group (P > 0.05), but the amplitude and latency in both groups were significantly higher than those in the positive control group (P < 0.05).(4) At 12 weeks after repair, the nerve injury site surrounded by necrotic tissues was visible in the positive control group; no injury was found in the autologous nerve group, and the surrounding necrotic area decreased, indicating good recovery; no injury was shown by toluidine blue staining in the type II collagen group, indicating good recovery. Overall, the type I collagen nerve conduit can be successfully prepared by the self-made mold, and it can be used for the reconstruction following median nerve injury in the rat forearm, helping nerve repair.
引文
[1]田野,邱逦.正中神经神经脂肪瘤病超声表现1例[J].中国医学影像技术,2017,33(3):467-467.
[2]Donoghoe N,Rosson GD,Dellon AL.Reconstruction of the human median nerve in the forearm with the neurotube.Microsurgery.2010;27(7):595-600.
[3]崔满意,田恒进,王志勇,等.小指展肌联合小指短屈肌转移重建拇指对掌功能[J].中华手外科杂志,2016,32(4):312-313.
[4]Gao KM,Lao J,Guan WJ,et al.Is it necessary to use the entire root as a donor when transferring contralateral C7nerve to repair median nerve?Neural Regen Res.2018;13(1):94-99.
[5]蔡东妙,王庆祥,陈振毅,等.连续股神经阻滞联合硬膜外吗啡镇痛在老年患者全膝关节置换术中的应用[J].临床麻醉学杂志,2016,32(3):234-236.
[6]Huang CT,Chen SH,Lue JH,et al.Neurosteroid allopregnanolone suppresses median nerve injury-induced mechanical hypersensitivity and glial extracellular signal-regulated kinase activation throughγ-aminobutyric acid type a receptor modulation in the rat cuneate nucleus.Anesthesiology.2016;125(6):1202.
[7]徐金东,郁丽娜,赵达强,等.超声引导胸椎旁神经阻滞在胸腔镜交感神经切断术中的应用[J].南方医科大学学报,2016,36(12):1655-1659.
[8]Aseem F,Williams JW,Walker FO,et al.Neuromuscular ultrasound in patients with carpal tunnel syndrome and normal nerve conduction studies.Muscle Nerve.2017;55(6):913-915.
[9]张会兰,易兵成,王先流,等.用高度取向石墨烯/聚乳酸(Gr/PLLA)复合超细纤维构建神经导管[J].高等学校化学学报,2016,37(5):972-982.
[10]贾赛雄,吴迪,利春叶,等.改良尺侧腕伸肌腱联合拇短伸肌腱转位重建拇对掌功能的临床应用[J].中华手外科杂志,2017,33(1):47-49.
[11]孟维锟,黄仲,谭振,等.血管支架支撑静脉神经导管修复兔周围神经缺损的实验研究[J].四川大学学报(医学版),2017,48(5):687-692.
[12]Flohr-Madsen S,Ytreb??LM,Valen K,et al.A randomised placebo-controlled trial examining the effect on hand supination after the addition of a suprascapular nerve block to infraclavicular brachial plexus blockade.Anaesthesia.2016;71(8):938-947.
[13]饶建伟,叶舟,占蓓蕾,等.丝胶蛋白对仿生丝素蛋白神经导管的改性研究[J].中华显微外科杂志,2016,39(3):251-257.
[14]王耿焕,沈和平,褚正民,等.神经外科重症监护室重型颅脑损伤患者医院感染的影响因素分析[J].中华神经外科杂志,2016,32(4):405-408.
[15]赵芹,林栋,刘海梅.冰岛刺参胶原蛋白多肽对PC12细胞氧化损伤的保护作用[J].食品工业科技,2016,37(18):354-358.
[16]Greening J,Dilley A.Posture-induced changes in peripheral nerve stiffness measured by ultrasound shear-wave elastography.Muscle Nerve.2016;55(2):213.
[17]邢冉,陈旭义,朱祥,等.神经干细胞在新型复合支架中的生长和分化[J].中国组织工程研究,2016,20(19):2857-2863.
[18]郭琦,刘灿,海宝,等.壳聚糖导管复合辛伐他汀/泊洛沙姆407水凝胶修复大鼠外周神经缺损的研究[J].中国比较医学杂志,2016,26(5):1-9.
[19]Goedee HS,van der Pol WL,van Asseldonk JH,et al.Diagnostic value of sonography in treatment-naive chronic inflammatory neuropathies.Neurology.2017;88(2):143.
[20]朱明珍,初红,卢祖能.腕管综合征患者口服及局部注射类固醇疗效的临床和神经超声评估研究[J].国际神经病学神经外科学杂志,2016,43(6):491-496.
[21]刘鐘阳,刘靓,黄景辉,等.复合雪旺细胞的神经组织工程材料联合脉冲电磁场促进大鼠坐骨神经缺损的再生[J].中华骨科杂志,2016,36(8):465-478.
[22]Schreiber S,Dannhardt-Stieger V,Henkel D,et al.Quantifying disease progression in ALS using peripheral nerve sonography.Muscle Nerve.2016;54(3):391-397.
[23]艾尔肯.热合木吐拉,彭峰,张莉,等.利用激光诱导光化学反应建立大鼠正中神经定量损伤模型[J].中华手外科杂志,2016,32(5):377-381.
[24]Stavros K,Paik D,Motiwala R,et al.Median nerve penetration by a persistent median artery and vein mimicking carpal tunnel syndrome.Muscle Nerve.2016;53(3):485-487.
[25]赵永青,雷利华,李瑞博,等.神经介入导管室应用精细护理预防医院感染的评价[J].中华医院感染学杂志,2017,27(14):3343-3345.
[26]Choi J,Kim JH,Jang JW,et al.Decellularized sciatic nerve matrix as a biodegradable conduit for peripheral nerve regeneration.Neural Regen Res.2018;13(10):1796-1803.
[27]方晨,吕圣龙,吴海英,等.电流感觉阈值在评估1型糖尿病患者早期周围神经受损中的价值[J].中国糖尿病杂志,2016,24(4):339-343.
[28]Jiang H,Wu Y,Dang Y,et al.Closed reduction using the percutaneous leverage technique and internal fixation with K-wires to treat angulated radial neck fractures in children-case report.Medicine.2017;96(1):e5806.
[29]陈军,沈华.神经导管内支架修复周围神经缺损的研究应用与进展[J].中国组织工程研究,2017,21(8):1273-1279.
[30]宋奇翔,叶宸,谭海颂,等.小鼠膀胱内压力与尿道外括约肌肌电图同步测定方法的建立与验证[J].第三军医大学学报,2017,39(2):179-184.
[31]王双,林斌,陈志达,等.抑制TNFR/RIPK信号通路对大鼠急性脊髓损伤后神经功能的影响[J].中国脊柱脊髓杂志,2017,27(4):353-360.
[2]Donoghoe N,Rosson GD,Dellon AL.Reconstruction of the human median nerve in the forearm with the neurotube.Microsurgery.2010;27(7):595-600.
[3]崔满意,田恒进,王志勇,等.小指展肌联合小指短屈肌转移重建拇指对掌功能[J].中华手外科杂志,2016,32(4):312-313.
[4]Gao KM,Lao J,Guan WJ,et al.Is it necessary to use the entire root as a donor when transferring contralateral C7nerve to repair median nerve?Neural Regen Res.2018;13(1):94-99.
[5]蔡东妙,王庆祥,陈振毅,等.连续股神经阻滞联合硬膜外吗啡镇痛在老年患者全膝关节置换术中的应用[J].临床麻醉学杂志,2016,32(3):234-236.
[6]Huang CT,Chen SH,Lue JH,et al.Neurosteroid allopregnanolone suppresses median nerve injury-induced mechanical hypersensitivity and glial extracellular signal-regulated kinase activation throughγ-aminobutyric acid type a receptor modulation in the rat cuneate nucleus.Anesthesiology.2016;125(6):1202.
[7]徐金东,郁丽娜,赵达强,等.超声引导胸椎旁神经阻滞在胸腔镜交感神经切断术中的应用[J].南方医科大学学报,2016,36(12):1655-1659.
[8]Aseem F,Williams JW,Walker FO,et al.Neuromuscular ultrasound in patients with carpal tunnel syndrome and normal nerve conduction studies.Muscle Nerve.2017;55(6):913-915.
[9]张会兰,易兵成,王先流,等.用高度取向石墨烯/聚乳酸(Gr/PLLA)复合超细纤维构建神经导管[J].高等学校化学学报,2016,37(5):972-982.
[10]贾赛雄,吴迪,利春叶,等.改良尺侧腕伸肌腱联合拇短伸肌腱转位重建拇对掌功能的临床应用[J].中华手外科杂志,2017,33(1):47-49.
[11]孟维锟,黄仲,谭振,等.血管支架支撑静脉神经导管修复兔周围神经缺损的实验研究[J].四川大学学报(医学版),2017,48(5):687-692.
[12]Flohr-Madsen S,Ytreb??LM,Valen K,et al.A randomised placebo-controlled trial examining the effect on hand supination after the addition of a suprascapular nerve block to infraclavicular brachial plexus blockade.Anaesthesia.2016;71(8):938-947.
[13]饶建伟,叶舟,占蓓蕾,等.丝胶蛋白对仿生丝素蛋白神经导管的改性研究[J].中华显微外科杂志,2016,39(3):251-257.
[14]王耿焕,沈和平,褚正民,等.神经外科重症监护室重型颅脑损伤患者医院感染的影响因素分析[J].中华神经外科杂志,2016,32(4):405-408.
[15]赵芹,林栋,刘海梅.冰岛刺参胶原蛋白多肽对PC12细胞氧化损伤的保护作用[J].食品工业科技,2016,37(18):354-358.
[16]Greening J,Dilley A.Posture-induced changes in peripheral nerve stiffness measured by ultrasound shear-wave elastography.Muscle Nerve.2016;55(2):213.
[17]邢冉,陈旭义,朱祥,等.神经干细胞在新型复合支架中的生长和分化[J].中国组织工程研究,2016,20(19):2857-2863.
[18]郭琦,刘灿,海宝,等.壳聚糖导管复合辛伐他汀/泊洛沙姆407水凝胶修复大鼠外周神经缺损的研究[J].中国比较医学杂志,2016,26(5):1-9.
[19]Goedee HS,van der Pol WL,van Asseldonk JH,et al.Diagnostic value of sonography in treatment-naive chronic inflammatory neuropathies.Neurology.2017;88(2):143.
[20]朱明珍,初红,卢祖能.腕管综合征患者口服及局部注射类固醇疗效的临床和神经超声评估研究[J].国际神经病学神经外科学杂志,2016,43(6):491-496.
[21]刘鐘阳,刘靓,黄景辉,等.复合雪旺细胞的神经组织工程材料联合脉冲电磁场促进大鼠坐骨神经缺损的再生[J].中华骨科杂志,2016,36(8):465-478.
[22]Schreiber S,Dannhardt-Stieger V,Henkel D,et al.Quantifying disease progression in ALS using peripheral nerve sonography.Muscle Nerve.2016;54(3):391-397.
[23]艾尔肯.热合木吐拉,彭峰,张莉,等.利用激光诱导光化学反应建立大鼠正中神经定量损伤模型[J].中华手外科杂志,2016,32(5):377-381.
[24]Stavros K,Paik D,Motiwala R,et al.Median nerve penetration by a persistent median artery and vein mimicking carpal tunnel syndrome.Muscle Nerve.2016;53(3):485-487.
[25]赵永青,雷利华,李瑞博,等.神经介入导管室应用精细护理预防医院感染的评价[J].中华医院感染学杂志,2017,27(14):3343-3345.
[26]Choi J,Kim JH,Jang JW,et al.Decellularized sciatic nerve matrix as a biodegradable conduit for peripheral nerve regeneration.Neural Regen Res.2018;13(10):1796-1803.
[27]方晨,吕圣龙,吴海英,等.电流感觉阈值在评估1型糖尿病患者早期周围神经受损中的价值[J].中国糖尿病杂志,2016,24(4):339-343.
[28]Jiang H,Wu Y,Dang Y,et al.Closed reduction using the percutaneous leverage technique and internal fixation with K-wires to treat angulated radial neck fractures in children-case report.Medicine.2017;96(1):e5806.
[29]陈军,沈华.神经导管内支架修复周围神经缺损的研究应用与进展[J].中国组织工程研究,2017,21(8):1273-1279.
[30]宋奇翔,叶宸,谭海颂,等.小鼠膀胱内压力与尿道外括约肌肌电图同步测定方法的建立与验证[J].第三军医大学学报,2017,39(2):179-184.
[31]王双,林斌,陈志达,等.抑制TNFR/RIPK信号通路对大鼠急性脊髓损伤后神经功能的影响[J].中国脊柱脊髓杂志,2017,27(4):353-360.