可注射型三维互锁导电神经黏合剂修复大鼠周围神经损伤的可行性及效果评价

Feasibility and efficacy evaluation of injectable three-dimensional interlocking conductive nerve adhesive for peripheral nerve injury repair

摘要

目的 探讨可注射型三维互锁导电神经黏合剂修复大鼠周围神经损伤的可行性及效果。方法 选取54只成年雄性SD大鼠，随机分为实验组（APT-C导电神经黏合剂）、阳性对照组（商用纤维蛋白胶）、传统对照组（11/0显微缝合线缝合），每组18只。以右侧坐骨神经横断修复为模型，实验组采用20 μL导电神经黏合剂黏合神经断端，阳性对照组用同等剂量纤维蛋白胶，传统对照组行神经外膜缝合。所有行为学、电生理、组织学及免疫荧光检测均采用单盲法评估。术后通过步行轨迹分析、电生理监测、腓肠肌恢复率检测及组织学、免疫荧光分析、生物力学测试及电镜检测评估修复效果。计量资料经Shapiro-Wilk检验正态性、Levene检验方差齐性，满足条件后以$\bar{x}\pm s$表示，多组比较采用单因素ANOVA，两两比较采用Tukey HSD校正。结果 实验组黏合剂剪切黏附强度为（62.7±4.5）kPa（为纤维蛋白胶的5~10倍），电导率为1.26×10^-3~8.72×10^-4 S/cm，生物相容性良好。术后2个月，实验组神经功能指数（SFI）为（−45.6±3.8），显著高于阳性对照组（−68.4±4.3）和传统对照组（−70.1±4.5）（t=4.21、5.36，P<0.05）；复合肌肉动作电位（CMAP）波幅更高、潜伏期更短，腓肠肌恢复率≥40%，均显著优于对照组（P<0.05）；组织学显示神经轴突排列有序、髓鞘完整，TUBB3、MBP阳性表达更强。结论 可注射型APT-C导电神经黏合剂可通过三维互锁网络结构实现较强黏附，并结合导电材料促进周围神经修复。在大鼠坐骨神经横断模型中，其修复效果优于商用纤维蛋白胶和传统显微缝合术，且操作较简便，为周围神经损伤修复提供了新的实验依据。

Abstract

Objective To investigate the feasibility and efficacy of injectable three-dimensional interlocking conductive nerve adhesive for repairing peripheral nerve injury in rats. Methods Fifty-four adult male SD rats were randomly divided into three groups (18 rats in each group): the experimental group (APT-C conductive nerve adhesive), the positive control group (commercial fibrin glue), and the traditional control group (11/0 microsuture). Using the right sciatic nerve transection repair as a model, the experimental group was treated with 20 μL conductive nerve adhesive to bond the nerve stumps, the positive control group with the same dose of fibrin glue, and the traditional control group with epineurial suture. All behavioral, electrophysiological, histological and immunofluorescence examinations were evaluated using a single-blind method. After surgery, the repair effect was evaluated by walking track analysis, electrophysiological monitoring, gastrocnemius muscle recovery rate detection, histopathological examination, and immunofluorescence analysis, biomechanical testing and electron microscopy examination. The normality of measurement data was verified by theShapiro-Wilk test, and the homogeneity of variance was tested by the Levene test. Data conforming to normal distribution and homogeneity of variance were expressed as mean±standard deviation. One-way analysis of variance (ANOVA) was used for multi-group comparison, and the Tukey HSD correction was adopted for pairwise comparison. Results The shear adhesive strength of the adhesive in the experimental group was (62.7±4.5) kPa, which was 5–10 times that of fibrin glue, with an electrical conductivity ranging from 1.26×10^-3 to 8.72×10^-4 S/cm and favorable biocompatibility. At 2 months postoperatively, the sciatic function index (SFI) of the experimental group was −45.6±3.8, which was significantly higher than those of the positive control group (−68.4±4.3) and the traditional control group (−70.1±4.5) (t=4.21, 5.36, P<0.05). The experimental group also exhibited higher compound muscle action potential (CMAP) amplitude, shorter latency period, and gastrocnemius muscle recovery rate ≥40%, all of which were significantly superior to those of the control groups (P<0.05). Histological results showed orderly arranged nerve axons and intact myelin sheaths, with stronger positive expression of TUBB3 and MBP. Conclusions The injectable APT-C conductive nerve adhesive can achieve strong adhesion through a three-dimensional interlocking network structure and promote peripheral nerve repair via conductive materials. In a rat sciatic nerve transection model, its repair effect was superior to that of commercial fibrin glue and traditional microsuture with simpler operation, providing new experimental evidence for peripheral nerve injury repair.

关键词

周围神经损伤 / 神经黏合剂 / 可注射水凝胶 / 导电聚合物 / 神经修复与再生

Key words

Peripheral nerve injury / Nerve adhesive / Injectable hydrogel / Conductive polymer / Nerve repair and regeneration

参考文献

[1] Liu Y, Zhang X, Xiao C, et al. Engineered hydrogels for peripheral nerve repair[J]. Materials Today Bio, 2023, 20: 100668. DOI: 10.1016/j.mtbio.2023.100668.

[2] Vijayavenkataraman S. Nerve guide conduits for peripheral nerve injury repair: a review on design, materials and fabrication methods[J]. Acta Biomaterialia, 2020, 106: 54-69. DOI: 10.1016/j.actbio.2020.02.003.

[3] Fekrazad R, Mortezai O, Pedram M, et al. Transected sciatic nerve repair by diode laser protein soldering[J]. J Photochem Photobiol B, 2017, 173: 441-447. DOI: 10.1016/j.jphotobiol.2017.06.008.

[4] 杨勇, 陈山林, 荣艳波, 等. 端侧吻合与自体神经移植治疗周围神经缺损的比较[J]. 北京大学学报(医学版), 2012, 44(6): 962-965.

[5] Qiu L, Qi See AA, Steele TWJ, et al. Bioadhesives in neurosurgery: a review[J]. J Neurosurg, 2019, 133(6): 1928-1938.

[6] Koopman JE, Duraku LS, de Jong T, et al. A systematic review and meta-analysis on the use of fibrin glue in peripheral nerve repair: can we just glue it?[J]. J Plast, Reconstr Aesthet Surg, 2022, 75(3): 1018-1033. DOI: 10.1016/j.bjps.2022.01.007.

[7] Roth J, Shtokman J, Shamir MH, et al. Regeneration of the transected rat sciatic nerve after suturing or adhesion with cyanoacrylate glue[J]. J Neurosurgery, 2011, 114(1): 245-252. DOI: 10.3171/2010.7.JNS091266.

[8] Sun Z, Hu H, Zhang X, et al. Recent advances in peptide-based bioactive hydrogels for nerve repair and regeneration: from material design to fabrication, functional tailoring and applications[J]. J Mater Chem B, 2024, 12(9): 2253-2273. DOI: 10.1039/d4tb00019f.

[9] Chow N, Miears H, Cox C, et al. Fibrin glue and its alternatives in peripheral nerve repair[J]. Ann Plast Surg, 2021, 86(1): 103-108. DOI: 10.1097/SAP.0000000000002408.

[10] Brennan M. Fibrin glue[J]. Blood Rev, 1991, 5(4): 240-244. DOI: 10.1016/0268-960x(91)90015-5.

[11] Wang H, Wang M, Wu J, et al. Nature-inspired gelatin-based adhesive hydrogel: a rapid and user-friendly solution for hemostatic applications[J]. Adv Healthc Mater, 2024, 13(3): e2304444. DOI: 10.1002/adhm.202304444.

[12] Lukin I, Erezuma I, Maeso L, et al. Progress in gelatin as biomaterial for tissue engineering[J]. Pharmaceutics, 2022, 14(6): 1177. DOI: 10.3390/pharmaceutics14061177.

[13] Jafari H, Ghaffari-Bohlouli P, Niknezhad SV, et al. Tannic acid: a versatile polyphenol for design of biomedical hydrogels[J]. J Mater Chem B, 2022, 10(31): 5873-5912. DOI: 10.1039/d2tb01056a.

[14] Ren X, Yang M, Yang T, et al. Highly conductive PPy−PEDOT:PSS hybrid hydrogel with superior biocompatibility for bioelectronics application[J]. ACS Appl Mater Interfaces, 2021, 13(21): 25374-25382. DOI: 10.1021/acsami.1c04432.

[15] Yan J, Wu R, Liao S, et al. Applications of polydopamine-modified scaffolds in the peripheral nerve tissue engineering[J]. Front Bioeng Biotechnol, 2020, 8: 590998. DOI: 10.3389/fbioe.2020.590998.

[16] 武竞衡, 王海花. 一种透明质酸/丝素蛋白双交联可注射水凝胶及其制备方法: CN202210302845.0[P]. 2022-05-02.

[17] Qiu P, Wang L, Wang J, et al. Adhesive chitosan-based hybrid biohydrogels for peripheral nerve injury repair[J]. Front Cell Dev Biol, 2024, 12: 1499766. DOI: 10.3389/fcell.2024.1499766.

[18] Tomić SL, Babić Radić MM, Vuković JS, et al. Alginate-based hydrogels and scaffolds for biomedical applications[J]. Mar Drugs, 2023, 21(3): 177. DOI: 10.3390/md21030177.

[19] Zhang F, Zhang M, Liu S, et al. Application of hybrid electrically conductive hydrogels promotes peripheral nerve regeneration[J]. Gels, 2022, 8(1): 41. DOI: 10.3390/gels8010041.

[20] Liu J, Guan X, Zheng S, et al. Neuritin accelerates Schwann cell dedifferentiation via PI3K/Akt/mTOR signalling pathway during Wallerian degeneration[J]. J Cell Mol Med, 2024, 28(16): e70012. DOI: 10.1111/jcmm.70012.

[21] Shao S, Xinqing Y, Xuehai Q, et al. Nanomaterial-enhanced conductive hydrogels for peripheral nerve repair: biomimetic design, mechanisms, and translational challenges[J]. Int J Nanomedicine, 2026, 21: 577139. DOI: 10.2147/IJN.S577139.

基金资助

北京市医院管理中心“登峰”计划专项经费资助(DFL20240402)