Have a personal or library account? Click to login
Protective Effect of the Human Epineural Patch Application after Sciatic Nerve Crush Injury Followed by Nerve Transection and End-to-End Repair Cover

Protective Effect of the Human Epineural Patch Application after Sciatic Nerve Crush Injury Followed by Nerve Transection and End-to-End Repair

Archivum Immunologiae et Therapiae Experimentalis
Volume 73 (2025): Issue 1 (January 2025)
By: Maria Siemionow,  Weronika Radecka,  Katarzyna Kozlowska,  Lucile Chambily,  Sonia Brodowska,  Dominika Kuc,  Gabrielle Filipek and  Katarzyna Budzynska  
Open Access
|Mar 2025

Figures & Tables

Fig 1.

Schematic illustration of the experimental design detailing the creation and application of the hEP as an innovative approach to enhance nerve regeneration following sciatic nerve crush injury, transection, and end-to-end repair. hEP, human epineural patch; hAM, human amniotic membrane.
Schematic illustration of the experimental design detailing the creation and application of the hEP as an innovative approach to enhance nerve regeneration following sciatic nerve crush injury, transection, and end-to-end repair. hEP, human epineural patch; hAM, human amniotic membrane.

Fig 2.

Surgical technique of the CTR procedure and application of the created hEP and hAM patch. (a) Creation of the sciatic nerve crush injury. (b) Sciatic nerve crush site 0.5 mm long. (c) End-to-end repair after resection of the sciatic nerve crush site. (d) Sciatic nerve repair site wrapped with the hEP. (e) Sciatic nerve repair site wrapped with the hAM. hAM, human amniotic membrane; hEP, human epineural patch.
Surgical technique of the CTR procedure and application of the created hEP and hAM patch. (a) Creation of the sciatic nerve crush injury. (b) Sciatic nerve crush site 0.5 mm long. (c) End-to-end repair after resection of the sciatic nerve crush site. (d) Sciatic nerve repair site wrapped with the hEP. (e) Sciatic nerve repair site wrapped with the hAM. hAM, human amniotic membrane; hEP, human epineural patch.

Fig 3.

Functional assessment of sciatic nerve regeneration was done using Toe-Spread and Pinprick tests in the 6-week study at 1, 3, and 6-week study points. (a) Motor recovery (Toe-Spread test) revealed no significant response at 1 week, but the hEP group showed significant improvement by 3 weeks and the highest motor response at 6 weeks. (b) Sensory recovery (Pinprick test) showed significantly better outcomes in the hEP group at 3 weeks and 6 weeks compared with the No protection and hAM groups, with the hEP group demonstrating the highest response at 6 weeks. Data are presented as mean ± SEM, with significance indicated as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.
Functional assessment of sciatic nerve regeneration was done using Toe-Spread and Pinprick tests in the 6-week study at 1, 3, and 6-week study points. (a) Motor recovery (Toe-Spread test) revealed no significant response at 1 week, but the hEP group showed significant improvement by 3 weeks and the highest motor response at 6 weeks. (b) Sensory recovery (Pinprick test) showed significantly better outcomes in the hEP group at 3 weeks and 6 weeks compared with the No protection and hAM groups, with the hEP group demonstrating the highest response at 6 weeks. Data are presented as mean ± SEM, with significance indicated as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.

Fig 4.

Functional assessment of sciatic nerve regeneration was done using Toe-Spread and Pinprick tests in the 12-week study at 1, 3, 6, and 12-week study points. (a) Motor recovery (Toe-Spread test) showed minimal response across all groups at 3 weeks. By 6 weeks, the hEP group maintained significantly higher response levels than the No protection group. By 9 weeks, the hEP group outperformed both groups, though statistical differences were observed for No protection. At 12 weeks, the hEP group achieved the highest recovery, significantly outperforming the No protection group. (b) Sensory recovery (Pinprick test) revealed that the hEP group had significantly higher responses at 6 weeks and 9 weeks than the other groups, with comparable recovery observed across all groups by 12 weeks. Data are mean ± SEM, with significance marked as *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.
Functional assessment of sciatic nerve regeneration was done using Toe-Spread and Pinprick tests in the 12-week study at 1, 3, 6, and 12-week study points. (a) Motor recovery (Toe-Spread test) showed minimal response across all groups at 3 weeks. By 6 weeks, the hEP group maintained significantly higher response levels than the No protection group. By 9 weeks, the hEP group outperformed both groups, though statistical differences were observed for No protection. At 12 weeks, the hEP group achieved the highest recovery, significantly outperforming the No protection group. (b) Sensory recovery (Pinprick test) revealed that the hEP group had significantly higher responses at 6 weeks and 9 weeks than the other groups, with comparable recovery observed across all groups by 12 weeks. Data are mean ± SEM, with significance marked as *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.

Fig 5.

Evaluation of muscle denervation atrophy by GMI. (a) At 6 weeks, no significant differences in the GMI were observed between groups. (b) In contrast, at 12 weeks, the hEP group exhibited a significantly higher GMI compared with the No protection group. Data are presented as mean ± SEM. Statistical significance was determined using a one-way ANOVA test for group comparisons, with significance levels as follows: *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. GMI, gastrocnemius muscle index; hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.
Evaluation of muscle denervation atrophy by GMI. (a) At 6 weeks, no significant differences in the GMI were observed between groups. (b) In contrast, at 12 weeks, the hEP group exhibited a significantly higher GMI compared with the No protection group. Data are presented as mean ± SEM. Statistical significance was determined using a one-way ANOVA test for group comparisons, with significance levels as follows: *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. GMI, gastrocnemius muscle index; hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.

Fig 6.

Histological assessment of the proximal, crushed, and distal parts of the injured segment at 6 weeks after sciatic nerve crush injury followed by nerve transection and end-to-end repair. (a) Proximal myelin thickness, (b) Crushed myelin thickness, (c) Distal myelin thickness, (d) Proximal fiber diameter, (e) Crushed fiber diameter, (f) Distal fiber diameter, (g) Percentage of proximal myelinated fibers, (h) Percentage of crushed myelinated fibers, (i) Percentage of distal myelinated fibers, (j) Proximal axonal density, (k) Crushed axonal density, and (l) Distal axonal density. The data are presented as mean ± SEM and statistical significance was determined using a one-way ANOVA test for group comparisons, with significance levels denoted as *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.
Histological assessment of the proximal, crushed, and distal parts of the injured segment at 6 weeks after sciatic nerve crush injury followed by nerve transection and end-to-end repair. (a) Proximal myelin thickness, (b) Crushed myelin thickness, (c) Distal myelin thickness, (d) Proximal fiber diameter, (e) Crushed fiber diameter, (f) Distal fiber diameter, (g) Percentage of proximal myelinated fibers, (h) Percentage of crushed myelinated fibers, (i) Percentage of distal myelinated fibers, (j) Proximal axonal density, (k) Crushed axonal density, and (l) Distal axonal density. The data are presented as mean ± SEM and statistical significance was determined using a one-way ANOVA test for group comparisons, with significance levels denoted as *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.

Fig 7.

Histological assessment of the proximal, crushed, and distal parts of the injured segment at 12 weeks after sciatic nerve crush injury followed by nerve transection and end-to-end repair. (a) Proximal myelin thickness, (b) Crushed myelin thickness, (c) Distal myelin thickness, (d) Proximal fiber diameter, (e) Crushed fiber diameter, (f) Distal fiber diameter, (g) Percentage of proximal myelinated fibers, (h) Percentage of crushed myelinated fibers, (i) Percentage of distal myelinated fibers, (j) Proximal axonal density, (k) Crushed axonal density, and (l) Distal axonal density. The data are presented as mean ± SEM and statistical significance was determined using a one-way ANOVA test for group comparisons, with significance levels denoted as *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.
Histological assessment of the proximal, crushed, and distal parts of the injured segment at 12 weeks after sciatic nerve crush injury followed by nerve transection and end-to-end repair. (a) Proximal myelin thickness, (b) Crushed myelin thickness, (c) Distal myelin thickness, (d) Proximal fiber diameter, (e) Crushed fiber diameter, (f) Distal fiber diameter, (g) Percentage of proximal myelinated fibers, (h) Percentage of crushed myelinated fibers, (i) Percentage of distal myelinated fibers, (j) Proximal axonal density, (k) Crushed axonal density, and (l) Distal axonal density. The data are presented as mean ± SEM and statistical significance was determined using a one-way ANOVA test for group comparisons, with significance levels denoted as *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. hAM, human amniotic membrane; hEP, human epineural patch; SEM, standard error of the mean.

Fig 8.

Expression evaluation of selected neurogenic, angiogenic, and immunogenic markers assessed at the crush injury sites and distal ends by immunofluorescence staining at 6 weeks following sciatic nerve crush injury, transection, and repair. (a) GFAP, (b) Laminin B, (c) NGF, (d) S-100, (e) VEGF, (f) vWF, (g) HLA-DR, and (h) HLA-I. Data were presented as mean ± SEM. A two-way ANOVA test for group comparison was used to define statistical significance at *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. GFAP, glial fibrillary acidic protein; HLA-DR, human leukocyte antigen-DR; HLA-I, human leukocyte antigen class I; NGF, nerve growth factor; SEM, standard error of the mean; VEGF, vascular endothelial growth factor; vWF, von Willebrand factor.
Expression evaluation of selected neurogenic, angiogenic, and immunogenic markers assessed at the crush injury sites and distal ends by immunofluorescence staining at 6 weeks following sciatic nerve crush injury, transection, and repair. (a) GFAP, (b) Laminin B, (c) NGF, (d) S-100, (e) VEGF, (f) vWF, (g) HLA-DR, and (h) HLA-I. Data were presented as mean ± SEM. A two-way ANOVA test for group comparison was used to define statistical significance at *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. GFAP, glial fibrillary acidic protein; HLA-DR, human leukocyte antigen-DR; HLA-I, human leukocyte antigen class I; NGF, nerve growth factor; SEM, standard error of the mean; VEGF, vascular endothelial growth factor; vWF, von Willebrand factor.

Fig 9.

Expression evaluation of selected neurogenic, angiogenic, and immunogenic markers assessed at the crush injury sites and distal ends by immunofluorescence staining at 12 weeks following sciatic nerve crush injury, transection, and repair. (a) GFAP, (b) Laminin B, (c) NGF, (d) S-100, (e) VEGF, (f) vWF, (g) HLA-DR, and (h) HLA-I. Data were presented as mean ± SEM. A two-way ANOVA test for group comparison was used to define statistical significance at *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. GFAP, glial fibrillary acidic protein; HLA-DR, human leukocyte antigen-DR; HLA-I, human leukocyte antigen class I; NGF, nerve growth factor; SEM, standard error of the mean; VEGF, vascular endothelial growth factor; vWF, von Willebrand factor.
Expression evaluation of selected neurogenic, angiogenic, and immunogenic markers assessed at the crush injury sites and distal ends by immunofluorescence staining at 12 weeks following sciatic nerve crush injury, transection, and repair. (a) GFAP, (b) Laminin B, (c) NGF, (d) S-100, (e) VEGF, (f) vWF, (g) HLA-DR, and (h) HLA-I. Data were presented as mean ± SEM. A two-way ANOVA test for group comparison was used to define statistical significance at *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. GFAP, glial fibrillary acidic protein; HLA-DR, human leukocyte antigen-DR; HLA-I, human leukocyte antigen class I; NGF, nerve growth factor; SEM, standard error of the mean; VEGF, vascular endothelial growth factor; vWF, von Willebrand factor.

Experimental group descriptions and evaluation methods

Experimental group numberRepair methodNumber of athymic nude rats per group
6 weeks study
1no protective wrappingn=6
2hEPn=6
3hAMn=6
12 weeks study
4no protective wrappingn=6
5hEPn=6
6hAMn=6
Total number of athymic nude rats (n=36)
DOI: https://doi.org/10.2478/aite-2025-0008 | Journal eISSN: 1661-4917
Journal RSS Feed
Language: English
Submitted on: Jan 11, 2025
Accepted on: Feb 25, 2025
Published on: Mar 26, 2025
Published by: Hirszfeld Institute of Immunology and Experimental Therapy
In partnership with: Paradigm Publishing Services
Publication frequency: 1 issue per year
Keywords:
Peripheral nerve repair,
Human epineural patch,
Human amniotic membrane,
Allografts,
Nerve regeneration,
Regenerative medicine
Related subjects:
Medicine,
Basic medical science,
Biochemistry,
Immunology,
Clinical medicine,
Clinical medicine, other,
Clinical chemistry

© 2025 Maria Siemionow, Weronika Radecka, Katarzyna Kozlowska, Lucile Chambily, Sonia Brodowska, Dominika Kuc, Gabrielle Filipek, Katarzyna Budzynska, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Previous articleVolume 73 (2025): Issue 1 (January 2025)Next article