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

Maria Siemionow

Maria Siemionow

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Siemionow, Maria
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Weronika Radecka

Weronika Radecka

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Radecka, Weronika
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Katarzyna Kozlowska

Katarzyna Kozlowska

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Kozlowska, Katarzyna
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Lucile Chambily

Lucile Chambily

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Chambily, Lucile
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Sonia Brodowska

Sonia Brodowska

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Brodowska, Sonia
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Dominika Kuc

Dominika Kuc

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Kuc, Dominika
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Gabrielle Filipek

Gabrielle Filipek

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Filipek, Gabrielle
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Katarzyna Budzynska

Katarzyna Budzynska

Department of Orthopaedics, University of Illinois at ChicagoChicago, USA
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Budzynska, Katarzyna
  
26 mar 2025
Protective Effect of the Human Epineural Patch Application after Sciatic Nerve Crush Injury Followed by Nerve Transection and End-to-End Repair's Cover Image
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Categoria dell'articolo: Original
Pubblicato online: 26 mar 2025
Ricevuto: 11 gen 2025
Accettato: 25 feb 2025
DOI: https://doi.org/10.2478/aite-2025-0008
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© 2025 Maria Siemionow et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

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 number Repair method Number of athymic nude rats per group
6 weeks study
1 no protective wrapping n=6
2 hEP n=6
3 hAM n=6
12 weeks study
4 no protective wrapping n=6
5 hEP n=6
6 hAM n=6
Total number of athymic nude rats (n=36)
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Medicina, Scienze medicali di base, Biochimica, Immunologia, Medicina clinica, Medicina clinica, altro, Chimica clinica
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