Autologous Nerve Grafts Research Articles

Nerve regeneration using the Bio 3D nerve conduit fabricated with spheroids.

Autologous nerve grafting is the gold standard method for peripheral nerve injury with defects. Artificial nerve conduits have been developed to prevent morbidity at the harvest site. However, the artificial conduit regeneration capacity is not sufficient. A Bio 3D printer is technology that creates three-dimensional tissue using only cells. Using this technology, a three-dimensional nerve conduit (Bio 3D nerve conduit) was created from several cell spheroids. We reported the first application of the Bio 3D nerve conduit for peripheral nerve injury. A Bio 3D nerve conduit that was created from several cells promotes peripheral nerve regeneration. The Bio 3D nerve conduit may be useful clinically to treat peripheral nerve defects.

Efficacy of Nerve-Derived Hydrogels to Promote Axon Regeneration Is Influenced by the Method of Tissue Decellularization.

Injuries to large peripheral nerves are often associated with tissue defects and require reconstruction using autologous nerve grafts, which have limited availability and result in donor site morbidity. Peripheral nerve-derived hydrogels could potentially supplement or even replace these grafts. In this study, three decellularization protocols based on the ionic detergents sodium dodecyl sulfate (P1) and sodium deoxycholate (P2), or the organic solvent tri-n-butyl phosphate (P3), were used to prepare hydrogels. All protocols resulted in significantly decreased amounts of genomic DNA, but the P2 hydrogel showed the best preservation of extracellular matrix proteins, cytokines, and chemokines, and reduced levels of sulfated glycosaminoglycans. In vitro P1 and P2 hydrogels supported Schwann cell viability, secretion of VEGF, and neurite outgrowth. Surgical repair of a 10 mm-long rat sciatic nerve gap was performed by implantation of tubular polycaprolactone conduits filled with hydrogels followed by analyses using diffusion tensor imaging and immunostaining for neuronal and glial markers. The results demonstrated that the P2 hydrogel considerably increased the number of axons and the distance of regeneration into the distal nerve stump. In summary, the method used to decellularize nerve tissue affects the efficacy of the resulting hydrogels to support regeneration after nerve injury.

ESWT Diminishes Axonal Regeneration following Repair of the Rat Median Nerve with Muscle-In-Vein Conduits but Not after Autologous Nerve Grafting.

Investigations reporting positive effects of extracorporeal shockwave therapy (ESWT) on nerve regeneration are limited to the rat sciatic nerve model. The effects of ESWT on muscle-in-vein conduits (MVCs) have also not been investigated yet. This study aimed to evaluate the effects of ESWT after repair of the rat median nerve with either autografts (ANGs) or MVCs. In male Lewis rats, a 7 mm segment of the right median nerve was reconstructed either with an ANG or an MVC. For each reconstructive technique, one group of animals received one application of ESWT while the other rats served as controls. The animals were observed for 12 weeks, and nerve regeneration was assessed using computerized gait analysis, the grasping test, electrophysiological evaluations and histological quantification of axons, blood vessels and lymphatic vasculature. Here, we provide for the first time a comprehensive analysis of ESWT effects on nerve regeneration in a rat model of median nerve injury. Furthermore, this study is among the first reporting the quantification of lymphatic vessels following peripheral nerve injury and reconstruction in vivo. While we found no significant direct positive effects of ESWT on peripheral nerve regeneration, results following nerve repair with MVCs were significantly inferior to those after ANG repair.

Muscle Grafts with Doxorubicin Pretreatment Produce "Empty Tubes" in the Basal Laminae, Promote Contentious Maturation of the Regenerated Axons, and Bridge 20-mm Sciatic Nerve Defects in Rats.

We newly developed a muscle graft that employs a doxorubicin pretreatment technique. The aims of this study were to reveal the biological and morphological features of the muscle tissue in the second week (Study I), to reveal the regeneration outcomes of functional and kinematic assessments of longer-term follow-up (16 weeks, Study II), and to make assessments of the muscle graft with doxorubicin pretreatment in the critical-sized nerve defect model (20 mm, Study III). A total of 26 adult rats were used in this study. Doxorubicin treatment was accomplished by immersion in a doxorubicin solution for 10 minutes followed by a rinsing procedure. The rats were divided into three groups: the muscle graft with and without doxorubicin pretreatment (M-graft-w-Dox and M-graft-w/o-Dox) groups and the autologous nerve graft (N-graft) group. Assays of apoptosis, immunofluorescent histochemistry including CD68 (macrophage marker), scanning electron microscopy (SEM), morphometrical studies of the regenerated axons, nerve conduction studies, and kinematic studies were performed. The M-graft-w-Dox group contained significantly larger numbers of apoptotic cells and CD68-positive cells. SEM revealed the existence of the basal lamina, so called "empty tubes," in the M-graft-w-Dox group. Study II showed contentious maturation of the regenerated axons, especially in the compound muscle action potentials. Study III showed that even at 20 mm, the M-graft-w-Dox group promoted axonal regeneration and functional regeneration. The M-graft-w-Dox group showed superior regeneration results, and this easy and short-term procedure can expand the muscle graft clinical indication for the treatment of peripheral nerve defects.

Physical Stimulation Combined with Biomaterials Promotes Peripheral Nerve Injury Repair.

Peripheral nerve injury (PNI) is a clinical problem with high morbidity that can cause severe damage. Surgical suturing or implants are usually required due to the slow speed and numerous factors affecting repair after PNI. An autologous nerve graft is the gold standard for PNI repair among implants. However, there is a potential problem of the functional loss of the donor site. Therefore, tissue-engineered nerve biomaterials are often used to bridge the gap between nerve defects, but the therapeutic effect is insufficient. In order to enhance the repair effect of nerve biomaterials for PNI, researchers are seeking to combine various stimulation elements, such as the addition of biological factors such as nerve growth factors or physical factors such as internal microstructural modifications of catheters and their combined application with physical stimulation therapy. Physical stimulation therapy is safer, is more convenient, and has more practical features than other additive factors. Its feasibility and convenience, when combined with nerve biomaterials, provide broader application prospects for PNI repair, and has therefore become a research hot spot. This paper will review the combined application of physical stimulation and biomaterials in PNI repair in recent years to provide new therapeutic ideas for the future use of physical stimulation in PNI repair.

Spinal accessory to suprascapular nerve transfer in brachial plexus injury: outcomes of anterior vs. posterior approach to the suprascapular nerve at associated ipsilateral spinal accessory nerve injury

Objective: The spinal accessory nerve (Acc) is susceptible to trauma in at least 6% of cases of brachial plexus injury (BPI). The impaired Acc function disables its utilization for transfer to the suprascapular nerve (SS). The selection of approach to SS is highly dependant on the anatomy of BPI. The purpose of this study was to determine the incidence of the anterior-posterior approach of Acc to SS transfer in BPI and associated functional outcomes. Methods. Twenty nine patients with BP/Acc associated injury were included. Ten patients underwent the transfer of Acc to SS by the anterior approach (AA), 19 patients – by the posterior approach (PA). Nine nerve transfers through AA and one nerve transfer through PA required the interposition of an autologous nerve graft. The functioning of the supra-/infraspinatus muscle was evaluated at 9 and 15mos. on the basis of the MRC and the external rotation (ER) range. ER more than +400 beyond the sagittal plane was regarded as effective recovery of function. Results. Impaired function (M3 or lower on MRC) of the lower trapezius muscle was associated with preserved anatomy of the SS in the supraclavicular region in 9 out of 10 cases. Eighteen patients (62%) recovered to M3 and higher (shoulder stability), 11of these (38%) showed recovery to M4-M5. Five of all patients recovered to M4-M5 and were able to produce ER within the effective ROM (+400-600 of ER). After the AA to the SS, shoulder stability was restored in 60% of cases (M4-M5 in 30%). After the PA to the SS, shoulder stability was restored in 74% of cases (M4-M5 in 42%). Only non-complete BPI showed effective recovery of power and function in terms of less than 6 mos. after injury. PA to SS with no graft provided shoulder stability in 72% of cases, AA to the SS and the graft interposition ensured shoulder stability in 50% of cases. Conclusions. The incidence of AA to the SS was 35%, PA – 65%; preserved anatomy of the SS in supraclavicular region was associated with an increased risk of trapezius muscle dysfunction; the PA to SS and consecutive direct end-to-end transfer of Acc showed better results compared to other combinations of nerve transfers in providing shoulder stability.

BIOPOLYMERIC NANOCOMPOSITE SCAFFOLDS FOR NERVE TISSUE ENGINEERING APPLICATIONS: A REVIEW

Human biological processes are heavily influenced by nervous system and it interacts in physiological processes, such as cognition and individual cell function therefore; injury of the peripheral or central nervous system (PNS or CNS) causes loss of sensory and motor functions, affecting the patients’ quality of life. The regeneration capability of the human adult nervous system is often limited, and its recovery is difficult due to the complex physiology system and limited regenerative capacity. Tissue engineering is a novel clinical treatment field utilizing a blend of polymeric scaffolds and cells, representing a promising methodology for nerve recovery. The engineered materials and the fabricated neural scaffolds have gained expanding consideration in the field of nerve repairs. Nerve scaffolds surpass autologous nerve grafts in terms of implantation rate and outcome. Additionally, they foster a conducive cellular environment enabling axonal proliferation, neurite extension and connection, and neural cell survivability, adhesion, and migration. Due to their exceptional chemical and physical capabilities, numerous natural and manmade polymeric materials have been preferentially used during neural scaffolds to date. This review article presents various natural and synthetic polymers being used in neural tissue engineering, along with their benefits and challenges for neural recovery.

Commentary: Corneal neurotization

Commentary: Corneal neurotization

Closing the Gap: Bridging Peripheral Sensory Nerve Defects with a Chitosan-Based Conduit a Randomized Prospective Clinical Trial.

Introduction: If tensionless nerve coaptation is not possible, bridging the resulting peripheral nerve defect with an autologous nerve graft is still the current gold standard. The concept of conduits as an alternative with different materials and architectures, such as autologous vein conduits or bioartificial nerve conduits, could not replace the nerve graft until today. Chitosan, as a relatively new biomaterial, has recently demonstrated exceptional biocompatibility and material stability with neural lineage cells. The purpose of this prospective randomized clinical experiment was to determine the efficacy of chitosan-based nerve conduits in regenerating sensory nerves in the hand. Materials and methods: Forty-seven patients with peripheral nerve defects up to 26 mm distal to the carpal tunnel were randomized to receive either a chitosan conduit or an autologous nerve graft with the latter serving as the control group. Fifteen patients from the conduit group and seven patients from the control group were available for a 12-month follow-up examination. The primary outcome parameter was tactile gnosis measured with two-point discrimination. The secondary outcome parameters were Semmens Weinstein Monofilament Testing, self-assessed pain, and patient satisfaction. Results: Significant improvement (in static two-point discrimination) was observed six months after trauma (10.7 ± 1.2 mm; p < 0.05) for chitosan-based nerve conduits, but no further improvement was observed after 12 months of regeneration (10.9 ± 1.3 mm). After six months and twelve months, the autologous nerve graft demonstrated comparable results to the nerve conduit, with a static two-point discrimination of 11.0 ± 2.0 mm and 7.9 ± 1.1 mm. Semmes Weinstein Filament Testing in the nerve conduit group showed a continuous improvement over the regeneration period by reaching from 3.1 ± 0.3 after three months up to 3.7 ± 0.4 after twelve months. Autologous nerve grafts presented similar results: 3.3 ± 0.4 after three months and 3.7 ± 0.5 after twelve months. Patient satisfaction and self-reported pain levels were similar between the chitosan nerve conduit and nerve graft groups. One patient required revision surgery due to complications associated with the chitosan nerve tube. Conclusion: Chitosan-based nerve conduits are safe and suitable for bridging nerve lesions up to 26 mm in the hand. Tactile gnosis improved significantly during the early regeneration period, and functional outcomes were similar to those obtained with an autologous nerve graft. Thus, chitosan appears to be a sufficient substitute for autologous nerve grafts in the treatment of small nerve defects in the hand.

A printability study of multichannel nerve guidance conduits using projection-based three-dimensional printing.

Multichannel nerve guidance conduits (NGCs) replicating the native architecture of peripheral nerves have emerged as promising alternatives to autologous nerve grafts. However, manufacturing multichannel NGCs is challenging in terms of desired structural stability and resolution. In this study, we systematically investigated the effects of photopolymer properties, inner diameter dimensions, printing parameters, and different conditions on multichannel NGCs printability using projection-based three-dimensional printing. Low viscosity and rapid photocuring properties were essential requirements. A standard model was generated to evaluate multichannel NGC printed quality. The results showed that printing deviations decreased with increased mechanical strength and inner diameter. Subsequently, gelatin methacrylate (GelMA) NGCs was selected as a representative. It was found that printing conditions, including printing temperature, peeling, and shrinkage affected final NGC accuracy and quality. PC-12 cells cultured with the GelMA NGCs displayed non-toxic and promoted cell migration. Our research provides an effective, time-saving, and high-resolution technology for manufacturing multichannel NGCs with high fidelity, which may be used as reference templates for biomedical applications.

Evaluation of Obturator Nerve Fascicular Topography Using Landmark Morphometrics ‐ a Pilot Study

IntroductionWhile the overall macroanatomy of peripheral nerves in the lower extremities are well researched, the intricacy of fascicular organization within larger polyfascicular nerves render them difficult to characterize. This obstacle is particularly salient considering the serviceability of these complex nerves in gross motor functions. Peripheral nerve injuries (PNI) occur in approximately 1.64% of people who have had trauma involving the upper or lower limb. Rates of effective distal reanimation using autologous nerve grafts remain low. The objective of this pilot research is to demonstrate the utility of landmark morphometric techniques in gaining a more comprehensive understanding of fascicular topography, leading to improved precision in clinical interventions.Material and methodsRight and left obturator nerves from the 25 formalin‐fixed whole‐body donors were isolated using typical dissection techniques. Sections of the nerves were collected from their immediate origin from the lumbar plexus to their exit from the obturator canal. Nerves were sectioned into nine equally divided segments and imaged (OptixCam Summit K2). Images of the nerve sections were evaluated for fascicle number and diameter (ImageJ). Nerves were then categorized by fascicular homology. Samples with matching fascicle numbers and arrangements were used for further analysis. Nine individuals were selected for landmark morphometric analyses and 130 landmarks were collected via semi‐landmark techniques (tpsdig232 v. 2w32). Generalized Procrustes analysis was performed to determine the mean landmark configuration, and a principal component analysis was used to explore variability in fascicular topography MorphoJ (v.1.07a). Wireframe graphs, lollipop diagrams, and transformation grids were generated to evaluate and visualize findings.ResultsThe average obturator nerve diameter of the original 25 donors was 2.29 cm (right) and 2.28 cm (left). Nerve length was 10.86 cm (right) and 10.83 cm (left). Principal component analysis revealed eight principal components, with the first two principal components explaining 64.6% of the overall shape variation. Both PC1 and PC2 show peripheral fascicles remaining more consistently located, while the direction and magnitude of variability demonstrated by the lollipop diagrams suggest much more variation in the relative locations of more interior fascicles.Conclusions and significanceThis pilot study successfully utilized a novel approach of examining the variability of peripheral nerve fascicles using landmark morphometric techniques. Peripheral fascicles of the intra‐abdominal obturator nerve were relatively consistent, while more interior fascicular bundles showed marked variability in their locations. Consequently, the relative location of non‐fascicular tissue was most variable in the central regions of the nerve. The application of these techniques in larger samples may inform grafting approaches which aim to provide adequate alignment and guidance of extraneural tissues supporting effective axon growth.

Alginate-Based Hydrogels and Tubes, as Biological Macromolecule-Based Platforms for Peripheral Nerve Tissue Engineering: A Review.

Unlike the central nervous system, the peripheral nervous system (PNS) has an inherent capacity to regenerate following injury. However, in the case of large nerve defects where end-to-end cooptation of two nerve stumps is not tension-free, autologous nerve grafting is often utilized to bridge the nerve gaps. To address the challenges associated with autologous nerve grafting, neural guidance channels (NGCs) have been successfully translated into clinic. Furthermore, hydrogel-based drug delivery systems have been extensively studied for the repair of PNS injuries. There are numerous biomaterial options for the production of NGCs and hydrogels. Among different candidates, alginate has shown promising results in PNS tissue engineering. Alginate is a naturally occurring polysaccharide which is biocompatible, non-toxic, non-immunogenic, and possesses modifiable properties. In the current review, applications, challenges, and future perspectives of alginate-based NGCs and hydrogels in the repair of PNS injuries will be discussed.

Brachial plexus bridging with specific extracellular matrix-modified chitosan/silk scaffold: a new expand of tissue engineered nerve graft

Objective. Brachial plexus injuries (BPIs) result in serious dysfunction, especially brachial plexus defects which are currently treated using autologous nerve graft (autograft) transplantation. With the development of tissue engineering, tissue engineered nerve grafts (TENGs) have emerged as promising alternatives to autografts but have not yet been widely applied to the treatment of BPIs. Herein, we developed a TENG modified with extracellular matrix generated by skin-derived precursor Schwann cells (SKP-SCs) and expand its application in upper brachial plexus defects in rats. Approach. SKP-SCs were co-cultured with chitosan neural conduits or silk fibres and subjected to decellularization treatment. Ten bundles of silk fibres (five fibres per bundle) were placed into a conduit to obtain the TENG, which was used to bridge an 8 mm gap in the upper brachial plexus. The efficacy of this treatment was examined for TENG-, autograft- and scaffold-treated groups at several times after surgery using immunochemical staining, behavioural tests, electrophysiological measurements, and electron microscopy. Main results. Histological analysis conducted two weeks after surgery showed that compared to scaffold bridging, TENG treatment enhanced the growth of regenerating axons. Behavioural tests conducted four weeks after surgery showed that TENG-treated rats performed similarly to autograft-treated ones, with a significant improvement observed in both cases compared with the scaffold treatment group. Electrophysiological and retrograde tracing characterizations revealed that the target muscles were reinnervated in both TENG and autograft groups, while transmission electron microscopy and immunohistochemical staining showed the occurrence of the superior myelination of regenerated axons in these groups. Significance. Treatment with the developed TENG allows the effective bridging of proximal nerve defects in the upper extremities, and the obtained results provide a theoretical basis for clinical transformation to expand the application scope of TENGs.

Three-Dimensional Microfilament Printing of a Decellularized Extracellular Matrix (dECM) Bioink Using a Microgel Printing Bath for Nerve Graft Fabrication and the Effectiveness of dECM Graft Combined with a Polycaprolactone Conduit.

Various synthetic and decellularized materials are being used to reconstruct peripheral nerve defects and replace autologous nerve grafts. In this study, we developed a microgel printing bath to three-dimensionally (3D) print a peripheral nervous system decellularized extracellular matrix nerve graft reinforced by a polycaprolactone (PCL) conduit. The straightforward fabrication method of an alginate microgel-supplemented printing bath allows a 30 μm filament resolution of a low viscous decellularized extracellular matrix hydrogel with neutral pH. When applied to a sciatic nerve defect model of rats, the total number of regenerated axons and relative gastrocnemius muscle weight ratio were comparable to those of the autologous nerve graft group. Meanwhile, the results were superior to those of the porcine decellularized nerve graft group or the 3D printed decellularized extracellular matrix graft group. This study will be the first step demonstrating that the 3D printed decellularized extracellular matrix (dECM) graft with a PCL conduit is an effective and reliable choice to replace an autologous nerve graft in the near future.

Evaluation of anatomical and histological characteristics of human peripheral nerves: as an effort to develop an efficient allogeneic nerve graft.

Management of peripheral nerve defects is a complicated problem in clinical contexts. Autologous nerve grafting, a gold standard for surgical treatment, has been well known to have several limitations, such as donor site morbidity, a limited amount of available donor tissue, and size mismatches. Acellular nerve allografts (ANAs) have been developed as an alternative and have been applied clinically with favorable outcomes. However, because of the limited availability of commercialized ANAs due to supplier-related issues and high costs, efforts continue to produce alternative sources for ANAs. The present study evaluated the anatomical and histological characteristics of human peripheral nerves using 25 donated human cadavers. The length, diameter, and branching points of various peripheral nerves (median, ulnar, tibial, lateral femoral cutaneous, saphenous, and sural nerves) in both the upper and lower extremities were evaluated. The cross-sectional area (CSA), ratio of fascicular area, and numbers of fascicles were also evaluated via histologic analysis. CSA, the ratio of fascicular area, and the number of fascicles were analyzed statistically in correlation with demographic data (age, sex, height, weight, BMI). The mean length of all evaluated nerves ranged from 17.1 to 41.4cm, and the mean diameter of all evaluated nerves ranged from 1.2 to 4.9mm. Multiple regression analysis revealed correlations between the ratio of fascicular area and sex (p = 0.005) and BMI (p = 0.024) (R2 = 0.051). The results of the present study will be helpful in selecting necessary nerve allograft sources while considering the characteristics of each nerve in the upper and lower extremities during ANAs production.

Advances and future directions in the care of patients with facial paralysis

Advances and future directions in the care of patients with facial paralysis

Sustainable release of nerve growth factor for peripheral nerve regeneration using nerve conduits laden with Bioconjugated hyaluronic acid-chitosan hydrogel

Peripheral nerve regeneration following injury is often incomplete, resulting in significant personal and socioeconomic costs. Nerve conduits have been developed to complement or replace autologous nerve grafts. In this study, repairing injured peripheral nerves in rats is investigated by fabricating poly(D, l-lactic acid) (PDLLA)/β-tricalcium phosphate (β-TCP) nerve conduits filled by injectable chitosan (CS)-hyaluronic acid (HA) hydrogels featuring sustained release of nerve growth factor (NGF). All CS-HA hydrogels have porosities higher than 80%. The mass loss of the CS-HA hydrogels reaches 70% in 8 weeks. In vitro experiments suggest that the CS-HA/NGF hydrogels are suitable for the adhesion, spreading, and differentiation of neuronal cells, which also allow for sustained release of NGF. In vivo studies on the repair of 10-mm sciatic nerve defects in rats are used to assess nerve repair rate through walking track analysis, gastrocnemius weight analysis, and histological analysis. It is shown that the PDLLA/β-TCP nerve conduits laden with CS-HA/NGF hydrogels have a significant positive effect on the axon regeneration and myelination, compared to the PDLLA/β-TCP hollow nerve conduits and the autograft group. These results show that the CS-HA/NGF injectable hydrogel can effectively promote nerve regeneration making it a good candidate in the field of neural tissue engineering.

Tissue Engineering Scaffold Slowly Releasing Neurotrophic Factors to Bridge Long Peripheral Nerve Defect

Consistent application of neurotropic factors is necessary in peripheral nerve regeneration, yet challenging to achieve. Here we used a novel neurotropic factor controlled release system consisted of fibrin, fibronectin and hydrogel to slowly release two neurotrophic factors. At the same time, physiological saline and reverse nerve suturing were used as negative and positive control. A year after surgery, animals which were treated by neurotrophic factor slow release system achieved far better neural regeneration and myelination, as well as superior recovery of hindfoot than the negative control group. In the meanwhile, the results in the experimental group are still inferior to the nerve allograft group. In can be concluded from those results that, consistent releasing of neurotrophic factors can significantly promote long peripheral nerve regeneration, but still short of achieving the results same as the gold standard of autologous nerve grafting.

Application of Hybrid Electrically Conductive Hydrogels Promotes Peripheral Nerve Regeneration.

Peripheral nerve injury (PNI) occurs frequently, and the prognosis is unsatisfactory. As the gold standard of treatment, autologous nerve grafting has several disadvantages, such as lack of donors and complications. The use of functional biomaterials to simulate the natural microenvironment of the nervous system and the combination of different biomaterials are considered to be encouraging alternative methods for effective tissue regeneration and functional restoration of injured nerves. Considering the inherent presence of an electric field in the nervous system, electrically conductive biomaterials have been used to promote nerve regeneration. Due to their singular physical properties, hydrogels can provide a three-dimensional hydrated network that can be integrated into diverse sizes and shapes and stimulate the natural functions of nerve tissue. Therefore, conductive hydrogels have become the most effective biological material to simulate human nervous tissue’s biological and electrical characteristics. The principal merits of conductive hydrogels include their physical properties and their electrical peculiarities sufficient to effectively transmit electrical signals to cells. This review summarizes the recent applications of conductive hydrogels to enhance peripheral nerve regeneration.

Loop nerve graft prefabrication for peripheral nerve defect reconstruction.

Delayed autologous nerve graft reconstruction is inevitable in devastating injuries. Delayed or prolonged repair time has deleterious effects on nerve grafts. We aimed improving and accelerating nerve graft reconstruction process in a rat long nerve defect model with loop nerve graft prefabrication particularly to utilize for injuries with tissue loss. Twenty-four Sprague-Dawley rats were allocated into three groups. 1.5 cm long peroneal nerve segment was excised, reversed in orientation, and used as autologous nerve graft. In conventional interpositional nerve graft group (Group 1), nerve defects were repaired in single-stage. In loop nerve graft prefabrication group (Group 2), grafts were sutured end-to-end (ETE) to the proximal peroneal nerve stumps. Distal ends of the grafts were sutured end-to-side to the peroneal nerve stumps 5 mm proximal to the ETE repair sites in first stage. In second stage, distal ends of the prefabricated grafts were transposed and sutured to distal nerve stumps. In staged conventional interpositional nerve graft group (Group 3), grafts were sutured ETE to proximal peroneal nerve stumps in first stage. Distal ends of the grafts and nerve stumps were tacked to the surrounding muscles until the final repair in second stage. Follow-up period was 4 weeks for each stage in Groups 2 and 3, and 8 weeks for Group 1. Peroneal function index (PFI), electrophysiology, and histological assessments were conducted after 8 weeks. P<0.05 was considered significant for statistical analysis. PFI results of Group 1 (-22.75±5.76) and 2 (-22.08±6) did not show statistical difference (p>0.05). Group 3 (-33.64±6.4) had a statistical difference compared to other groups (p<0.05). Electrophysiology results of Group 1 (16.19±2.15 mV/1.16±0.21 ms) and 2 (15.95±2.82 mV/1.17±0.16 ms) did not present statistical difference (p>0.05), whereas both groups had a statistical difference compared to Group 3 (10.44±1.96 mV/1.51±0.15 ms) (p<0.05). Axon counts of Group 1 (2227±260.4) and 3 (2194±201.1) did not have statistical difference (p>0.05), whereas both groups had significantly poor axon counts compared to Group 2 (2531±91.18) (p<0.05). Loop nerve graft prefabrication improved axonal regeneration without delay. Loop prefabrication can accelerate prolonged regeneration time for the injuries indicating a delayed nerve reconstruction. Higher axon counts derived with loop nerve prefabrication may even foster its investigation in immediate long nerve defect reconstructions in further studies.