Tissue-engineered rhesus monkey nerve grafts for the repair of long ulnar nerve defects:similar outcomes to autologous nerve grafts

Acellular nerve allografts can help preserve normal nerve structure and extracellular matrix composition. These allografts have low immunogenicity and are more readily available than autologous nerves for the repair of long-segment peripheral nerve defects. In this study, we repaired a 40-mm ulnar nerve defect in rhesus monkeys with tissue-engineered peripheral nerve, and compared the outcome with that of autograft. The graft was prepared using a chemical extract from adult rhesus monkeys and seeded with allogeneic Schwann cells. Pathomo- rphology, electromyogram and immunohistochemistry findings revealed the absence of palmar erosion or ulcers, and that the morphology and elasticity of the hypothenar eminence were normal 5 months postoperatively. There were no significant differences in the mean peak compound muscle action potential, the mean nerve conduction velocity, or the number of neurofilaments between the experimental and control groups. However, outcome was significantly better in the experimental group than in the blank group. These findings suggest that chemically extracted allogeneic nerve seeded with autologous Schwann cells can repair 40-mm ulnar nerve defects in the rhesus monkey. The outcomes are similar to those obtained with autologous nerve graft.

Chitosan tube bridging autologous nerve segments for the repair of long-segmental sciatic nerve defects

BACKGROUND：Previous tissue-engineered nerve studies have focused on artificial nerve and nerve cell cultures.The effects of regeneration chambers with autologous nerve bridging for the repair of nerve defects remain unclear.OBJECTIVE：To explore the feasibility and advantages of chitosan tube bridging autologous nerve segments for repairing 12-mm sciatic nerve defects in rats.DESIGN,TIME AND SETTING：A randomized,controlled,animal study using nerve tissue engineering was performed at the Animal Laboratory and Laboratory of Histology and Embryology,Liaoning Medical University from June 2008 to March 2009.MATERIALS：Chitosan powder was purchased from Jinan Haidebei Marine Bioengineering,China.METHODS：A sciatic nerve segment of approximately 8 mm was excised from the posterior margin of the piriformis muscle of Sprague Dawley rats.The two nerve ends shrank to form a 12-mm defect,and the nerve defect was repaired using a chitosan tube bridging autologous nerve segment （bridge group）,a chitosan tube-encapsulated autologous nerve segment （encapsulation group）,and a chitosan tube alone （chitosan tube alone group）,respectively.MAIN OUTCOME MEASURES：Histological and ultrastructural changes of the injured sciatic nerve;number of regenerated myelinated nerve fibers; nerve conduction velocity; leg muscle atrophy; and sciatic nerve functional index.RESULTS：At 4 months after implantation,the chitosan tube was absorbed.The tube was thin,but maintained the original shape,and vascular proliferation was observed around the tube.In the bridge group,regenerative myelinated nerve fibers were thick and orderly,with a thick myelin sheath and intact axonal structure.The number of myelinated nerve fibers and nerve conduction velocity were significantly greater compared with the other groups （P〈 0.01）.Moreover,nerve and muscle function was significantly improved following chitosan tube bridging autologous nerve segment treatment compared with the other groups （P〈 0.05 or P 〈 0.01）.CONCLUSION：Chitosan tube bridging autologous nerve segments exhibited better repair effects on nerve defects compared with chitosan tubeencapsulated autologous nerve segments and a chitosan tube alone.This method provided a simple and effective treatment for long-segmental nerve defects.

Autologous nerve anastomosis versus human amniotic membrane anastomosisA rheological comparison following simulated sciatic nerve injury

The sciatic nerve is biological viscoelastic solid,with stress relaxation and creep characteristics.In this study,a comparative analysis of the stress relaxation and creep characteristics of the sciatic nerve was conducted after simulating sciatic nerve injury and anastomosing with autologous nerve or human amniotic membrane.The results demonstrate that,at the 7 200-second time point,both stress reduction and strain increase in the human amniotic membrane anastomosis group were significantly greater than in the autologous nerve anastomosis group.Our findings indicate that human amniotic membrane anastomosis for sciatic nerve injury has excellent rheological characteristics and is conducive to regeneration of the injured nerve.

Autologous nerve graft repair of different degrees of sciatic nerve defect:stress and displacement at the anastomosis in a three-dimensional finite element simulation model

In the repair of peripheral nerve injury using autologous or synthetic nerve grafting, the mag- nitude of tensile forces at the anastomosis affects its response to physiological stress and the ultimate success of the treatment. One-dimensional stretching is commonly used to measure changes in tensile stress and strain; however, the accuracy of this simple method is limited. There- fore, in the present study, we established three-dimensional finite element models of sciatic nerve defects repaired by autologous nerve grafts. Using PRO E 5.0 finite element simulation software, we calculated the maximum stress and displacement of an anastomosis under a 5 N load in 10-, 20-, 30-, 40-mm long autologous nerve grafts. We found that maximum displacement increased with graft length, consistent with specimen force. These findings indicate that three-dimensional finite element simulation is a feasible method for analyzing stress and displacement at the anas- tomosis after autologous nerve grafting.

Autologous transplantation with fewer fibers repairs large peripheral nerve defects

Peripheral nerve injury is a serious disease and its repair is challenging. A cable-style autologous graft is the gold standard for repairing long peripheral nerve defects; however, ensuring that the minimum number of transplanted nerve attains maximum therapeutic effect remains poorly understood. In this study, a rat model of common peroneal nerve defect was established by resecting a 10-mm long right common peroneal nerve. Rats receiving transplantation of the common peroneal nerve in situ were designated as the in situ graft group. Ipsilateral sural nerves（10–30 mm long） were resected to establish the one sural nerve graft group, two sural nerves cable-style nerve graft group and three sural nerves cable-style nerve graft group. Each bundle of the peroneal nerve was 10 mm long. To reduce the barrier effect due to invasion by surrounding tissue and connective-tissue overgrowth between neural stumps, small gap sleeve suture was used in both proximal and distal terminals to allow repair of the injured common peroneal nerve. At three months postoperatively, recovery of nerve function and morphology was observed using osmium tetroxide staining and functional detection. The results showed that the number of regenerated nerve fibers, common peroneal nerve function index, motor nerve conduction velocity, recovery of myodynamia, and wet weight ratios of tibialis anterior muscle were not significantly different among the one sural nerve graft group, two sural nerves cable-style nerve graft group, and three sural nerves cable-style nerve graft group. These data suggest that the repair effect achieved using one sural nerve graft with a lower number of nerve fibers is the same as that achieved using the two sural nerves cable-style nerve graft and three sural nerves cable-style nerve graft. This indicates that according to the ‘multiple amplification＇ phenomenon, one small nerve graft can provide a good therapeutic effect for a large peripheral nerve defect.

Strain and stress variations in the human amniotic membrane and fresh corpse autologous sciatic nerve anastomosis in a model of sciatic nerve injury

A 10-mm long sciatic nerve injury model was established in fresh normal Chinese patient cadavers. Amniotic membrane was harvested from healthy maternal placentas and was prepared into multilayered,coiled,tubular specimens.Sciatic nerve injury models were respectively anastomosed using the autologous cadaveric sciatic nerve and human amniotic membrane.Tensile test results showed that maximal loading,maximal displacement,maximal stress,and maximal strain of sciatic nerve injury models anastomosed with human amniotic membrane were greater than those in the autologous nerve anastomosis group.The strain-stress curves of the human amniotic membrane and sciatic nerves indicated exponential change at the first phase,which became elastic deformation curves at the second and third phases,and displayed plastic deformation curves at the fourth phase,at which point the specimens lost their bearing capacity.Experimental findings suggested that human amniotic membranes and autologous sciatic nerves exhibit similar stress-strain curves, good elastic properties,and certain strain and stress capabilities in anastomosis of the injured sciatic nerve.

Hypoxic pre-conditioned adipose-derived stem/progenitor cells embedded in fibrin conduits promote peripheral nerve regeneration in a sciatic nerve graft model

Recent results emphasize the supportive effects of adipose-derived multipotent stem/progenitor cells(ADSPCs)in peripheral nerve recovery.Cultivation under hypoxia is considered to enhance the release of the regenerative potential of ADSPCs.This study aimed to examine whether peripheral nerve regeneration in a rat model of autologous sciatic nerve graft benefits from an additional custom-made fibrin conduit seeded with hypoxic pre-conditioned(2%oxygen for 72 hours)autologous ADSPCs(n=9).This treatment mode was compared with three others:fibrin conduit seeded with ADSPCs cultivated under normoxic conditions(n=9);non-cell-carrying conduit(n=9);and nerve autograft only(n=9).A 16-week follow-up included functional testing(sciatic functional index and static sciatic index)as well as postmortem muscle mass analyses and morphometric nerve evaluations(histology,g-ratio,axon density,and diameter).At 8 weeks,the hypoxic pre-conditioned group achieved significantly higher sciatic functional index/static sciatic index scores than the other three groups,indicating faster functional regeneration.Furthermore,histologic evaluation showed significantly increased axon outgrowth/branching,axon density,remyelination,and a reduced relative connective tissue area.Hypoxic pre-conditioned ADSPCs seeded in fibrin conduits are a promising adjunct to current nerve autografts.Further studies are needed to understand the underlying cellular mechanism and to investigate a potential application in clinical practice.

Chitin scaffold combined with autologous small nerve repairs sciatic nerve defects

Although autologous nerve transplantation is the gold standard for treating peripheral nerve defects,it has many clinical limitations.As an alternative,various tissue-engineered nerve grafts have been developed to substitute for autologous nerves.In this study,a novel nerve graft composed of chitin scaffolds and a small autologous nerve was used to repair sciatic nerve defects in rats.The novel nerve graft greatly facilitated regeneration of the sciatic nerve and myelin sheath,reduced atrophy of the target muscle,and effectively restored neurological function.When the epineurium of the small autogenous nerve was removed,the degree of nerve regeneration was similar to that which occurs after autogenous nerve transplantation.These findings suggest that our novel nerve graft might eventually be a new option for the construction of tissue-engineered nerve scaffolds.The study was approved by the Research Ethics Committee of Peking University People's Hospital(approval No.2019 PHE27)on October 18,2019.

Chemically extracted acellular allogeneic nerve graft combined with ciliary neurotrophic factor promotes sciatic nerve repair

A chemically extracted acellular allogeneic nerve graft can reduce postoperative immune rejection, similar to an autologous nerve graft, and can guide neural regeneration. However, it remains poorly understood whether a chemically extracted acellular allogeneic nerve graft combined with neurotrophic factors provides a good local environment for neural regeneration. This study investigated the repair of injured rat sciatic nerve using a chemically extracted acellular allogeneic nerve graft combined with ciliary neurotrophic factor. An autologous nerve anastomosis group and a chemical acellular allogeneic nerve bridging group were prepared as controls. At 8 weeks after repair, sciatic functional index, evoked potential amplitude of the soleus muscle, triceps wet weight recovery rate, total number of myelinated nerve fibers and myelin sheath thickness were measured. For these indices, values in the three groups showed the autologous nerve anastomosis group 〉 chemically extracted acellular nerve graft ＋ ciliary neurotrophic factor group 〉 chemical acellular allogeneic nerve bridging group. These results suggest that chemically extracted acellular nerve grafts combined with ciliary neurotrophic factor can repair sciatic nerve defects, and that this repair is inferior to autologous nerve anastomosis, but superior to chemically extracted acellular allogeneic nerve bridging alone.

Validation of a novel animal model for sciatic nerve repair with an adipose-derived stem cell loaded fibrin conduit

Despite the regenerative capabilities of peripheral nerves, severe injuries or neuronal trauma of critical size impose immense hurdles for proper restoration of neuro-muscular circuitry. Autologous nerve grafts improve re-establishment of connectivity, but also comprise substantial donor site morbidity. We developed a rat model which allows the testing of different cell applications, i.e., mesenchymal stem cells, to improve nerve regeneration in vivo. To mimic inaccurate alignment of autologous nerve grafts with the injured nerve, a 20 mm portion of the sciatic nerve was excised, and sutured back in place in reversed direction. To validate the feasibility of our novel model, a fibrin gel conduit containing autologous undifferentiated adipose-derived stem cells was applied around the coaptation sites and compared to autologous nerve grafts. After evaluating sciatic nerve function for 16 weeks postoperatively, animals were sacrificed, and gastrocnemius muscle weight was determined along with morphological parameters（g-ratio, axon density ＆ diameter） of regenerating axons. Interestingly, the addition of undifferentiated adipose-derived stem cells resulted in a significantly improved re-myelination, axon ingrowth and functional outcome, when compared to animals without a cell seeded conduit. The presented model thus displays several intriguing features： it imitates a certain mismatch in size, distribution and orientation of axons within the nerve coaptation site. The fibrin conduit itself allows for an easy application of cells and, as a true critical-size defect model, any observed improvement relates directly to the performed intervention. Since fibrin and adipose-derived stem cells have been approved for human applications, the technique can theoretically be performed on humans. Thus, we suggest that the model is a powerful tool to investigate cell mediated assistance of peripheral nerve regeneration.

Fascial pedicle artificial nerve tissue compared with silicone tube bridging to repair sciatic nerve defects in rats

BACKGROUND： Silicone tube bridging for peripheral nerve defects has been shown to be successful in guiding neural regeneration. However, this method is accompanied by complications. Because materials for bridging nerve fibers should exhibit biocompatibility, the development of novel artificial tissues to bridge nerve grafts has become important in the field of nerve tissue engineering for the repair of peripheral nerve defects. OBJECTIVE： To investigate effectiveness and feasibility of fascial pedicle artificial nerve tissue to repair peripheral nerve defects, and to compare to autologous nerve grafts and silicone tube bridging methods. DESIGN, TIME AND SETTING： Randomized, controlled, neural tissue engineering-based, animal experiments were performed at the Laboratory of Human Anatomy in Qingdao University Medical College from March 2006 to March 2007. MATERIALS： Medical absorbable collagen sponge was purchased from Henan Province Tiangong BJo-Material, China. Cantata 2-track 4-trace EMG-evoked potential instrument was purchased from Dantec, Denmark. Medical silicone tube was purchased from Shenzhen Legend Technology, China. METHODS： Forty healthy, adult, male, Sprague Dawley rats were randomly assigned to four groups fascial pedicle nerve, autologous nerve, silicone tube, and normal, with 10 rats in each group. A 10-mm defective sciatic nerve section was produced in rats following the removal of the fascial pedicle. The fascial flap surrounding the defect was harvested; one side of the nerve pedicle was maintained and then sutured into a tube with the fascia surface as the pipe inner wail. The tube was filled with a medical absorbable collagen （Bodyin） to construct a bridge between the artificial tissue nerve graft and the damaged sciatic nerve. The sciatic nerve defects in the autologous nerve and silicone tube groups were bridged using autologous nerve grafts and a medical silicone tube with matched specifications. MAIN OUTCOME MEASURES： At 4 months after transplantation, electromyogram was used to detect sciatic nerve conduction velocity and action potential amplitude. Hematoxylin-eosin and Nissl staining were used to determine the number of spinal cord anterior horn motor neurons and neurites Osmium tetroxide staining of the sciatic nerve bridge section was performed to detect the number and diameter of nerve fibers. RESULTS： There were no differences in sciatic nerve conduction velocity, action potential amplitude, the number of spinal cord anterior horn motor neurons and neurites, sciatic nerve fiber number, and diameter between the autologous nerve graft and normal groups （P 〉 0.05）. However, these values were significantly greater than in the silicone tube group （P 〈 0.05）. CONCLUSION： Quantitative results suggested that artificial nerve tissue, with an autologous tissue fascia flap as a nerve conduit, could be used to repair peripheral nerve defects. The regenerated fascial pedicle artificial nerve tissue was similar to an autologous nerve graft in terms of morphology and functional recovery and was superior to results from silicone tube bridging transplants.