Prolonged cold-preservation of nerve allografts

The goal of this study was to determine the effect of varying durations of cold-preservation on the immunogenicity of nerve allografts and their subsequent ability to facilitate neuroregeneration across a short nerve gap. Allografts preserved for 1, 4, and 7 weeks were compared to untreated allografts and isografts. There was a shift from an interferon-gamma-producing cellular response (untreated allografts) to an absence of response (7-week cold-preserved allografts and isografts). There were no detectable alloantibodies by flow cytometry. Histomorphometry distal to the graft showed robust regeneration in the isograft and 7-week cold-preserved groups when compared to the untreated allograft group. Increasing duration of cold-preservation diminished the cellular immune response. This cold-preservation does not preclude subsequent nerve regeneration across a short nerve graft. Prolonged cold-preservation of nerve allograft tissue could serve as a means to produce unlimited graft material for use in peripheral nerve reconstruction.     

Acellular nerve allografts in peripheral nerve regeneration: a comparative study

Introduction: Processed nerve allografts offer a promising alternative to nerve autografts in the surgical management of peripheral nerve injuries where short deficits exist. Methods: Three established models of acellular nerve allograft (cold‐preserved, detergent‐processed, and AxoGen‐processed nerve allografts) were compared with nerve isografts and silicone nerve guidance conduits in a 14‐mm rat sciatic nerve defect. Results: All acellular nerve grafts were superior to silicone nerve conduits in support of nerve regeneration. Detergent‐processed allografts were similar to isografts at 6 weeks postoperatively, whereas AxoGen‐processed and cold‐preserved allografts supported significantly fewer regenerating nerve fibers. Measurement of muscle force confirmed that detergent‐processed allografts promoted isograft‐equivalent levels of motor recovery 16 weeks postoperatively. All acellular allografts promoted greater amounts of motor recovery compared with silicone conduits. Conclusion: These findings provide evidence that differential processing for removal of cellular constituents in preparing acellular nerve allografts affects recovery in vivo. Muscle Nerve, 2011     

Nerve allograft regeneration and immunological tolerance of rats with sciatic nerve transplantation after intragastric pretreatment with ultraviolet B-irradiated donor splenocytes★

BACKGROUND: Nerve allograft rejection is an unavoidable problem for nerve allografts. Traumatic peripheral nerve injuries are commonly reconstructed using autogenous nerve grafts. However, this form of reconstruction is limited by insufficient autologous nerves for large gap repairs and by morbidity at the nerve donor site. OBJECTIVE: To examine sciatic nerve regeneration and immune tolerance reaction after intragastric administration of ultraviolet B-irradiated (UVB) donor splenocytes. DESIGN, TIME AND SETTING: A complete randomized grouping design and controlled experiment. The experiments were conducted in the Department of Orthopedics, the First Affiliated Hospital to Shanxi Medical University, China, between March and October 2007. MATERIALS: Fourteen adult male SD rats and fourteen male Wistar rats, weighing 250–300 g, were randomly matched as donors and acceptors. A further seven male SD rats (weight 250–300 g, age 12–16 weeks) were used for nerve isografts. Immune preparations and the Epics XL flow cytometer were purchased from B-D Company, USA. A computer-assisted electromyograph machine was provided by Keypoint P, Dantel Company, Denmark. METHODS: Splenocytes from Wistar rats were isolated, purified, and cultured, and then irradiated with ultraviolet B. In the first control group (Group 1), the SD rats received a syngeneic SD nerve isograft. In the second control group (Group 2), the SD rats received a nerve allograft from Wistar rats without pretreatment. In the oral-tolerance treated group (Group 3), the SD recipient rats were inoculated with 2.5×107 Lewis UVB-irradiated donor splenocyte cells by intestinal tract administration at seven days before transplantation. MAIN OUTCOME MEASURES: (1) The recent end and the middle and distal end of the transplanted nerve were cut at 8 and 12 weeks after operation. Recovery of nerve regeneration was measured with HE staining using the total number, density, and diameter of the nerve fibers. (2) The level of CD25+T lymphocytes in peripheral blood was detected with the Epics XL flow cytometer at one week after operation. (3) The bilateral sciatic nerves were exposed from the sciatic notch up to 0.5 mm beyond the distal graft site at eight weeks post-engraftment. Bipolar platinum stimulating electrodes were placed under the sciatic nerve proximally and the mean conduction velocity was recorded with recording electrodes on the posterior tibial nerve at 0.3 cm distal to the nerve graft. RESULTS: Eight weeks after operation, total axon number and fiber density in Group 3 were higher than that in Group 1 (P < 0.05), neural regeneration in Group 3 was lower than that in Group 1 (P < 0.05) , The level of CD25+T lymphocytes in peripheral blood of Group 3 was significantly lower than that of Group 2 (P < 0.05). There was no significant difference between Group 3 and Group 1 (P > 0.05). At eight weeks post-engraftment the mean conduction velocity of Group 3 approximated that of Group 1. The untreated allografts in Group 2 demonstrated no measurable recovery of conduction velocity. CONCLUSION: Pretreatment with a single intragastric dose of UVB-modified donor antigen specifically induced tolerance to peripheral nerve allografts in rats. Key Words: sciatic nerve; transplantation, homologous; immune tolerance     

Nerve stepping stone has minimal impact in aiding regeneration across long acellular nerve allografts

Introduction: Acellular nerve allografts (ANAs) yield less consistent favorable outcomes compared with autografts for long gap reconstructions. We evaluated whether a hybrid ANA can improve 6‐cm gap reconstruction. Methods: Rat sciatic nerve was transected and repaired with either 6‐cm hybrid or control ANAs. Hybrid ANAs were generated using a 1‐cm cellular isograft between 2.5‐cm ANAs, whereas control ANAs had no isograft. Outcomes were assessed by graft gene and marker expression (n = 4; at 4 weeks) and motor recovery and nerve histology (n = 10; at 20 weeks). Results: Hybrid ANAs modified graft gene and marker expression and promoted modest axon regeneration across the 6‐cm defect compared with control ANA (P < 0.05), but yielded no muscle recovery. Control ANAs had no appreciable axon regeneration across the 6‐cm defect. Discussion: A hybrid ANA confers minimal motor recovery benefits for regeneration across long gaps. Clinically, the authors will continue to reconstruct long nerve gaps with autografts. Muscle Nerve 57 : 260–267, 2018     

Immunohistochemical assessment of rat nerve isografts and immunosuppressed allografts

Objective: Autologous peripheral nerve grafts are commonly used clinically as a treatment for peripheral nerve injuries. However, in research using an autologous graft is not always feasible due to loss of function, which in many cases is assessed to determine the efficacy of the peripheral nerve graft. In addition, using allografts for research require the use of an immunosuppressant, which creates unwanted side effects and another variable within the experiment that can affect regeneration. The objective of this study was to analyze graft rejection in peripheral nerve grafts and the effects of cyclosporine A (CSA) on axonal regeneration.

Methods: Peripheral nerve grafts in inbred Lewis rats were compared with Sprague-Dawley (SD) rats to assess graft rejection, CSA side effects, immune responses, and regenerative capability. Macrophages and CD8+ cells were labeled to determine graft rejection, and neurofilaments were labeled to determine axonal regeneration.

Results: SD rats without CSA had significantly more macrophages and CD8+ cells compared to Lewis autografts, Lewis isografts, and SD allografts treated with CSA. Lewis autografts, Lewis isografts, and SD autografts had significantly more regenerated axons than SD rat allografts. Moreover, allografts in immunosuppressed SD rats had significantly less axons than Lewis rat autograft and isografts.

Discussion: Autografts have long been the gold standard for treating major nerve injuries and these data suggest that even though CSA is effective at reducing graft rejection, axon regeneration is still superior in autografts versus immunosuppressed allografts.     

Immunomodulation by costimulation blockade inhibits rejection of nerve allografts

The aim of this study was to investigate if costimulation blockade could be used to modulate the immune response, to prevent rejection, and to stimulate regeneration into nerve allografts. Nerve allografts from Balb/C mice, and isogenic nerve grafts (isografts) from C57/BL6 mice, were used to bridge a 7-mm gap of the sciatic nerve in C57/BL6 mice. Allograft recipients were treated with either a triple treatment with anti-lymphocyte function antigen-1 (anti-LFA), anti-CD40 ligand (anti-CD40L), and cytotoxic T-lymphocyte antigen 4 immunoglobulin (anti-CTLA4Ig) or isotype antibodies (placebo) at postoperative days 0, 2, 4, and 6 (intraperitoneal). After 5 or 9 days, the nerve grafts, together with the proximal and the distal nerve segments, were evaluated by histology and immunocytochemistry for inflammatory cells [CD4-positive (CD4+) and CD8-positive (CD8+) staining cells] and axonal outgrowth (neurofilaments). The immune response was inhibited by costimulation blockade with less extensive inflammation and a lower number of CD4+ staining cells in triple-treated allografts at 9 days. The regeneration rate was significantly faster in isografts (0.75 mm/day) compared with allografts with placebo treatment (0.39 mm/day), but not when compared with triple-treated allografts (0.49 mm/day). At 9 days, the axons were significantly longer in nerve isografts than in nerve allografts, irrespective of treatment. Hence, costimulation blockade neither increased the regeneration rate nor the outgrowth length in triple-treated allografts. We conclude that costimulation blockade inhibits the immune response in nerve allografts without deterring early axonal outgrowth.     

Schwann cells seeded in acellular nerve grafts improve functional recovery

Introduction: This study evaluated whether Schwann cells (SCs) from different nerve sources transplanted into cold‐preserved acellular nerve grafts (CP‐ANGs) would improve functional regeneration compared with nerve isografts. Methods: SCs isolated and expanded from motor and sensory branches of rat femoral and sciatic nerves were seeded into 14mm CP‐ANGs. Growth factor expression, axonal regeneration, and functional recovery were evaluated in a 14‐mm rat sciatic injury model and compared with isografts. Results: At 14 days, motor or sensory‐derived SCs increased expression of growth factors in CP‐ANGs versus isografts. After 42 days, histomorphometric analysis found CP‐ANGs with SCs and isografts had similar numbers of regenerating nerve fibers. At 84 days, muscle force generation was similar for CP‐ANGs with SCs and isografts. SC source did not affect nerve fiber counts or muscle force generation. Conclusions: SCs transplanted into CP‐ANGs increase functional regeneration to isograft levels; however SC nerve source did not have an effect. Muscle Nerve 49 : 267–276, 2014     

The impact of motor and sensory nerve architecture on nerve regeneration

Sensory nerve autografting is the standard of care for injuries resulting in a nerve gap. Recent work demonstrates superior regeneration with motor nerve grafts. Improved regeneration with motor grafting may be a result of the nerve's Schwann cell basal lamina tube size. Motor nerves have larger SC basal lamina tubes, which may allow more nerve fibers to cross a nerve graft repair. Architecture may partially explain the suboptimal clinical results seen with sensory nerve grafting techniques. To define the role of nerve architecture, we evaluated regeneration through acellular motor and sensory nerve grafts. Thirty-six Lewis rats underwent tibial nerve repairs with 5 mm double-cable motor or triple-cable sensory nerve isografts. Grafts were harvested and acellularized in University of Wisconsin solution. Control animals received fresh motor or sensory cable isografts. Nerves were harvested after 4 weeks and histomorphometry was performed. In 6 animals per group from the fresh motor and sensory cable graft groups, weekly walking tracks and wet muscle mass ratios were performed at 7 weeks. Histomorphometry revealed more robust nerve regeneration in both acellular and cellular motor grafts. Sensory groups showed poor regeneration with significantly decreased percent nerve, fiber count, and density (p < 0.05). Walking tracks revealed a trend toward improved functional recovery in the motor group. Gastrocnemius wet muscle mass ratios show a significantly greater muscle mass recovery in the motor group (p < 0.05). Nerve architecture (size of SC basal lamina tubes) plays an important role in nerve regeneration in a mixed nerve gap model.     

Functional characterization of optimized acellular peripheral nerve graft in a rat sciatic nerve injury model

Abstract

Objectives: Acellular grafts are a viable option for use in nerve reconstruction surgeries. Recently, our lab created a novel optimized decellularization procedure that removes immunological material while leaving the majority of the extracellular matrix structure intact. The optimized acellular (OA) graft has been shown to elicit an immune response equal to or less than that elicited by the isograft, the analog of the autograft in the rat model. We investigated the performance of the OA graft to provide functional recovery in a long-term study.

Methods: We performed a long-term functional regeneration evaluation study using the sciatic functional index to quantify recovery of Lewis rats at regular time intervals for up to 52 weeks after graft implantation following 1 cm sciatic nerve resection. OA grafts were compared against other decellularized methods (Sondell treatment and thermal decellularization), as well as the isograft and primary neurorrhaphy.

Results: The OA graft supported comparable functional recovery to the isograft and superior regeneration to thermal and Sondell decellularization methods. Furthermore, the OA graft promoted early recovery to a greater degree compared to acellular grafts obtained using either the thermal or the Sondell methods.

Discussion: Equivalent functional recovery to the isograft suggests that the OA nerve graft may be a future clinical alternative to the current autologous tissue graft.     

Polyethylene glycol treated allografts not tissue matched nor immunosuppressed rapidly repair sciatic nerve gaps,maintain neuromuscular functions,and restore voluntary behaviors in female rats

Many publications report that ablations of segments of peripheral nerves produce the following unfortunate results: ( 1 ) Immediate loss of sensory signaling and motor control; ( 2 ) rapid Wallerian degeneration of severed distal axons within days; ( 3 ) muscle atrophy within weeks; ( 4 ) poor behavioral (functional) recovery after many months, if ever, by slowly‐regenerating (～1mm/d) axon outgrowths from surviving proximal nerve stumps; and (5) Nerve allografts to repair gap injuries are rejected, often even if tissue matched and immunosuppressed. In contrast, using a female rat sciatic nerve model system, we report that neurorrhaphy of allografts plus a well‐specified‐sequence of solutions (one containing polyethylene glycol: PEG) successfully addresses each of these problems by: ( a ) Reestablishing axonal continuity/signaling within minutes by nonspecific ally PEG‐fusing (connecting) severed motor and sensory axons across each anastomosis; ( b ) preventing Wallerian degeneration by maintaining many distal segments of inappropriately‐reconnected, PEG‐fused axons that continuously activate nerve‐muscle junctions; ( c ) maintaining innervation of muscle fibers that undergo much less atrophy than otherwise‐denervated muscle fibers; ( d ) inducing remarkable behavioral recovery to near‐unoperated levels within days to weeks, almost certainly by CNS and PNS plasticities well‐beyond what most neuroscientists currently imagine; and ( e ) preventing rejection of PEG‐fused donor nerve allografts with no tissue matching or immunosuppression. Similar behavioral results are produced by PEG‐fused autografts. All results for Negative Control allografts agree with current neuroscience data 1‐5 given above. Hence, PEG‐fusion of allografts for repair of ablated peripheral nerve segments expand on previous observations in single‐cut injuries, provoke reconsideration of some current neuroscience dogma, and further extend the potential of PEG‐fusion in clinical practice.     

Costimulation blockade in transplantation of nerve allografts: long-term effects

Costimulation blockade can prevent rejection of nerve allografts in short-term studies. We tested if costimulation blockade also prevented rejection of nerve allografts in long-term experiments, thereby improving functional recovery. A 7-mm sciatic nerve defect in C57/BL6 mice was bridged either by nerve allografts from Balb/C mice or by isogenic nerve grafts (isografts) from C57/BL6 mice. Costimulation blockade in the form of a triple treatment with anti-LFA-1, anti-CD40L, and CTLA4Ig was given at post-operative days 0, 2, 4, and 6 (intraperitoneal). Control mice (placebo; allografts) with nerve grafts were treated with isotype antibodies during the same time period. After 49 days, tetanic muscle force, wet weight of gastrocnemius muscle, histology, and morphometry in the tibial nerve were evaluated. Costimulation blockade diminished rejection of the nerve allografts. Axons bridged the graft. Treatment increased wet weight of the gastrocnemius muscle and resulted in a higher mean myelin area/nerve fiber in the tibial nerve distal to the nerve grafts. Tetanic muscle force and number of axons in tibial nerve showed no differences between groups. We conclude that rejection is suppressed by costimulation blockade. Treatment improves recovery of target muscle and myelination after nerve allografting.     

Vascularization is delayed in long nerve constructs compared with nerve grafts

Introduction: Nerve regeneration across nerve constructs, such as acellular nerve allografts (ANAs), is inferior to nerve auto/isografts especially in the case of long defect lengths. Vascularization may contribute to poor regeneration. The time course of vascular perfusion within long grafts and constructs was tracked to determine vascularization. Methods: Male Lewis rat sciatic nerves were transected and repaired with 6 cm isografts or ANAs. At variable days following grafting, animals were perfused with Evans Blue albumin, and grafts were evaluated for vascular perfusion by a blinded observer. Results: Vascularization at mid‐graft was re‐established within 3–4 days in 6 cm isografts, while it was established after 10 days in 6 cm ANAs. Conclusions: Vascular perfusion is reestablished over a shorter time course in long isografts when compared with long ANAs. The differences in vascularization of long ANAs compared with auto/isografts suggest regenerative outcomes across ANAs could be affected by vascularization rates. Muscle Nerve 54 : 319–321, 2016     

Regeneration across preserved peripheral nerve grafts

The potential to store nerve grafts for a prolonged period of time was assessed in a rat sciatic nerve model. Three-centimeter syngeneic nerve grafts were stored in Belzer/University of Wisconsin cold storage solution at different temperatures (5°C, 22°C, or 37°C) for varying time periods (6 h, 24 h, or 3 weeks) prior to transplantation. Functional assessment using serial walking track analyses revealed no difference between storage times and temperatures. At 14 months postengraftment, the conduction velocities and the number of myelinated fibers that had regenerated across all grafts stored at 5°C for all time periods tested were superior to grafts stored at either 22°C or 37°C. Nerve grafts stored for up to 3 weeks at 5°C acted as effective conduits for proximal regenerating fibers and resulted in histologic, electrophysiologic, and functional results equivalent to fresh nerve grafts. Nerve graft storage may be applicable to nerve allografts and potentially provide allograft material that requires reduced or no associated host immunosuppression.© 1995 John Wiley & Sons, Inc.     

Controlled release of nerve growth factor enhances sciatic nerve regeneration

Based on previous studies demonstrating the potential of growth factors to enhance peripheral nerve regeneration, we developed a novel growth factor delivery system to provide sustained delivery of nerve growth factor (NGF). This delivery system uses heparin to immobilize NGF and slow its diffusion from a fibrin matrix. This system has been previously shown to enhance neurite outgrowth in vitro, and in this study, we evaluated the ability of this delivery system to enhance nerve regeneration through conduits. We tested the effect of controlled NGF delivery on peripheral nerve regeneration in a 13-mm rat sciatic nerve defect. The heparin-containing delivery system was studied in combination with three doses of NGF (5, 20, or 50 ng/mL) and the results were compared with positive controls (isografts) and negative controls (fibrin alone, NGF alone, and empty conduits). Nerves were harvested at 6 weeks postoperatively for histomorphometric analysis. Axonal regeneration in the delivery system groups revealed a marked dose-dependent effect. The total number of nerve fibers at both the mid-conduit level and in the distal nerve showed no statistical difference for NGF doses at 20 and 50 ng/mL from the isograft (positive control). The results of this study demonstrate that the incorporation of a novel delivery system providing controlled release of growth factors enhances peripheral nerve regeneration and represents a significant contribution toward enhancing nerve regeneration across short nerve gaps.     

An efficient stereological sampling approach for quantitative assessment of nerve regeneration

Aims: The aim of the present study was to find the most efficient sampling strategy for stereological analysis of peripheral nerve, including the number of myelinated nerve fibres, axon cross‐sectional area and myelin sheet thicknesses of nerve fibres. Methods: Two groups of rats underwent experimental resection of the tibial and peroneal nerves. The first group received tibial‐peroneal end to end autograft repair (n = 6). Tibial and peroneal nerves were isolated, transected, and separated 1 cm distal to the trifurcation, where they lay adjacent to each other by a 1‐cm gap, then repaired with an autologous nerve graft taken from the tibial nerve. The proximal stump of the tibial nerve and distal stump of the peroneal nerve were connected to each other by means of the nerve graft. The second group received tibial‐peroneal repair with a flexible collagen tube (n = 6). After 90 days of recovery, animals were sacrificed and nerve segments were removed and sectioned for microscopy. Three different sampling strategies, that is, small, medium and large step sizes were applied to obtain each quantitative parameter. Results: There are no significant differences between these sampling strategies with respect to total number of myelinated nerve fibres, axon cross‐sectional area and myelin sheet thicknesses of nerve fibres. Conclusion: Findings show that one can achieve the desired estimate precisely with a rather large and less time‐consuming sampling approach. In addition, it was observed that the size discrepancy of nerve regeneration can be improved by collagen tube conduit even with a 1‐cm gap.     

Clinical applications of peripheral nerve allotransplantation. Present medical experience and review of the literature

Nerve autograft has become recently the gold standard for the reconstruction of surgical nerve gap. Functional results of using this technique continue to improve with the advances in micro-neurosurgery and with greater understanding of neurobiology and nerve regeneration. Surgical reconstruction of extensive nerve injuries frequently exhausts the patient's own source of expandable autogenous nerve grafts. Nerve allografts would offer a limitless supply of graft material. The requirement of only transient immunosuppression to support the regeneration of host axons across the nerve allograft toward distal host receptors renders the nerve allograft an advantageous option for these otherwise irreparable nerve injuries. The methods of graft harvesting, preservation and patients' immunosuppressive regimen as well as the clinical outcomes following the nerve allotransplantation are discussed.     

Peripheral nerve transplantation: the effects of predegenerated grafts and immunosuppression.

Research involving nerve transplantation has shown that tissue rejection limits the neurologic recovery unless the host is immunosuppressed. This study investigates an alternative to permanent or temporary immunosuppression using a rat model with nerve transplants from Brown-Norway rat donors to bridge defects in the sciatic nerve of Lewis rat recipients as these two inbred strains differ at both major and minor histocompatibility loci. The specific aim of this study was to evaluate if predegenerated nerve grafts decreased the tissue rejection and improved the neurologic recovery of animals with allogenic nerve grafts to avoid the problems associated with either short- or long-term immunosuppression. The animals in the experimental groups received cyclosporin-A, predegenerated grafts, both, or neither. The predegenerated grafts were produced by division of the nerve three weeks prior to grafting to allow for Wallerian degeneration to occur. The outcome was assessed by measurements stressing functional recovery (sensory testing, gait analysis, joint flexion contracture), studies of muscle recovery (muscle weight and hydroxyproline concentration), and histologic studies (axonal counts and inflammatory reaction). The animals receiving the predegenerated grafts without cyclosporin did have an improved recovery (joint flexion contracture 35 degrees +/- 8 degrees and hydroxyproline ratio 1.52 +/- 0.16) as compared to the joint flexion contractures and hydroxyproline ratios of the allograft group of animals without either cyclosporin-A or pretreatment and the ungrafted control group (47 degrees +/- 18 degrees, 1.68 +/- 0.34, and 53 degrees +/- 15 degrees, 4.50 +/- 0.27, respectively, p less than 0.01). However, all the isograft groups and allograft groups with cyclosporin-A, regardless of whether the graft had been predegenerated or not, had greater neurologic recovery than the allograft group with predegenerated grafts but without cyclosporin-A by the same parameters (p less than 0.01). Allograft groups with short-term immunosuppression with cyclosporin-A did as well as isograft groups, and isograft groups with predegenerated grafts did not do any better than isografts without pretreatment (p less than 0.01).     

The dose-related effect of monoclonal antibodies against adhesion molecules ICAM-1 and LFA-1 on peripheral nerve allograft rejection in a rat model

Donor-specific immunosuppression using anti-intercellular adhesion molecule-1 (ICAM-1) and anti-lymphocyte function-associated antigen-1 (LFA-1) has been shown to inhibit nerve allograft rejection without side effects. This dose-response study evaluated several dosing regimens using a 2-week course of three monoclonal antibodies (mAbs) against ICAM-1 and LFA-1 in combination on peripheral nerve allograft rejection in a rat model. Assessments of regeneration included walking track, electrophysiological, and histomorphologic analyses. Donor (ACI)-specific tolerance induction was assessed. Toxicity and mAb serum levels were monitored. At 18 weeks post engraftment, intermediate and high-dose groups were histologically indistinguishable from isograft controls, and superior to the untreated allograft group which demonstrated a significantly lower percent nerve tissue than all other groups. There were no differences in print length factor after 12 weeks or conduction velocity at sacrifice between any groups. Tolerance induction was not demonstrated. During mAb administration, animals in higher dose groups experienced temporary systemic side effects. This study demonstrated that a short course of mAb therapy directed against ICAM-1/LFA-1 inhibits rejection in rat peripheral nerve allografts by an unknown mechanism. The use of immune modulation in nerve transplantation may eliminate the need for systemic immunosuppression.     

Indefinite survival of peripheral nerve allografts after temporary Cyclosporine A immunosuppression

It is hypothesized that unlike solid organ transplants immunosuppression of peripheral nerve allografts is needed only for the finite time period required for regeneration of proximal host nerve axons through the allograft and subsequent re-establishment of host end-organ connections. The aim of this study was to explore the consequences of temporary and continuous systemic Cyclosporine A (CsA) immunosuppression upon peripheral nerve allograft survival. Buffalo rats received Lewis nerve allografts under CsA immunosuppression (5 mg/kg/day) either continuously for 20 weeks, or for only 10 weeks followed by abrupt withdrawal. At 20 weeks, the nerve segments from both groups were regrafted into na?ve Buffalo or Lewis recipients without further immunosuppression. These grafts were compared with isografts, unimmunosuppressed allografts and allografts immunosuppressed for 10 weeks in situ. By eight weeks following regrafting, the secondary Lewis recipients had rejected the temporarily immunosuppressed allografts and accepted the continuously immunosuppressed allograft, while the secondary Buffalo recipients accepted both the temporarily and continuously immunosuppressed allografts as assessed by histology and morphometry. Functional recovery was earlier in secondary recipient strain animals that received temporarily immunosuppressed allografts in comparison to those that received continuously immunosuppressed allografts. Analysis of secondary recipients of temporarily immunosuppressed allografts demonstrated greater in vitro MLR and LDA reactivity than did those receiving continuously immunosuppressed allografts. These findings support the hypothesis that donor alloantigens are lost or replaced by the recipient after immunosuppression withdrawal. Moreover, the change to recipient antigenicity in the nerve allograft is retarded and incomplete under continuous CsA immunosuppression, resulting in acceptance by both secondary donor and recipient strains upon regraftment.     

神经桥接与导管套接修复猫动眼神经形态学观察

目的探求导管修复颅内段动眼神经的可行性.方法 20只健康家猫随机分为2组.将右侧动眼神经于脑池段切断后,分组采用自体神经桥接和导管套接的方法修复.术后14周末光镜、电镜观察神经纤维的连续性、再生纤维数目和直径.结果神经修复14周后,神经桥接组67%、导管套接组75%的动物其动眼神经功能均有一定程度的恢复.形态学显示两组均取得较好的神经再生效果,两组间再生纤维直径差异无显著性(P>0.05),但导管套接组再生神经纤维数目较多,差异有显著性(P<0.05).结论导管套接法可作为颅内段动眼神经损伤后的一种修复方法,其疗效近似或稍优于神经桥接法.