Low-Intensity Pulsed Ultrasound Prompts Both Functional and Histologic Improvements While Upregulating the Brain-Derived Neurotrophic Factor Expression after Sciatic Crush Injury in Rats.

The aim of this study was to determine that low-intensity pulsed ultrasound (LIPUS) at an intensity of 140 mW/cm2 promotes functional and histologic improvements in sciatic nerve crush injury in a rat model and to investigate changes over time in relevant growth factors and receptors, exploring the mechanism of LIPUS in the recovery process after injury. Toe angle in the toe-off phase, regenerative axonal length, myelinated nerve fiber density, diameter of myelinated nerve fiber, axon diameter and myelin sheath thickness were significantly higher in the LIPUS group than in the sham group. Gene and protein expression of brain-derived neurotrophic factor (BDNF) was upregulated in the LIPUS group. In conclusion, LIPUS contributed to rapid functional and histologic improvement and upregulated BDNF expression after sciatic nerve crush injury in rats.

Effect of photobiomodulation on mitochondrial dynamics in peripheral nervous system in streptozotocin-induced type 1 diabetes in rats.

There is no effective treatment to halt peripheral nervous system damage in diabetic peripheral neuropathy. Mitochondria have been at the center of discussions as important factors in the development of neuropathy in diabetes. Photobiomodulation has been gaining clinical acceptance as it shows beneficial effects on a variety of nervous system disorders. In this study, the effects of photobiomodulation (904 nm, 45 mW, 6.23 J/cm2, 0.13 cm2, 60 ns pulsed time) on mitochondrial dynamics were evaluated in an adult male rat experimental model of streptozotocin-induced type 1 diabetes. Results presented here indicate that photobiomodulation could have an important role in preventing or reversing mitochondrial dynamics dysfunction in the course of peripheral nervous system damage in diabetic peripheral neuropathy. Photobiomodulation showed its effects on modulating the protein expression of mitofusin 2 and dynamin-related protein 1 in the sciatic nerve and in the dorsal root ganglia neurons of streptozotocin-induced type 1 diabetes in rats.

UVB irradiation induces contralateral changes in galanin, substance P and c-fos immunoreactivity in rat dorsal root ganglia, dorsal horn and lateral spinal nucleus.

The selection of control group is crucial, as the use of an inadequate group may strongly affect the results. In this study we examine the effect on contralateral tissue protein levels, in a model of unilateral UVB irradiation, as the contralateral side is commonly used as a control. Previous studies have shown that UVB irradiation increases immunoreactivity for inflammatory regulated neuropeptides. Unilateral UVB irradiation of rat hind paw was performed and corresponding contralateral spinal cord and dorsal root ganglia (DRG) were collected 2-96 h after and investigated for changes in galanin, substance P and c-fos immunoreactivity. Control tissue was collected from naïve rats. Measurement of skin blood flow from contralateral heel hind paws (Doppler), revealed no change compared to naïve rats. However, UVB irradiation caused a significant reduction in the contralateral proportion of galanin immunopositive DRG neurons, at all-time points, as well as an increase in the contralateral spinal cord dorsal horn, around the central canal and in the lateral spinal nucleus (2-48 h). The contralateral proportion of SP positive DRG neurons and dorsal horn immunoreactivity was unchanged, whereas the lateral spinal nucleus area showed increased immunoreactivity (48 h). UVB irradiation also induced a slight contralateral upregulation of c-fos in the dorsal horn/central canal area (24 and 48 h). In summary, unilateral UVB irradiation induced contralateral changes in inflammatory/nociceptive neuropeptides in spinal cord and afferent pathways involved in pain signaling already within 24 h, a time point when also ipsilateral neurochemical/physiological changes have been reported for rats and humans.

Low-Level Laser Therapy in Different Wavelengths on the Tibialis Anterior Muscle of Wistar Rats After Nerve Compression Injury.

OBJECTIVE: Traumatic injuries are common and may promote disruption of neuromuscular communication, triggering phenomena that lead to nerve degeneration and affect muscle function. A laser accelerates tissue recovery; however, the parameters used are varied, making it difficult to compare studies. The purpose of this study was to evaluate the effect of low-level laser therapy, at 660- and 830-nm wavelengths, on the tibialis anterior muscle of Wistar rats after sciatic nerve compression. METHODS: Twenty animals were separated into 4 groups: control, sciatic nerve injury, lesion + 660-nm laser, and lesion + 830-nm laser. In the lesion groups, the right sciatic nerve was surgically exposed and compressed with hemostatic forceps for 30 seconds. After the third postoperative day, the groups with laser therapy were submitted to treatment for 2 weeks totaling 10 applications, performed directly on the surgical scar of the nerve injury. Grip strength was analyzed before and after the nerve injury and during the treatment period. The tibialis anterior muscle was processed for light microscopy, area measurement, smaller diameter, number of fibers, nuclei, and connective tissue. RESULTS: The animals submitted to the injury experienced muscular atrophy and morphological changes in the number of muscle fibers and nuclei. In the connective tissue morphometry, there was a decrease in the treated groups compared with the untreated groups. CONCLUSION: The laser treatment at different wavelengths showed no improvement in the tibialis anterior muscle of Wistar rats within the morphological and functional aspects evaluated.

Dose-dependent effect of the pulsed Nd:YAG laser in the treatment of crushed sciatic nerve in Wister rats: an experimental model.

The objective of the study was to investigate the efficacy of three energy densities 4, 10, and 50 J/cm2 of pulsed Nd:YAG laser for the treatment of crushed sciatic nerve in Wister rats by evaluating changes in the sciatic functional index and the electrophysiology.A total of 180 Wistar rats were involved in the study. Rats were randomly assigned to five groups. Rats were subjected to the sciatic nerve crushing. Control negative (CONT-ve), which received no crushing; control positive (CONT+ve), which received crushing with no laser; and HILT-4, HILT-10, and HILT-50 groups, which received pulsed Nd:YAG laser (10 Hz, 360 mJ/cm2) with energy densities 4, 10, and 50 J/cm2, respectively. The SFI, the amilitude of compound motor action potential (CMAP) and sciatic motor nerve conduction velocity (MNCV) were measured before and after seven, 14, and 21 days after crushing. For the SFI and electrophysiological analysis, repeated measures ANOVA is used, followed by Bonferroni's repeated-measures test. Statistical significance was set at p < 0.05. After one week, there was no significant difference in SFI, CMAP, and MNCV among the three laser groups with significant changes between them and CONT-ve and CONT+ve groups. There was a significant increase in either CMAP amplitude or MNCV after 14 days with significant decrease in the SFI after 21 days among all treatment groups. The pulsed Nd:YAG laser applied with energy densities 4, 10, and 50 J/cm2 significantly decreased the SFI and increased the CMAP and MNCV of the crushed sciatic nerve in Wister rats. Among laser doses, the difference in the rate of recovery in the electrophysiology was found after two weeks while in the SFI after three weeks. The improvement after the nerve injury was time and dose dependent.

Effects of Photobiomodulation with Low-level Laser Therapy on Muscle Repair Following a Peripheral Nerve Injury in Wistar Rats.

Peripheral nerve injury (PNI) can lead to sensory and/or motor impairment. As a treatment photobiomodulation (PBM) has demonstrated positive effects in terms of the maintenance of muscle activation and trophism. Wistar rats were divided into five groups: control, injury, injury + PBMn (irradiation over injured nerve), injury + PBMm (irradiation over affected muscle) and injury + PBMnm (irradiation over nerve and muscle). The left sciatic nerve was submitted to a crushing injury. Treatment was administered with low-level laser (780 nm, 0.04 cm2 , 1 W cm-2 , 3.2 J) over the injured nerve and/or the tibialis anterior muscle. The effects of PBM were favorable on muscle morphology and gene expression of calcineurin, myogenin and acetylcholine receptors. PBM led to an acceleration on muscle repair process, and effects were more evident in 2 weeks after PNI. Thus, PBM is indicated for the area over both the injured nerve and the affected muscle.

Selective stimulation of bullfrog sciatic nerve by gold nanorod assisted combined electrical and near-infrared stimulation.

Selective stimulation of the nervous system is an important way to improve the therapeutic efficacy and minimize side effects. This paper introduces an improved method using combined electrical and near-infrared stimulation to realize selective excitation and inhibition of different sciatic nerve branches. Both the electrical stimulation and the near-infrared laser are added to the main trunk of the sciatic nerve, and gold nanorods are injected into the light irradiation point of the nerve to increase the absorption of light. Two cuff recording electrodes are added to the two sciatic nerve branches, respectively. The compound nerve action potential recorded by the cuff electrode is transmitted to the physiological signal instrument. In the experiment, selective activation and inhibition of the two nerve branches are achieved by adjusting the electrical stimulation parameters, the light stimulus parameters and the location of the light. These results demonstrate that combined electrical and near-infrared stimulation, which can effectively activate or suppress the different nerve fibers in the nerve fiber bundle, is suitable for selective regulation of peripheral nerve. Meanwhile, the photoelectric combined stimulation can reduce both the electrical energy and light energy needed for the stimulation, and reduce the electrical damage and light damage to the nerve.

A shape-memory and spiral light-emitting device for precise multisite stimulation of nerve bundles.

We previously demonstrated that for long-term spastic limb paralysis, transferring the seventh cervical nerve (C7) from the nonparalyzed side to the paralyzed side results in increase of 17.7 in Fugl-Meyer score. One strategy for further improvement in voluntary arm movement is selective activation of five target muscles innervated by C7 during recovery process. In this study, we develop an implantable multisite optogenetic stimulation device (MOSD) based on shape-memory polymer. Two-site stimulation of sciatic nerve bundles by MOSD induces precise extension or flexion movements of the ankle joint, while eight-site stimulation of C7 nerve bundles induce selective limb movement. Long-term implant of MOSD to mice with severed and anastomosed C7 nerve is proven to be both safe and effective. Our work opens up the possibility for multisite nerve bundle stimulation to induce highly-selective activations of limb muscles, which could inspire further applications in neurosurgery and neuroscience research.

Mechanical allodynia induced by optogenetic sensory nerve excitation activates dopamine signaling and metabolism in medial nucleus accumbens.

The mesolimbic dopaminergic signaling, such as that originating from the ventral tegmental area (VTA) neurons in the medial part of the nucleus accumbens (mNAc), plays a role in complex sensory and affective components of pain. To date, we have demonstrated that optogenetic sensory nerve stimulation rapidly alters the dopamine (DA) content within the mNAc. However, the physiological role and biochemical processes underlying such rapid and regional dynamics of DA remain unclear. In this study, using imaging mass spectrometry (IMS), we observed that sensitized pain stimulation by optogenetic sensory nerve activation increased DA and 3-Methoxytyramine (3-MT; a post-synaptic metabolite obtained following DA degradation) in the mNAc of the experimental mice. To delineate the mechanism associated with elevation of DA and 3-MT, the de novo synthesized DA in the VTA/substantia nigra terminal areas was evaluated using IMS by visualizing the metabolic conversion of stable isotope-labeled tyrosine (13C15N-Tyr) to DA. Our approach revealed that at steady state, the de novo synthesized DA occupied >10% of the non-labeled DA pool in the NAc within 1.5 h of isotope-labeled Tyr administration, despite no significant increase following pain stimulation. These results suggested that sensitized pain triggered an increase in the release and postsynaptic intake of DA in the mNAc, followed by its degradation, and likely delayed de novo DA synthesis. In conclusion, we demonstrated that short, peripheral nerve excitation with mechanical stimulation accelerates the mNAc-specific DA signaling and metabolism which might be associated with the development of mechanical allodynia.

Biochemical changes in injured sciatic nerve of rats after low-level laser therapy (660 nm and 808 nm) evaluated by Raman spectroscopy.

The aim of this study was to identify biochemical changes in sciatic nerve (SN) after crush injury and low-level laser therapy (LLLT) with 660 nm and 808 nm by Raman spectroscopy (RS) analysis. A number of 32 Wistar rats were used, divided into four groups (control 1, control 2, LASER 660 nm, and LASER 808 nm). All animals underwent surgical procedure of the SN and groups control 2, LASER 660 nm, and LASER 808 nm were submitted to SN crush damage (axonotmesis). The LLLT in the groups LASER 660 nm and LASER 808 nm was applied daily for 21 consecutive days (100 mW, 30 s, 133 J/cm2 fluence). The hind paw was removed and the SN was dissected and positioned on an aluminum support to collect dispersive Raman spectra (830 nm excitation, 30 s accumulation). To estimate the biochemical changes in the SN associated with LLLT, the principal component analysis (PCA) was applied. The Raman spectra of the sciatic nerve fragments showed peaks of the major biochemical components of the nerve, especially sphingolipids, phospholipids, glycoproteins, and collagen. The spectral features identified in some of the principal component loading vectors are referred to the biochemical elements present on the SN and were increased in the groups treated with LLLT, mainly lipids (sphingo and phospholipids) and proteins (collagen)-constituents of the myelin sheath. The RS was effective in identifying the biochemical differences in the SN after the crush injury, and LASER 660 nm was more efficient than the LASER 808 nm in cell proliferation and repair of the injured SN.

Towards Safe Infrared Nerve Stimulation: A Systematic Experimental Approach.

Neural activation by infrared nerve stimulation (INS) gains growing interest as a potential alternative to conventional electric nerve stimulation, since unambiguous advantages like contact-free operation, enhanced spatial selectivity and lack of (electrical) stimulation artifacts are promising for both future electrophysiological research and clinical application. For the systematic investigation of laser nerve activation, we recently introduced a novel experimental approach. Comprising a defined focused beam profile, it enables remote controlled, contact-free pulsed laser stimulation of the rat sciatic nerve, simultaneous to high-speed temperature measurement in vivo. Up to now, successful neural activation with single laser pulses (2 - 6 mJ) was observed in all performed experiments, however, it strongly depended on the particular nerve location. Hence, we depict the investigation of spatial dependency of the nerve response and identify `regions of excitability' on the nerve surface, that are highly susceptible to INS. By means of thermal imaging, we simultaneously monitored the nerve surface temperature, where we observed progressing temperature build-up during single pulse stimulation with repetition rates above 4 Hz. In this work, we present current results of our ongoing research.

Effect of low-level 1800 MHz radiofrequency radiation on the rat sciatic nerve and the protective role of paricalcitol.

The nervous system is an important target of radiofrequency (RF) radiation exposure since it is the excitable component that is potentially able to interact with electromagnetic fields. The present study was designed to investigate the effects of 1,800 MHz RF radiation and the protective role of paricalcitol on the rat sciatic nerve. Rats were divided into four groups as control, paricalcitol, RF, and RF + paricalcitol. In RF groups, the rats were exposed to 1,800 MHz RF for 1 h per day for 4 weeks. Control and paricalcitol rats were kept under the same conditions without RF application. In paricalcitol groups, the rats were given 0.2 µg/kg/day paricalcitol, three times per week for 4 weeks. Amplitude and latency of nerve compound action potentials, catalase activities, malondialdehyde (MDA) levels, and ultrastructural changes of sciatic nerve were evaluated. In the RF group, a significant reduction in amplitude, prolongation in latency, an increase in the MDA level, and an increase in catalase activity and degeneration in the myelinated nerve fibers were observed. The electrophysiological and histological findings were consistent with neuropathy, and the neuropathic changes were partially ameliorated with paricalcitol administration. Bioelectromagnetics. 39:631-643, 2018. © 2018 Wiley Periodicals, Inc.

Photobiomodulation induces antinociception, recovers structural aspects and regulates mitochondrial homeostasis in peripheral nerve of diabetic mice.

Diabetic peripheral neuropathy (DPN) is a nervous disorder caused by diabetes mellitus, affecting about 50% of patients in clinical medicine. Chronic pain is one of the major and most unpleasant symptoms developed by those patients, and conventional available treatments for the neuropathy, including the associated pain, are still unsatisfactory and benefit only a small number of patients. Photobiomodulation (PBM) has been gaining clinical acceptance once it is able to promote early nerve regeneration resulting in significant improvement in peripheral nerves disabilities. In this work, the effects of PBM (660 nm, 30 mW, 1.6 J/cm2 , 0.28 cm2 , 15 s in a continuous frequency) on treating DPN-induced pain and nerve damage were evaluated in an experimental model of diabetic-neuropathy induced by streptozotocin in mice. PBM-induced antinociception in neuropathic-pain mice was dependent on central opioids release. After 21 consecutive applications, PBM increased nerve growth factor levels and induced structural recovery increasing mitochondrial content and regulating Parkin in the sciatic nerve of DPN-mice. Taking together, these data provide new insights into the mechanisms involved in the effects of PBM-therapy emphasizing its therapeutic potential in the treatment of DPN.

Comparative effect of photobiomodulation associated with dexamethasone after sciatic nerve injury model.

To analyze the effect of photobiomodulation and dexamethasone on nerve regeneration after a sciatic nerve crushing model. Twenty-six Swiss mice were divided into the following groups: naive; sham; injured, low-level laser therapy (LLLT) (660 nm, 10 J/cm2, 0.6 J, 16.8 J total energy emitted during the 28 days of radiation, 20 s, for 28 days); dexamethasone (Dex) (local injection of 2 mg/kg for 10 consecutive days); and LLLT group associated with Dex (LLLT/Dex), with the same parameters of the other groups. For nerve injury, a portable adjustable pinch was used. The animals were evaluated using the Sciatic Functional Index (SFI) and Sciatic Static Index (SSI). The results obtained were evaluated with Image J™ and Kinovea™. Data and images were obtained at baseline and after 7, 14, 21, and 28 days after surgery. The evaluation of hyperalgesia, using Hargreaves, and behavior through the open field was also performed. In functional and static analysis, all groups presented significant differences when compared to the injured group. In the analysis of the SSI results, the group treated with both LLLT and dexamethasone was more effective in improving the values of this parameter, and in the SFI, the laser-treated group obtained better results. In the evaluation through the open field and the Hargreaves, there was no difference. The application of LLLT and dexamethasone was effective in nerve regeneration according to the results and was more effective when LLLT was associated with dexamethasone than in LLLT alone for the SSI.

A histopathological and biochemical evaluation of oxidative injury in the sciatic nerves of male rats exposed to a continuous 900-megahertz electromagnetic field throughout all periods of adolescence.

The effects on human health of the electromagnetic field (EMF) emitted by mobile phones, used by approximately 7 billion people worldwide, have become an important subject for scientific research. Studies have suggested that the EMF emitted by mobile phones can cause oxidative stress in different tissues and age groups. Young people in adolescence, a time period when risky behaviors and dependences increase, use mobile phones more than adults. The EMF emitted by mobile phones, which are generally carried in the pocket or in bags when not in use, will very probably affect the sciatic nerve. No previous study has investigated the effect of mobile phone use in adolescence on peripheral nerve. This study was planned accordingly. Twenty-four male Sprague Dawley rats aged 21 days were divided equally into control (CGr), Sham (SGr) and EMF (EMFGr) groups. No procedure was performed on CGr rats. EMFGr were exposed to the effect of a 900-megahertz (MHz) EMF for 1 h at the same time every day between postnatal days 21-59 (the entire adolescent period) inside a cage in the EMF apparatus. SGr rats were placed inside the cage for 1 h every day without being exposed to EMF. All rats were sacrificed at the end of the study period, and 1 cm sections of sciatic nerve were extracted. Malondialdehyde (MDA), glutathione, catalase (CAT) superoxide dismutase (SOD) values were investigated biochemically in half of the right sciatic nerve tissues. The other halves of the nerve tissues were subjected to routine histopathological tissue procedures, sectioned and stained with hematoxylin and eosin (H&E) and Masson's trichrome. Histopathological evaluation of slides stained with Masson's trichrome and H&E revealed a normal appearance in Schwann cells and axons in all groups. However, there was marked thickening in the epineurium of sciatic nerves from EMFGr rats. MDA, SOD and CAT levels were higher in EMFGr than in CGr and SGr at biochemical analyses. Apoptotic index (AI) analysis revealed a significant increase in the number of TUNEL (+) cells when EMFGr was compared with CGr and SGr. In conclusion, our study results suggest that continuous exposure to a 900-MHz EMF for 1 h throughout adolescence can cause oxidative injury and thickening in the epineurium in the sciatic nerve in male rats.

Effects of Localized X-Ray Irradiation on Peripheral Nerve Regeneration in Transected Sciatic Nerve in Rats.

Low-dose radiation has been used in clinical and experimental models for the prevention of scarring and for fracture healing. There is evidence that low-dose radiation improves the hormesis of various cell types but little is known about its effects on peripheral nerve tissue. In this study, we investigated the beneficial effects of low-dose radiation on the regeneration of transectional peripheral nerve injury in an experimental rat model. Seventy-two male Sprague-Dawley rats received transection injury to the left sciatic nerves, and the nerves were subsequently sutured by epineurium end-to-end anastomosis to restore continuity. Animals were randomly assigned to one of two treatment groups (n = 36/group): 1 Gy X-ray irradiation or control (sham irradiation). Gait analysis, electrophysiological examination and morphological investigations were performed. In addition, Western blot and qRT-PCR were performed to determine the level of vascular endothelial growth factor (VEGF) and growth-associated protein-43 (GAP-43). Content of VEGF and GAP-43 in the regenerated sciatic nerve of the irradiated group was higher than the control group. At 4 to 12 weeks after surgery, the irradiated animals exhibited a significantly improved functional recovery relative to controls. At 12 weeks after surgery, amplitude and conduction velocity of the irradiated group were higher than the control group (P < 0.05). The number of nerve fibers, diameter of axons and morphological structure of the myelin sheath in the irradiated group were superior to those of the control group. These results suggest that low-dose radiation contributed to regeneration and functional recovery after transverse peripheral nerve injury by inducing increased production of VEGF and GAP-43, which promote the axonal regeneration and myelination.

Model study of combined electrical and near-infrared neural stimulation on the bullfrog sciatic nerve.

This paper implemented a model study of combined electrical and near-infrared (808 nm) neural stimulation (NINS) on the bullfrog sciatic nerve. The model includes a COMSOL model to calculate the electric-field distribution of the surrounding area of the nerve, a Monte Carlo model to simulate light transport and absorption in the bullfrog sciatic nerve during NINS, and a NEURON model to simulate the neural electrophysiology changes under electrical stimulus and laser irradiation. The optical thermal effect is considered the main mechanism during NINS. Therefore, thermal change during laser irradiation was calculated by the Monte Carlo method, and the temperature distribution was then transferred to the NEURON model to stimulate the sciatic nerve. The effects on thermal response by adjusting the laser spot size, energy of the beam, and the absorption coefficient of the nerve are analyzed. The effect of the ambient temperature on the electrical stimulation or laser stimulation and the interaction between laser irradiation and electrical stimulation are also studied. The results indicate that the needed stimulus threshold for neural activation or inhibition is reduced by laser irradiation. Additionally, the needed laser energy for blocking the action potential is reduced by electrical stimulus. Both electrical and laser stimulation are affected by the ambient temperature. These results provide references for subsequent animal experiments and could be of great help to future basic and applied studies of infrared neural stimulation (INS).

Effect of photobiomodulation therapy (808 nm) in the control of neuropathic pain in mice.

Neuropathic pain can be defined as the pain initiated or caused by a primary lesion or dysfunction of the central or peripheral nervous system. Photobiomodulation therapy (PBM) stands out among the physical therapy resources used for analgesia. However, application parameters, especially the energy density, remain controversial in the literature. Therefore, this study aimed to investigate the PBM effect, in different energy densities to control neuropathic pain in mice. Fifty (50) mice were induced to neuropathy by chronic constriction surgery of the sciatic nerve (CCI), treated with PBM (808 nm), and divided into five groups: GP (PBM simulation), GS (sham), GL10, GL20, GL40 (energy density of 10, 20, and 40 J/cm2, respectively). The evaluations were carried out using the hot plate test and Randall and Selitto test, before and after the CCI surgery, every 15 days during the 90 days experiment. ß-Endorphin blood dosage was also tested. For both the hot plate and Randall and Selitto tests, the GL20 and GL40 groups presented reduction of the nociceptive threshold from the 30th day of treatment, the GL10 group only after day 75, and the GP group did not show any improvement throughout the experiment. The ß-endorphin dosage was higher for all groups when compared to the GP group. However, only the GL20 group and GL40 presented a significant increase. This study demonstrates that PBM in higher energy density (20, 40 J/cm2) is more effective in the control of neuropathic pain.

Analysis of the variation in low-level laser energy density on the crushed sciatic nerves of rats: a morphological, quantitative, and morphometric study.

The objective of this study was to evaluate three energy densities of low-level laser therapy (LLLT, GaAlAs, 780 nm, 40 mW, 0.04 cm2) for the treatment of lesions to peripheral nerves using the sciatic nerve of rats injured via crushing model (15 kgf, 5.2 MPa). Thirty Wistar rats (♂, 200-250 g) were divided into five groups (n = 6): C-control, not injured, and irradiated; L0-injured nerve without irradiation; L4-injured nerve irradiated with LLLT 4 J/cm2 (0.16 J); L10-injured nerve irradiated with LLLT 10 J/cm2 (0.4 J); and L50-injured nerve irradiated with LLLT 50 J/cm2 (2 J). The animals were sacrificed 2 weeks after the injury via perfusion with glutaraldehyde (2.5%, 0.1 M sodium cacodylate buffer). The nerve tissue was embedded in historesin, cut (3 µm), mounted on slides, and stained (Sudan black and neutral red). The morphological and quantitative analysis (myelin and blood capillary densities) and morphometric parameters (maximum and minimum diameters of nerve fibers, axon diameter, G-ratio, myelin sheath thickness) were assessed using the ImageJ software. ANOVA (parametric) or Kruskal-Wallis (nonparametric) tests were used for the statistical analysis. Groups L0, L4, L10, and L50 exhibited diminished values of all the quantitative and morphometric parameters in comparison to the control group. The morphological, quantitative, and morphometric data revealed improvement after injury in groups L4, L10, and L50 (irradiated groups) compared to the injured-only group (L0); the best results, in general, were observed for the L10 group after 15 days of nerve injury.

Light-Activated Sealing of Acellular Nerve Allografts following Nerve Gap Injury.

Introduction Photochemical tissue bonding (PTB) uses visible light to create sutureless, watertight bonds between two apposed tissue surfaces stained with photoactive dye. In phase 1 of this two-phase study, nerve gaps repaired with bonded isografts were superior to sutured isografts. When autograft demand exceeds supply, acellular nerve allograft (ANA) is an alternative although outcomes are typically inferior. This study assesses the efficacy of PTB when used with ANA. Methods Overall 20 male Lewis rats had 15-mm left sciatic nerve gaps repaired using ANA. ANAs were secured using epineurial suture (group 1) or PTB (group 2). Outcomes were assessed using sciatic function index (SFI), gastrocnemius muscle mass retention, and nerve histomorphometry. Historical controls from phase 1 were used to compare the performance of ANA with isograft. Statistical analysis was performed using analysis of variance and Bonferroni all-pairs comparison. Results All ANAs had signs of successful regeneration. Mean values for SFI, muscle mass retention, nerve fiber diameter, axon diameter, and myelin thickness were not significantly different between ANA + suture and ANA + PTB. On comparative analysis, ANA + suture performed significantly worse than isograft + suture from phase 1. However, ANA + PTB was statistically comparable to isograft + suture, the current standard of care. Conclusion Previously reported advantages of PTB versus suture appear to be reduced when applied to ANA. The lack of Schwann cells and neurotrophic factors may be responsible. PTB may improve ANA performance to an extent, where they are equivalent to autograft. This may have important clinical implications when injuries preclude the use of autograft.