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.

Simultaneous inferior alveolar nerve regeneration and osseointegration with a nerve growth factor-supplying implant: a preliminary study.

PURPOSE: Although nerve growth factor (NGF) has been proved to enhance inferior alveolar nerve (IAN) regeneration, its clinical application remains a challenging issue. This study investigated the functional regeneration of IAN injury by supplying NGF using an NGF-supplying implant and its effect on the osseointegration. MATERIALS AND METHODS: In canine IAN transection-and-repair models (n = 9), NGF-supplying implants connected to osmotic pumps were installed just above the transection site. In the right IAN, NGF 300 µg in phosphate buffered saline (PBS) 2 mL was loaded in the pump and pure PBS 2 mL was loaded in the left IAN. The gross clinical finding was evaluated by wound healing, inflammation, implant exposure, and loss of fixture. To evaluate IAN regeneration, electrophysiologic (amplitude, latency, conduction velocity, and peak voltage) and histomorphometric (axon count and density, myelin thickness, and ratio of axon diameter to fiber diameter) analyses were performed. Implant stability quotient, bone-to-implant contact ratio, and new bone area were measured to assess the osseointegration of the NGF-supplying implant. RESULTS: The conduction velocity (2.675 m/second) and peak voltage (1.940 µV) of the NGF group at 6 weeks were considerably higher than those of the PBS group (1.892 m/second and 1.300 µV, respectively). The same results were observed for axon count (NGF vs PBS, 4,576.107 ± 270.413 vs 3,606.972 ± 242.876), axon density (10,707.458 ± 638.835 vs 7,899.781 ± 1,063.625/mm(2)), and myelin thickness (1.670 ± 0.555 vs 1.173 ± 0.388 µm). There were no meaningful differences for the other parameters. CONCLUSIONS: Supplying NGF with specially designed dental implants can be a new therapeutic approach to enable IAN regeneration and osseointegration simultaneously.

Ocular effects of pegylated interferon-α in patients with chronic hepatitis B.

PURPOSE: To evaluate the early retinal changes and its reflection on the visual field examination in chronic hepatitis B (CHB) patients using pegylated interferon-α (PEGIFN-α) monotherapy. PATIENTS AND METHODS: Thirty eyes of fifteen patients with CHB were examined prospectively for changes in the fundus examination and visual field examination (both Humphrey Perimetry and Frequency Doubling Perimetry). The patients were examined before and in 3 months intervals after starting the PEGIFN-α treatment. The changes in the fundus examination were noted and the visual field examinations, retinal nerve fiber thickness, Schirmer scores and color vision before and at 3 months of the treatment were compared. The statistical evaluation was performed with paired-t test, using SPSS 16.0 Inc. (Chicago, IL). RESULTS: The mean age of the 15 patients (seven male, eight female) was 52.5 ± 12.4 years. There was no significant retinal change in none of the patients. Neither the visual field examination with Humphrey Field Analyzer nor the Frequency Doubling Perimetry results has demonstrated any significant change during 3 months follow-up. There was a statistically significant increase in the retinal nerve fiber layer (RNFL) thickness; while Schirmer test scores for dry eye assessment was significantly decreased. CONCLUSION: PEGIFN-α monotherapy, which is used for treatment of CHB, may cause some changes in the thickness of RNFL that may necessitate the close follow-up for further morphological changes of the optic disc in these patients.

Biofabrication of nerve fibers with mimetic myelin sheath-like structure and aligned fibrous niche.

Nerve tissues contain hierarchically ordered nerve fibers, while each of the nerve fibers has nano-oriented fibrous extracellular matrix and a core-shell structure of tubular myelin sheath with elongated axons encapsulated. Here, we report, for the first time, a ready approach to fabricate biomimetic nerve fibers which are oriented and have a core-shell structure to spatially encapsulate two types of cells, neurons and Schwann cells. A microfluidic system was designed and assembled, which contained a coaxial triple-channel chip and a stretching loading device. Alginate was used first to assist the fabrication, which was washed away afterwards. The orientation of the biomimetic nerve fibers was optimized by the control of the compositions of methacrylate hyaluronan and fibrin, together with the parameters of microfluidic shearing and external stretching. Also, neurons and Schwann cells, which were respectively located in the core and shell of the fibers, displayed advanced biologic functions, including neurogenesis and myelinating maturation. We demonstrate that the neural performance is relatively good, compared to that resulted from individually encapsulated in single-layer microfibers. The present study brings insights to fabricate biomimetic nerve fibers for their potential in neuroscience research and nerve regeneration. Moreover, the present methodology on the fabrication of oriented fibers with different types of cells separately encapsulated should be applicable to biomimetic constructions of various tissues.

Role of intracellular calcium in NI-35-evoked collapse of neuronal growth cones.

A myelin-associated protein from the central nervous system, the neurite growth inhibitor NI-35, inhibits regeneration of lesioned neuronal fiber tracts in vivo and growth of neurites in vitro. Growth cones of cultured rat dorsal root ganglion neurons arrested their growth and collapsed when exposed to liposomes containing NI-35. Before morphological changes, the concentration of free intracellular calcium ([Ca2+]i) showed a rapid and large increase in growth cones exposed to liposomes containing NI-35. Neither an increase in [Ca2+]i nor collapse of growth cones was detected in the presence of antibodies to NI-35. Dantrolene, an inhibitor of calcium release from caffeine-sensitive intracellular calcium stores, protected growth cones from collapse evoked by NI-35. Depletion of these caffeine-sensitive intracellular calcium stores prevented the increase in [Ca2+]i evoked by NI-35. The NI-35-evoked cascade of intracellular messengers that mediates collapse of growth cones includes the crucial step of calcium release from intracellular stores.

Adjunctive use of the non-ionic surfactant Poloxamer 188 improves fetal dopaminergic cell survival and reinnervation in a neural transplantation strategy for Parkinson's disease.

Although neural transplantation of fetal dopaminergic cells is a promising therapy for Parkinson's disease, poor transplanted cell survival limits its efficacy. In the present study it was hypothesized that the use of Poloxamer 188 (P188), a non-ionic surfactant, during cell preparation and transplantation may protect cells from associated mechanical injury and thus improve transplanted cell survival in a rat model of Parkinson's disease. Fetal rat dopaminergic tissue was dissociated in media with or without P188 and then cultured for 1 week or transplanted into the striatum of rats with unilateral 6-hydroxydopamine lesions of the nigrostriatal dopaminergic pathway. Fetal dopaminergic cell survival and reinnervation of the host brain were examined using tyrosine hydroxylase immunohistochemistry and stereological quantification. The number of surviving tyrosine hydroxylase-immunoreactive cells in vitro and in vivo was significantly increased by 2.2-fold by incubating fetal dopaminergic cells with P188 during tissue dissociation. Furthermore, the striatal reinnervation in parkinsonian rats that received intrastriatal transplants of P188-exposed dopaminergic cells was significantly enhanced (1.8-fold increase) compared with rats that received non-P188-treated cells. In conclusion, P188 protects fetal dopaminergic cells from mechanical injury by increasing cell survival and enhances dopaminergic fibre outgrowth into the transplanted striatum. Use of P188 may thus be an important adjunct to improve the clinical efficacy of neural transplantation for Parkinson's disease.

Qualitative Histologic Assessment vs. Geomorphometric Analysis of Nerve Fiber Shape after the Intraneural Application of Liposomal Bupivacaine / Evaluación Histológica Cualitativa Versus Análisis Geomorfométrico de la Forma de la Fibra Nerviosa Después de la Aplicación Intraneural de Bupivacaína Liposomal

SUMMARY: The preserved form of all components of the nerve fiber is a prerequisite for the proper conduction of the nerve impulse. various factors can change the shape of nerve fibers. In everyday practice, qualitative histological analysis is the gold standard for detecting changes in shape. Geometric morphometry is an innovative method that objectively enables the assessment of changes in nerve fibers' shape after local anesthetics action. A total of sixty sciatic nerves were used as material, which was intraneural injected with saline solution in the control group (n=30), and a solution of 1.33 % liposomal bupivacaine (n=30) in the test group. After the animals were sacrificed, nerve samples were taken and histological preparations were made. The preparations were first described and examined using a qualitative histological method, after which digital images were made. The images were entered into the MorphoJ program and processed using the method of geometric morphometry. Qualitative histological examination revealed no differences in nerve fibers after intraneurally applied physiological solution and liposomal bupivacaine. Using the method of geometric morphometry, a statistically significant change in the shape of axons was found after intraneurally applied saline solution and liposomal bupivacaine (p=0.0059). No significant differences in histological changes were found after the qualitative histological analysis of nerve fiber cross-section preparations. A statistically significant change in the shape of nerve fiber axons was observed after geometric morphometric analysis of digital images after intraneural application of saline and liposomal bupivacaine.

La forma conservada de todos los componentes de la fibra nerviosa es un requisito previo para la conducción correcta del impulso nervioso. Varios factores pueden cambiar la forma de las fibras nerviosas. En la práctica diaria, el análisis histológico cualitativo es el estándar de oro para detectar cambios de forma. La morfometría geométrica es un método innovador que permite evaluar objetivamente los cambios en la forma de las fibras nerviosas después de la acción de los anestésicos locales. Se utilizó como material un total de sesenta nervios ciáticos, que se inyectaron intraneuralmente con solución salina en el grupo control (n=30), y una solución de bupivacaína liposomal al 1,33 % (n=30) en el grupo de prueba. Después de sacrificados los animales, se tomaron muestras de nervios y se realizaron preparaciones histológicas. Primero se describieron y examinaron las preparaciones utilizando un método histológico cualitativo, después de lo cual se tomaron imágenes digitales. Las imágenes fueron ingresadas al programa MorphoJ y procesadas mediante el método de morfometría geométrica. El examen histológico cualitativo no reveló diferencias en las fibras nerviosas después de la aplicación intraneural de solución fisiológica y bupivacaína liposomal. Usando el método de morfometría geométrica, se encontró un cambio estadísticamente significativo en la forma de los axones después de la aplicación intraneural de solución salina y bupivacaína liposomal (p = 0,0059). No se encontraron diferencias significativas en los cambios histológicos después del análisis histológico cualitativo de las preparaciones de secciones transversales de fibras nerviosas. Se observó un cambio estadísticamente significativo en la forma de los axones de las fibras nerviosas después del análisis de morfometría geométrica de imágenes digitales después de la aplicación intraneural de solución salina y bupivacaína liposomal.

Neurotrophin-3 released from implant of tissue-engineered fibroin scaffolds inhibits inflammation, enhances nerve fiber regeneration, and improves motor function in canine spinal cord injury.

Spinal cord injury (SCI) normally results in cell death, scarring, cavitation, inhibitory molecules release, etc., which are regarded as a huge obstacle to reconnect the injured neuronal circuits because of the lack of effective stimulus. In this study, a functional gelatin sponge scaffold was used to inhibit local inflammation, enhance nerve fiber regeneration, and improve neural conduction in the canine. This scaffold had good porosity and modified with neurotrophin-3 (NT-3)/fibroin complex, which showed sustained release in vitro. After the scaffold was transplanted into canine spinal cord hemisection model, hindlimb movement, and neural conduction were improved evidently. Migrating host cells, newly formed neurons with associated synaptic structures together with functional blood vessels with intact endothelium in the regenerating tissue were identified. Taken together, the results demonstrated that using bioactive scaffold could establish effective microenvironment stimuli for endogenous regeneration, providing a potential and practical strategy for treatment of spinal cord injury. © 2018 The Authors Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2158-2170, 2018.

Linezolid-induced optic neuropathy: a mitochondrial disorder?

We report a case of bilateral mitochondrial optic neuropathies secondary to long-term linezolid treatment, show the nature of recovery, review the findings in the literature and propose a potential mitochondrial mechanism for linezolid-induced mitochondrial optic neuropathy. This is an observational case report and literature review with presentation of the clinical course of linezolid mitochondrial optic neuropathies through clinical and psychophysical documentation. Main outcome measures included: visual acuity, funduscopical examinations and peripapillary retinal nerve fibre layer (PRNFL) optical coherence tomography (OCT). A 6-year-old boy presented with bilateral optic neuropathies secondary to 1 year of linezolid treatment for osteomyelitis of the mandible. On presentation, visual acuities were 20/400 in both eyes, with considerable optic disc oedema, hyperaemia and PRNFL swelling confirmed by OCT. 2 weeks after the discontinuation of linezolid, visual acuities returned to 20/25 in both eyes, with reduction in the optic disc oedema, hyperaemia and PRNFL swelling. 3 months after the discontinuation of linezolid treatment, visual acuities were stable at 20/20 in both eyes, with a marked decrease in PRNFL swelling confirmed by OCT, and the development of mild temporal optic disc pallor in both eyes. Doctors should be aware of impairments of vision among patients on long-term linezolid treatment and promptly discontinue treatment to prevent irreversible vision loss. The development and resolution of bilateral optic neuropathies with considerable PRNFL swelling in this patient provide insight into the more general rubric of mitochondrial optic neuropathies.

Assessing the effects of three dental impression materials on the isolated sciatic nerve of rat and frog.

The effects on nerve tissue of three dental impression pastes were compared in this study. Two of the impression pastes, Examix and Express 3M, contained vinyl polysiloxane while the other, Xanthopren, did not. An in vitro model based on the isolated sciatic nerve of the frog and rat was used. As an indication of the proper functioning of the fibres in the nerve, the amplitude of evoked compound action potential (CAP) was monitored continuously. The results clearly showed that the number of active nerve fibres in the isolated sciatic nerves of either rat or frog exposed directly to impression pastes containing vinyl polysiloxane, decreased much faster than those of the nerves in contact to impression material without vinyl polysiloxane. When the nerve of the frog was exposed to Xanthopren there was a decrease in the CAP to 50% of the control values within 56.87+/-2.42 h (n=6). This value was called inhibition time to 50%, IT(50) and for Examix it was found to be 9.97+/-1.53 h. When the nerve of the rat was exposed to Xanthopren, the IT(50) was 15.34+/-2.97 h (n=6) for the Xanthopren and only 2.86+/-1.20 h for Examix and 2.76+/-0.48 h for Express 3M (n=6). There was no significant difference between the action of the last two compounds (P=0.85). This fast nerve fibre inactivation could be caused either by the chemical used for the synthesis of the two impression pastes, Examix and Express 3M, or by the unusual constriction of the nerve when it is embedded in the materials with vinyl polysiloxane. There is strong evidence to support the first case, since the incubation of the nerve in the presence of Examix, Express 3M and Xantopren in a way so the nerve was not in contact with the impression pastes, shows a much faster decrease of the CAP in the presence of the first two pastes. The decrease is caused by the death of nerve fibres, since there is no recovery in the CAP after the removal of Examix from the incubating saline.

A multi-walled silk fibroin/silk sericin nerve conduit coated with poly(lactic-co-glycolic acid) sheath for peripheral nerve regeneration.

The linearly oriented multi-walled silk fibroin/silk sericin (SF/SS) nerve conduits (NCs) can provide physical cues similar to native peripheral nerve fasciculi, but the mechanical properties of which are not excellent enough. In this study, NCs with a novel and bionic design with dual structures were developed. The important features of our NCs is that the internal skeleton (the multi-walled SF/SS conduits) has a bionic structure similar to the architecture of native peripheral nerve fasciculi, which is beneficial for nerve regeneration, and the outer sheath (the hollow poly(lactic-co-glycolic acid) [PLGA] conduits) could provide strong mechanical protection for the internal skeleton. The linearly oriented multi-walled SF/SS conduit was fabricated and inserted in the hollow PLGA sheath lumen and then used for the bridge across the sciatic nerve defect in rats. The outcome of the peripheral nerve repair post implantation was evaluated. The functional and morphological parameters were examined and showed that the novel PLGA-coated SF/SS NCs could promote peripheral nerve regeneration, approaching those elicited by nerve autografts that are the first candidate for repair of peripheral nerve defects. Thus, these updated NCs have potential usefulness to enhance functional recovery after repair of peripheral nerve defect.

Functional Recovery of Carbon Nanotube/Nafion Nanocomposite in Rat Model of Spinal Cord Injury.

BACKGROUND: Acute spinal cord injury (ASCI) can lead to paraplegia or quadriplegia, the treatment of which has been a major problem. New therapeutic approaches in developing carbon nanotubes (CNT) functionalized with the Nafion nanocomposite, a sulfonated tetrafluoroethylene copolymer, have been shown to increase the length of selected neurites in vitro. OBJECTIVE: We hypothesized that the administration of the CNT/Nafion nanocomposite after experimental SCI will promote regeneration of axons into the lesion cavity and the functional recovery of the hind limbs in a rat model. METHODS: To evaluate this hypothesis through this experimental research paper, transection SCI was induced at the T9-T10 vertebral level in adult female rats. One week after transection, the epicenter of the lesion was injected with 25 lL of vehicle (saline), or 1 lg/mL, 10 lg/mL, or 100 lg/mL of CNT/Nafion nanocomposite. Behavioral analysis was carried out by assessing tail flick, chronic pain or mechanical allodynia, motor coordination, and the results of the rotarod test performed pre- and post-surgery, on days 3, 7, 14, 21 and 28, using the tail flick analysis, Noldus CatWalk gait analysis, open-field locomotor test, and Rotarod test. At 28 days post-injection, the rats were euthanized and spinal cord tissue was extracted. RESULTS: We found that post-SCI, administration of the CNT/Nafion nanocomposite resulted in decreased lesion volume, increased neurofilament-positive fibers and corticospinal tract fibers in the lesion, and no increase in reactive gliosis (P < 0.001). Additionally, post-SCI administration of CNT/Nafion nanocomposite induced a modest improvement in hind limb locomotor recovery without inducing hyperalgesia. CONCLUSION: These data suggest that the CNT/Nafion nanocomposite may be an effective material to promote axonal repair and regeneration after SCI.

GABAergic control of action potential propagation along axonal branches of mammalian sensory neurons.

The main axons of mammalian sensory neurons are usually viewed as passive transmitters of sensory information. However, the spindle afferents of jaw-closing muscles behave as if action potential traffic along their central axons is phasically regulated during rhythmic jaw movements. In this paper, we used brainstem slices containing the cell bodies, stem axons, and central axons of these sensory afferents to show that GABA applied to the descending central (caudal) axon often abolished antidromic action potentials that were elicited by electrical stimulation of the tract containing the caudal axons of the recorded cells. This effect of GABA was most often not associated with a change in membrane potential of the soma and was still present in a calcium-free medium. It was mimicked by local applications of muscimol on the axons and was blocked by bath applications of picrotoxin, suggesting activation of GABA(A) receptors located on the descending axon. Antidromic action potentials could also be blocked by electrical stimulation of local interneurons, and this effect was prevented by bath application of picrotoxin, suggesting that it results from the activation of GABA(A) receptors after the release of endogenous GABA. We suggest that blockage is caused mainly by shunting within the caudal axon and that motor command circuits use this mechanism to disconnect the rostral and caudal compartments of the central axon, which allows the two parts of the neuron to perform different functions during movement.

Periosteum Metabolism and Nerve Fiber Positioning Depend on Interactions between Osteoblasts and Peripheral Innervation in Rat Mandible.

The sympathetic nervous system controls bone remodeling by regulating bone formation and resorption. How nerves and bone cells influence each other remains elusive. Here we modulated the content or activity of the neuropeptide Vasoactive Intestinal Peptide to investigate nerve-bone cell interplays in the mandible periosteum by assessing factors involved in nerve and bone behaviors. Young adult rats were chemically sympathectomized or treated with Vasoactive Intestinal Peptide or Vasoactive Intestinal Peptide10-28, a receptor antagonist. Sympathectomy depleted the osteogenic layer of the periosteum in neurotrophic proNerve Growth Factor and neurorepulsive semaphorin3a; sensory Calcitonin-Gene Related Peptide-positive fibers invaded this layer physiologically devoid of sensory fibers. In the periosteum non-osteogenic layer, sympathectomy activated mast cells to release mature Nerve Growth Factor while Calcitonin-Gene Related Peptide-positive fibers increased. Vasoactive Intestinal Peptide treatment reversed sympathectomy effects. Treating intact animals with Vasoactive Intestinal Peptide increased proNerve Growth Factor expression and stabilized mast cells. Vasoactive Intestinal Peptide10-28 treatment mimicked sympathectomy effects. Our data suggest that sympathetic Vasoactive Intestinal Peptide modulate the interactions between nervous fibers and bone cells by tuning expressions by osteogenic cells of factors responsible for mandible periosteum maintenance while osteogenic cells keep nervous fibers at a distance from the bone surface.

Evaluation of the anesthetic effect of epinephrine-free articaine and mepivacaine through quantitative sensory testing.

INTRODUCTION: Long lasting anesthesia of the soft tissue beyond the dental treatment affects patients in daily routine. Therefore a sophisticated local anesthesia is needed. The purpose of this study was an evaluation of the clinical use of epinephrine-free local anesthetic solutions in routine short-time dental treatments. MATERIALS AND METHODS: In a prospective, single-blind, non-randomized and controlled clinical trial, 31 patients (16 male, 15 female patients) undergoing short-time dental treatment under local anesthesia (plain solutions of articaine 4% and mepivacaine 3%) in area of maxillary canine were tested with quantitative sensory testing QST. Paired-Wilcoxon-testing (signed-rank-test) and Mc Nemar tests have been used for statistical results. RESULTS: Significant differences in all tested parameters to the time of measurements were found. Mepivacaine showed a significantly stronger impact for the whole period of measurement (128 min) on thermal and mechanical test parameters and to the associated nerve fibers. CONCLUSION: Plain articaine shows a faster onset of action associated with a shorter time of activity in comparison to plain mepivacaine. In addition to this articaine shows a significant low-graded effect on the tested nerve-fibers and therefore a least affected anesthesia to the patient. The clinical use of an epinephrine-free anesthetic solution can be stated as possible option in short dental routine treatments to the frequently used vasoconstrictor containing local anesthetics. Patients may benefit from shorter numbness.

Dexamethasone treatment reduces sensory neuropeptides and nerve sprouting reactions in injured teeth.

Dental injuries have been shown to generate extensive structural and cytochemical changes in sensory fibers that contain neuropeptides such as calcitonin gene-related peptide (CGRP) or substance P (SP). The present study was designed to test whether the anti-inflammatory drug dexamethasone (DEX) can alter neural responses to dental injuries. DEX (20 micrograms/100 g body weight) was given to adult rats (n = 10) prior to dental surgery and daily thereafter for 4 days. Control animals received sterile saline vehicle (n = 6) or no injection (n = 1). Each rat was then anesthetized for dental surgery and a cavity was drilled partway through dentin on the anterior side of the right maxillary first molar. Pulp exposure injuries were also made on two right mandibular molars in 14 of 17 rats. After 4 days of daily drug treatment, the rats were anesthetized and fixed by perfusion with formaldehyde-picric acid, and their jaws were prepared for immunocytochemistry. Neural CGRP immunoreactivity near the maxillary cavity injury site of DEX-treated rats was reduced more than 50% compared to controls, as determined both qualitatively and by digital analysis. The SP immunoreactive (IR) fibers in molar pulp also had extensive inhibition of neural reactions to cavity injury. DEX also reduced the immunoreactivity for CGRP and SP in normal contralateral rat molars of all treated rats, and it caused a postoperative loss of weight. Pretreatment for 1-5 days prior to the 4 day injury gave the same results as pretreatment for 1 h. The mandibular pulp exposure injuries induced a chronic abscess and advancing pulpal necrosis but did not show differences in nerve reactions between DEX-treated rats and the controls. In conclusion, the synthetic steroid dexamethasone suppressed the CGRP and SP neuropeptide immunoreactivity in normal dental nerves and it reduced nerve-sprouting responses to dentin cavity injuries; however, sensory nerve reactions to pulpal exposure injuries were not affected by DEX in these experiments.

Effects of neonatal sympathectomy and capsaicin treatment on bone remodeling in rats.

Bone metabolism may be influenced by the innervation of skeletal tissues. Neuropeptides such as vasoactive intestinal peptide, from sympathetic nerves, and calcitonin gene-related peptide, from sensory nerves, have been implicated as local modulators of bone metabolism. The effect of neonatal sympathectomy and of capsaicin-induced sensory denervation in rats was studied on the following: (i) the radial bone growth and apposition rate in tibiae (normal growth and modeling) and (ii) the percentage of periosteal surface of the mandible occupied by osteoclasts during induced remodeling. Neonate rats were treated with guanethidine, capsaicin, or appropriate vehicle. At seven weeks, maxillary molars were removed to induce remodeling on the buccal surface of the mandible. Animals were killed four days after surgery. Cross-sectional cortical area, medullary area, and periosteal apposition rate were measured by histomorphometry in ground sections of tibiae. The percentage of periosteal surface at the remodeling site occupied by osteoclasts (stained for acid phosphatase) was measured in frozen, undecalcified sections. There was no significant difference in cortical or medullary area or periosteal apposition rate in tibiae between each drug treatment and its control. However, the mandibular bone surface occupied by osteoclasts was increased 45.5% (P less than or equal to 0.005) in animals treated neonatally with guanethidine compared to controls. In contrast, the mandibular surface occupied by osteoclasts was decreased 21.2% (P less than or equal to 0.04) in animals treated neonatally with capsaicin compared to controls. The alteration of bone remodeling (osteoclast surface) by both treatments indicates that sensory and sympathetic nerves play a role in focal metabolism of bone.

Polyol metabolism of retrograde axonal transport in diabetic rat large optic nerve fiber.

PURPOSE: The role of the polyol pathway metabolism in progressive impairment of retrograde axonal transport was evaluated in the optic nerve of rats with streptozotocin-induced diabetes. METHODS: Rats with streptozotocin-induced diabetes received a low (3 mg/kg body weight) or high dose (10 mg/kg body weight) of oral aldose reductase inhibitor (ARI). At 1 and 3 months after induction of diabetes, Fluoro-Gold (FG, Chemicon, Temecula, CA) was injected into the dorsal lateral geniculate nucleus. Percentages of FG-labeled large, medium, and small retinal ganglion cells (RGCs) per total population were calculated in the retinas of ARI-treated diabetic, untreated diabetic, and normal control rats. RESULTS: Mean percentages of FG-labeled large RGCs per total population were significantly decreased in nontreated diabetic rats compared with control animals at 1 month of induced diabetes. This decrease in FG labeling was not observed in both the low- and high-dose ARI-treated diabetic rats. At 3 months of induced diabetes, FG labeling of both large and medium RGCs was significantly decreased. This decrease was completely ameliorated by high-dose ARI treatment. CONCLUSIONS: These results indicate that diabetes affects retrograde axonal transport progressively through selective impairment of RGCs and that the polyol pathway metabolism is involved in such impairment.

Transmitter-loaded polymeric microspheres induce regrowth of dopaminergic nerve terminals in striata of rats with 6-OH-DA induced parkinsonism.

Biodegradable controlled-release microspheres made with the biocompatible biodegradable polyester excipient poly (DL-lactide-co-glycolide) represent a new technology for drug delivery to the central nervous system (CNS). A suspension of 3 microliters of dopamine (DA) or NE containing microspheres, or empty microspheres, was implanted in 2 sites of the DA denervated striatum of rats previously unilaterally lesioned with 6-OH-DA. Contralateral rotational behavior induced by apomorphine was used as an index of lesion success. Following implantation of the microspheres, rotational behavior was also used as an index of functional recovery. Both DA and NE microsphere implanted rats displayed a 30-50% reduction in the number of apomorphine induced rotations up to 12 weeks post implantation. Empty microspheres caused no changes in rotational behavior. Implantation of a mixture of DA/NE microspheres resulted in an 80% decrease in the number of apomorphine induced rotations, registered up to 4 weeks. Immunocytochemical examination revealed growth of DA and tyrosine hydroxylase immunoreactive fibers in the striatum of DA and NE microsphere implanted rats. Interestingly, functional behavior correlated with the degree of fiber ingrowth. This method to deliver substances to the CNS will be tested for therapeutic usefulness in patients with Parkinson's disease, in a recently approved clinical trial in Göteborg.

An in vitro temporomandibular joint-nerve preparation for pain study in rats.

A novel in vitro TMJ-nerve preparation was developed to quantitatively study peripheral sensory mechanisms of temporomandibular joint (TMJ). The TMJ region on one side (including mandibular head, disc, retrodiscal tissue and mandibular fossa) of adult Wistar albino rats was excised together with the auriculo-temporal nerve. The block was preserved in a modified Krebs-Henseleit solution saturated with O(2)/CO(2) (95/5%) gas mixture. Using a calibrated von Frey type apparatus, mechanical noxious stimulation was applied directly to various sites within the TMJ region. In addition, thermal and chemical noxious stimuli were also attempted. Stable recordings of single unit activities from the auriculo-temporal nerve could be obtained for as long as 5 h, which was sufficient to analyze the response properties of the TMJ units to various stimuli. This new preparation would be useful for investigating TMJ peripheral sensory mechanisms, especially pain, and potentially makes it possible to reveal neural mechanisms of temporomandibular arthralgia, a syndrome that has recently shown an increased incidence in clinical dentistry.