BACKGROUND AND OBJECTIVES: The object of this study was to evaluate the effect of platelet rich plasma (PRP) on facial nerve regeneration from an axotomy injury in the guinea pig model. MATERIALS AND METHOD: Experiments involved the transection and repair of right facial nerve. The right facial nerve of 14 albino guinea pigs were completely transected and immediately sutured, followed by fibrin glue only (control group) or fibrin glue +PRP (PRP group). Western blot assay was used to detect neurotrophic factors secreted by PRP. Nerve regeneration was assessed by motor function, electrophysiology, and histology studies. RESULTS: High levels of neurotrophin-3, angiopoietin-1, glial cell line derived neurotrophic factors, nerve growth factors and brain derived neurotrophic factors were demonstrated in PRP. Motor function recovery, compound motor action potentials, and axon count showed significant improvement in guinea pig treated with PRP. CONCLUSION: There was an improved functional outcome with the use of PRP in comparison with control. The increased nerve regeneration found in this study may be due to the neurotrophic factors secreted by PRP.
Action Potentials ; Angiopoietin-1 ; Animals ; Axons ; Axotomy ; Blood Platelets ; Blotting, Western ; Brain-Derived Neurotrophic Factor ; Electrophysiology ; Facial Nerve ; Fibrin Tissue Adhesive ; Glial Cell Line-Derived Neurotrophic Factor ; Glial Cell Line-Derived Neurotrophic Factors ; Guinea Pigs ; Nerve Growth Factor ; Nerve Growth Factors ; Nerve Regeneration ; Platelet-Rich Plasma ; Recovery of Function ; Regeneration

Parkinson's disease is the second most common neurodegenerative disorder characterized by the progressive degeneration of dopaminergic neurons and a biochemical reduction of striatal dopamine levels. Despite the lack of fully understanding of the etiology of Parkinson's disease, accumulating evidences suggest that Parkinson's disease may be caused by the insufficient support of neurotrophic factors, and by microglial activation, resident immune cells in the brain. Naringin, a major flavonone glycoside in grapefruits and citrus fruits, is considered as a protective agent against neurodegenerative diseases because it can induce not only anti-oxidant effects but also neuroprotective effects by the activation of anti-apoptotic pathways and the induction of neurotrophic factors such as brain-derived neurotrophic factor and vascular endothelial growth factor. We have recently reported that naringin has neuroprotective effects in a neurotoxin model of Parkinson's disease. Our observations show that intraperitoneal injection of naringin induces increases in glial cell line-derived neurotrophic factor expression and mammalian target of rapamycin complex 1 activity in dopaminergic neurons of rat brains with anti-inflammatory effects. Moreover, the production of glial cell line-derived neurotrophic factor by naringin treatment contributes to the protection of the nigrostriatal dopaminergic projection in a neurotoxin model of Parkinson's disease. Although the effects of naringin on the nigrostriatal dopaminergic system in human brains are largely unknown, these results suggest that naringin may be a beneficial natural product for the prevention of dopaminergic degeneration in the adult brain.
Adult ; Animals ; Antioxidants ; Brain ; Brain-Derived Neurotrophic Factor ; Citrus ; Citrus paradisi ; Dopamine ; Dopaminergic Neurons ; Glial Cell Line-Derived Neurotrophic Factor ; Humans ; Injections, Intraperitoneal ; Nerve Growth Factors ; Neurodegenerative Diseases ; Neuroprotective Agents ; Parkinson Disease ; Rats ; Sirolimus ; Vascular Endothelial Growth Factor A

OBJECTIVE: Cellular diversity in the mammalian central nervous system is originated from precursor cells present in the neural ectoderm. The multipotent neural stem cells(NSCs) rapidly proliferate to give rise to transiently dividing progenitors that eventually differentiate into several cell types of neural cells. The authors investigate whether NSCs could differentiate neurons and glia and express neurotrophic factor. METHODS: To establish human neural cell lines, we isolated neural stem cells from human fetal telencephalon. Secondly, to investigate the expression of neurotrophic factor, basic fibroblast growth factor(bFGF), brain-derived neurotrophic factor(BDNF) and glial derived neurotrophic factor(GDNF) in rat and human cell, mRNA expressions of bFGF, BDNF and GDNF were detected by the reverse transcripted polymerase chain reaction(RT-PCR) analysis. RESULTS: In the NSCs cultures of embryonic rat striata and human fetal telencephalon, we demonstrated that bFGF induces the proliferation of stem cell, which give rise to spheres of undifferentiated cell that generate neurons and glia. Also, neurotrophic factor transcripts were identified using PCR in rat and human NSCs. CONCLUSION: These results demonstrate that human NSCs derived from human fetal telencephalon could differentiate neurons and glia and express neurotrophic factors. Therefore, NSCs may be an important key for the therapeutic application of neurotrophic factors.
Animals ; Brain-Derived Neurotrophic Factor ; Cell Line ; Central Nervous System ; Ectoderm ; Fibroblasts ; Glial Cell Line-Derived Neurotrophic Factor ; Humans* ; Nerve Growth Factors* ; Neural Stem Cells* ; Neuroglia ; Neurons ; Polymerase Chain Reaction ; Rats ; RNA, Messenger* ; Stem Cells ; Telencephalon*

STUDY DESIGN: Experimental, prospective study OBJECTIVES: To examine the changes in the variable factors after an acute spinal cord injury(SCI) in rats and dogs simultaneously. SUMMARY OF LITERATURE REVIEW: No study has examined the variations of several factors in a SCI model in different species. MATERIALS AND METHODS: In rats, a laminectomy was performed at the T10 level and the injured spinal cord was extracted. In Beagle dogs, the laminectomy level was T10 and T11. The motor function was evaluated using a modified Tarlov's scale. A RT2 profiler PCR array was used to examine each factor (inflammatory cytokines, factors-related with apoptosis, neurotrophic factors, factors-related with extraceullar matrix). RESULTS: IL-2, TNF, TNFRSF11B increased with time and showed no statistical difference between two species, but TNFSF13B showed a significant difference. BDNF decreased with time in both species, and GDNF was significantly lower in dogs. NGFbeta, CTNF and its receptors showed no significant changes in the two species. MMP1 increased in both species but MMP7 decreased in rats and increased in dogs with time, and showed a significant difference between species. CONCLUSION: The change in inflammatory cytokines and extracellular matrix correlates with each factor in the combined patterns. Moreover, during the first week after SCI, inflammatory cytokines, apoptosis, neutrophic factors, and extracellular matrix factors may show a partial difference between experimental animals, which means that an animal model can be selected according to the particular experimental plan.
Animals ; Apoptosis ; Brain-Derived Neurotrophic Factor ; Cytokines ; Dogs ; Extracellular Matrix ; Glial Cell Line-Derived Neurotrophic Factor ; Interleukin-2 ; Laminectomy ; Models, Animal ; Nerve Growth Factors ; Polymerase Chain Reaction ; Prospective Studies ; Rats ; Spinal Cord ; Spinal Cord Injuries

OBJECTIVETo investigate the effects of hyperbaric oxygen (HBO) treatment on the activation of astrocytes and the expression of glia-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) in the brain after traumatic brain injury (TBI).

METHODS54 male SD rats were randomly divided into three groups (n = 18): sham-operated, TBI and HBO treatment groups. TBI was induced with Feeney's method, bone window was opened without strike on the brain tissue in the sham-operated group. HBO group rats received HBO treatment for 60 min in the hyperbaric chamber containing O2 100% at 3 ATA. When neurological functions were measured 48 h after TBI, rats were decapitated, the brain water content of 18 rats was measured, 18 brains were sliced for the morphological observation after Nissl staining and for the immunohistochemistry staining of astrocyte markers glial fibrillary acidic protein (GFAP), vimentin and S100, and the other 18 brains of injured side were used for Western blot analysis of GDNF and NGF.

RESULTSHBO treatment reduced the neurological deficit, brain water content and hippocampal neuronal loss. In the observed cortex and hippocampal area astrocytes were activated, the cell number of positive expression of astrocyte markers GFAP, vimentin and S100 was increased, and the expression of GDNF and NGF was elevated after TBI. However, these indices were all enhanced further after the HBO treatment.

CONCLUSIONIt is suggested that HBO may be an effective therapy for TBI and upregulation of the expression of GDNF and NGF may underly the effect of HBO.

Animals ; Astrocytes ; metabolism ; Brain Injuries ; metabolism ; therapy ; Disease Models, Animal ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Glial Fibrillary Acidic Protein ; metabolism ; Hyperbaric Oxygenation ; methods ; Male ; Nerve Growth Factors ; metabolism ; Rats ; Rats, Sprague-Dawley ; S100 Proteins ; metabolism ; Vimentin ; metabolism

Advances in the neurobiology of growth factors, neural development, and prevention of cell death have resulted in a heightened clinical interest for the development of protective and regenerative neuromodulatory strategies for the cavernous nerves (CNs), as therapies for prostate cancer and other pelvic malignancies often result in neuronal damage and debilitating loss of sexual function. Nitric oxide released from the axonal end plates of these nerves within the corpora cavernosa causes relaxation of smooth muscle, initiating the haemodynamic changes of penile erection as well as contributing to maintained tumescence; the loss of CN function is primarily responsible for the development of erectile dysfunction (ED) after pelvic surgery and serves as the primary target for potential neuroprotective or regenerative strategies. Evidence from pre-clinical studies for select neuromodulatory approaches is reviewed, including neurotrophins, glial cell line-derived neurotrophic factors (GDNF), bone morphogenic proteins, immunophilin ligands, erythropoetin (EPO), and stem cells.
Animals ; Bone Morphogenetic Proteins ; therapeutic use ; Brain-Derived Neurotrophic Factor ; therapeutic use ; Erectile Dysfunction ; drug therapy ; etiology ; therapy ; Erythropoietin ; therapeutic use ; Glial Cell Line-Derived Neurotrophic Factor ; therapeutic use ; Growth Differentiation Factor 5 ; Humans ; Immunophilins ; Male ; Nerve Growth Factors ; therapeutic use ; Neurotransmitter Agents ; therapeutic use ; Penis ; innervation ; Peripheral Nerve Injuries ; Postoperative Complications ; Stem Cell Transplantation

OBJECTIVE: To evaluate the changes of GDNF after severe brain injury. METHODS: Changes GDNF in cortex, thalamus and pontine was observed by using immunohistochemistry and image analysis technique. RESULTS: The lever of GDNF was increasing at 1 d, reaching peak at 3 d, not obviously decreasing at 5 d and still higher than normal at 7 d after severe brain injury. CONCLUSION As an objective indication, the time sequence regularity of GDNF after brain injury may be applied in brain injury time estimation.
Animals ; Biomarkers ; Brain/metabolism* ; Brain Injuries/metabolism* ; Female ; Forensic Medicine ; Glial Cell Line-Derived Neurotrophic Factor ; Image Processing, Computer-Assisted ; Male ; Nerve Growth Factors/genetics* ; Rats ; Rats, Sprague-Dawley ; Time Factors

PURPOSE OF STUDY: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. MATERIALS AND METHODS: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with beta-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells(1x10(6)) or BDNF-Ad infected Schwann cells(1x10(6)) were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. RESULTS: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were 1.54+/-4.0*10(6) and 9.66+/-9.6*10(6). 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell 0.69 microgram/microliter of DNA was detected and in BDNF-Adenovirus transfected Schwann cell 0.795 microgram/microliter of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. CONCLUSION: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.
Adenoviridae ; Animals ; beta-Galactosidase ; Brain-Derived Neurotrophic Factor* ; Calcium* ; Cell Adhesion ; Cell Culture Techniques ; Cytarabine ; DNA ; DNA, Complementary ; Fibroblasts ; Gait ; Ganglia, Spinal ; Gene Library ; Genetic Therapy ; Genetic Vectors ; Glial Cell Line-Derived Neurotrophic Factor ; Humans ; Kidney ; Micropore Filters ; Nerve Growth Factors ; Nerve Regeneration ; Neurons ; Peripheral Nerves ; Polymerase Chain Reaction ; Rats* ; Regeneration* ; RNA, Messenger ; Schwann Cells ; Sciatic Nerve*

Vectors used to carry foreign genes play an important role in gene therapy, among which, the adeno-associated virus (AAV) has many advantages, such as nonpathogenicity, low immunogenicity, stable and long-term expression and multiple-tissue-type infection, etc. These advantages have made AAV one of the most potential vectors in gene therapy, and widely used in many clinical researches, for example, Parkinson's disease. This paper introduces the biological characteristics of AAV and the latest research progress of AAV carrying neurotrophic factor, dopamine synthesis related enzymes and glutamic acid decarboxylase gene in the gene therapy of Parkinson's disease.
Animals ; Aromatic-L-Amino-Acid Decarboxylases ; genetics ; Dependovirus ; genetics ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; Glutamate Decarboxylase ; genetics ; Humans ; Nerve Growth Factors ; genetics ; Neurturin ; genetics ; Parkinson Disease ; therapy

BACKGROUND AND OBJECTIVES: Sensorineural hearing loss is caused by ototoxic drugs, radiation therapy, noise exposure and trauma, etc. They make irreversible changes in cochlear hair cells and degeneration of spiral ganglion neurons. It is known that neurotrophins and other growth factors have an important role in protectingcochlear hair cells and spiral ganglion neurons. We designed this study to analyze the effect of neurotrophins and growth factors delivered to the inner ear of deafened guinea pig. MATERIALS AND METHOD: Healthy 15 guinea pigs with normal Preyer's reflex were chosen, and were made deaf by infusion of kanamycin and ethacrynic acid. Myringotomy was done to both ears, normal saline injected into the left ear, and BDNF (brainderived neurotrophic factor), GDNF (glial cell derived neurotrophic factor), NT-3 (neurotrophin-3), IGF (insulin-like growth factor), EGF (epidermal growth factor), FGF (fibroblast growth factor) were injected in the right ear. RESULTS: Statistically significant hearing gain was obtained up to 35.00+/-13.78 dB in the group 1 (BDNF, GDNF, NT-3 treated group), and up to 34.0+/-5.47 dB in the group 2 (IGF treated group). However, no statistically significant hearing gain was observed in the group 3 (EGF, FGF treated group). CONCLUSION: We observed statistically important improvement of hearing threshold in the BDNF, GDNF, NT-3 treated group and IGF treated group.
Animals ; Brain-Derived Neurotrophic Factor ; Ear ; Ear, Inner ; Epidermal Growth Factor ; Ethacrynic Acid ; Glial Cell Line-Derived Neurotrophic Factor ; Guinea ; Guinea Pigs ; Hair ; Hearing ; Hearing Loss, Sensorineural ; Intercellular Signaling Peptides and Proteins ; Kanamycin ; Models, Animal ; Nerve Growth Factors ; Neurons ; Noise ; Reflex ; Spiral Ganglion