The success of regeneration attempt is based on an ideal combination of stem cells, scaffolding and growth factors. Tissue constructs help to maintain stem cells in a required area for a desired time. There is a need for easily obtainable cells, potentially autologous stem cells and a biologically acceptable scaffold for use in humans in different difficult situations. This study aims to address these issues utilizing a unique combination of stem cells from gingiva and a hydrogel scaffold, based on a natural product for regenerative application. Human gingival mesenchymal stem cells (HGMSCs) were, with due induction, differentiated to neuronal lineages to overcome the problems associated with birth tissue-related stem cells. The differentiation potential of neuronal lineages was confirmed with suitable specific markers. The properties of mesenchymal stem cells in encapsulated form were observed to be similar to free cells. The encapsulated cells (3D) were then subjected to differentiation into neuronal lineages with suitable inducers, and the morphology and gene expression of transient cells were analyzed. HGMSCs was differentiated into neuronal lineages as both free and encapsulated forms without any significant differences. The presence of Nissl bodies and the neurite outgrowth confirm the differentiation. The advantages of this new combination appear to make it a promising tissue construct for translational application.
Gene Expression ; Gingiva ; Humans* ; Hydrogel* ; Intercellular Signaling Peptides and Proteins ; Mesenchymal Stromal Cells* ; Neurites ; Neurons* ; Nissl Bodies ; Parturition ; Regeneration ; Stem Cells

The development and differentiation of cells in the spinal ganglion were studied by electron microscopy in human fetuses ranging from 12 mm to 260 mm crown rump length. At 12 mm embryo the primitive neuroblasts which had a single process, contained a large numbers of free ribosome and mitochondria but very little rough endoplasmic reticulum. At 30 mm fetus, the primitive spinal ganglion consisted of bipolar neuroblasts, satellite cells and undifferentiated cells. Spindle-shaped bipolar neuroblasts formed spinal ganglion of loosely grouped cells at 50 mm fetus. Two neuroblast cell types, a small cell contained large clumps of rough endoplasmic reticulum at periphery, could be distinguished. At 80 mm fetus, the spinal ganglion constituted of bipolar neuroblast with apparently random distribution of small and large neurons with processes, together with satellite cells and blood vessels. The presences of a large numbers of neurotubules in the Golgi-central region were one of the first sign of further maturation of the neuroblast. During next prenatal stage from 120 mm on fetus, the ganglion cells were large and contained much rough endoplasmic reticulum, neurotubules and extensive Golgi complex. A large number of neuroblasts became transformed into unipolar cells from 180 mm to 260 mm feuts. Nissl bodies appeared during this stage. The ganglion-satellite cell boundary became complicated with increasing age, then enlarging in parallel with the increase in volume of the nerve cell. During next prenatal stage up to 180 mm fetus, the unipolar ganglion cell increased in number and size, and the cytoplsm contained all intracytoplasmic structures which were also found in mature spinal ganglion except for large pigment granules.
Blood Vessels ; Crown-Rump Length ; Embryonic Structures ; Endoplasmic Reticulum, Rough ; Fetus* ; Ganglia, Spinal* ; Ganglion Cysts ; Golgi Apparatus ; Humans* ; Microscopy, Electron ; Mitochondria ; Neurons ; Nissl Bodies ; Ribosomes

Cerebrospinal fluid (CSF) contains several molecules which are essential for neurogenesis. Human dental pulp stem cells (hDPSCs) are putatively neural crest cell-derived that can differentiate into neurons and glial cells under appropriate neurotrophic factors. The aim of this study was to induce differentiation of hDPSCs into neuroglial phenotypes using retinoic acid (RA) and CSF. The hDPSCs from an impacted third molar were isolated by mechanical and digestion and cultured. The cells have treated by 10⁻⁷µM RA (RA group) for 8 days, 10% CSF (CSF group) for 8 days and RA with CSF for 8 days (RA/CSF group). Nestin, microtubule-associated protein 2 (MAP2), and glial fibrillary acidic protein immunostaining were used to examine the differentiated cells. Axonal outgrowth was detected using Bielschowsky's silver impregnation method and Nissl bodies were stained in differentiated cells by Cresyl violet. The morphology of differentiated cells in treated groups was significantly changed after 3–5 days. The results of immunocytochemistry showed the presence of neuroprogenitor marker nestin was seen in all groups. However, the high percentage of nestin positive cells and MAP2, as mature neural markers, were observed at the pre-induction and induction stage, respectively. Nissl bodies were detected as dark-blue particles in the cytoplasm of treated cells. Our findings showed the RA as pre-inducer and CSF as inducer for using in vitro differentiation of neuron-like cells and neuroglial cells from hDPSCs.
Axons ; Cerebrospinal Fluid* ; Cytoplasm ; Dental Pulp* ; Digestion ; Glial Fibrillary Acidic Protein ; Humans* ; Immunohistochemistry ; In Vitro Techniques ; Methods ; Microtubule-Associated Proteins ; Molar, Third ; Nerve Growth Factors ; Nestin ; Neural Crest ; Neurogenesis ; Neuroglia* ; Neurons ; Nissl Bodies ; Phenotype ; Silver ; Stem Cells* ; Tretinoin* ; Viola

OBJECTIVETo study the effect of erythropoietin (EPO) on the activities of antioxidant enzymes, namely catalase (CAT) and glutathione peroxidase (GSH-Px) in the brain tissues of aged rats.

METHODSThirty SD rats were randomly divided into normal control, aging model, and recombinant human erythropoietin (rhEPO) treatment groups (n=10). Morris water maze was used to compare the behavioral indexes. The rats were then sacrificed to observe Nissl bodies in the hippocampal neurons with Nissl staining and test the activities of CAT and GSH-Px in the brain tissues.

RESULTSCompared with the control group, the aging rats showed significantly deteriorated learning and memory abilities (P<0.05), which were improved obviously by rhEPO treatment (P<0.05). The number of Nissl bodies in the neurons was reduced in the aging rats compared with that in the control group, and rhEPO treatment increased the number of Nissle bodies but failed to restore the control level. The aging rats also showed significantly lowered activities of CAT and GSH-Px in the brain tissue (P<0.05), which were increased significantly after rhEPO treatment (P<0.05).

CONCLUSIONEPO can enhance the activities of the antioxidant enzymes in the brain tissues of aged rats to increase the antioxidant capacity and produces an anti-aging effect.

Aging ; Animals ; Brain ; enzymology ; Catalase ; metabolism ; Epoetin Alfa ; Erythropoietin ; pharmacology ; Glutathione Peroxidase ; metabolism ; Learning ; drug effects ; Male ; Memory ; drug effects ; Nissl Bodies ; drug effects ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacology

AIMTo study the effects of noise on event-related potential(ERP) and its mechanism in hippocampus in rats.

METHODSMale SD rats were divided into 2 groups: control group (CG) and noise exposure group(NG). The rats in NG were exposed to white noise 105 dB SPL for 2.5 h/d x 20 d. P300 were recorded at parietal bone in rats. The Nissl body, NMDAR2B and [Ca2+]i of neurons in hippocampus were analyzed.

RESULTSThe peak latency (PL) of ERP P3a, P3 and P3b in NG were significantly longer than that in CG in the 14th and 20th exposure day. The amount of Nissl body in dentate gyrus (DG) and CA1 region and NMDAR2B in DG, CA1 and CA3 region of hippocampus of NG were significantly decreased than those of CG as well, while the concentration of Ca2+ in neurons increased markedly in NG.

CONCLUSIONDecreased Nissl body and NMDAR2B and increased [Ca2+]i in hippocampus in long-term noise exposed rats might cause the change of ERP P300.

Animals ; Calcium ; metabolism ; Environmental Exposure ; adverse effects ; Event-Related Potentials, P300 ; physiology ; Hippocampus ; metabolism ; physiology ; Male ; Neurons ; metabolism ; physiology ; Nissl Bodies ; metabolism ; Noise ; adverse effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism

The development of the cardiac ganglion was studied by electron microscopy in human fetuses ranging from 30mm to 270mm crown rump length. At 40mm fetus, the cardiac ganglia were observed in the adventitia of both the aorta and pulmonary artery, superior aspect of the left and right atrium, and interatrial septum. The cardiac ganglia were comprised of clusters of undifferentiated cells, neuroblasts, and unmyelinated nerve fibers. The ganglia were small and uncapsulated until 70mm fetus. At 70mm fetus, the cardic ganglia consisted of neuroblasts, satellite cells, and unmyelinated nerve fibers. Each ganglion was ensheathed in a connective tissue capsule. The cytoplasm of neuroblast contained Nissl bodies, mitochondria, coated vesicles, extensive Golgicomplex, and rough endoplasmic reticulum. Synaptic contacts between the cholinergic preganglionic axon and dendrites of postganglionic neuron were first observed. At 100mm fetus, the cardiac ganglia consisted of small clusters of ganglion cells and dendrites, together with supporting elements and blood vessels. During next prenatal stage from 170mm fetus, the ganglion cells were large and each contained a large nucleus with one or more nucleoli. The cytoplasm of ganglion cells contained much rough endoplasmic reticulum and extensive Golgi complex. Cholinergic preganglionic axons were numerous and interposed between the satellite cells. Adrenergic axons were rarely observed. A great number of synaptic junctions between the cholinergic preganglionic axon terminals and the dendrites of postganglinic neuron were found, and a few axosomatic synapses were also observed. Adrenergic nerve terminals did not seem to be involved in the synaptic transmission. The cardiac ganglion cells of the human fetal heart were innervated only by cholinergic nerve.
Adventitia ; Aorta ; Axons ; Blood Vessels ; Coated Vesicles ; Connective Tissue ; Crown-Rump Length ; Cytoplasm ; Dendrites ; Endoplasmic Reticulum, Rough ; Fetal Heart ; Fetus* ; Ganglia ; Ganglion Cysts* ; Golgi Apparatus ; Heart Atria ; Humans* ; Microscopy, Electron ; Mitochondria ; Nerve Fibers, Unmyelinated ; Neurons ; Nissl Bodies ; Presynaptic Terminals ; Pulmonary Artery ; Synapses ; Synaptic Transmission

OBJECTIVE: To estimate the detrimental effects of shortwave exposure on rat hippocampal structure and function and explore the underlying mechanisms. METHODS: One hundred Wistar rats were randomly divided into four groups (25 rats per group) and exposed to 27 MHz continuous shortwave at a power density of 5, 10, or 30 mW/cm2 for 6 min once only or underwent sham exposure for the control. The spatial learning and memory, electroencephalogram (EEG), hippocampal structure and Nissl bodies were analysed. Furthermore, the expressions of N-methyl-D-aspartate receptor (NMDAR) subunits (NR1, NR2A, and NR2B), cAMP responsive element-binding protein (CREB) and phosphorylated CREB (p-CREB) in hippocampal tissue were analysed on 1, 7, and 14 days after exposure. RESULTS: The rats in the 10 and 30 mW/cm2 groups had poor learning and memory, disrupted EEG oscillations, and injured hippocampal structures, including hippocampal neurons degeneration, mitochondria cavitation and blood capillaries swelling. The Nissl body content was also reduced in the exposure groups. Moreover, the hippocampal tissue in the 30 mW/cm2 group had increased expressions of NR2A and NR2B and decreased levels of CREB and p-CREB. CONCLUSION Shortwave exposure (27 MHz, with an average power density of 10 and 30 mW/cm2) impaired rats' spatial learning and memory and caused a series of dose-dependent pathophysiological changes. Moreover, NMDAR-related CREB pathway suppression might be involved in shortwave-induced structural and functional impairments in the rat hippocampus.
Animals ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Dose-Response Relationship, Radiation ; Electroencephalography ; radiation effects ; Hippocampus ; radiation effects ; Male ; Memory ; radiation effects ; Nissl Bodies ; physiology ; radiation effects ; Radio Waves ; adverse effects ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, N-Methyl-D-Aspartate ; genetics ; metabolism ; Spatial Learning ; radiation effects