Effect of intranasal administration of NeuroEPO in the histological structure of the olfactory mucosa of Wistar rats / Efecto de la administración intranasal de NeuroEPO en la estructura histológica de la mucosa olfatoria de ratas Wistar

Introduction: Strokes and neurodegenerative diseases are major global health problems, not only because they cause high mortality and disability, but due to the lack of effective therapies. NeuroEPO, a variant of recombinant human erythropoietin (rHu-EPO) with a low sialic acid content, has shown encouraging results as a potential neuroprotective agent when administered intranasally. Objective: To determine the effect of intranasal administration of NeuroEPO on the histological structure of the olfactory mucosa of Wistar rats. Materials and Methods: An experimental, prospective, and longitudinal study was conducted on Wistar rats. Ten healthy animals were randomly distributed into two groups of five each. The control group received a vehicle (0.3 μl/g/day) and the treated group received NeuroEPO (300 μg/kg/day). Both treatments were administered intranasally for 28 days. The histological characteristics of the olfactory mucosa were evaluated. The medians between the study groups were compared using the Mann-Whitney U test. Results: There were no alterations in the histological characteristics of the olfactory epithelium. However, slight hypertrophy and hyperplasia of the Bowman's glands were observed at the level of the lamina propria in the group treated with NeuroEPO. Conclusions: The administration of the nasal formulation of NeuroEPO did not induce histological alterations of the olfactory mucosa of Wistar rats under the experimental conditions of this research.

Introducción: Los accidentes cerebrovasculares y las enfermedades neurodegenerativas constituyen un importante problema de salud mundial. No solo porque causan una alta mortalidad y discapacidad, sino por la falta de terapias eficaces para tratarlos. La NeuroEPO, una variante de la eritropoyetina humana recombinante (rHu-EPO) con bajo contenido en ácido siálico, ha mostrado resultados alentadores como potencial agente neuroprotector al ser administrada por vía intranasal. Objetivo: Determinar el efecto de la administración intranasal de NeuroEPO en la estructura histológica de la mucosa olfatoria de ratas Wistar. Materiales y Métodos: Se realizó un estudio experimental, prospectivo y de corte longitudinal en ratas Wistar. Se utilizaron diez animales sanos distribuidos aleatoriamente en dos grupos de cinco cada uno. El grupo control recibió vehículo (0,3 μl/g/día) y el grupo tratado recibió NeuroEPO (300 μg/kg/día). Ambos tratamientos fueron administrados por vía intranasal durante 28 días. Fueron evaluadas las características histológicas de la mucosa olfatoria. Las medianas de los grupos del estudio fueron comparadas mediante la prueba U de Mann-Whitney. Resultados: No se evidenciaron alteraciones en las características histológicas del epitelio olfatorio. Sin embargo, a nivel de la lámina propia en el grupo tratado con NeuroEPO, se observó una ligera hipertrofia e hiperplasia de las glándulas de Bowman. Conclusiones: La administración de la formulación nasal de NeuroEPO no indujo alteraciones histopatológicas de la mucosa olfatoria de ratas Wistar en las condiciones experimentales de esta investigación.

Biopsy of the olfactory epithelium from the superior nasal septum: is it possible to obtain neurons without damaging olfaction? / Biópsia do epitélio olfatório do septo nasal superior: é possível obter neurônios sem prejudicar o olfato?

Abstract Introduction Olfactory epithelium biopsy has been useful for studying diverse otorhinolaryngological and neurological diseases, including the potential to better understand the pathophysiology behind COVID-19 olfactory manifestations. However, the safety and efficacy of the technique for obtaining human olfactory epithelium are still not fully established. Objective This study aimed to determine the safety and efficacy of harvesting olfactory epithelium cells, nerve bundles, and olfactory epithelium proper for morphological analysis from the superior nasal septum. Methods During nasal surgery, 22 individuals without olfactory complaints underwent olfactory epithelium biopsies from the superior nasal septum. The efficacy of obtaining olfactory epithelium, verification of intact olfactory epithelium and the presence of nerve bundles in biopsies were assessed using immunofluorescence. Safety for the olfactory function was tested psychophysically using both unilateral and bilateral tests before and 1 month after the operative procedure. Results Olfactory epithelium was found in 59.1% of the subjects. Of the samples, 50% were of the quality necessary for morphological characterization and 90.9% had nerve bundles. There was no difference in the psychophysical scores obtained in the bilateral olfactory test (University of Pennsylvania Smell Identification Test [UPSIT®]) between means before biopsy: 32.3 vs. postoperative: 32.5, p= 0.81. Also, no significant decrease occurred in unilateral testing (mean unilateral test scores 6 vs. 6.2, p= 0.46). None out of the 56 different odorant identification significantly diminished (p> 0.05). Conclusion The technique depicted for olfactory epithelium biopsy is highly effective in obtaining neuronal olfactory tissue, but it has moderate efficacy in achieving samples useful for morphological analysis. Olfactory sensitivity remained intact.

Resumo Introdução A biópsia do epitélio olfatório tem sido útil para estudar diversas doenças otorrinolaringológicas e neurológicas, incluindo seu potencial para melhor compreender a fisiopatologia por trás das manifestações olfatórias na COVID‐19. No entanto, a segurança e eficácia da técnica de obtenção de epitélio olfatório humano ainda não estão totalmente estabelecidas. Objetivos Este estudo teve como objetivo determinar a segurança e eficácia da coleta de células do epitélio olfatório, feixes nervosos e epitélio olfatório adequados para análise morfológica, no septo nasal superior. Método Durante a cirurgia nasal, 22 indivíduos sem queixas olfatórias foram submetidos a biópsias de epitélio olfatório do septo nasal superior. A eficácia da obtenção de epitélio olfatório, a verificação de epitélio olfatório íntegro e a presença de feixes nervosos nas biópsias foram avaliadas por imunofluorescência. A segurança da função olfatória foi testada psicofisicamente usando testes unilaterais e bilaterais antes e um mês após o procedimento cirúrgico. Resultados Epitélio olfatório foi encontrado em 59,1% dos sujeitos. Das amostras, 50% apresentaram a qualidade necessária para a caracterização morfológica e 90,9% continham feixes nervosos. Não houve diferença nos escores psicofísicos obtidos no teste olfatório bilateral (University of Pennsylvania Smell Identification Test [UPSIT®]) entre as médias antes da biópsia: 32,3 vs. pós‐operatório: 32,5, p = 0,81. Além disso, nenhuma diminuição significante ocorreu no teste unilateral (escore médio do teste unilateral 6 vs. 6,2, p = 0,46). Não houve redução significante na identificação de nenhum dos 56 odorantes diferentes (p > 0,05). Conclusão A técnica descrita para biópsia de epitélio olfatório é altamente eficaz na obtenção de tecido olfatório neuronal, mas tem eficácia moderada na obtenção de amostras adequadas para análise morfológica. A capacidade olfativa permaneceu intacta.

Evaluation of the olfactory bulb volume and morphology in patients with coronavirus disease 2019: can differences create predisposition to anosmia?

SUMMARY OBJECTIVE: This study aimed to investigate whether the volume and morphology of the olfactory bulb are effective in the occurrence of anosmia in patients after COVID-19 infection. METHODS: The olfactory bulbus volume was calculated by examining the brain magnetic resonance imaging of cases with positive (+) COVID-19 polymerase chain reaction test with and without anosmia. Evaluated magnetic resonance imaging images were the scans of patients before they were infected with COVID-19. The olfactory bulbus and olfactory nerve morphology of these patients were examined. The brain magnetic resonance imaging of 59 patients with anosmia and 64 controls without anosmia was evaluated. The olfactory bulb volumes of both groups were calculated. The olfactory bulb morphology and olfactory nerve types were examined and compared between the two groups. RESULTS: The left and right olfactory bulb volumes were calculated for the anosmia group and control group as 47.8±15/49.3±14.3 and 50.5±9.9/50.9±9.6, respectively. There was no statistically significant difference between the two groups. When the olfactory bulb morphology was compared between the two groups, it was observed that types D and R were dominant in the anosmia group (p<0.05). Concerning olfactory nerve morphology, type N was significantly more common in the control group (p<0.05). CONCLUSIONS: According to our results, the olfactory bulb volume does not affect the development of anosmia after COVID-19. However, it is striking that the bulb morphology significantly differs between the patients with and without anosmia. It is clear that the evaluation of COVID-19-associated smell disorders requires studies with a larger number of patients and a clinicoradiological approach.

Human olfactory mesenchymal stromal cells co-expressing horizontal basal and ensheathing cell proteins in culture / Las células mesenquimales del estroma olfatorio humano coexpresan proteínas de las células basales horizontales y de recubrimiento neural en cultivo

Introduction: The olfactory neuro-epithelium has an intrinsic capability of renewal during lifetime provided by the existence of globose and horizontal olfactory precursor cells. Additionally, mesenchymal stromal olfactory cells also support the homeostasis of the olfactory mucosa cell population. Under in vitro culture conditions with Dulbecco modified eagle/F12 medium supplemented with 10% fetal bovine serum, tissue biopsies from upper turbinate have generated an adherent population of cells expressing mainly mesenchymal stromal phenotypic markers. A closer examination of these cells has also found co-expression of olfactory precursors and ensheathing cell phenotypic markers. These results were suggestive of a unique property of olfactory mesenchymal stromal cells as potentially olfactory progenitor cells. Objective: To study whether the expression of these proteins in mesenchymal stromal cells is modulated upon neuronal differentiation. Materials and methods: We observed the phenotype of olfactory stromal cells under DMEM/F12 plus 10% fetal bovine serum in comparison to cells from spheres induced by serum-free medium plus growth factors inducers of neural progenitors. Results: The expression of mesenchymal stromal (CD29+, CD73+, CD90+, CD45-), horizontal basal (ICAM-1/CD54+, p63+, p75NGFr+), and ensheathing progenitor cell (nestin+, GFAP+) proteins was determined in the cultured population by flow cytometry. The determination of Oct 3/4, Sox-2, and Mash-1 transcription factors, as well as the neurotrophins BDNF, NT3, and NT4 by RT-PCR in cells, was indicative of functional heterogeneity of the olfactory mucosa tissue sample. Conclusions: Mesenchymal and olfactory precursor proteins were downregulated by serum-free medium and promoted differentiation of mesenchymal stromal cells into neurons and astroglial cells.

Introducción. El recambio celular del neuroepitelio olfatorio ocurre durante la vida del individuo gracias a precursores olfatorios. Además, las células mesenquimales del estroma también contribuyen a la homeostasis de la mucosa. Cuando un explante de una biopsia de mucosa se cultiva en un medio esencial mínimo, se genera una población predominante de células adherentes que expresan proteínas típicas de las células mesenquimales del estroma. La coexpresión de marcadores fenotípicos de precursores olfatorios y de células del recubrimiento del nervio olfatorio constituiría una propiedad única de las células mesenquimales del estroma. Objetivo. Determinar si la diferenciación celular de las células mesenquimales hacia fenotipos neurales modula la expresión de los marcadores mesenquimales característicos. Materiales y métodos. Se compararon las células aisladas de la mucosa olfatoria en un medio de cultivo con suplemento de 10 % de suero fetal bovino con esferas generadas en un medio sin suero más factores de crecimiento. Resultados. Se determinó la expresión de proteínas de las células mesenquimales del estroma (CD29+, CD73+, CD90+, CD45-), de las basales horizontales (ICAM-1/CD54+, p63+, p75NGFr+), y de las del recubrimiento del nervio olfatorio (nestin+, GFAP+) en la misma población cultivada. La determinación de Oct 3/4, Sox-2 y Mash-1, así como de las neurotrofinas BDNF, NT3 y NT4, sugirió que las células del estroma son funcionales. La expresión de las proteínas de las células mesenquimales y los precursores olfatorios, disminuyó en las células de las mesenesferas inducidas por ausencia de suero en el medio de cultivo. Conclusión. Las células mesenquimales del estroma de la mucosa olfatoria presentan una tendencia dominante hacia la diferenciación neural.

Reversal of Olfactory Disturbance in Allergic Rhinitis Related to OMP Suppression by Intranasal Budesonide Treatment

PURPOSE: We evaluated the severity of olfactory disturbance (OD) in the murine model of allergic rhinitis (AR) and local allergic rhinitis (LAR) in mice. We also investigated the therapeutic effect of an intranasal steroid on OD. METHODS: Forty BALB/c mice were divided into 5 groups (n = 8 for each). The control group was sensitized intraperitoneally (i.p.) and challenged intranasally (i.n.) with saline. Mice in the AR group got i.p. and i.n. ovalbumin (OVA) administration for AR induction. The LAR group was challenged i.n. with 1% OVA for inducing local nasal allergic inflammation, without inducing the systemic allergy. The OD group got an i.p. methimazole administration (75 mg/kg) to induce total destruction of olfactory mucosa. Mice in the intranasal budesonide group received i.n. budesonide (12.8 μg per time, 30 minutes after the i.n. OVA challenge) while using OVA to cause systemic allergies. We conducted a buried-food pellet test to functionally assess the degree of OD in each group by measuring the time taken until finding hidden food. We evaluated the damage to olfactory epithelium using histopathologic evaluation and compared the degree of olfactory marker protein (OMP) expression in olfactory epithelium using immunofluorescent staining. RESULTS: Mice of the AR (81.3 ± 19.8 seconds) and LAR groups (66.2 ± 12.7 seconds) spent significantly more time to detect the pellets than the control group (35.6 ± 12.2 seconds, P < 0.01). After treatment, the intranasal budesonide group exhibited significantly better results (35.8 ± 11.9 seconds) compared with the AR and LAR groups (P < 0.01). The AR and LAR groups showed considerable olfactory epithelial damage and suppression of OMP expression compared with the control group. In the intranasal budesonide group, the olfactory lesions and OMP expression had improved substantially. CONCLUSIONS: OD may be caused by olfactory epithelial damage and suppression of OMP expression in nasal allergic inflammation and could be reversed using an intranasal steroid.

Mechanisms and Therapeutic Effects of Human Olfactory Mucosa-Derived Mesenchymal Stem Cells on Experimental Autoimmune Encephalomyelitis in Mice / 中山大学学报(医学科学版)

@#【Objective】 To study the mechanisms and therapeutic effects of human olfactory mucosa-derived mesenchymal stem cells（OMSC）on experimental autoimmune encephalomyelitis（EAE）in mice.【Methods】Under local anesthesia by using nasal endoscopy，olfactory epithelia of healthy donors were obtained，digested and cultured up to the 5th passage. OMSC were identified，differentiated and stained. EAE models were induced in C57 female mice by myelin oligodendrocyte glycoprotein（MOG35- 55）and pertussis toxin（PT）. Neurological function was documented daily. On day 16 after immunization（peak of incidence），the mice were divided randomly into two groups and treated with OMSC and PBS via tail vein injection respectively. On day 24 after immunization ，blood was collected from angular vein and levels of IL-10，IL-17，IFN-γ and IL-6 were determined by cytometric beads array（CBA）. The size of the spinal cord lesion in mice was observed and measured by using HE and LFB staining. Peripheral blood lymphocytes（PBL）of healthy donors were obtained and then co-cultured with OMSC. The proportions of CD4+ T cells secreting IFN- γ（Th1 cells）in lymphocyte group and co-culture group were compared after 2 days of cultivation. Adding IDO or COX pathway inhibitor to co- culture group and cultivating for 2 days，we observed and compared the proportions of Th1 cells in lymphocyte group，co-culture group and inhibitor treatment group respectively.【Results】OMSC exhibited certain mesenchymal stem cell-like characteristics with respect to expression of stem cell surface markers and multilineage differentiation potentials. After induced by MOG35- 55 and PT，EAE models showed different levels of neurological damage. Compared with those in PBS treatment group，in OMSC treatment group，the severity of neural dysfunction in mice was significantly reduced（P =0.002），the level of IFN-γ in serum was lower（P = 0.032），but no significant differences in the levels of IL-10，IL-17 and IL-6 were found between two groups. HE and LFB staining revealed that the inflammatory infiltration and demyelinating areas in OMSC treatment group were less than those in PBS treatment group. The proportion of Th1 cells was lower in co-culture group than that in lymphocyte group（P = 0.001），higher in IDO inhibitor group than that in co-culture group（P = 0.01），but no significant difference was found between IDO inhibitor group and lymphocyte group or between COX inhibitor group and co-culture group.【Conclusions】OMSC may regulate the proportion of Th1 lymphocytes through IDO pathway so as to inhibit the demyelinating injuries of EAE in mice. This study provides a new idea for the clinical treatment of multiple sclerosis.

The role of transcription factors in differentiation and development of the olfactory epithelium stem cells / 中华耳鼻咽喉头颈外科杂志

As a sensor of the olfactory system, olfactory epithelium plays an important role in the olfactory system. In addition, olfactory epithelium is the only neuroepithelial epithelium in mammals that can maintain its self-renewal all along. There are of great significance in researching regenerating and repairing neural tissues, transplanting and treatment neural stem cells as well as the occurrence of olfactory disorders and intervention. This review will describe the characteristics of olfactory epithelial stem cells, and mainly summerize the function and significance of each transcription factor in the process of olfactory epithelial stem cell development and differentiation, in order to provide new ideas for the study of olfactory epithelial stem cells.

Olfactory epithelium as an infinitive source of neural stem cells for derivation of inner ear hair cells / El epitelio olfatorio como fuente infinita de células madre neurales para la derivación de los vestibulocitos del oído interno

Surgical techniques for treatment of sensory neural hearing loss (SNHL) have unpredictable outcomes and in recent years cell therapy investigated for treatment of SNHL. Olfactory epithelium proceed neurogenesis during life time and provide an easily accessible source of neural stem cells. So the aim of this study was isolating neural stem cells from olfactory epithelium of rat and differentiation of these cells into hair cells of inner ear in vitro. The epithelium tissue of olfactory mucosa of rats were removed and digested by collagenase H. The digested tissue was cultured in flasks in suspension forms to create spheres. Spheres were passaged and from passage 2 spheres selected for differentiation. At this stage cells of spheres isolated from each other and placed in flask containing defined differentiation medium. Cells at this stage cultured in adhesive form. Immunohistochemistry and RT-PCR were used for neural stem cells and hair cells identification. Spheres formed from olfactory epithelium culture and immunohistochemistry revealed that cells of spheres from passage one and two expressed the neural stem cells markers. After culture of isolated cells in differentiation medium, the morphology of cells begun to change. The cells presented neural cells projections and after 10 days the projections elongated more and interact to each other in multi layers. RT-PCR and immunohistochemistry revealed that differentiated cells expressed hair cells specific genes. In this study we showed that neural stem cells of olfactory epithelium can differentiate into hair cells of inner ear and therefore can be used for treatment of SNHL.

Las técnicas quirúrgicas para el tratamiento de la pérdida auditiva neural sensorial (PANS) tienen resultados impredecibles y en los últimos años la terapia celular ha sido investigada para su tratamiento. El epitelio olfatorio se forma durante la neurogénesis y proporciona una fuente fácilmente accesible de células madre neurales. El objetivo de este estudio fue aislar las células madre neurales del epitelio olfativo de la rata y la diferenciación de estas células en vestibulocitos del oído interno in vitro. Se retiró el tejido del epitelio de la mucosa olfatoria de ratas y fue digerido con colagenasa H. El tejido se cultivó en forma de suspensión para crear esferas. Se seleccionaron dos esferas para la diferenciación. En esta fase, las células de esferas fueron aisladas unas de otras y colocadas en un medio de diferenciación definido. Células en esta etapa fueron cultivadas en forma adhesiva. Inmunohistoquímica y RT-PCR se utilizó para las células madre neurales y la identificación de células ciliadas. Las esferas formadas a partir del cultivo del epitelio olfatorio y la inmunohistoquímica revelaron que las células de esferas en etapas uno y dos expresaban los marcadores de células madre neurales. Se observaron cambios en la morfología de las células después del cultivo de células aisladas. RT-PCR e inmunohistoquímica revelaron que las células diferenciadas expresaron células específicas de gen de vestibulocitos. Se observó que las células madre neuronales de epitelio olfatorio puede diferenciarse en células en forma de cabello del oído interno y por lo tanto puede ser utilizado para el tratamiento de PANS.

Rabbit olfactory stem cells. Isolation protocol and characterization

PURPOSE: To describe a new technique for isolation of a mesenchymal stem cells (MSCs) population from the olfactory mucosa in rabbits. METHODS: Olfactory stem cells (OSCs) were retrieved from under the cribriform plate of the Ethmoid bone. Several assays were accomplished to characterize the cell population and attest its viability in vitro. The cells were submitted to flow cytometry with the antibodies CD34, CD45, CD73, CD79, CD90 and CD105 and also they were induced to differentiate in three lineages. Functional evaluation involved analysis of in vitro growth behavior, colony forming unit like fibroblasts (CFU-f) and cryopreservation response. Further transduction with Green Fluorescent Protein (GFP) was also performed. RESULTS: The OSCs showed mesenchymal features, as positive response to CD34, CD73 and CD90 antibodies and plasticity. Additionally, these cells have high proliferated rate, and they could be cultured through many passages and kept the ability to proliferate and differentiate after cryopreservation. The positive response to the transduction signalizes the possibility of cellular tracking in vivo. This is a desirable feature in case those cells are used for pre-clinical trials. CONCLUSION: The cells harvested were mesenchymal stem cells and the technique described is therefore efficient for rabbit olfactory stem cells isolation.

Olfaction in Neurologic and Neurodegenerative Diseases: A Literature Review

Introduction Loss of smell is involved in various neurologic and neurodegenerative diseases, such as Parkinson disease and Alzheimer disease. However, the olfactory test is usually neglected by physicians at large. Objective The aim of this study was to review the current literature about the relationship between olfactory dysfunction and neurologic and neurodegenerative diseases. Data Synthesis Twenty-seven studies were selected for analysis, and the olfactory system, olfaction, and the association between the olfactory dysfunction and dementias were reviewed. Furthermore, is described an up to date in olfaction. Conclusion Otolaryngologist should remember the importance of olfaction evaluation in daily practice. Furthermore, neurologists and physicians in general should include olfactory tests in the screening of those at higher risk of dementia. .

Influence of different blood serum concentration and growth time on olfactory ensheathing cells from the olfactory mucosa of adult rats / 中国组织工程研究

BACKGROUND:Preliminary experiments have reported the influence of serum and nerve growth factor on olfactory ensheathing cells proliferation in vitro, but there are less studies concerning choice of serum concentration and growth time for in vitro culture of olfactory ensheathing cells. OBJECTIVE:To find out the influence of different blood serum concentration and growth time on olfactory ensheathing cells from the olfactory mucosa of adult rats based on the growth curve of olfactory ensheathing cells. METHODS:The olfactory ensheathing cells from the olfactory mucosa of adult rats were separated, culture and identified in vitro. Sulforhodamine B and microplate reader were employed to measure absorbance values and plot growth curve of olfactory ensheathing cells. RESULTS AND CONCLUSION:When cultured for the same time in blood serum of different concentrations, absorbance values, especial y in the groups 10%, 20%, 30%, 40%, tended to increase with time except the 0%group. When cultured in the same serum for different time, absorbance values increased within the first 9 days, then promoted rapidly in the groups 10%, 20%, 30%, 40%at 13 days, entered the plateau phase at 19 days, and decreased at 23 days;meanwhile, in the other groups (50%, 60%, 70%, 80%, 90%) the absorbance values peaked at the 13th day and then decreased gradual y. These findings indicate that different serum concentrations and different growth time in vitro affect cellgrowth and survival of olfactory ensheathing cells significantly, which should be ful y considered when cells are cultured in an in vitro condition.

Understanding the Human Sensory Conduction of Smell / 한양의대학술지

The olfactory epithelium is the main end organ for the sense of smell in humans and vertebrates. Specially differenciated neuronal cells called olfactory receptor neurons (ORNs) play a key role in the olfactory epithelium by expressing the olfactory receptors (ORs) on their apical surface membrane. The ORs are G-protein coupled receptors that transmit signals from odorants to ORNs by molecular cascades using cyclic adenosine monophosphate, calcium ions and other molecules, which result in the depolarization of ORN. Unlike other mammalian animals, only about 30% of OR genes in the human genome are expressed. The Nobel Prize was awarded to the scientists who cloned these ORs for the first time. Each ORN expresses only a single type of OR, and ORNs which express the same type of OR converge together into the same glomeruli in the olfactory bulb. A single OR recognizes multiple odorants, and a single odorant is recognized by multiple ORs with varying affinities. At the higher neurons beyond the bulb, neuronal connections are divergent. The combinatorial model of odor identification and discrimination is well established at the convergence level, but little is known about the action mechanisms of neuronal divergence for odor identification and discrimination and further study is required.

Evaluación del olfato en pacientes con patología tumoral en la región selar, intervenidos mediante abordaje endoscópico transesfenoidal transnasal / Assessment of smell in patients with tumor pathology in the sellar region, operated by transnasal transsphenoidal endoscopic approach

Introducción: Para acceder a la región selar, podemos utilizar las técnicas transcraneal, transeptal, o transnasal endoscópica, pudiendo provocar diferentes grados de hiposmia. Se ha descrito menor morbilidad al utilizar la técnica endoscópica, pero faltan estudios dirigidos a los resultados olfatorios. Objetivo: Determinar la presencia de deterioro olfatorio en los pacientes sometidos a un abordaje transnasal endoscópico. Material y método: Se reclutaron 12 pacientes con tumores en la región selar durante 8 meses. Se les realizó un test de olfato preoperatorio, fueron intervenidos mediante abordaje transnasal endoscópico y controlados al mes posoperatorio. Resultados: Se logró seguimiento a 10 pacientes. Seis (60%) presentaron un test de olfato preoperatorio normal. Al mes posoperatorio, se constató mejoría olfatoria en 1 (10%) paciente, 8 (80%) se mantuvieron en la misma categoría y 1 (10%) presentó deterioro olfatorio. En suma, 9 de 10 pacientes (90%) mantienen o mejoran su olfato al mes posoperatorio. Conclusión: Nuestros resultados sugieren que el abordaje transnasal endoscópico utilizado en este estudio no produce deterioro olfatorio. Dado que además es una técnica de abordaje efectiva y relativamente segura, consideramos que constituye una alternativa factible para utilizar en pacientes con patología tumoral en la región selar.

Introduction: To access the sellar region we can use the transcranial, transeptal, or transnasal endoscopic approaches, which may cause different degrees of hyposmia. It has described less morbidity to use the endocopic technique, however, there are few studies directed at olfactory outcomes. Aim: To determine the presence of olfactory impairment secondary to endoscopic transnasal approach. Material and method: 12 patients with tumors in the sellar region were enrolled during 8 months. They were underwent a smell test preoperatively, operated by endoscopic transnasal approach, and controlled with postoperative retesting, after one month. Results: Follow-up was achieved to 10 patients. 6 (60%) presented a normal preoperative smell test. Within one postoperative month, olfactory improvement was found in 1 (10%) patient, 8 (80%) remained in the same category and 1 (10%) had olfactory impairment. In all, 9 out of 10 patients (90%) maintain or improve their sense of smell after surgery. Conclusions: Our results suggest that the transnasal endoscopic approach used in this study, doesn't produce olfactory impairment. Given that is also an effective and relatively safe approach, we believe that is a feasible alternative for use in patients with tumor pathology in the sellar region.

Olfactory neuroepithelium in the superior and middle turbinates: which is the optimal biopsy site?

INTRODUCTION: Olfactory neuroepithelium (ON) biopsy has several therapeutic applications for both disorders of olfaction and neurodegenerative diseases. Successful collection of ON is still anything but routine due to a dearth of studies on the distribution of ON in the superior and middle turbinates. AIM: To determine the location in which ON is most likely to be present in endoscopically removed cadaver superior and middle turbinates as well as the influences of gender, age, and naris side on the presence of ON and the extent to which it is present. METHODS: We conducted a prospective anatomical study. The superior and middle turbinates on both sides endoscopically removed from 25 fresh cadavers (less than 12 h post-mortem). The turbinates were halved into anterior and posterior segments for a total of 200 specimens, which were analyzed after hematoxylin and eosin and immunohistochemical staining. Hematoxylin and eosin-stained slides were subjected to blind examination by 3 independent pathologists, and the presence of ON was graded on a 5-point scale from 0 to 4. Kappa measurement was used to determine the agreement between pairs of observers. RESULTS: ON was present in 82.9% of superior turbinate samples and in 17.1% of middle turbinate samples. Immunohistochemistry detected ON in superior turbinates only by S-100 staining and only in 15 fragments. Gender, age, and naris side had no statistically significant effects on the presence of ON. CONCLUSION: When biopsying ON, the posterior portion of the superior turbinate should be targeted whenever possible because it has the highest concentration of ON among the nasal structures.

Expression and function of aquaporin-4 in olfactory system of mice / 第二军医大学学报

Objective To investigate the expression and function of aquaporin-4(AQP4) in the olfactory system of mice. Methods The differences of AQP4 expression in olfaction system between wild-type and AQP4-null mice were studied by immunoblotting and immunofluorescence methods. The differences of mouse olfactory functions in the two groups were examined by two olfactory behavioral assays: the busied food pellet test and olfaction maze test, and the odorant-stimulated electroolfactogram (EOG) recording. Results The results of immunoblotting and immunofluorescence showed no AQP4 expression in the olfactory system in AQP4-null mice. Immunofluorescence result also indicated that AQP4 was mainly distributed in the membrane of support cells, duct cells of Bowmann's gland, basal cells of olfactory epithelium, membrane of Bowmann's gland epithelial cells, olfactory sheath cells surrounding the olfactory bundles, and the membrane of cells in the olfactory bundle layer and glomeruler layer. The results of olfactory behavioral assay were significantly different between the two groups at all time points tested in both the olfaction maze test and the buried food pellet test (P<0.05). It was showed that the EOGs under different pressures of saturated trimethylamine had a similar shape in both groups, and the amplitude of EOGs increased with the increase of pressure. While under the same pressure, the EOG amplitude of AQP4-null mice was significantly lower than that of wild-type mice(P<0.05). Conclusion AQP4 is widely distributed in the olfactory system of mice, including the olfactory mucosa, olfactory nerve, and olfactory bulb, which can protect the olfactory neural bundle and facilitate neural signal transfer.

Compound Betamethasone enhances the expression of olfactory marker protein in olfactory mucosa of mice injured by influenza virus / 基础医学与临床

Objective To investigate the effects of compound Betamethasone on the expression of olfactory marker protein(OMP)in murine olfactory mucosa injured by influenza virus.Methods An animal model was developed by intranasal application of influenza virus to mice.Compound Betamethasone was injected i.p.(3.5 mg/kg)on day 2 and day 4 after the insult.The expression of OMP was tested by immunohistochemistry and Western blot.Results The expression of OMP was significantly downregnlated in the olfactory mucosa of influenza virus control group 1 and influenza virus control group 2;the expression of OMP was significantly upregulated in the olfactory mucosa of post-infection compound Betamethasone group 1 and post-infection compound Betamethasone group 2.Conclusion Compound Betamethasone can enhance the expression of OMP in the olfactory mucosa injured by influenza virus.

Compared the abilities of repairing nerve defect between glial cells in lamina propria and in olfactory bulb / 中华骨科杂志

Objective To compare their competence to repair peripheral nerve defect between lami-na propria glial cells and olfactory bulb glial cells. Methods Glial cells in nasal lamina propria and olfac-tory bulb had been cultured in vitro for 2 weeks, then purified and condensed for later transplantation. 60 adult wistar rats were randomized into three groups of 20 rats each: A (control), B (glial cells in olfactory bulb were transplanted into epineuria lumen) and C (glial cells in lamina propria were transplanted into epineuria lumen). Rats' left sciatic nerves were excised 25 mm long axons and retained epineuria lumen anastomosed to proximal ends. Culture mediums, glial cells in olfactory bulb and in lamina propria were transplanted into epineuria lumen of A, B, and C groups respectively. At 3 months postoperative,the regeneration of injured sciatic nerves were evaluated by methods of gross observation, microscopy, transmission electron microscopy, the retrograde fluorescence gold, the immunofluorescence assay of glial fibre acid protein and nerve growth factors, the enzyme linked immunoassay of myelin basic protein and neurofilament protein, and the extremity function scores. Results The regeneration of injured sciatic nerve and the function of injured limbs were su-perior in C group to in B group and in A group. The transportation distance of retro-marked fluorescence red and the concentrations of glial fibre acid protein, nerve growth factors, myelin basic protein and neurofila-ment protein in injured sciatic nerve were high in C group, middle in B group, and low in A group. The dif-ference between groups had statistical significance. Conclusion Olfactory ensheathing cells could repairinjured nerve defect and the competence that the olfactory cells in lamina propria were superior to in olfacto-ry bulb.

Experimental study of the effects of olfactory lamina propria transplantation and ganglioside GMI treatment on spinal curd injury / 中华老年医学杂志

Objective To observe the effects of olfactory lamina propria (OLP) transplantation and ganglioside GM1 treatment on spinal cord injury (SCI) in rats.Methods Totally 50 healthy pure breed female Sprague-Dawley (SD) rats after spinal cord hemiseetion were randomly divided into 5 groups and were given different treatments: (OLP + GM1) treatment group (group A), GM1 treatment group (group B), OLP treatment group (group C), spinal cord injury but without treatment group (group D) and healthy control group (group E). The recovery of neurological function was evaluated by somatosensory evoked potential (SEP) and pathological examination after surgery. Results In group A, in some rats an escaping response in right hind leg occurred, but in other groups, the motor function was not significantly improved. Histological examination showed that transplanted olfactory lamina propria survived in the transplantation area and expanded on certain routes. NF positive nerve fibers passed through the transplantation area. Compared with group B, C, D, the N1-wave latency was(4.71±0. 72)ms 4 weeks after operation(P<0. 01), and the NF density was(7. 31±0. 26) ×104/mm28 weeks after operation in group A(P<0. 05). Conclusions Olfactory lamina propria (OLP) transplantation and ganglioside GM1 treatment have a synergistic effect on SCI.

Histopathological Changes of Olfactory Epithelium in the Rats Exposed to Cigarette Smoke / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Although exposure to cigarette smoke has been reported to be associated with olfactory dysfunction, the pathophysiology is poorly understood. The purpose of this study is to evaluate the histopathological change of olfactory epithelium according to exposure duration of cigarette smoke and to investigate the effects of cigarette smoke on the olfactory epithelium. SUBJECTS AND METHOD: Thirty-six healthy Sprague-Dawley rats were divided into 2 groups. Experimental groups (n=28) were exposed to cigarette smoke during 2.5 hours (total 5 cigarettes) per one day for 5 days, 11 days and 3, 4, 5, 6, 9 weeks. Control group (n=8) was placed at the same room without smoke exposure and sacrificed at 4 and 9 weeks. Histopathological changes of olfactory epithelium through light microscope and immunohistochemistric findings of olfactory marker protein (OMP) through confocal microscope were observed. The numbers of cells in olfactory epithelium were counted at each period. RESULTS: From 5 days of cigarette smoke exposure, defection of epithelial cells, random arrangement of olfactory receptor cells and decreased thickness of olfactory epithelium were noticed. Numbers of olfactory receptor cells were significantly decreased at 11 days and 3 weeks after smoke exposure, and this finding of decreased number of olfactory receptor cells were continued until 9 weeks of exposure. Numbers of OMP-positive olfactory receptor cells were continuously decreased according to exposure duration. CONCLUSION: The olfactory epithelial cells could be directly affected by cigarette smoke. The numbers of olfactory epithelial cells including olfactory receptor cells were continuously decreased until 9 weeks after cigarette smoke exposure.

ISOLATION AND CHARACTERIZATION OF CULTURED ENSHEATHING CELLS FROM ADULT RAT OLFACTORY BULBS AND NASAL OLFACTORY MUCOSA / 解剖学报

Objective On the basis that olfactory ensheathing cells (OECs) transplanted into injured spinal cord may facilitate axonal regeneration, the OECs were cultured from olfactory bulb and nasal olfactory mucosa in the present study, in order to explore if the olfactory mucosa could be a new donation for transplanting the olfactory ensheathing cells. Methods OECs were harvested from olfactory bulb and mucosa based on the differing rates of attachment of the various cell types, following GFAP and NGFRp75 immunocytochemistry. Results Three morphological and immunohistochmically distinct types of cell which appeared bipolar,tripolar and flat morphology were present in primary cultures of adult rat olfactory bulb and olfactory mucosa.Conclusion The method of purification for OECs based on the different rates of attachment among the various cell types is simple, inexpensive and practical. The OECs from nasal olfactory mucosa like ones from the olfactory bulb is an accessible source of tissue for autologous grafting in human spinal paralegia in the future.