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Objective · To investigate ciliary beat frequency (CBF), and estrogen and progesterone receptor expression levels of human fallopian tubes after mifepristone treating in vitro. Methods · Human fallopian tube mucosa explants (n=25) were treated with different concentrations of mifepristone (0.1,1 and 10 μmol/L) or progesterone (10 μmol/L) separately, or both mifepristone and progesterone. After 24 h of treatment, CBF was measured. Quantitative real-time PCR and immunohistochemistry were used to research the expression of estrogen receptor-α (ERα) and progesterone receptor (PR) of human fallopian tubes after mifepristone treating. The ultrastructure of epithelial cells of fallopian tube after mifepristone (10 μmol/L) treating were observed with transmission electron microscopy. Results · The CBF at the concentrations of 0.1, 1 and 10 μmol/L was not affected by mifepristone (P=0.728,P=0.405 and P=0.941). The CBF decreased markedly in the group of 10 μmol/L progesterone compared to control group (P=0.000). Mifepristone (0.1,1 and 10 μmol/L) dose dependently antagonized the progesterone-induced CBF decrease (P=0.484, P=0.000 and P=0.000). Mifepristone upregulated the expression levels of ERα and PR in the fallopian tubes, but the ultrastructure of the cilia had no significant change. Conclusion · Mifepristone acts as progesterone antagonist in the human fallopian tube, which may explain the tubal contraceptive mechanism when used as an emergency contraceptive.
ABSTRACT
Objective · To investigate ciliary beat frequency (CBF), and estrogen and progesterone receptor expression levels of human fallopian tubes after mifepristone treating in vitro. Methods · Human fallopian tube mucosa explants (n=25) were treated with different concentrations of mifepristone (0.1,1 and 10 μmol/L) or progesterone (10 μmol/L) separately, or both mifepristone and progesterone. After 24 h of treatment, CBF was measured. Quantitative real-time PCR and immunohistochemistry were used to research the expression of estrogen receptor-α (ERα) and progesterone receptor (PR) of human fallopian tubes after mifepristone treating. The ultrastructure of epithelial cells of fallopian tube after mifepristone (10 μmol/L) treating were observed with transmission electron microscopy. Results · The CBF at the concentrations of 0.1, 1 and 10 μmol/L was not affected by mifepristone (P=0.728,P=0.405 and P=0.941). The CBF decreased markedly in the group of 10 μmol/L progesterone compared to control group (P=0.000). Mifepristone (0.1,1 and 10 μmol/L) dose dependently antagonized the progesterone-induced CBF decrease (P=0.484, P=0.000 and P=0.000). Mifepristone upregulated the expression levels of ERα and PR in the fallopian tubes, but the ultrastructure of the cilia had no significant change. Conclusion · Mifepristone acts as progesterone antagonist in the human fallopian tube, which may explain the tubal contraceptive mechanism when used as an emergency contraceptive.
ABSTRACT
OBJECTIVE:We compared the adverse effects of two types of real ambient particles; i.e., total suspended particles from an electrostatic precipitator of a steel mill and fine air particles from an urban ambient particulate matter of 2.5 µm, on mucociliary clearance.METHOD:Mucociliary function was quantified by mucociliary transport, ciliary beating frequency and the amount of acid and neutral mucous in epithelial cells through morphometry of frog palate preparations. The palates were immersed in one of the following solutions: total suspended particles (0.1 mg/mL), particulate matter 2.5 µm 0.1 mg/mL (PM0.1) or 3.0 mg/mL (PM3.0) and amphibian Ringer’s solution (control). Particle chemical compositions were determined by X-ray fluorescence and gas chromatography/mass spectrometry.RESULTS:Exposure to total suspended particles and PM3.0 decreased mucociliary transport. Ciliary beating frequency was diminished by total suspended particles at all times during exposure, while particulate matter of 2.5 µm did not elicit changes. Particulate matter of 2.5 µm reduced epithelial mucous and epithelium thickness, while total suspended particles behaved similarly to the control group. Total suspended particles exhibited a predominance of Fe and no organic compounds, while the particulate matter 2.5 µm contained predominant amounts of S, Fe, Si and, to a lesser extent, Cu, Ni, V, Zn and organic compounds.CONCLUSION:Our results showed that different compositions of particles induced different airway epithelial responses, emphasizing that knowledge of their individual characteristics may help to establish policies aimed at controlling air pollution.
Subject(s)
Animals , Mucociliary Clearance , Mucus , Particulate Matter/chemistry , Particulate Matter/toxicity , Steel/chemistry , Anura , Cilia , Epithelium , Gas Chromatography-Mass Spectrometry , Mucus/chemistry , Palate/cytology , Spectrometry, X-Ray EmissionABSTRACT
Introducción: Secreciones sinonasales patológicas y elevados niveles de factor de necrosis tumoral alfa (TNF alfa) se han encontrado en mucosa sin usal de pacientes con sinusitis crónica. Las células ciliadas respiratorias tienen una reserva funcional que les permite autorregular su frecuencia de batido ciliar (FBC) en respuesta a cambios en la viscosidad, modificando los niveles de calcio intraacelular [Ca+²]ic. Objetivo: Nuestro objetivo es determinóar si TNFalfa afecta el mecanismo de autorregulación y la homeostasis del calcio intraacelular frente a cambios en la viscosidad. Material y método: Cultivos primarios de explantes de tejido adenoideo. Registro de FBC mediante microfotodensitometría. Cultivos tratados con TNFalfa (10 ng/ml) o control durante 24 y 48 horas. Se in crementó la viscosidad agregando dextrano 500 al 10 por ciento y 20 por ciento. Se midió [Ca+²]ic en células cargadas con Fura 2AM. Resultados: El tratamiento con TNFalfa por 48 horas produjo una significativa disminución de la FBC a baja viscosidad, aumento significativo de [Ca+²]ic y caída mayor de FBC en cultivos tratados con tapsigargina (bloqueador bomba calcio-ATPasa retículo). in o se encontró diferencia a alta viscosidad. Conclusión: Después de 48 horas de exposición a TNFalfa se observa un efecto negativo en el mecanismo de adaptación de las células ciliadas a un medio con baja viscosidad, probablemente secundario a cambios en la homeostasis del [Ca+²]ic.
Introduction: Pathologic sinonasal secretions and elevated levels of tumor necrosis factor alpha (TNFalpha) have been in oted in sin us mucosa ofpatients with chronic sinusitis. Respiratory ciliated cells have a functional reserve that allows them to autoregulate their ciliary beat in response to the changesin viscosity, modify in g intraacellular calcium levels [Ca+²]ic. Aim: Our goal was to determinate if TNFalpha affect this autoregulation to viscosity and calcium homeostasis. Material and Method: Primary cultures from adenoid tissue expiants. Ciliary beat frequency (CBF) was recorded usin g microphotodensitometry Cultures viere treated with TNF alpha (10 ng/ml) or control during 24 and 48 hours. Viscosity was increased by add in g dextran 500 10 percent and 20 percent. [Ca+²]ic was determined in cells loaded with Fura-2AM. Results: 48 hours treatment with TNFalpha produced a significant decrease in CBF at low viscosity significant increase in [Ca+²]ic and greater decrese in CBF in cultures treated with thapsigargin (endoplasmic calcium-ATPase pump blocker). Conclusions: After 48 hours of exposure to TNF alpha a negative effect in the adaptation mechanism to a low viscous media is observed in ciliated cells, probably secondary to changesin homeostasis of [Ca+²]ic.
Subject(s)
Humans , Child, Preschool , Child , Calcium/physiology , Cilia , Cilia/physiology , Tumor Necrosis Factor-alpha/pharmacology , Nasal Mucosa , Paranasal Sinuses , Cells, Cultured , Densitometry , Time Factors , Photomicrography , Homeostasis , Fluorescent Antibody Technique , ViscosityABSTRACT
BACKGROUND AND OBJECTIVES: There are conflicting results about ciliary activity in chronic rhinosinusitis (CRS). And dynamic movements reacting to various stimuli in sinusitis mucosa have been studied rarely. This study was designed to investigate the ciliary activity according to the severity of sinusitis and clinical symptoms. We aimed to identify the dynamic ciliary activity in response to purinergic stimulation in sinusitis. SUBJECTS AND METHOD: Nasal mucosal samples were obtained from 44 CRS and 20 normal subjects. We measured ciliary beat frequency (CBF) in the basal and activated status. For evaluating the correlation of CBF with clinical findings, nasal symptoms, endoscopic findings, CT findings, allergy tests, and olfactory tests were reviewed. RESULTS: No significant differences were found in basal CBF between normal and CRS mucosa. However, the CBF stimulated by ATP in normal and CRS mucosa showed marked differences: both the maximal increase and duration of increased CBF by ATP were significantly reduced in CRS mucosa. The olfactory test showed significant correlation with basal CBF, but the presence of allergy or nasal symptoms did not show significant correlation with the basal CBF. The CT score and presence of nasal polyposis showed negative correlation with the basal CBF (p<0.05). CONCLUSION: The basal CBF of normal and CRS mucosa showed no difference. However, dynamic ciliary activity responding to purinergic stimulation was markedly attenuated in sinusitis mucosa. The severity of sinusitis and presence of polyposis negatively affected CBF.
Subject(s)
Adenosine Triphosphate , Hypersensitivity , Mucous Membrane , SinusitisABSTRACT
BACKGROUND AND OBJECTIVES: Although sinusitis is commonly caused by nasal diseases that interfere with ostiomeatal unit, non-rhinogenic sinusitis, for example, post-traumatic or odontogenic, is often encountered. However, it remains unclear whether non-rhinogenic sinusitis affects the function and morphology of the nasal mucosa. The aim of the current study is to observe effects of experimentally induced maxillary sinusitis on ciliary beat frequency and histopathology of the nasal mucosa in rabbits. MATERIALS AND METHOD: Maxillary sinusitis was induced in the right sides of 10 rabbits by partially occluding the natural ostia and inoculating the maxillary sinus with P. aeruginosa. After 4 weeks, the nasal mucosa was obtained from the inferior turbinate and the septum in the right (experimental group) and the left side (control group A). Both sides of the nasal mucosa in another 5 normal rabbits were used as the control group B. Ciliary beat frequency (CBF) was measured by video-computerized analysis and degree of inflammation in histopathology was graded using a 100-point scale of inflammation score. CBF and inflammation score were compared between the three groups. RESULTS: CBFs of the experimental group were significantly lower than those of the control groups A and B. Inflammation scores of the experimental group were significantly higher than those of the control groups. In the experimental group, there were no correlations between CBFs of the maxillary and nasal mucosa, but weak correlations existed between inflammation scores of the maxillary and nasal mucosa. CONCLUSION: Primary maxillary sinusitis can locally induce dysfunction and inflammation of the nasal mucosa.
Subject(s)
Rabbits , Inflammation , Maxillary Sinus , Maxillary Sinusitis , Nasal Mucosa , Nose Diseases , Sinusitis , TurbinatesABSTRACT
BACKGROUND AND OBJECTIVES: Osmolarity is known to affect ciliary beat frequency (CBF) ; however, little is known about the effects of saline spray or irrigation on ciliary activity in human nasal epithelium. The aim of this study was to assess whether CBF is affected by hypertonic, isotonic or hypotonic saline solution in vitro and whether histologic changes are associated with the alteration of ciliary movement. MATERIALS AND METHODS: Normal turbinate mucosa was exposed to five different concentrations including 0.06%, 0.12%, 0.9%, 3.0%, and 7.0% of phosphate-buffered saline solutions. CBF was measured up to 60 minutes after exposure to each concentration of saline solution and histologic changes were examined by transmission electron microscopy. RESULTS: Isotonic and hypotonic saline solutions produced no ciliary slowing ; however, ciliostasis was observed within a few minutes in 3.0% or 7.0% saline solution. Immediately after complete ciliostasis in the hypertonic solutions, the solutions were replaced with 0.12% or 0.9% saline solution in order to observe the reversibility of CBF. Only 0.12% hypotonic saline induced the recovery of ciliary movement. The ultrastructural changes demonstrated that the ciliary slowing might be attributed to epithelial damage by fluid transport toward the surrounding medium. CONCLUSION: Our results suggest that isotonic and hypotonic saline solutions may be more appropriate for nasal irrigation than hypertonic saline solutions.
Subject(s)
Humans , Hypertonic Solutions , Microscopy, Electron, Transmission , Mucous Membrane , Nasal Lavage , Nasal Mucosa , Osmolar Concentration , Saline Solution, Hypertonic , Sodium Chloride , TurbinatesABSTRACT
BACKGROUND AND OBJECTIVES: This study was designed to investigate the relationship between the sinus mucosal hypertrophy and ciliary beat frequency (CBF) in patients with chronic sinusitis in vitro. MATERIALS AND METHODS: In 14 patients with chronic sinusitis, mucosal samples were obtained from the superior, inferior, and lateral walls of the maxillary sinus. Using a video-computerized analysis technique, we measured two parameters for ciliary activity at five different sites selected randomly from each sample=the ciliated area (%) showing CBF equal to or higher than 10 Hz (A10) and the weighted frequency (Fw). Mucosal hypertrophy of the maxillary sinus was calculated using a computer program on CT scans. RESULTS: A(>or=10) and Fw were 91% and 11.3 Hz in control group, respectively, whereas they were 47% and 9.1 Hz in chronic sinusitis group, respectively (p=0.03). There was a significant inverse correlation between A(>or=10) or Fw and mucosal hypertrophy of the maxillary sinus (p=0.001). CONCLUSION: The results of this study suggest that the severity of mucosal hypertrophy on CT scan may correlate with the ciliary activity.
Subject(s)
Humans , Hypertrophy , Maxillary Sinus , Sinusitis , Tomography, X-Ray ComputedABSTRACT
BACKGROUND AND OBJECTIVES: The aim of this study is to analyze ciliary beat frequency (CBF)and ciliary morphology of the middle ear bulla and the upper respiratory tract in mice. MATERIALS AND METHODS: Fifteen mice with 37-42 gram weight were dissected and specimens were taken from the middle ear bulla, the eustachian tube, the inferior turbinate, the nasopharynx, and the upper trachea. The CBF of each specimen was measured using a video-computerized analysis system and the morphology of cilia was also studied using scanning electron microscopy (SEM). RESULTS: The measured CBF ranged from 9.38 Hz to 13.59 Hz, and there were no significant differences in the CBF values among the middle ear bulla, the eustachian tube, the inferior turbinate and the nasopharynx, except for the upper trachea. The mean CBF value of the upper trachea was 11.6 Hz, which was significantly lower than those of other parts. SEM showed that the cilia of the upper tracheal region were shorter than those of other parts. CONCLUSION: The CBF value of the middle ear bulla is similar to those of other parts in the upper respiratory tract except for the upper trachea. In the upper trachea, the CBF is lower and ciliary lenghth of epithelium is shorter than those of other parts.
Subject(s)
Animals , Mice , Cilia , Ear, Middle , Epithelium , Eustachian Tube , Microscopy, Electron, Scanning , Nasopharynx , Respiratory System , Trachea , TurbinatesABSTRACT
BACKGROUND AND OBJECTIVES: The in vitro effects of pneumococcal neuraminidase and staphylococcal beta-toxin on ciliary activity were investigated at different concentrations and lengths of exposure. MATERIALS AND METHODS: The ciliated epithelial cells were taken from the maxillary sinus mucosa of rabbits. Ciliary beat frequency (CBF) was measured at concentrations of 0.01, 0.1 and 1.0 U/mL of neuraminidase and 0.1, 1.0, 2.0, 5.0 and 10 U/mL of beta-toxin using a video-computerized analysis technique. The CBF was measured 2, 4, 6, 12, 24 and 48 hours after administration of the neuraminidase and beta-toxin. In the control group, normal saline was percutaneously applied to the right maxillary sinus. In the experimental group, 2 U/mL of beta- toxin was applied to the left maxillary sinus using the same technique. At 7 days, all of the mucosae were taken from the inferomedial wall of the maxillary sinus for light microscopy. RESULTS: There was no change in CBF during a 48-hour incubation at 0.01, 0.1 and 1.0 U/mL of neuraminidase. However, the CBF dropped significantly after an 8-hour incubation at 2.0 U/mL of beta-toxin (p<0.05, repeated measures ANOVA). No ciliary activity was observed after a 12-hour incubation at 10 U/mL of beta-toxin. The mucoid, purulent discharge was observed in the maxillary sinuses of the experimental group. Prominent epithelial disruption and infiltration of inflammatory cells into the epithelium and lamina propria were observed in the beta-toxin-applied group. CONCLUSION: The results of this study suggest that staphylococcal beta-toxin may reduce ciliary activity and induce sinusitis without occlusion of the natural ostium of the maxillary sinus in rabbits. This study provides another animal model of sinusitis for understanding the pathogenesis of sinusitis caused by bacterial exotoxins.
Subject(s)
Rabbits , Epithelial Cells , Epithelium , Exotoxins , Maxillary Sinus , Microscopy , Models, Animal , Mucous Membrane , Neuraminidase , Sinusitis , Staphylococcus aureus , Staphylococcus , Streptococcus pneumoniae , StreptococcusABSTRACT
Few studies have attempted a systematic comparison of ciliary beat frequency (CBF) and ciliary ultrastructure across different species and different levels of the respiratory tract. The aim of this study was to observe the CBF and ciliary ultrastructure of mice, rats and guinea pigs according to varying sites of the respiratory tract. Balb/c mice, Wistar rats and Dunkin-Hartley guinea pigs were used. We measured CBF using a video-computerized analysis technique at the middle of the maxilloturbinal, the nasopharynx, the upper trachea and the main bronchus in vitro. Ciliary length and the proportion of ciliated epithelium were assessed with scanning electron microscopy (SEM). In the rat, CBF was lower in the main bronchus (9.7+/-0.4 Hz) than at other sites, but there was no difference in CBF values across different airway sites in mice and guinea pigs. The CBF in the main bronchus was higher in guinea pigs than in rats and mice. SEM showed that the cilia of the rat were significantly shorter in the upper trachea and the main bronchus than in the maxilloturbinal and the nasopharynx. The respiratory epithelia of guinea pigs were more ciliated than those of mice and rats, especially in the upper trachea and the main bronchus. The guinea pig may be a superior experimental animal for ciliary function studies because the guinea pig has a less variable CBF and more uniform distribution of ciliated cells along different levels of the airway. These results provide valuable data relevant to ciliary functional studies using animal models.
Subject(s)
Animals , Mice , Rats , Bronchi , Cilia , Epithelium , Guinea Pigs , Guinea , Microscopy, Electron, Scanning , Models, Animal , Nasopharynx , Rats, Wistar , Respiratory System , TracheaABSTRACT
OBJECTIVES: The aim of this study was to investigate whether the ciliary activity of respiratory epithelium is affected in allergic rhinitis. METHODS: Twenty Wistar rats were divided into an unsensitized control group and sensitized allergic group. The sensitized group was immunized intraperitoneally with ovalbumin, followed by intranasal administration of ovalbumin. Allergy was determined by an increase in nasal symptoms, the number of tissue eosinophils and a positive result to a passive cutaneous anaphylaxis (PCA) test. Nasal, nasopharyneal, tracheal, and bronchial epithelial cells were obtained from both the control and allergic groups. Ciliary beat frequency (CBF) was measured using a video-computerized analysis technique in vitro. We compared the CBF of two groups in each site. We also evaluated the findings of the nasal mucosa of both groups with an scanning electron microscope. RESULTS: In vitro CBF measurement demonstrated that the CBF of the control and allergic groups did not differ significantly (p>0.05). CONCLUSION: CBF is not affected by respiratory allergy.
Subject(s)
Animals , Rats , Administration, Intranasal , Eosinophils , Epithelial Cells , Hypersensitivity , Nasal Mucosa , Ovalbumin , Passive Cutaneous Anaphylaxis , Rats, Wistar , Respiratory Mucosa , RhinitisABSTRACT
The mucociliary system has primary defence mechanism in the respiratory tract. The effects of various drugs used clinically in the treatment of disease of the nasal cavity have not been fully elucidated. The aim of this study was to investigate the effects of alpha1 receptor agonist, phenylephrine hydrochloride on ciliary beat frequency in vitro using a video computerized analysis technique. The ciliated epithelial cells from the nasal mucosa in four volunteers were collected in a culture medium and exposed to 0.125%, 0.25%, 0.5%, 1.0%, and 2.5% phenylephrine hydrochloride solution according to 0.5 hour, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, and 6 days. There was a significant decrease in ciliary beat frequency with exposure to 0.125% phenylephrine hydrochloride solution for 12 hours, and 0.25% phenylephrine hydrochloride solution for 8 hours(p<0.05). There were cilioinhibitory effects with concentration dependent response by phenylephrine hydrochloride solution. After substitution of the culture medium with phenylephrine hydrochloride free one showed no ciliary recovery in all groups. The results of this study suggest that phenylephrine hydrochloride may have phamacologically a cilioinhibitory effect in vitro on ciliated epithelium.