Fibrin-Genipin Hydrogel for Cartilage Tissue Engineering in Nasal Reconstruction.

OBJECTIVES: Nasal reconstruction is limited by the availability of autologous cartilage. The aim is to investigate an adhesive biomaterial for tissue engineering of nasal cartilage by evaluating mechanical properties of hydrogels made of fibrin crosslinked with genipin as compared to native tissue. METHODS: Hydrogels of fibrin, fibrin-genipin, and fibrin-genipin with extracellular matrix (ECM) particles were created and evaluated with mechanical testing to determine compression, tensile, and shear properties. Rabbit nasal septal cartilage was harvested and tested in these modalities for comparison. Transmission electron microscopy characterized hydrogel structure. RESULTS: Fibrin-genipin gels had higher compressive, tensile, and shear moduli compared to fibrin alone or fibrin-genipin with ECM. However, all hydrogel formulations had lower moduli than the rabbit nasal septal cartilage. Electron microscopy showed genipin crosslinking increased structural density of the hydrogel and that cartilage ECM created larger structural features with lower crosslinking density. CONCLUSION: The addition of genipin significantly improved mechanical properties of fibrin hydrogels by increasing the compressive, tensile, and shear moduli. The addition of cartilage ECM, which can add native structure and composition, resulted in decreased moduli values. Fibrin-genipin is a bioactive and biomechanically stable hydrogel that may offer promise as a scaffold for cartilage tissue engineering in nasal reconstruction, yet further augmentation is required to match material properties of native nasal cartilage.

Decellularization of Human Nasal Septal Cartilage for the Novel Filler Material of Vocal Fold Augmentation.

OBJECTIVES: The clinical application of allogenic and/or xenogenic cartilage for vocal fold augmentation requires to remove the antigenic cellular component. The objective of this study was to assess the effect of cartilage decellularization and determine the change in immunogenicity after detergent treatment in human nasal septal cartilage flakes made by the freezing and grinding method. METHODS: Human nasal septal cartilages were obtained from surgical cases. The harvested cartilages were treated by the freezing and grinding technique. The obtained cartilage flakes were treated with 1% Triton X-100 or 2% sodium dodecyl sulfate (SDS) for decellularization of the cartilage flakes. Hematoxylin and eosin stain (H&E stain), surface electric microscopy, immunohistochemical stain for major histocompatibility complex I and II, and ELISA for DNA contents were performed to assess the effect of cartilage decellularization after detergent treatment. RESULTS: A total of 10 nasal septal cartilages were obtained from surgical cases. After detergent treatment, the average size of the cartilage flakes was significantly decreased. With H&E staining, the cell nuclei of decellularized cartilage flakes were not observed. The expression of major histocompatibility complex (MHC)-I and II antigens was not identified in the decellularized cartilage flakes after treatment with detergent. DNA content was removed almost entirely from the decellularized cartilage flakes. CONCLUSION: Treatment with 2% SDS or 1% Triton X-100 for 1 hour appears to be a promising method for decellularization of human nasal septal cartilage for vocal fold augmentation.

Microvascular remodeling of nasal mucosa in allergic rhinitis induced by an allergen in Sprague-Dawley rats.

This study aimed to observe microvascular changes in the nasal mucosa of Sprague-Dawley (SD) rats with allergic rhinitis (AR) after persistent exposure to an allergen with fluticasone propionate (FP) treatment. Ninety healthy SD rats were randomly distributed into the control group (A, N = 30), the group with continued exposure to an allergen (B, N = 30), and FP treatment group (C, N = 30). The animals of the persistence group were subjected to persistent exposure to an allergen after 7 weeks of modeling of ovalbumin (OVA) provocation in the nasal mucosa for 16 weeks. At the 8th, 12th, and 16th week after OVA provocation, each group was euthanized at each time point: the FP treatment after OVA provocation, and animals of the control group were not stimulated with OVA and were sacrificed at the same time point. The nasal mucosa of 5 animals from each group was analyzed for the expression of vascular endothelial growth factor (VEGF), and another 5 animals were used to make micro vascular corrosion casts for a scanning electron microscope. The results demonstrate that FP has a strong inhibitory effect on angiogenesis in AR. Inhalation of FP had an antiangiogenic effect through inhibition of VEGF expression but does not completely reverse the remodeling of the nasal mucosa in the short term nor does it have complete control over the expression of VEGF mRNA.

Nasal gel and olfactory cleft.

OBJECTIVE: To evaluate whether a nasal gel, administrated using a radial-hole inhaler, reaches the olfactory cleft and if a different administration method influences distribution. MATERIAL AND METHOD: Sixteen healthy volunteers underwent a nasal endoscopy at 1 and 7minutes after the administration of a intranasal gel, with a different method in each fossa. RESULTS: No dye deposition was identified at the olfactory cleft, middle turbinate or middle meatus. In all cases the gel was identified at the nasal vestibule. On the right side, the second most frequent dye identification area was the inferior turbinate, with a rate of 87% at the first minute and 75% at 7 minutes. It was followed by the septum (75 and 62%) and the inferior meatus (6.2 and 12.5%). On the left side, the second most frequent stained area was the septum (18.7 and 13.5%), followed by the inferior meatus (6.5 and 65%). No inferior turbinate staining was found in the left side. There was a significant difference in the deposition rate at the septum (P<.01) and inferior turbinate (P<.001), when both administration methods were compared. CONCLUSIONS: No nasal gel, administrated using a radial-hole inhaler, was found at the olfactory cleft, middle turbinate or middle meatus. Gel distribution was located at the anterior and inferior portion of the nose, independent of the administration method used. Significantly different gel distribution rates were found at the septum and inferior turbinate when the 2 administration methods were compared.

Isolation of human nasoseptal chondrogenic cells: a promise for cartilage engineering.

In cartilaginous tissues, perichondrium cambium layer may be the source of new cartilage. Human nasal septal perichondrium is considered to be a homogeneous structure in which some authors do not recognize the perichondrium internal zone or the cambium layer as a layer distinct from adjacent cartilage surface. In the present study, we isolated a chondrogenic cell population from human nasal septal cartilage surface zone. Nasoseptal chondrogenic cells were positive for surface markers described for mesenchymal stem cells, with exception of CD146, a perivascular cell marker, which is consistent with their avascular niche in cartilage. Although only Sox-9 was constitutively expressed, they also revealed osteogenic and chondrogenic, but not adipogenic, potentials in vitro, suggesting a more restricted lineage potential compared to mesenchymal stem cells. Interestingly, even in absence of chondrogenic growth factors in the pellet culture system, nasoseptal chondrogenic cells had a capacity to synthesize sulfated glycosaminoglycans, large amounts of collagen type II and to a lesser extent collagen type I. The spontaneous chondrogenic potential of this population of cells indicates that they may be a possible source for cartilage tissue engineering. Besides, the pellet culture system using nasoseptal chondrogenic cells may also be a model for studies of chondrogenesis.

Comparison of ultrastructure and lectin histochemistry on the anterior medial gland of nasal septum in rat and gerbil.

The anterior medial gland (AMG), located in the submucosa of rodent nasal septum, is suggested to provide fluid for humidification of inspired air. Tremendous variation of the environmental air humidity, on which rats and gerbils depend to live, leads us to expect a multiplicity on ultrastructure and various subcellular glycoconjugate distribution within the AMG acinar cells between these two species. Electron microscopy revealed that: (1) The nucleus of AMG acinar cells in rat was irregular-shaped, but that in gerbil was round or elliptical; (2) Secretory granules in rat AMG acinar cells contained homogenous content with various electron density. However, two types of secretory granules in gerbil AMG acinar cells were found: one with lamellated-structure and high electron density, while the others had particulate materials; (3) Myoepithelial cells were present in the acinus of medial and posterior regions in rat AMG, but absent in gerbil; and (4) Nerve terminals were present only in the medial and posterior rat AMG, but in all three regions of the gerbil AMG. Lectin histochemistry demonstrated that: (1) Rat and gerbil AMG acinar cells expressed strong affinity toward Con A and WGA, but neither showed any reactivity toward UEA and PNA; and (2) Varying degrees of reactivity toward different lectins, including DBA, PNA, SBA and EBL, were recognized in rat and gerbil AMG acinar cells. We confirm the species variation on the ultrastructure and lectin histochemistry of AMG in rats and gerbils, and speculate that these variations may be due to the different living environment.

The origin and early development of the nasal septum in human embryos.

Based on scanning electron microscopic dissections of human embryos and fetuses of the sixth to the twelfth week (Carnegie stages 16-23 and early fetus), the origin of the nasal septum was studied. The findings show that the nasal septum does not grow downwards. It is derived from the tissue between the primary choanae: as such, its anlage is present from the very beginning. Its contact and fusion with the palatal shelves is made possible by the elevation of the palatal shelves from the vertical into the horizontal position, as the tongue descends.

The acid-sensing ion channel 2 (ASIC2) of ciliated cells in the developing rat nasal septum.

The airway epithelium is exposed to an acidic environment in certain conditions. The acid-sensing ion channel 2 (ASIC2) belongs to the epithelial amiloride-sensitive sodium channel and degenerin (ENaC/DEG) family and is expressed on cilia of the respiratory epithelium. The aim of this study was to detect the expression of ASIC2 in the nasal septum in the embryonic stage of the rat. ASIC2 expression was not observed in the primary cilium but was found in some cilia on embryonic day 17 (E17). After E18, all cilia showed ASIC2 immunoreactivity. RT-PCR analysis revealed that ASIC2b, a subtype of ASIC2, was expressed in the nasal septum while ASIC2a was not. Quantitative Real-time RT-PCR studies indicated that the expression level of ASIC2 mRNA was highest on E21, just before birth. These results imply that ASIC2 plays little part in the development of the nasal septum epithelium. On the other hand, ASIC2, especially ASIC2b, may function for the survival and retention of ciliated cells of the nasal septum against dynamic changes in the pH environment at birth.

Eosinophilic substance is "not amyloid" in the mouse nasal septum.

An eosinophilic substance (ES) is usually observed in the mouse nasal septum and increases in volume with aging. It has been described as amyloid in textbooks and one report. However, it has been described as "not amyloid" in other reports because there was a negative reaction to Congo red. In this study, the ES was investigated histopathologically and electron microscopically to determine whether it was amyloid or not. The ES was only observed at the interstitium of clear HE-stained nasal glands in the septum, in which 2 kinds of glands were present (dark and clear stained by HE). The volume of the ES was small in young mice and large in older ones. Neither nasal gland degeneration nor inflammation resulted, even if a large amount of the ES was observed. The ES reacted negatively to Congo red but was strongly positive to periodic acid-Schiff reaction with prior diastase treatment. In the electron microscope observation, the ES consisted of amorphous material and collagen, but no nonbranching fibrils. Similar amorphous material was also observed in the nasal gland epithelial cells and was connected to the material in the rough endoplasmic reticulum. The above-mentioned findings indicated that the ES was not amyloid and suggested the ES might consist of not only collagen but also complex carbohydrate, which was produced by the nasal gland epithelial cells.

[The existence vomeronasal organ in adult humans]. / Wystepowanie narzadu lemieszowo-nosowego u ludzi doroslych.

The influence of chemical substances (feromones) on human emotional and physical condition has fascinated psychologists, sexuologists and laryngologists since centurie. Literature conveys inconsistent information on vomeronasal organ (VNO) occurrence in humans. This organ is often called Jacobson's, and 2 symmetrical openings leading into it, located on both sides of septum, are called Ruyasch's ducts. The aim of the study was to analyze vomeronasal organ occurrence in humans in relation to age and sex. The study was conducted in a group of 634 patients, aged 18-80 years. All patients underwent routine ENT examination including rhinoscopy, nasal cavity examination with usage of 2.5x magnification lens (surgical glasses) and surgical microscope with 10x magnification. All persons had nasal cavities examined endoscopically. Every time presence of vomeronasal organ openings, along with localization, size and symmetry of these was noted. Subjects, who presented Jacobson's organ, were asked to fill a questionnaire concerning influence of smells on erotic sensations. Vomeronasal organ was fund in 312 persons, that is 49.21%. In 83.65% of cases vomeronasal organ opening size was smaller than 0.2 mm, what restricted its visibility to usage of magnifying lens, microscope, or endoscope. In 16.34% of cases only vomeronasal organ ducts openings were well visible in routine rhinoscopy without magnification. Vomeronasal organ was found more often in men than women. VNO was significantly more rare in patients with nasal septal deviation. In these cases, vomeronasal organ was usually found unilaterally, in all the cases on the concave side of deviated nasal septum.

Estudo clínico e ultraestrutural pós-turbinectomia inferior parcial / Clinical and ultrastructural study after partial inferior turbinectomy

O presente trabalho refere-se ao estudo dos resultados clínicos e histológicos obtidos após a turbinectomia inferior parcial (TIP), cirurgia indicada no tratamento da obstrução nasal crônica causada pela hipertrofia das conchas nasais inferiores. MATERIAL E MÉTODOS: Foram estudados vinte pacientes, divididos em dois grupos de dez cada (grupos A e B), submetidos à TIP, associada à septoplastia ou não. Os pacientes foram reavaliados clinicamente e histologicamente (com biópsia das áreas regeneradas das conchas inferiores), em dois períodos pós-operatórios diferentes: um grupo após oito a doze meses (grupo A) e outro após dois anos de TIP (grupo B). RESULTADOS: Os resultados clínicos mostraram-se satisfatórios para o alívio da obstrução nasal no grupo A, e insatisfatórios no grupo B. Entretanto, melhores resultados histológicos, com maior recuperação e diferenciação epitelial da mucosa regenerada das conchas inferiores após a TIP foram observados no grupo B, com sua ultraestrutura ciliar normal. CONCLUSÕES: A cirurgia revelou ser eficaz a curto, mas não em médio prazo, apesar da recuperação histológica ter sido importante.

We report clinical and histological results obtained after partial inferior turbinectomy (PIT), surgery indicated for the treatment of chronic nasal obstruction. METHODS: Twenty patients were divided into two groups submitted to PIT plus septoplasty and PIT alone. The patients were reassessed clinically and histologically by means of a biopsy of the regenerated areas in the inferior turbinates at two different times after PIT, i.e., after 8 to 12 months (group A) and after two years (group B). RESULTS: The clinical results proved to be satisfactory for the relief of nasal obstruction in group A and unsatisfactory in group B. However, better histological results with better recovery and epithelial differentiation of the regenerated mucosa of the inferior turbinates after PIT were observed in group B. CONCLUSION: Surgery proved to be effective on a short-term but not on a long-term basis, and histological recovery did not accompany improvement of clinical signs and symptoms.

Scanning electron microscopy of ciliae and saccharine test for ciliary function in septal deviations.

OBJECTIVE: To investigate the difference in mucociliary clearance and surface mucosal structure of the nasal septum and lateral nasal wall in patients with and without septal deviation. METHOD: The saccharine-dye test was used to measure the mucociliary clearance time in both nasal cavities of 20 patients with nasal septal deviation (study group) and was compared with that of 30 patients without septal deviation (control group). Bilateral septal and lateral nasal wall mucosal biopsies were taken from the study group during septoplasty, and unilateral biopsies were taken from 10 of the control group. These biopsies were studied under the scanning electron microscope. RESULTS: In the study group, mucociliary clearance on the side opposite the septal deviation was significantly slower than on the other side. Mucociliary clearance on both sides of the deviated septum of the study group was significantly slower than clearance in the control group. There was no statistically significant difference in the distribution of mucosal cilia of the cavities on either side of the deviated septum in the study group, nor between the distribution in the study group and controls. CONCLUSION: Patients with septal deviation display no change in mucosal surface anatomy but have decreased mucociliary activity on both sides of the deviation, the least activity being on the side opposite the deviation.

Clinical and ultrastructural study after partial inferior turbinectomy.

UNLABELLED: We report clinical and histological results obtained after partial inferior turbinectomy (PIT), surgery indicated for the treatment of chronic nasal obstruction. METHODS: Twenty patients were divided into two groups submitted to PIT plus septoplasty and PIT alone. The patients were reassessed clinically and histologically by means of a biopsy of the regenerated areas in the inferior turbinates at two different times after PIT, i.e., after 8 to 12 months (group A) and after two years (group B). RESULTS: The clinical results proved to be satisfactory for the relief of nasal obstruction in group A and unsatisfactory in group B. However, better histological results with better recovery and epithelial differentiation of the regenerated mucosa of the inferior turbinates after PIT were observed in group B. CONCLUSION: Surgery proved to be effective on a short-term but not on a long-term basis, and histological recovery did not accompany improvement of clinical signs and symptoms.

Biomechanical strength of human nasal septal lining: comparison of the constituent layers.

OBJECTIVE/HYPOTHESIS: Nasal septal perforation is a common complication following surgery involving the nasal septum. Septoplasty, septorhinoplasty, and submucosal resection may result in the inadvertent resection of perichondrium, which may predispose the patient to septal perforations. STUDY DESIGN: Controlled human cadaver study testing the biomechanical strength of the constituent layers of nasal septal lining. METHODS: Uniform samples of nasal septal mucosa, perichondrium, and a composite of both layers were obtained from five fresh human cadavers. The mechanical tensile strength of these layers was evaluated and compared with the Instron 4301 Mechanical Testing System (Canton, MA). RESULTS: Mixed-effects regression analysis demonstrated a significant difference in the tensile strength of the three groups (mean values +/- SD: mucosa, 662 +/- 308 g; perichondrium, 1370 +/- 798 g; composite, 2340 +/- 1252 g). All three pairwise comparisons among the three groups showed a significant difference in tensile strength. CONCLUSION: The perichondrial layer imparts the majority of the biomechanical strength to septal lining. Lining flaps containing both perichondrium and mucosa are stronger than flaps with either perichondrium or mucosa alone. Dissection in the subperichondrial plane during septal surgery provides a stronger septal flap and may prevent the development of nasal septal perforation during nasal surgery.

The effect of silver nitrate on nasal septal cartilage.

Epistaxis from the anterior septum is frequently treated with a topical application of silver nitrate, which cauterizes the bleeding vessel. However, this treatment causes a septal perforation in a small percentage of patients. We report our study of the histologic effect of topical silver nitrate on samples of septal tissue obtained from 11 patients. We found that 30 seconds of exposure allowed silver nitrate to penetrate to a depth of approximately 1 mm. Longer exposure (45 and 60 sec) resulted in no significant additional penetration. Similarly, the amount of silver nitrate deposition into the chondrocytic lacunae did not vary significantly with the length of exposure. On the other hand, the depth of deposition into the extracellular matrix was positively associated with the duration of exposure. We found no direct evidence that silver nitrate exerted any damaging effect on septal cartilage. Instead, the development of septal perforations in patients who receive topical silver nitrate may be attributable to necrosis of the septal cartilage following damage to the overlying perichondrium, from which it derives its blood supply.

The effects of water-soluble cyclodextrins on the histological integrity of the rat nasal mucosa.

The aim of this study was to investigate the effect of highly water-soluble cyclodextrins (CDs) on the histological integrity of the nasal mucosa. In order to evaluate their effects, the in vivo single and repeated nasal exposure studies were performed using male Wistar rats. The rat nasal cavity was excised after an application of various CD solutions at different times. The morphological appearances of the rat nasal mucosae were analyzed with the light microscopic and the scanning electron microscopic studies. By utilizing 5-min exposure of each CD solution to the nasal mucosa, no tissue damage was visible for 1.5% w/v beta-CD and 5 and 20% w/v hydroxypropyl beta-CD (HP beta-CD), and the effects were quite similar to controls. However, using 20% w/v randomly methylated beta-CD (RM beta-CD) showed severe damage on the integrity of nasal mucosa. The severity was similar to 1% w/v polyoxyethylene-9-lauryl ether or l% w/v sodium deoxycholate. Meanwhile, 30 or 60 min exposure to 10% w/v HP beta-CD or RM beta-CD resulted in no obvious mucosal damage. In addition, in vivo repeated dosing of RM beta-CD did not show any toxicity up to 20% w/v. These results suggest that at least, less than 10% w/v CD solutions do not induce gross tissue damage and can keep the histological integrity of the nasal mucosa.

The human vomeronasal organ: part IV. Incidence, topography, endoscopy, and ultrastructure of the nasopalatine recess, nasopalatine fossa, and vomeronasal organ.

BACKGROUND: Previous reports on the human vomeronasal organ (VNO) have been inconsistent. Observations of fossae on the nasal septum have been reported as the VNO. METHODS: Adult human subjects (210) and cadavers (31) were examined using rigid nasal endoscopy, serial histology, and biopsy ultrastructure (5). RESULTS: The nasopalatine fossa (NPF) and the nasopalatine recess (NPR) are discrete, but variable, structures located adjacent to the VNO region. The NPF is not a vomeronasal pit. A septal mucosal pit could hide the vomeronasal duct opening. The VNO is a submucosal structure located 2-8 mm superior to the NPR and cannot be positively identified either macroscopically or endoscopically. CONCLUSION: The VNO has long been mistaken for the NPF and septal mucosal pits. We show that serial histology is the correct method for identifying the VNO.

Two-photon excitation laser scanning microscopy of human, porcine, and rabbit nasal septal cartilage.

Two-photon excitation laser scanning microscopy (TPM) was used to image human, porcine, and rabbit nasal septal cartilage. TPM provides optical sections of thick tissue specimens in situ without the use of exogenous dyes or need for tissue fixation. The cartilage tissue was imaged using near-infrared light generated by a mode-locked titanium/sapphire laser that was raster-scanned and coupled to an inverted microscope. Absorption of two photons by endogenous molecules and subsequent fluorescence was filtered to specific spectral bandwidths and detected with photomultiplier tubes. Two-photon stimulated fluorescence was detected with photomultiplier tubes optimized to specific spectral bandwidths. Signal intensity corresponds to the concentration of fluorophores, principally NADH, NADPH, and flavoproteins hence providing a means of redox imaging the cellular metabolic state. Specimens were scanned from the surface to a depth of about 150 microm. Image size was 50 x 50 microm with a diffraction limited pixel size of 0.4 microm. Cell membranes, nuclei, and matrix structures were identified in human, pig, and rabbit tissues. TPM provides a means to study three dimensional chondrocyte structure and matrix organization in situ at substantial depths, and permits longitudinal examination of cultured tissue explants without the need for exogenous dyes, tissue preparation, or fixation.

Hereditary angioedema: report of a case.

Hereditary angioedema is caused by an absolute deficiency or the functional inactivity of C1 esterase inhibitor in plasma. A precise diagnosis is important because, unlike allergic forms of mucocutaneous edema, this condition does not respond to epinephrine, antihistamines, or corticosteroids. We report the case of a 24-year-old man who experienced an acute attack after he had stopped taking his prophylactic medication.