Rat Trachea Research Articles

Immunohistochemical analysis and distribution of epithelial mast cells in the rat larynx and trachea.

Mast cells (MCs) in rat airways have been classified into two subtypes: epithelial MCs and connective tissue MCs (CTMCs). However, the immunohistochemical characteristics, cellular morphology, and distribution of epithelial MCs in the upper airways remain unclear. The present study investigated the morphological characteristics and distribution of epithelial MCs using 5-hydroxytryptamine (5-HT) and other immunohistochemical markers in sectioned or whole-mount preparations of the rat larynx and trachea. A double immunofluorescence analysis revealed the colocalization of 5-HT immunoreactivity with c-kit, a stem cell factor receptor commonly used as a MC marker, in both epithelial MCs and CTMCs. Dopa decarboxylase, an enzyme involved in 5-HT synthesis, was detected in both subtypes, suggesting their ability to synthesize and release 5-HT. Tryptase and histidine decarboxylase (a biosynthetic enzyme of histamine), which are well-known mediators of MCs, were exclusive to CTMCs. Epithelial MCs were pleomorphic with long cytoplasmic processes, whereas CTMCs were round and lacked cytoplasmic processes. The density of epithelial MCs was significantly higher in the glottis and cranial part of the trachea than in the epiglottis and other parts of the trachea. The present results showed that the morphology and immunohistochemical characteristics of epithelial MCs were different from those of CTMCs in the rat larynx and trachea, and variform epithelial MCs were predominantly located at the entrance of the upper airways. Epithelial MCs may release 5-HT to regulate innate immune responses by modulating epithelial cell functions at the entrance gate of the upper airways.

Lentiviral vector gene therapy and CFTR modulators show comparable effectiveness in cystic fibrosis rat airway models

Mutation-agnostic treatments such as airway gene therapy have the potential to treat any individual with cystic fibrosis (CF), irrespective of their CF transmembrane conductance regulator (CFTR) gene variants. The aim of this study was to employ two CF rat models, Phe508del and CFTR knockout (KO), to assess the comparative effectiveness of CFTR modulators and lentiviral (LV) vector-mediated gene therapy. Cells were isolated from the tracheas of rats and used to establish air-liquid interface (ALI) cultures. Phe508del rat ALIs were treated with the modulator combination, elexacaftor-tezacaftor-ivacaftor (ETI), and separate groups of Phe508del and KO tracheal epithelial cells were treated with LV-CFTR followed by differentiation at ALI. Ussing chamber measurements were performed to assess CFTR function. ETI-treated Phe508del ALI cultures demonstrated CFTR function that was 59% of wild-type level, while gene-addition therapy restored Phe508del to 68% and KO to 47% of wild-type level, respectively. Our findings show that rat Phe508del-CFTR protein can be successfully rescued with ETI treatment, and that CFTR gene-addition therapy provides significant CFTR correction in Phe508del and KO ALI cultures to levels that were comparable to ETI. These findings highlight the potential of an LV vector-based gene therapy for the treatment of CF lung disease.

Promoting successful healing of artificial trachea by intratracheal administration of basic fibroblast growth factor†.

This study evaluated the effect of intratracheal administration of basic fibroblast growth factor (bFGF) on tracheal healing following implantation of a novel layered polyglycolic acid (PGA) material to replace a critical-size defect in rat trachea. A critical-size defect in the rat cervical trachea was covered with PGA. Distilled water (DW) or 3.125, 6.25, 12.5 or 25 µg bFGF was administered into the trachea for 2 weeks (n = 6 for each of 5 groups). Regenerated areas of cilia, ciliary beat frequency and ciliary transport function (CTF) in the centre of the PGA were measured. To examine potential side effects of intratracheal administration of bFGF, the right lower lobe was pathologically evaluated. All rats survived during the study period. Histological examination showed ciliated epithelization on the PGA material after 2 weeks. Bronchoscopy revealed stenosis due to granulation following administration of high concentrations of bFGF (12.5 and 25 µg). Compared with the DW group, groups administered 3.125, 6.25, 12.5 and 25 µg bFGF had significantly larger areas of regenerated cilia (15.2%, 27.0%, 41.3%, 33.1% and 31.0%, respectively; P = 0.00143), improved ciliary beat frequency (7.10, 8.18, 10.10, 9.50 and 9.50 Hz, respectively), and improved CTS (6.40, 9.54, 16.89, 16.41 and 14.29 µm/sec, respectively). Pathological examination of the right lower lobe revealed pulmonary fibrosis and hyperplasia with high concentrations of bFGF (12.5 and 25 µg). Intratracheal administration of bFGF effectively promoted tracheal regeneration at an optimal dose of 6.25 µg following implantation of an artificial trachea.

Relaxant Activity of 4H-Pyran and 1,6-Dihydropyridine Derivatives on Isolated Rat Trachea

Derivatives of 4H-pyrans and 1,6-dihydropyridines have generated considerable attention due to their interesting biological and therapeutic values. Their pharmacological activities include vasorelaxant, anticarcinogenic, antimicrobial, and antioxidant activities. Thus, the aim of the current work is to determine the relaxant effect of synthesized 4H-pyran and 1,6-dihydropyridine derivatives with potential anti-asthmatic properties on the smooth muscle airway, with a possible Ca2+-channel blockade as a mechanism of action due to their analogy with 1,4-dihidropyridines. 4H-pyrans and 1,6-dihydropyridines were achieved using multicomponent reactions by microwave and conventional heating. Also, test samples were evaluated ex vivo to determine their relaxant effect on isolated rat tracheal rings pre-contracted with carbachol. All compounds evaluated showed a significant relaxant effect on carbachol-induced contraction in tracheal rat rings. Compounds 4b, 4e, 7a, and 8d were the most potent from the entire series and were also more potent than theophylline, used as a positive control. In conclusion, in the current work some relaxant compounds of the airway smooth muscle with potential to be developed as anti-asthmatic drugs were obtained.

Bisphenol A promotes cell death in healthy respiratory system cells through inhibition of cell proliferation and induction of G2/M cell cycle arrest.

Bisphenol A (BPA) is a substance that can harm the environment and human health by interfering with the normal functioning of the body's hormonal system. It is commonly found in various plastic-based products such as cosmetics, canned foods, beverage containers, and medical equipment and as well as it can also be absorbed by inhalation. There have been limited studies on the effects of BPA on lung fibroblasts, and it is still unclear how high levels of BPA can impact respiratory system cells, particularly the lungs and trachea. In this research, we aimed to investigate the cell cycle disruption potential of BPA on respiratory system cells by examining healthy trachea and lung cells together for the first time. The findings indicated that BPA exposure can alter the healthy cells' morphology, leading to reduced cellular viability that has been assessed by MTT and SRB assays. BPA treatment was able to activate caspase3 as expected, which could cause apoptosis in treated cells. Although the highest dose of BPA did not increase the apoptotic rate of rat trachea cells, it remarkably caused them to become necrotic (52.12%). In addition to quantifying the induction of apoptosis and necrosis by BPA, cell cycle profiles were also determined using flow cytometry. Thereby, BPA treatment unexpectedly inhibited the cell cycle's progression by causing G2/M cell cycle arrest in both lung and tracheal cells, which hindered cell proliferation. The findings of the study suggested that exposure to BPA could lead to serious respiratory problems, even respiratory tract cancers via alterations in the cell cycle.

Computed Tomographic Anatomy and Topography of the Lower Respiratory System of the Mature Rat (Rattus norvegicus)

Among various mammals, the laboratory rat is widely used for experimental purposes and belongs to the order Rodents, the family Muridae, and the genus Rattus. The absence of a thorough study of the topography and CT anatomy of the thoracic structures necessitates such a study. This study aims to accurately describe the topography of the lungs, trachea, and heart in rats based on CT scan findings compared with the anatomical findings in this animal. CT scan images were prepared from 10 adult male rats at a distance of 1 mm using a Siemens Somatom Spirit CT Scanner, followed by anatomical studies. Morphometric factors were measured on CT scan images, and then samples were studied anatomically by autopsy. The thorax, lungs, heart, and intrathoracic trachea, as well as the locations of organs, were carefully examined in anatomical studies, and detailed descriptions were obtained from CT scan images and autopsy observations. The tracheal bifurcation was fixed in the fourth and fifth ribs of all samples. The right lung was larger and more voluminous than the left one. The heart was tilted to the left, and the right bronchus was shorter than the left one. According to the obtained results, the right lung was more voluminous than the left one in R. norvegicus, which was confirmed by accurate measurements. It was also longer than the left lung but with the same width and height. It should be noted that the use of CT scans allows the anatomical examination of body structures when being alive and active, and this can be very effective practically. In general, the lungs continue from the second rib to the last rib, but the right lung occupies a little more space from the front and the dorsal. Regarding the lobulation of the lungs, the left lung has one lobe and the right lung has four lobes. It should be noted that the use of CT scans allows the anatomical examination of body structures when being alive and active, and this can be very effective practically.

A 3D printable tissue adhesive

Tissue adhesives are promising alternatives to sutures and staples for joining tissues, sealing defects, and immobilizing devices. However, existing adhesives mostly take the forms of glues or hydrogels, which offer limited versatility. We report a direct-ink-write 3D printable tissue adhesive which can be used to fabricate bioadhesive patches and devices with programmable architectures, unlocking new potential for application-specific designs. The adhesive is conformable and stretchable, achieves robust adhesion with wet tissues within seconds, and exhibits favorable biocompatibility. In vivo rat trachea and colon defect models demonstrate the fluid-tight tissue sealing capability of the printed patches, which maintained adhesion over 4 weeks. Moreover, incorporation of a blood-repelling hydrophobic matrix enables the printed patches to seal actively bleeding tissues. Beyond wound closure, the 3D printable adhesive has broad applicability across various tissue-interfacing devices, highlighted through representative proof-of-concept designs. Together, this platform offers a promising strategy toward developing advanced tissue adhesive technologies.

Investigating the relaxant effect of aqueous-alcoholic extract of Melissa Officinalis on smooth muscle of isolated rat trachea and its potential mechanisms

Investigating the relaxant effect of aqueous-alcoholic extract of Melissa Officinalis on smooth muscle of isolated rat trachea and its potential mechanisms

Sodium Selenite Ameliorates Silver Nanoparticles Induced Vascular Endothelial Cytotoxic Injury by Antioxidative Properties and Suppressing Inflammation Through Activating the Nrf2 Signaling Pathway

Silver nanoparticles (AgNP) are the dominant nanomaterials in commercial products and the medical field, but the widespread occurrence of AgNP has become a global threat to human health. Growing studies indicate that AgNP exposure can induce vascular endothelial toxicity by excessive oxidative stress and inflammation, which is closely related to cardiovascular disease (CVD), but the potential intrinsic mechanism remains poorly elucidated. Thus, it has been crucial to control the toxicological effects of AgNP in order to improve their safety and increase the outcome of their applications.Multiple researches have demonstrated that sodium selenite (Se) possesses the capability to counteract the toxicity of AgNP, but the functional role of Se in AgNP-induced CVD is largely unexplored. The aim of this study was to explore the potential protective effect of Se on AgNP-induced vascular endothelial lesion and elucidate the underlying mechanisms. An in vivo model of toxicity in animals was established by the instillation of 200 µL of AgNP into the trachea of rats both with (0.2 mg/kg/day) and without Se treated. In vitro experiments, human umbilical vein endothelial cells (HUVECs) were incubated with AgNP (0.3 µg/mL ) and Se for a duration of 24 h. Utilizing transmission electron microscopy, we observed that the internalization of AgNP-induced endothelial cells was desquamated from the internal elastic lamina, the endoplasmic reticulum was dilated, and the medullary vesicle formed. Se treatment reduced the levels of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1), inhibited the release of pro-inflammatory cytokines (specifically tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6), improved endothelial cell permeability, integrity, and dysfunction, and prevented damage to the aortic endothelium caused by AgNP. Importantly, we found that Se showed the capacity against AgNP with biological functions in guiding the intracellular reactive oxygen species (ROS) scavenging and meanwhile exhibiting anti-inflammation effects. Se supplementation decreased the intracellular ROS release and suppressed NOD-like receptor protein 3 (NLRP3) and nuclear factor kappa-B (NF-κB) mediated inflammation within AgNP-intoxicated rats and HUVECs. The anti-oxidant stress and anti-inflammatory effects of Se were at least partly dependent on nuclear factor erythroid 2-related factor 2 (Nrf2). Overall, our results indicated that the protectiveness of Se against AgNP-induced vascular endothelial toxicity injury was at least attributed to the inhibition of oxidative ROS and pro-inflammatory NF-κB/NLRP3 inflammasome by activating the Nrf2 and antioxidant enzyme (HO-1) signal pathway.

An ex vivo rat trachea model reveals abnormal airway physiology and a gland secretion defect in cystic fibrosis.

Cystic fibrosis (CF) is a genetic disease hallmarked by aberrant ion transport that results in delayed mucus clearance, chronic infection, and progressive lung function decline. Several animal models have been developed to study the airway anatomy and mucus physiology in CF, but they are costly and difficult to maintain, making them less accessible for many applications. A more available CFTR-/- rat model has been developed and characterized to develop CF airway abnormalities, but consistent dosing of pharmacologic agents and longitudinal evaluation remain a challenge. In this study, we report the development and characterization of a novel ex vivo trachea model that utilizes both wild type (WT) and CFTR-/- rat tracheae cultured on a porcine gelatin matrix. Here we show that the ex vivo tracheae remain viable for weeks, maintain a CF disease phenotype that can be readily quantified, and respond to stimulation of mucus and fluid secretion by cholinergic stimulation. Furthermore, we show that ex vivo tracheae may be used for well-controlled pharmacological treatments, which are difficult to perform on freshly excised trachea or in vivo models with this degree of scrutiny. With improved interrogation possible with a durable trachea, we also established firm evidence of a gland secretion defect in CFTR-/- rat tracheae compared to WT controls. Finally, we demonstrate that the ex vivo tracheae can be used to generate high mucus protein yields for subsequent studies, which are currently limited by in vivo mucus collection techniques. Overall, this study suggests that the ex vivo trachea model is an effective, easy to set up culture model to study airway and mucus physiology.

Allyl isothiocyanate mitigates airway inflammation and constriction in a house dust mite-induced allergic asthma model via upregulation of tight junction proteins and the TRPA1 modulation

Asthma is a chronic inflammatory disease that has been associated with insufficient vegetable intake. Allyl Isothiocyanate (AITC) is a natural isothiocyanate found in cruciferous plants with anti-inflammatory and antioxidant abilities. Our study aimed to investigate the potential effect of AITC on tracheal constriction in a house dust mite (HDM)-induced asthma animal model, and explore the underlying mechanisms. To investigate the effects of AITC on HDM-induced allergic asthma model, established by intranasally administering extracts of HDM and AITC or DEX was given orally for four weeks. Flexivent SCIREQ, H&E staining, ELISA were employed to evaluate the lung function and the cytokine secretion. Possible mechanisms were determined by Western blot. Rat tracheae contraction was measured by Labscribe. We utilized lung epithelial cells (BEAS-2B) to assess the adhesion response to the combination of inflammatory factors TNF-α and IL-4. The results of the study showed that AITC significantly reduced tracheal constriction in ex vivo experiments and improved lung function in in vivo experiments compared to HDM-induced mice. Additionally, AITC decreased cytokine secretion, inflammatory cell infiltration in the lung, and constriction-related proteins expression in both lung and tracheae. Moreover, AITC increased tight junction-related protein expression in lung tissues. In vitro experiments showed that AITC had a protective effect through TRPA1 channel without affecting cell viability. Our results demonstrate that AITC has potential anti-asthma effects in HDM-induced asthma models by alleviating airway inflammation and airway constriction through increasing tight junction-related protein expression and suppressing Ca2+ signaling. These findings suggest that AITC may be a beneficial adjuvant therapy in asthma treatment.

Impact of the cAMP efflux and extracellular cAMP-adenosine pathway on airway smooth muscle relaxation induced by formoterol and phosphodiesterase inhibitors

β2-adrenoceptors agonists and phosphodiesterase (PDE) inhibitors are effective bronchodilators, due to their ability to increase intracellular cyclic AMP (cAMP) levels and induce airway smooth muscle (ASM) relaxation. We have shown that increment of intracellular cAMP induced by β2-adrenoceptors agonist fenoterol is followed by efflux of cAMP, which is converted by ecto-PDE and ecto-5′-nucleotidases (ecto-5′NT) to adenosine, leading to ASM contraction. Here we evaluate whether other classical bronchodilators used to treat asthma and chronic obstructive pulmonary disease (COPD) could induce cAMP efflux and, as consequence, influence the ASM contractility. Our results showed that β2-adrenoceptor agonists formoterol and PDE inhibitors IBMX, aminophylline and roflumilast induced cAMP efflux and a concentration-dependent relaxation of rat trachea precontracted with carbachol. Pretreatment of tracheas with MK-571 (MRP transporter inhibitor), AMP-CP (ecto-5′NT inhibitor) or CGS-15943 (nonselective adenosine receptor antagonist) potentiated the relaxation induced by β2-adrenoceptor agonists but did not change the relaxation induced by PDE inhibitors. These data showed that all bronchodilators tested were able to induce cAMP efflux. However, only β2-adrenoceptor-induced relaxation of tracheal smooth muscle was affected by cAMP efflux and extracellular cAMP-adenosine pathway.

Rosuvastatin relaxes rat thoracic aorta, pulmonary artery, and trachea via nitric oxide, prostanoids, and potassium channels

Purpose: This study aimed to determine the functional effects and mechanisms of the action of rosuvastatin on vascular and tracheal smooth muscle tissues. 
 Materials and Methods: Vascular and tracheal rings (2-3 mm) isolated from the thoracic aortas, pulmonary arteries, and tracheas of Wistar Albino male rats (250-300 g) were placed in chambers in the isolated tissue bath system. As the resting tension, 1 g was selected. Vascular rings contracted with 10-6 M phenylephrine after a 90-minute equilibration period. Tracheal rings contracted with 10-5 M acetylcholine. After the contraction was steady, rosuvastatin (10-8-10-4 M) was cumulatively applied to the vascular and tracheal rings. The defined experimental methodology was repeated following the incubation of selective inhibitors of signaling pathways and K+ channel blockers to ascertain rosuvastatin's functional effect mechanisms. 
 Results: In the precontracted rat vascular and tracheal rings, rosuvastatin induced concentration-dependent relaxation. The maximal relaxation level in vessel samples was 96%. On the other hand, the maximal relaxation level in tracheal samples was found to be 75%. The vasorelaxant effects of rosuvastatin were dramatically attenuated by endothelium removal, L-NAME treatment, and indomethacin incubation (up to 27%). With the incubation of tetraethylammonium, glyburide, 4-Aminopyridine, and anandamide, rosuvastatin-mediated vascular smooth muscle relaxation levels were significantly decreased (up to 38%). Moreover, With the incubation of tetraethylammonium, glyburide, and 4-Aminopyridine rosuvastatin-mediated tracheal smooth muscle relaxation levels were significantly decreased (up to 30%). 
 Conclusion: Rosuvastatin has a noticeable relaxing effect on the vascular and tracheal smooth muscles. The vasorelaxant effect of rosuvastatin involves intact endothelium, nitric oxide, prostanoids, and K+ channels (BKCa, KV, and KATP channels). Furthermore, nitric oxide, prostanoids, BKCa channels, KV channels, and KATP channels play a role in rosuvastatin-induced tracheal smooth muscle relaxation

Incompletely Decellularized Tracheal Matrix Scaffold for Tissue Engineering.

Dense cartilaginous extracellular matrix makes decellularization and repopulation of tracheal cartilage difficult. However, the dense matrix isolates cartilaginous antigens from the recipient's immune system. Therefore, allorejection may be avoided by removing antigens from noncartilaginous tissues. In this study, incompletely decellularized tracheal matrix scaffolds were developed for tracheal tissue engineering. Brown Norway rat tracheae were decellularized with 4% sodium deoxycholate treatment. The cell and antigen removal efficacy, histoarchitecture, surface ultrastructure, glycosaminoglycan, collagen contents, mechanical properties, and chondrocyte viability of the scaffold were evaluated in vitro. Brown Norway rat tracheal matrix scaffolds ( n = 6) were implanted subcutaneously into Lewis rats and observed for 4 weeks. Brown Norway rat tracheae ( n = 6) and Lewis rat scaffolds ( n = 6) were implanted as controls. Histologic analysis of macrophage and lymphocyte infiltration was performed. One decellularization cycle removed all cells and antigens from noncartilaginous tissue. Incomplete decellularization preserved the structural integrity of the tracheal matrix and chondrocyte viability. Except for 31% glycosaminoglycan loss, the scaffold had comparable collagen content and tensile and compressive mechanical properties to those of the native trachea. The allogeneic scaffold showed remarkably reduced CD68 + , CD8 + , and CD4 + cell infiltration compared with the allografts and demonstrated similar cell infiltration to the syngeneic scaffold. It also maintained the three-dimensional tracheal structure and cartilage viability in vivo. Incompletely decellularized trachea did not induce immunorejection and maintained the integrity and viability of cartilage in vivo. Tracheal decellularization and repopulation can be simplified for urgent tracheal replacement. The present study describes the development of an incomplete decellularization protocol that creates a decellularized matrix scaffold for tracheal tissue engineering, aiming to provide preliminary data that this method may generate suitable tracheal scaffolds for use in tracheal replacement.

Effectıveness of Curcumın on Tracheal Anastomosıs Created in the Rat Model

Objective: We aimed to investigate the effect of curcumin on tracheal wound healing in the anastomosis model following tracheal incision in rats. Methods: Twenty-one healthy male Wistar Albino rats were included in the study. A horizontal incision was made between the 2nd-3th rings of the rat trachea and damage was created in the inner tracheal mucosa. Tracheal incision area was sutured and anastomosis was performed. Rats were divided into three groups. Ringer was given to the first group, corticosteroids to the 2nd group, and curcumin to the 3rd group for 28 days. The rats were sacrificed on the 28th day and the tracheal anastomosis line samples were sent for histological examination. Wound healing parameters, tracheal lumen width, wall thickness, and stenosis index were evaluated. Results: Statistically significant difference was detected in tracheal lumen width and wall thickness in the curcumin group (p<0.05). Statistically significant differences were observed in the curcumin group in parameters of inflammation, collagen production and fibrosis. (p<0.05). Conclusion: It is important to prevent the formation of tracheal stenosis, which is difficult to treat in clinical practice. Desired results cannot be achieved in the side-effect profile of some medical drugs used in this regard. We found the positive effect of Curcumin in the prevention of stenosis following tracheal injury.

IMPACT OF PARTIAL STENOSIS OF TRACHEA ON ANTIOXIDANT SYSTEM IN YOUNG RATS

Introduction. Recently, there has been a significant rise in the attention of researchers and clinicians towards the issue of complex hypoxic trauma affecting various organs and body systems. This is particularly important when considering the cells that are most vulnerable to oxygen deprivation, including brain tissue, lungs, certain organs of the gastrointestinal tract, systemic immunity. The significance of this problem becomes even more relevant when examining the impact of hypoxia on the cells of sexually immature mammals and humans, as well as exploring potential treatments for acute hypoxic conditions including laryngomalacia and stridor observed in children under the age of one year. Moreover, there are a number of associated complex and unresolved issues in experimental clinical practice. The aim of the study is to investigate changes in the enzyme activity of the pro-oxidant-antioxidant system of the lung tissue cells and the central nervous system (cerebrocortex) during the experimental modelling of respiratory pathology (by ligating the upper trachea of rats), the features of their recovery and the dynamics of this process after the surgical removal of the tracheal ligature.
 Materials and methods. The studies were performed on 37 sexually immature male Wistar rats, aged 2528 days. The test animals were divided into four groups: the first group served as the control (intact) rats, the second group consisted of rats with experimentally induced tracheal stenosis, which were removed from the experiment in 7 days after the operation, the third group consisted of rats with experimentally induced tracheal stenosis who were removed from the experiment in 21 days after the operation, and the fourth group consisted of rats whose tracheal ligature was removed on the 7th day after the operation and then they were removed from the experiment on the 21st day. The following biochemical parameters were studied: malondialdehyde content, activity of catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase.
 Results. The findings obtained demonstrate an increase in malondialdehyde content and antioxidant enzyme activity in the lung and brain tissues of rats on the 7th and 21st days following partial tracheal stenosis. Restoring airway ventilation did not fully normalize these indicators to the control values.
 In conclusion, the findings indicate that the impaired pro-oxidant-antioxidant balance in lung and central nervous system cells did not completely restore even two weeks after relieving the compressive effect on the trachea and resolving the hypoxic respiratory system disorders partially. This suggests the necessity to continue therapy with pharmacological antioxidant agents following surgical intervention. However, further research is required to explore this aspect of the problem in depth.

Development of novel layered polyglycolic acid sheet for regeneration of critical-size defect in rat trachea.

Polyglycolic acid (PGA) sheets are difficult to adapt to the central airway because of poor durability against high air pressure. Therefore, we developed a novel layered PGA material to cover the central airway and examined its morphologic traits and functional performance as a potential tracheal replacement. A critical-size defect in rat cervical tracheas was covered with the material. Morphologic changes were bronchoscopically and pathologically evaluated. Functional performance was evaluated by regenerated ciliary area, ciliary beat frequency and ciliary transport function determined by measuring the moving distance of microspheres dropped onto the trachea (µm/s). The evaluation time points were 2 weeks, 1 month, 2 months and 6 months after surgery (n = 5, respectively). Forty rats underwent implantation, and all survived. Histological examination confirmed ciliated epithelization on the luminal surface after 2 weeks. Neovascularization was observed after 1 month, tracheal glands after 2 months and chondrocyte regeneration after 6 months. Although the material was gradually replaced by self-organization, tracheomalacia was not bronchoscopically observed at any time point. The area of regenerated cilia significantly increased between 2 weeks and 1 month (12.0% vs 30.0%; P = 0.0216). The median ciliary beat frequency significantly improved between 2 weeks and 6 months (7.12 vs 10.04 Hz; P = 0.0122). The median ciliary transport function was significantly improved between 2 weeks and 2 months (5.16 vs 13.49 µm/s; P = 0.0216). The novel PGA material showed excellent biocompatibility and tracheal regeneration both morphologically and functionally 6 months after tracheal implantation.

Elabela relaxes rat pulmonary artery and trachea via BKCa, KV, and KATP channels

ObjectiveElabela is a newly discovered peptide hormone. This study aimed to determine the functional effects and mechanisms of action of elabela in rat pulmonary artery and trachea. Materials and methodsVascular rings isolated from the pulmonary arteries of male Wistar Albino rats were placed in chambers in the isolated tissue bath system. The resting tension was set to 1 g. After the equilibration period, the pulmonary artery rings were contracted with 10-6 M phenylephrine. Once a stable contraction was achieved, elabela was applied cumulatively (10-10-10-6 M) to the vascular rings. To determine the vasoactive effect mechanisms of elabela, the specified experimental protocol was repeated after the incubation of signaling pathway inhibitors and potassium channel blockers. The effect and mechanisms of action of elabela on tracheal smooth muscle were also determined by a similar protocol. ResultsElabela exhibited a concentration-dependent relaxation in the precontracted rat pulmonary artery rings (p < .001). Maximal relaxation level was 83% (pEC50: 7.947 CI95(7.824–8.069)). Removal of the endothelium, indomethacin incubation, and dideoxyadenosine incubation significantly decreased the vasorelaxant effect levels of elabela (p < .001). Elabela-induced vasorelaxation levels were significantly reduced after iberiotoxin, glyburide, and 4-Aminopyridine administrations (p < .001). L-NAME, methylene blue, apamin, TRAM-34, anandamide, and BaCl2 administrations did not cause a significant change in the vasorelaxant effect level of elabela (p = 1.000). Elabela showed a relaxing effect on precontracted tracheal rings (p < .001). Maximal relaxation level was 73% (pEC50: 6.978 CI95(6.791–7.153)). The relaxant effect of elabela on tracheal smooth muscle was decreased significantly after indomethacin, dideoxyadenosine, iberiotoxin, glyburide, and 4-Aminopyridine incubations (p < .001). ConclusionsElabela exerted a prominent relaxant effect in the rat pulmonary artery and trachea. Intact endothelium, prostaglandins, cAMP signaling pathway, and potassium channels (BKCa, KV, and KATP channels) are involved in the vasorelaxant effect of elabela. Prostaglandins, cAMP signaling pathway, BKCa channels, KV channels, and KATP channels also contribute to elabela-induced tracheal smooth muscle relaxant effect.

2023 TERMIS – AMERICAS Conference &amp; Exhibition Boston Marriott Copley Place April 11–14, 2023

2023 TERMIS – AMERICAS Conference &amp; Exhibition Boston Marriott Copley Place April 11–14, 2023

Effect of the Hydroalcoholic Extract from the Leafy Stems of Waltheria indica L. (Malvaceae) on Acetylcholine and Barium Chloride-Induced Contractions on Isolated Rat Tracheal Tissue

A previous study has reported the interesting relaxant effect of the hydroalcoholic extract from Waltheria indica L. (Malvaceae) leafy stems, a plant with several therapeutic uses. The present study aimed to investigate the preventive effect of this plant using an ex vivo model of the rat trachea. Two agonists, acetylcholine (10-6 - 1.5×10-5 M) and barium chloride (10-5- 10-1 M) were used to induce contractions. The preventive effect was assessed on rat tracheal rings pretreated with hydroalcoholic extract (1.92 mg/mL), glibenclamide, atropine, and papaverine, all at 10 µM. Acetylcholine and barium chloride provoke contractions in a concentration-dependent manner, with a maximum contractile response of 3.953 ± 0.692 g and 2.999 ± 0.326 g, respectively. The EC50 values were 3.711 ± 0.823 µM and 9.502 ± 12.354 mM, respectively, for acetylcholine- and barium chloride-induced contraction. Glibenclamide caused a rightward shift of the acetylcholine-response curve, followed by a reduction of the maximum contraction (from 3.953 ± 0.692 g to 3. 116 ± 0.244 g). The hydroalcoholic extract, atropine, papaverine, and their combinations induced a complete suppression of the contractile response to acetylcholine (p &lt; 0.0001) and barium chloride (p &lt; 0.0001). The hydroalcoholic extract exhibited a potent relaxant effect comparable to that of atropine and papaverine. It can be concluded that the hydroalcoholic extract of W. indica can potently prevent acetylcholine-and barium chloride-induced contractions. The possible mechanisms by which the extract exerts its relaxant effect may involve the blockade of muscarinic receptors, the inhibition of phosphodiesterase activity, and/or the calcium channel.