Tracheal Cartilage Development Research Articles

Study of developing tracheal cartilage under light microscope

Background: Airway formation is one of the earliest events in development of respiratory system. However, maturation of airway continues well in post-natal life. The trachea has framework of incomplete ring of hyaline cartilage united by fibrous tissue and smooth muscle through which air conduction takes place. The trachea-bronchial mucosa is target for host of airborne toxic agents; bacterial and viral agents, allergens, reactive gases, aerosols, and irritant particles. This fascinates many workers for detailed study of epithelium or lining air passage. Aims and Objectives: The present study was undertaken to document normal developmental microscopic structure of tracheal cartilage in fetuses of different age group (gestational ages between 12th and 40th weeks) and to compare the difference between different age groups. Materials and Methods: Tissues from trachea of various age group fetuses were collected, processed, and stained with H and E and Masson’s trichrome. Results: Development of tracheal cartilage was observed into three trimesters. The 2nd and 3rd trimesters were further divided in the early 2nd (13th–16th weeks), mid-2nd (17th–20th weeks), late 2nd (21st–24th weeks), early 3rd (25th–28th weeks), mid-3rd (29th–32nd weeks), and late 3rd (33rd–40th weeks) phases. Conclusion: Perichondrium of cartilage had shown well-marked outer fibrous layer with increased vascularity by late 2nd to early 3rd trimester. Well-marked inner cellular layer of perichondrium was noticed by 3rd trimester, while cartilage had shown active chondrocytes by late 2nd to early 3rd trimester. At the late 3rd trimester, typical hyaline cartilage had shown chondrocytes in cell nest surrounded by territorial matrix.

Disruption of a hedgehog-foxf1-rspo2 signaling axis leads to tracheomalacia and a loss of sox9+ tracheal chondrocytes.

ABSTRACTCongenital tracheomalacia, resulting from incomplete tracheal cartilage development, is a relatively common birth defect that severely impairs breathing in neonates. Mutations in the Hedgehog (HH) pathway and downstream Gli transcription factors are associated with tracheomalacia in patients and mouse models; however, the underlying molecular mechanisms are unclear. Using multiple HH/Gli mouse mutants, including one that mimics Pallister–Hall Syndrome, we show that excessive Gli repressor activity prevents specification of tracheal chondrocytes. Lineage-tracing experiments show that Sox9+ chondrocytes arise from HH-responsive splanchnic mesoderm in the fetal foregut that expresses the transcription factor Foxf1. Disrupted HH/Gli signaling results in (1) loss of Foxf1, which in turn is required to support Sox9+ chondrocyte progenitors, and (2) a dramatic reduction in Rspo2, a secreted ligand that potentiates Wnt signaling known to be required for chondrogenesis. These results reveal an HH-Foxf1-Rspo2 signaling axis that governs tracheal cartilage development and informs the etiology of tracheomalacia.This article has an associated First Person interview with the first author of the paper.

Incoherent Feedforward Regulation via Sox9 and ERK Underpins Mouse Tracheal Cartilage Development.

Tracheal cartilage provides architectural integrity to the respiratory airway, and defects in this structure during embryonic development cause severe congenital anomalies. Previous genetic studies have revealed genes that are critical for the development of tracheal cartilage. However, it is still unclear how crosstalk between these proteins regulates tracheal cartilage formation. Here we show a core regulatory network underlying murine tracheal chondrogenesis from embryonic day (E) 12.5 to E15.5, by combining volumetric imaging of fluorescence reporters, inhibitor assays, and mathematical modeling. We focused on SRY-box transcription factor 9 (Sox9) and extracellular signal-regulated kinase (ERK) in the tracheal mesenchyme, and observed a synchronous, inverted U-shaped temporal change in both Sox9 expression and ERK activity with a peak at E14.5, whereas the expression level of downstream cartilage matrix genes, such as collagen II alpha 1 (Col2a1) and aggrecan (Agc1), monotonically increased. Inhibitor assays revealed that the ERK signaling pathway functions as an inhibitory regulator of tracheal cartilage differentiation during this period. These results suggest that expression of the cartilage matrix genes is controlled by an incoherent feedforward loop via Sox9 and ERK, which is supported by a mathematical model. Furthermore, the modeling analysis suggests that a Sox9-ERK incoherent feedforward regulation augments the robustness against the variation of upstream factors. The present study provides a better understanding of the regulatory network underlying the tracheal development and will be helpful for efficient induction of tracheal organoids.

Development pattern of tracheal cartilage in human embryos.

IntroductionCongenital tracheal anomalies are associated with high morbidity and mortality. The etiology of congenital tracheal anomalies is not well understood, but often attributed to malformed tracheal cartilage. The development of tracheal cartilage has not been described in detail. In this study, we aimed to investigate the development pattern and timing of normal tracheal cartilage to better understand the etiology of tracheal anomalies.Materials and methodsThe development of tracheal cartilage was examined by studying the trachea in histological sections of 14 healthy human embryos from the Carnegie collection. Two specimens for Carnegie Stages 17–23 (42–60 days of embryological development) were studied.ResultsAt Carnegie Stages 17–19 (42–51 days), a continuous mesenchymal condensation was observed ventral to the tracheal lumen. At Stages 20 and 21 (51–54 days), this pre‐tracheal mesenchyme showed sites of increased condensation indicative of future tracheal rings. Furthermore, growth centers were identified both proximally and distally in the trachea. Characteristic horseshoe shaped tracheal rings were apparent at Carnegie Stages 22 and 23 (54–60 days).ConclusionsIn human embryos, tracheal rings arise from growth centers in the ventral mesenchyme at approximately 51–54 days of embryological development. The observation of proximal and distal growth centers suggests a centripetal growth gradient, potentially contributing to occurrence of complete tracheal ring deformity (CTRD). Although this study shows new insights on tracheal cartilage development, the exact origin of congenital tracheal defects has yet to be elucidated.

Notum balances Wnt signaling during tracheal cartilage development

Notum balances Wnt signaling during tracheal cartilage development

Correlation of Symptoms with Bronchoscopic Findings in Children with a Prenatal Diagnosis of a Right Aortic Arch and Left Arterial Duct.

A right aortic arch (RAA) with a left arterial duct (LAD) together encircle the trachea and have the potential to cause tracheobronchial compression and published guidelines recommend bronchoscopy in symptomatic patients. The aim of the study was to describe the incidence of tracheal compression in a cohort of prenatally diagnosed RAA and LAD. Retrospective review of clinical course and imaging of prenatal cases of RAA and LAD assessed with flexible bronchoscopy over an 11-year period. 34 cases of prenatally diagnosed RAA with LAD underwent bronchoscopy at median age of 9months (range 0.4-123) of whom 11 had respiratory symptoms and 23 were asymptomatic. In the neonatal period, three cases demonstrated respiratory symptoms. An aberrant left subclavian artery (ALSA) was identified in 29 cases. Pulsatile tracheal compression was identified in 32/34 (94%) cases and two cases showed normal tracheal appearances. Significant tracheal compression (> 70% occlusion) was present in 25/34 (74%) cases of which 16 were asymptomatic. Significant carinal compression (> 70% occlusion) was identified in 14/34 (42%) cases, an ALSA was observed in 13/14. Surgical relief of a vascular ring has been performed in 27 (79%) cases at a median age of 15months (range 0.6-128months). At surgery, a fibrous remnant of an atretic left aortic arch was identified in 11/27 (41%) cases. Significant tracheal compression may be present in infants even without symptoms. If early relief of airway compression is to be achieved to promote normal development of tracheal cartilage, early bronchoscopy should be considered.

The anoctamin (TMEM16) gene family: calcium‐activated chloride channels come of age

The anoctamin <i>(TMEM16)</i> gene family: calcium‐activated chloride channels come of age

Regulation of Sox9 by Sonic Hedgehog (Shh) is essential for patterning and formation of tracheal cartilage

We report that Sonic Hedgehog (Shh) regulates both formation and patterning of tracheal cartilage by controlling the expression pattern and level of the chondrogenic gene, Sox9. In Shh(-/-) tracheo-esophageal tubes, Sox9 expression is transient and not restricted ventrally to the site of chondrogenesis, and is absent at the time of chondrogenesis, resulting in the failure of tracheal cartilage formation. Inhibition of Hedgehog signalling with cyclopamine in tracheal cultures prevents tracheal cartilage formation, while treatment of Shh(-/-) tracheal explant with exogenous Shh peptide rescues cartilage formation. Both exogenous Bmp4 and Noggin rescue cartilage phenotype in Shh(-/-) tracheal culture, while promoting excessive cartilage development in wild-type trachea through induction of Sox9 expression. The ventral and segmented expression of Sox9 in tracheal primordia under Shh modulated by Bmp4 and Noggin thus determine where and when tracheal cartilage develops. These results indicate that Shh signalling is a critical determinant in tracheal cartilage development.

A simple in vitro culture system for tracheal cartilage development

Semi-circular tracheal cartilage is a critical determinant of maintaining architectural integrity of the respiratory airway. The current effort to understand the morphogenesis of tracheal cartilage is challenged by the lack of appropriate model systems. Here we report an in vitro tracheal cartilage system using embryonic tracheal–lung explants to recapitulate in vivo tracheal cartilage developmental processes. With modifications of a current lung culture protocol, we report a consistent in vitro technique of culturing tracheal cartilage from primitive mouse embryonic foregut for the first time. This tracheal culture system not only induces the formation of tracheal cartilage from the mouse embryonic foregut but also allows for the proper patterning of the developed tracheal cartilage. Furthermore, we show that this culture technique can be applied to culturing other types of cartilage in vertebrae, limbs, and ribs. We believe that this novel application of our in vitro culture system will facilitate the manipulation of cartilage development under various conditions and thus enabling us to advance our current limited knowledge on cartilage biology and development.

GROWTH HORMONE THERAPY IN TRACHEOMALACIA

Growth hormone may be involved in normal cartilage production. Two patients with GH deficient states were treated with GH. Both patients, one with panhypopituitarism (patient 1) and another with pseudohypoparathyroidism (patient 2) presented with severe tracheomalacia. Both patients required tracheostomy. Patient 1 had his tracheostomy at age 3 years and 9 years later, despite numerous ENT procedures to reduce granulation tissue, could not be extubated due to collapse of the tracheal cartilage. Patient 1 was treated for hypothyroidism and hypoadrenalism. An insulin-glucagon test showed no GH values greater than 2 ng/ml (Hybritech IRMA). At one month of life, patient 2 had a tracheostomy and was diagnosed with pseudohypoparathyroidism with no significant rise in phosphate excretion, cyclic-AMP, or calcium (urinary or serum ionized) after PTH administration. This patient had a low IGF-1 and IBP-3. Patient 1 was treated with GH at a dose of 0.05 mg/kg/dose SC 6X per week; patient 2 at a dose of 0.1 mg/kg/day SC (the higher dose due to resistance to peptide hormones), each for six months. After treatment, the tracheal cartilage had become more firm, allowing extubation in both patients. This experience suggests a role for GH in the development of tracheal cartilage.