Anatomic features of the fetal round and oval windows, and their relations with the tympanic nerve.

OBJECTIVE: The study aimed to examine morphometric properties of the round window (RW) and oval window (OW) and to show their relation with the tympanic nerve (the Jacobson's nerve, JN) in human fetuses from the otologic surgeon's perspective. METHODS: Thirty temporal bones of 15 fetal cadavers (8 males, 7 females) aged with 24.40 ± 3.71 weeks were included in the study. The height, width and surface area of the RW and OW and also distance from the JN to the OW and RW were measured. RESULTS: The height, width and surface area of the RW in this work were measured as 1.48 ± 0.25 mm, 1.57 ± 0.37 mm, and 2.05 ± 0.69 mm2, respectively. The RW was detected as round-shaped (8 cases, 26.7%), oval-shaped (15 cases, 50%), and dome-shaped (7 cases, 23.3%). The height, width and surface area of the OW were measured as 1.42 ± 0.26 mm, 2.90 ± 0.44 mm, and 3.63 ± 0.74 mm2, respectively. The OW was observed as oval-shaped (15 cases, 50%), kidney-shaped (10 cases, 33.3%), D-shaped (4 cases, 13.3%), and trapezoid-shaped (1 case, 3.3%). The JN was found 1.21 ± 0.60 and 1.18 ± 0.54 mm away from the RW and OW, respectively. CONCLUSION: This study containing morphological data about the shapes, diameters and area of the RW and OW may be useful to predict surgical difficulty, and to select implants of suitable size preoperatively for the windows. Knowing the relationship between the JN and the windows can be helpful to avoid iatrogenic injuries of the nerve.

Developmental aspects of the tympanic membrane: Shedding light on function and disease.

The ear drum, or tympanic membrane (TM), is a key component in the intricate relay that transmits air-borne sound to our fluid-filled inner ear. Despite early belief that the mammalian ear drum evolved as a transformation of a reptilian drum, newer fossil data suggests a parallel and independent evolution of this structure in mammals. The term "drum" belies what is in fact a complex three-dimensional structure formed from multiple embryonic cell lineages. Intriguingly, disease affects the ear drum differently in its different parts, with the superior and posterior parts being much more frequently affected. This suggests a key role for the developmental details of TM formation in its final form and function, both in homeostasis and regeneration. Here we review recent studies in rodent models and humans that are beginning to address large knowledge gaps in TM cell dynamics from a developmental biologist's point of view. We outline the biological and clinical uncertainties that remain, with a view to guiding the indispensable contribution that developmental biology will be able to make to better understanding the TM.

Differing contributions of the first and second pharyngeal arches to tympanic membrane formation in the mouse and chick.

We have proposed that independent origins of the tympanic membrane (TM), consisting of the external auditory meatus (EAM) and first pharyngeal pouch, are linked with distinctive middle ear structures in terms of dorsal-ventral patterning of the pharyngeal arches during amniote evolution. However, previous studies have suggested that the first pharyngeal arch (PA1) is crucial for TM formation in both mouse and chick. In this study, we compare TM formation along the anterior-posterior axis in these animals using Hoxa2 expression as a marker of the second pharyngeal arch (PA2). In chick, the EAM begins to invaginate at the surface ectoderm of PA2, not at the first pharyngeal cleft, and the entire TM forms in PA2. Chick-quail chimera that have lost PA2 and duplicated PA1 suggest that TM formation is achieved by developmental interaction between a portion of the EAM and the columella auris in PA2, and that PA1 also contributes to formation of the remaining part of the EAM. By contrast, in mouse, TM formation is highly associated with an interdependent relationship between the EAM and tympanic ring in PA1.

Explicación ontogénica para la asociación entre dehiscencia del tegmen tympani y dehiscencia del canal semicircular superior / Ontogenetic explanation for tegmen tympani dehiscence and superior semicircular canal dehiscence association

Objetivos: Analizar la ontogenia del canal semicircular superior y del tegmen tympani y determinar si hay factores embriológicos comunes que expliquen la dehiscencia asociada de ambos. Métodos: Se han analizado 77 series embriológicas humanas de edades comprendidas entre las 6 semanas y recién nacidos. Las preparaciones estaban cortadas en serie y teñidas con la técnica de tricrómico de Martins. Resultados: La prolongación tegmentaria del tegmen tympani y el canal semicircular superior se originan de la misma estructura, la cápsula ótica, y poseen el mismo tipo de osificación endocondral; mientras que la prolongación escamosa del tegmen tympani se desarrolla desde la escama del temporal y su osificación es de tipo directa o intramembranosa. En la osificación de la prolongación tegmentaria colaboran los núcleos de osificación de los canales semicirculares superior, externo y accesorio del tegmen, los cuales por crecimiento se extienden hasta la prolongación tegmentaria, este hecho sumado a que ambas estructuras comparten una capa común de periostio externo podría explicar la coexistencia de falta de cobertura ósea en el tegmen y en el canal. Conclusión: El desarrollo del canal semicircular y tegmen tympani podrían explicar las causas de la asociación de ambas dehiscencias (AU)

Objectives: To analyze the ontogeny of the superior semicircular canal and tegmen tympani and determine if there are common embryological factors explaining both associated dehiscence. Methods: We analyzed 77 human embryological series aged between 6 weeks and newborn. Preparations were serially cut and stained with Masson's trichrome technique. Results: The tegmental prolongation of tegmen tympani and superior semicircular canal originate from the same structure, the otic capsule, and have the same type of endochondral ossification; while the extension of the squamous prolongation of tegmen tympani runs from the temporal squama and ossification is directly of intramembranous type. The nuclei of ossification of the superior and external semicircular canals and accessory of tegmen collaborate in the ossification of the tegmental extension and by growth extend to the tegmental prolongation. This fact plus the fact that both structures share a common layer of external periosteum could explain the coexistence of lack of bone coverage in tegmen and superior semicircular canal. Conclusion: The development of the semicircular canal and tegmen tympani could explain the causes of the association of both dehiscences (AU)

Developmental mechanisms of the tympanic membrane in mammals and non-mammalian amniotes.

The tympanic membrane is a thin layer that originates from the ectoderm, endoderm, and mesenchyme. Molecular-genetic investigations have revealed that interaction between epithelial and mesenchymal cells in the pharyngeal arches is essential for development of the tympanic membrane. We have recently reported that developmental mechanisms underlying the tympanic membrane seem to be different between mouse and chicken, suggesting that the tympanic membrane evolved independently in mammals and non-mammalian amniotes. In this review, we summarize previous studies of tympanic membrane formation in the mouse. We also discuss its formation in amniotes from an evolutionary point of view.

Developmental genetic bases behind the independent origin of the tympanic membrane in mammals and diapsids.

The amniote middle ear is a classical example of the evolutionary novelty. Although paleontological evidence supports the view that mammals and diapsids (modern reptiles and birds) independently acquired the middle ear after divergence from their common ancestor, the developmental bases of these transformations remain unknown. Here we show that lower-to-upper jaw transformation induced by inactivation of the Endothelin1-Dlx5/6 cascade involving Goosecoid results in loss of the tympanic membrane in mouse, but causes duplication of the tympanic membrane in chicken. Detailed anatomical analysis indicates that the relative positions of the primary jaw joint and first pharyngeal pouch led to the coupling of tympanic membrane formation with the lower jaw in mammals, but with the upper jaw in diapsids. We propose that differences in connection and release by various pharyngeal skeletal elements resulted in structural diversity, leading to the acquisition of the tympanic membrane in two distinct manners during amniote evolution.

Developmental origin and fate of middle ear structures.

Results from developmental and phylogenetic studies have converged to facilitate insight into two important steps in vertebrate evolution: (1) the ontogenetic origin of articulating elements of the buccal skeleton, i.e., jaws, and (2) the later origins of middle ear impedance-matching systems that convey air-borne sound to the inner ear fluids. Middle ear ossicles and other skeletal elements of the viscerocranium (i.e., gill suspensory arches and jaw bones) share a common origin both phylogenetically and ontogenetically. The intention of this brief overview of middle-ear development is to emphasize the intimate connection between evolution and embryogenesis. Examples of developmental situations are discussed in which cells of different provenance, such as neural crest, mesoderm or endoderm, gather together and reciprocal interactions finally determine cell fate. Effects of targeted mutagenesis on middle ear development are described to illustrate how the alteration of molecularly-controlled morphogenetic programs led to phylogenetic modifications of skeletal development. Ontogenetic plasticity has enabled the diversification of jaw elements as well as middle ear structures during evolution. This article is part of a special issue entitled "MEMRO 2012".

Closure of the middle ear with special reference to the development of the tegmen tympani of the temporal bone.

Closure of the middle ear is believed to be closely related to the evolutionary development of the mammalian jaw. However, few comprehensive descriptions are available on fetal development. We examined paraffin-embedded specimens of 20 mid-term human fetuses at 8-25 weeks of ovulation age (crown-rump length or CRL, 38-220 mm). After 9 weeks, the tympanic bone and the squamous part of the temporal bone, each of which was cranial or caudal to Meckel's cartilage, grew to close the lateral part of the tympanosquamosal fissure. At the same time, the cartilaginous tegmen tympani appeared independently of the petrous part of the temporal bone and resulted in the petrosquamosal fissure. Subsequently, the medial part of the tympanosquamosal fissure was closed by the descent of a cartilaginous inferior process of the tegmen tympani. When Meckel's cartilage changed into the sphenomandibular ligament and the anterior ligament of the malleus, the inferior process of the tegmen tympani interposed between the tympanic bone and the squamous part of the temporal bone, forming the petrotympanic fissure for the chorda tympani nerve and the discomalleolar ligament. Therefore, we hypothesize that, in accordance with the regression of Meckel's cartilage, the rapidly growing temporomandibular joint provided mechanical stress that accelerated the growth and descent of the inferior process of the tegmen tympani via the discomalleolar ligament. The usual diagram showing bony fissures around the tegmen tympani may overestimate the role of the tympanic bone in the fetal middle-ear closure.

Prenatal growth of the human tympanic membrane.

The questions connected with the morphology of developing tympanic membrane are rather inadequately dealt with in the relevant literature. The aim of this article has been the morphometric analysis of the prenatal growth of human tympanic membrane. The experiment was conducted on 33 fetuses aged from the 4th to 8th month of gestation. Significant individual variability of the tympanic membrane measurements was revealed in the material studied. The growth of the structure discussed was bilaterally symmetric during the whole period investigated. The fourth and 7th month of gestation seem to be the crucial stages for the tympanic membrane development. The measurements of the membrane exhibited the highest variability, simultaneously being negatively correlated with the fetal age in that periods. Relatively more intensive growth of the vertical diameter of the membrane was noted in the 5th and 8th months of gestation. In the 8th month of gestation the tympanic membrane reached a vertically elongated shape, typical of the postnatal period. On the basis of our results it is possible to conclude that the quantitative developmental process takes place in the tympanic membrane till the end of the prenatal period, determining the final functional capacity of the structure discussed.

[Contribution to the development of the union between the manubrium of the malleus and the tympanic membrane in human fetus]. / Contribución al desarrollo de la unión del mango del martillo y la membrana del tímpano en fetos humanos.

The development of the union between the manubrium of the malleus and the tympanic membrane was studied in human embryo. For that purpose 25 temporal bones of human foetus, aged from 36 days (14 mm) to 29 weeks (270 mm) were analyzed. Samples were fixed in a 10% formaldehyde solution, decalcified with 2% nitric acid, embedded in Paraplast, cut in sections of 7 microns thick and stained with Martin's trichrome method. During the development a pseudojoint between the malleus and the tympanic membrane several stages were seen. In the first stage, the manubrium was adhered to membrane mesenchyme of primitive tympanum, in the second one this mesenchyme was loose and there appeared capillars in it, in the third one there were collagen fibers in a radial disposition, and in the forth stage, the hollow in the meatal plug gave independence to the tympanic membrane for the external acoustic meatus. The distal portion of the manubrium, included in tympanic membrane, has remained with a cartilaginous structure during end of the time it has been studied by us. The collagen fibers at the level of the umbo surround the manubrium, while the rest of it remains in its anterior edge.

Contribución al desarrollo de la unión del mango del martillo y la membrana del tímpano en fetos humanos / Contribution to the development of the union between the manubrium of the malleus and the tympanic membrane in human fetus

Se ha estudiado el desarrollo de la unión entre el mango del martillo y la membrana del tímpano en embriones y fetos humanos.Para ello se han utilizado 25 huesos temporales de fetos humanos, cuyas edades cronológicas van desde los 36 días (14 mm) hasta las 29 semanas (270 mm). Las muestras fueron fijadas en formol al 10 por ciento, descalcificadas con ácido nítrico al 2 por ciento, incluidas en parafina, cortadas en serie a 7 µm, y teñidas con el método Tricrómico de Martins. Se han identificado varias etapas en el desarrollo de la pseudoarticulación entre el martillo y la membrana del tímpano. En la primera el mango se adhiere íntimamente al mesénquima de la membrana del tímpano primitiva, en la segunda dicho mesénquima se hace laxo y en él aparecen capilares, en la tercera aparecen fibras de colágeno de disposición radial, y en la cuarta el ahuecamiento del tapón meatal da independencia a la membrana del tímpano del conducto auditivo externo. La porción distal del mango, incluida en la membrana del tímpano, ha permanecido con una estructura cartilaginosa durante todo el tiempo estudiado. Las fibras de colágeno a nivel del umbo rodean al extremo terminal del mango mientras que en el resto de él lo hacen solamente en su arista anterior (AU)

The development of the union between the manubrium of the malleus and the tympanic membrane was studied in human embryo. For that purpose 25 temporal bones of human foetus, aged from 36 days (14 mm) to 29 weeks (270 mm) were analyzed. Samples were fixed in a 10% formaldehyde solution, decalcified with 2% nitric acid, embedded in Paraplast, cut in sections of 7 microns thick and stained with Martin's trichrome method. During the development a pseudojoint between the malleus and the tympanic membrane several stages were seen. In the first stage, the manubrium was adhered to membrane mesenchyme of primitive tympanum, in the second one this mesenchyme was loose and there appeared capillars in it, in the third one there were collagen fibers in a radial disposition, and in the forth stage, the hollow in the meatal plug gave independence to the tympanic membrane for the external acoustic meatus. The distal portion of the manubrium, included in tympanic membrane, has remained with a cartilaginous structure during end of the time it has been studied by us. The collagen fibers at the level of the umbo surround the manubrium, while the rest of it remains in its anterior edge (AU)

Assembling a functional tympanic membrane: signals from the external acoustic meatus coordinate development of the malleal manubrium.

In terrestrial mammals, hearing starts with the perception of acoustic pressure by the tympanic membrane. Vibrations in this membrane are then transduced into the inner ear by the ossicle chain of the middle ear, composed of the malleus, incus and stapes. The proper connection of the ossicle chain with the tympanic membrane, provided by the insertion of the manubrium of the malleus into the eardrum, is essential for the functionality of the hearing apparatus. We describe here the mechanisms regulating the development of the manubrium and its integration into the tympanic membrane. We show that the external acoustic meatus (EAM), which eventually forms the outer epithelium of the tympanic membrane, plays an essential role in this developmental process. Histological and expression analyses indicate that the manubrium develops close to the EAM with a similar temporal sequence. In addition, when the middle ear ossicles are allowed to develop in vitro under conditions that do not support further EAM development, the manubrium develops only up to the stage of its induction at the time of explantation. Moreover, genetically or teratogenically derived alterations in the EAM also have an effect on manubrial development. Finally, we show that the EAM is the source of two quite opposite activities, one that induces chondrogenesis and another that represses it. The combination of these two activities results in the proper positioning of the manubrium.

Maturation of tympanic membrane layers and collagen in the embryonic and post-hatch chick (Gallus domesticus).

The surface area of the chick (Gallus domesticus) tympanic membrane (TM) increases by as much as 400% from hatching to 70 days of age (Cohen et al. [1992] J. Morphol. 212:187-193). The present study is concerned with the processes that contribute to this remarkable size increase. Middle-ear specimens were harvested in embryos aged between E10 and E18, and in post-hatch animals between days P1 and P57. Specimens were embedded in paraffin, cut in serial sections, stained for collagen fibers, and examined with light microscopy. Four locations were examined in each specimen: the anterior and posterior perimeter of the TM, the TM, over the extra-stapedius, and the TM at the tip of the extra-columella. The thickness of the epithelial, respiratory, and lamina propria layers was measured at each location. The radial collagen fibers in the lamina propria were also counted at each location, and fiber density per square micrometer was determined at each age. Thickness of the epithelial and respiratory layers remains relatively constant throughout development in all areas of the TM, whereas the lamina propria at the extra-columella and extra-stapedius continues to thicken with increasing age. Collagen density also increases during development, and this is attributed to an increase in fiber number and a reduction in the space between fibers. The results suggest that collagen may be synthesized first in the central regions of the TM and then later in more peripheral areas of the TM.

Development of the mammalian ear: coordinate regulation of formation of the tympanic ring and the external acoustic meatus.

The tympanic membrane in mammals is a trilaminar structure formed by the apposition of two epithelial cell layers, along with an intervening layer of cells derived from pharyngeal arch mesenchyme. One epithelial layer is contributed by the external acoustic meatus, a derivative of the first pharyngeal cleft. The other epithelial layer is contributed by the tubotympanic recess, a derivative of the first pharyngeal pouch. We demonstrate here an absolute correlation between formation of the external acoustic meatus and formation of the tympanic ring, a first arch-derived membrane bone that anchors the tympanic membrane. Experimental loss of the tympanic ring by retinoic acid treatment, or duplication of the ring in Hoxa-2 null mutant embryos, resulted in corresponding alterations in formation of the external acoustic meatus. We suggest that the tympanic ring primordium induces formation and morphogenesis of the external acoustic meatus, and that expression of the Hoxa-2 and goosecoid genes may be involved in regulating the formation and morphogenesis of these structures.

Applied embryology and post-natal morphology of the human tympanic membrane.

This article deals with the morphological as well as histological changes of the tympanic membrane and bone from early embryological life until mature age. An overview of the continuous changes is given with reference to pertinent clinical implications.

Micropathologic changes of pars tensa in children with otitis media with effusion.

Clinical studies have supported a relationship between otitis media with effusion in children and chronic otitis media in adults. Although clinical studies are numerous, relatively little is known about the histopathologic changes of the tympanic membrane in otitis media with effusion. Tympanic membranes were taken from the intermediate zone of the anteroinferior quadrant of the tympanic membrane during surgery for placement of tympanostomy tubes in 30 children (age range, 4 to 10 years) who did not show any improvement after 3 months of conservative treatment for otitis media with effusion. Control specimens were taken from normal temporal bones at autopsy. All specimens were observed with light and electron microscopy. Histologic degeneration of the tympanic membrane of patients with otitis media with effusion occurred most often in the lamina propria and the submucosal layer. There was an increase in the thickness of the tympanic membrane as a result of edema and fibrosis of the submucosal layer. A decrease in the thickness of the outer and inner fibrous layers occurred in the lamina propria. The histopathologic changes observed in the lamina propria may result in a change in the elastic properties of the tympanic membrane.

Middle-ear development: II. Morphometric changes in the conducting apparatus of the chick.

The ontogeny of various middle-ear structures was examined in 11 groups of chicks between 10 days embryonic and adult. Measurements of the tympanic membrane surface area and height, columella length, and that of the columella footplate, annular ligament, and oval window area were obtained using video micrographs and computer digitization techniques. The oval window matures first at 53 days post-hatching, whereas the columella achieves adult size at 74 days. The tympanic membrane surface area is the last middle-ear variable studied to reach adult size (79 days post-hatch). The columella increases its length from 0.63 mm (10 days embryonic) to 2.73 mm in the adult. The tympanic membrane area expands by 280% whereas the columellar footplate area increases by 11x. As a result, the pressure amplification of the middle ear due to the tympanic membrane/columellar footplate area ratio improves by over 400%. These data further contribute to our understanding of the functional development of the middle ear.