CC BY
2SOME MICROSTRUCTURAL FEATURES CARTILAGE OF THE NOSE SEPTUM AND THEIR IMPORTANCE IN
SEPTOPLASTY
1Abzalova Sh. R., 2Bobokhonov M.G.
1,2 Tashkent Pediatric Medical Institute https://doi.org/10.5281/zenodo.13168887
Abstract. Currently, information about the cellular, biochemical and mechanical properties of the structures of the nasal septum is scattered. For example, the structure of cartilage is often based on studies done on other types of cartilage, such as knee hyaline cartilage. This study by the authors provides an opportunity for a detailed understanding of the properties of nasal septum cartilage, which is important for rhinosurgeons and future researchers of tissue engineers in the development of cartilage scaffolds (production of bioengineered scaffolds) for septoplasty.
Keywords: nasal septum; microstructure, perichondrium; collagen fibers; biomechanical properties.
Actuality. Deformation (according to ICD-10: J34. 2 - mixed septum) of the septum is one of the most common circumstances of surgical intervention in rhinosurgery. According to the literature, from 70.0 to 96.5% of the population has some degree of curvature of the nasal septum, but up to 3% of them are subject to septoplasty [2.,3,4].
A detailed understanding of the clinical microstructures of the nasal septum cartilage is important for otorhinolaryngologists and future scientific researchers in this field.
Although the standard basic septoplasty technique is often sufficient for patients with mild to moderate mid-posterior septal deviation, problems arise with caudal septal deviation and associated dentofacial anomalies [1].
Secondly, modern methods of histological research make it possible not only to study tissues as a whole, but also to isolate individual types of cells from them (to study their life activity over a long time), to isolate individual cellular organelles and their constituent macromolecules (for example, deoxyribonucleic acid molecules - DNA, collagen), explore their functional features. Such opportunities have opened up in connection with the creation of new modern medical devices and technologies - various types of microscopes, computer equipment, X-ray diffraction analysis, the use of nuclear magnetic resonance (NMR), radioactive isotopes and autoradiography, electrophoresis and chromatography, fractionation of cellular contents using ultracentrifugation, separation and cultivation of cells, production of hybrids (using biotechnological methods - production of hybridomas), monoclonal antibodies, recombinant DNA, etc. [2]. As is known, the implementation of the main functions of the nose, such as respiratory, protective, informational, calorific, olfactory, excretory, and suction, is ensured by the clear interaction of all microanatomical structures and physiological mechanisms [3,4].
Oral and tracheal breathing cannot replace nasal breathing! In modern conditions, almost any surgical intervention on the microstructures of the nasal cavity is accompanied by surgical tissue trauma of varying severity. Measures aimed at quickly restoring mucociliary clearance are
becoming important, which is impossible without restoring the morphofunctional structure of the nasal mucosa [3,4,5,6,7].
The solid support of the nasal septum is formed by the cartilage of the nasal septum, the vomer and the perpendicular plate of the ethmoid bone and is covered with a mucoperichondrial flap. The mucoperichondrial flap of the nasal septum normally in humans consists of 3 layers: 1. Covering epithelium; 2. The lamina propria of the mucous membrane; 3. Perichondrium, and in its bony part - periosteum (periosteum); 4. Cartilage (Figure 1)
» A i «
¿-a' /- . • *
Jf 1r ** \ ^ ^ 50 Mm
Figure 1. Sample of the nasal septum. In the hyaline cartilage of the septum, perichondrium (P), chondroblasts, chondrocytes and blood vessels (asterisks) are distinguished. Beneath the pseudostratified columnar ciliated epithelium (white arrow) with goblet cells (black arrow) are glands (dashed line). Hematoxylin-eosin staining *400.
The supporting basis of the external nose and nasal septum is hyaline cartilage. The septum is bordered below by the median palatine suture (Sutura palatina mediana). The growth and development, pathology of these structures are interdependent and are reflected in the formation of the "rhinomaxillary complex" [9,15,16].
Cartilage is a connective tissue consisting of collagen fibers embedded in a strong matrix of chondroitin sulfates. (Figures 2,3)
The purpose of our work is, based on an analysis (review) of literature data and our own research, to unify (standardize) the existing accumulated knowledge about the microstructural properties of the cartilage of the nasal septum and to identify important gaps (directions) necessary for future research. Electron microscopy technique.
Cartilage from septoplasty patients was immediately placed in 10% phosphate-buffered formalin. Samples were fixed in this solution for 24 hours and then transferred to phosphate buffered saline and postfixed with 1% aqueous osmium tetroxide and then dehydrated using progressive concentrations of ethanol and water, followed by progressive concentrations of ethanol and hexamethyldisilazane.
Samples were mounted on substrates, coated with gold using a PAC-1 Pelco Advanced Coater, and imaged using a Philips Optics scanning electron microscope.
The resulting images were viewed and analyzed using photo editing software (Adobe Photoshop 7.0). Fiber width measurements of all samples (where present) were taken from each micrograph at ><3000 magnification and analyzed using spreadsheet software (Microsoft Excel).
Figure 2. Schematic histological structure of hyaline cartilage covered with perichondrium: 1 - fibrous layer of perichondrium; 2 - chondrogenic layer of the perichondrium; 3 - young chondrocytes; 4 - chondrocyte in the lacuna; 5 - intercellular substance (cartilage matrix); 6 - interstitial growth; 7 - isogenic groups of chondrocytes (mature cartilage cells) (according to A. Ham and D. Cormack, 1983) [10]. Results. Using optical microscopy, we studied the ultrastructure of the cartilage of the nasal septum in 6 children and found:
Figure 3. Hyaline cartilage of the nasal septum under a light microscope.
Using scanning electron microscopy, the ultrastructure of the nasal septal cartilage in 3 children was studied and it was found that the collagen fibers of the septal cartilage are located in a mesh frame with larger lacunae and thicker fibers measuring 3.18 p,m (standard deviation = 0.75 p,m) with a confidence interval 99.9% from 2.74 to 3.54 |im, corresponds to the data of Holden PK, Liaw LH, Wong BJ (2008) [10,11].
According to Chinese otorhinolaryngologists Jia H, Sun K, Liu X, Li Y, Liu Z, Guo Y (2022), using scanning electron microscopy, they studied the ultrastructure of the cartilage of the nasal septum measuring 5x5 mm in 8 patients with a deviated nasal septum who had undergone septoplasty and It was found that the cartilage of the nasal septum looks the same in patients of different ages [12].
Figure 4. Scanning electron micrographs of human nasal septum cartilage. Each subsequent image from A to C has increasing magnifications of1000x, 2000x, and 3000x. (A) General arrangement of tightly interconnected sheets of collagen fibers. (B and C) At higher magnification, thick collagen fibers are visible, clearly arranged in a dense linear array
around the wider and more round lacunae.
Brown W.E., Lavernia L, Bielajew B.J., Hu J.C., Athanasiou K.A. (2023) [13] studied the mechanical and biochemical properties of cartilage obtained from several (7) sites - the human nasal septum. The septal cartilage contained the most type II collagen and less collagen I and III, making it more hyaline (very similar to human articular cartilage, despite the lack of articulations in the nose) (Figure 5). When comparing the mechanical properties of the seven points of the nasal septum with each other, the authors did not identify any statistical differences. They showed that the septum contains thick sheets of broad, highly organized collagen fibers that can resist exudation of water, resulting in decreased tissue permeability and a higher aggregate compressive modulus.
When comparing the biochemical composition of septal cartilage across 7 regions, no statistical differences were also revealed and were consistent with the distribution of collagen and GAG in articular cartilage. Given the similarity of nasal septal cartilage to articular cartilage, multiple biomimetic implantation and tissue engineering strategies are possible.
Conclusion. There is insufficient information about the microstructure of the nasal septum cartilage. In fact, the microstructure of this cartilage differs slightly from normal hyaline cartilage. The authors stained sections with hematoxylin and eosin (H&E) to illustrate tissue and cell morphology; for histochemical visualization of GAGs - safranin-O/fast green and for histochemical visualization of collagen fibers - picrosirius red (PSR). To determine collagen directions, picrosirius red-stained slides were also imaged under a microscope under polarized light [13]. As can be seen from (Figure 5), in all regions the peripheral zones contained flat, elongated cells oriented parallel to the edges, and in the central zones - round chondrocytes in the lacunae. The nasal septum S stained most strongly for GAG. ULC showed less zonal cell organization but higher cell density and more intense collagen staining. Strong pericellular GAG staining in all regions suggests the presence of chondrons. The black dotted lines indicate the boundary between cartilage and mucoperichondrium, indicated by the letter "M" [13].
Total
H&E GAG GAG Col
TO
CD CD
O r-J
CD Q_
CO
CO
CD CD
C_>
CD Q_
o rvj
53
C_5 ^
TO
CD CD
CD Q_
o r-vl
C_>
tr CD
c_>
-C• .- .. - - • *"
3SP . :> 1 •
■ ■ ■ ■ } • ivj M M
* ' * J ' £ « c - - r, CM- Ä1
M „ \ >\ f * j*/.. _ _ Ml
to - ' JT - d 200 |jm 0M 200 |_im
Figure 5. Histology by region. Lines: S (septum) — cartilage of the nasal septum; ULC (upper lateral cartilage) - upper lateral cartilage and LLC (over lateral cartilage) - lower lateral cartilage. Columns: H&E (hematoxylin & eosin) — hematoxylin and eosin staining; GAG 1 and 2 (glycosaminoglycan) — glycosaminoglycans — stained with safranin-O/fast green; Total Col (total collagen) - total collagen - stained with Picrosirius red (PSR). "M" mark the border
between cartilage and mucoperichondrium.
ULC showed less zonal cell organization but higher cell density. Cells were grouped into lacunae with dense pericellular GAG staining. GAG staining was slightly more intense in the central zone than in the peripheral zones. However, this pattern was not as pronounced as in the septum. The color of collagen was found to be uniform throughout the thickness of the ULC. In addition, the transition between the cartilage and the surrounding mucoperichondrial tissue was less distinct than in the septum, as shown by the dotted line in (Figure 5) [13].
LLC had similar cell organization and morphology as ULC. However, in the central zone the cell density was lower than in the ULC. Cells were grouped into lacunae with intense GAG staining. GAG staining was slightly more intense in the peripheral zones compared to the central zone. Collagen staining was intense and uniform throughout the cartilage. The transition from cartilage to mucoperichondrium in the ULC is also not as pronounced as in the septum [13].
REFERENCES
1. Babakhanov G.K. Diagnosis and treatment of deviated nasal septum in children. Dissertation for the degree of Doctor of Medical Sciences (Doctor of Science). 14.00.04 -Otorhinolaryngology. Tashkent, 2020. 203 pp. [PDF dissertation]
2. Afanasyev Yu.I., Yurina N.A., Vinnikov Ya.A., Radostina A.I. Chentsov Yu.S. Histology, embryology, cytology. Textbook. M.: GEOTAR-Media, 2014: 800 pp.
3. Piskunov G.Z. Physiology and pathophysiology of the nose and paranasal sinuses. Russian rhinology. 2017; 25(3): 51-57. doi:10.17116/rosrino201725351-57 https://mediasphera.ru/issues/rossijskaya rinologiya/2017/3/downloads/ru/1086954742017031051
4. Shilenkova V.V., Fedoseeva O.V. Nasal cycle in healthy adults. Russian rhinology. 2016;(2):20-25.
5. Khasanov S.A., Makhsudov S.N., Babakhanov G.K. Otorhinolaryngological and orthodontic issues of the pathology of the rhinomaxillary complex in children. [Edited by S.A. Khasanova]: Tashkent Pediatric Medical Institute of the Ministry of Health of the Republic of Uzbekistan. - T.: "University", 2022. 284 p.
6. Karpishchenko S. A., Aleksandrov A. N., Shakhnazarov A. E., Fatalieva A. F., Kucherenko M. E. Functional state of the nasal cavity after endoscopic septoplasty. Russian otorhinolaryngology. 2020;19(3):16-21. https://doi.org/10.18692/1810-4800-2020-3 -16-21
7. Moldavskaya A.A., Khrappo N.S., Levitan B.N., Petrov V.V. Features of the organization of the mucous membrane and the vascular system of the nasal cavity: morphofunctional and clinical aspects (review). Advances of modern natural science. Number: 5 Year: 2006 Pages: 18-22 [eLIBRARY] [PDF]
8. Ozdemir S, Celik H, Cengiz C, Zeybek ND, Bahador E, Aslan N. Histopathological effects of septoplasty techniques on nasal septum mucosa: an experimental study. Eur Arch Otorhinolaryngol. 2019 Feb;276(2):421-427. doi: 10.1007/s00405-018-5226-7. Epub 2018 Dec 1. PMID: 30506412 [DOI] [PubMed] [sci-hub]
9. Babakhanov G.K. Diagnosis and treatment of deviated nasal septum in children. Dissertation for the degree of Doctor of Medical Sciences (Doctor of Science). 14.00.04 -Otorhinolaryngology. Tashkent, 2020. 203 pp. [PDF dissertation]
10. Ham A., Cormack D. Histology: In volume 3, 11 pages. Transl. from English / A. Ham, D. Cormack. - M.: Mir, 1983. [Ham A., Cormack D. Histology: In 3 volumes. Trans. from English / - Moscow: Mir, 1983.] (rus.)
11. Holden PK, Liaw LH, Wong BJ. Human nasal cartilage ultrastructure: characteristics and comparison using scanning electron microscopy. Laryngoscope. 2008 Jul; 118(7):1153-1156. doi: 10.1097/MLG.0b013e31816ed5ad. [PMC free article] [PubMed] [CrossRef] [Google Scholar]
12. Jia H, Sun K, Liu X, Li Y, Liu Z, Guo Y. [Scanning electron microscope of the human nasal septum]. Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi [Journal of clinical otorhinolaryngology, head, and neck surgery]. 2022 May;36(5):335-337;342. doi: 10.13201/j.issn.2096-7993.2022.05.002. PMID: 35483681 Free PMC article. Chinese.[PubMed] [PMC]
13. Brown WE, Lavernia L, Bielajew BJ, Hu JC, Athanasiou KA. Human nasal cartilage: Functional properties and structure-function relationships for the development of tissue engineering design criteria. Acta Biomater. 2023 Sep 15;168:113-124. doi: 10.1016/j.actbio.2023.07.011. Epub 2023 Jul 16. PMID: 37454708 Free article. [PubMed] [open access] [Google Scholar] [CrossRef]
14. Ustunel I, Cayli S, Guney K, Celik-Ozenci C, Tanriover G, Sahin Z, Balkan E, Demir R. Immunohistochemical distribution patterns of collagen type II, chondroitin 4-sulfate, laminin and fibronectin in human nasal septal cartilage. Acta Histochem. 2003;105(2):109-14. doi: https://pubmed.ncbi.nlm.nih.gov/12831162/. PMID: 12831162 [PubMed]
15. Tursunov, E. A., Kornilova, O. A., Umarkhodzhaev, A. M., Saidaliyev, M. S., & Abzalova, Sh. R. (2008). Introduction to the study of private cytology in the course of histology. Morphology, 133(2), 139-139.
16. Abzalova Sh. R., Ajimuratov X. /Histologiya atlasi/ in Korakolpok language, Tashkent, pages -26-30.
