Immunohistochemical characterization of the rabbit tracheal cartilages
ABSTRACT
The objective of this study was to immunohistochemically elucidate the major extracellular matrix constituents of rabbit tracheal cartilage. The impetus for this project is the need for crucial design and validation criteria for tissue engineering juxtaposed with the conspicuous lack of trachea extracellular matrix data in the literature. Tracheal tissue specimens were harvested from New Zealand White rabbits, and were immunostained for collagen I, collagen II, aggrecan and decorin; and a Verhoeff-Van Gieson stain was performed to visualize elastin. The most striking result was the highly organized relationship between distinct fibrous (containing collagen I, decorin and elastin) and hyaline-like (containing collagen II and aggrecan) regions of the tracheal wall. The tracheal cartilage stained strongly with collagen II throughout, with periodic bands of aggrecan in the tracheal arches, meaning that there were areas void of aggrecan immunostaining alternating with areas with strong aggrecan immunostaining. In contrast, the periphery of the cartilage and the perichondrium itself exhibited strong collagen I staining and no collagen II staining. Elastin fibers and decorin were also detected along the periphery of the cartilage in the perichondrium and corresponded highly with the distribution of collagen I staining. The body of the rabbit trachea is therefore composed of a hyaline-cartilage structure primarily made of collagen II and bands of aggrecan, surrounded by a fibrous region composed of elastin and collagen I, indicative of a flexible tissue with distinct regions of compressive integrity. This information will be a valuable reference to future tissue engineering efforts in the creation of a biosynthetic substitute for laryngotracheal reconstruction.
The objective of this study was to immunohistochemically elucidate the major extracellular matrix constituents of rabbit tracheal cartilage. The impetus for this project is the need for crucial design and validation criteria for tissue engineering juxtaposed with the conspicuous lack of trachea extracellular matrix data in the literature. Tracheal tissue specimens were harvested from New Zealand White rabbits, and were immunostained for collagen I, collagen II, aggrecan and decorin; and a Verhoeff-Van Gieson stain was performed to visualize elastin. The most striking result was the highly organized relationship between distinct fibrous (containing collagen I, decorin and elastin) and hyaline-like (containing collagen II and aggrecan) regions of the tracheal wall. The tracheal cartilage stained strongly with collagen II throughout, with periodic bands of aggrecan in the tracheal arches, meaning that there were areas void of aggrecan immunostaining alternating with areas with strong aggrecan immunostaining. In contrast, the periphery of the cartilage and the perichondrium itself exhibited strong collagen I staining and no collagen II staining. Elastin fibers and decorin were also detected along the periphery of the cartilage in the perichondrium and corresponded highly with the distribution of collagen I staining. The body of the rabbit trachea is therefore composed of a hyaline-cartilage structure primarily made of collagen II and bands of aggrecan, surrounded by a fibrous region composed of elastin and collagen I, indicative of a flexible tissue with distinct regions of compressive integrity. This information will be a valuable reference to future tissue engineering efforts in the creation of a biosynthetic substitute for laryngotracheal reconstruction.
Cite this paper
nullWemer, R. , Detamore, M. and Weatherly, R. (2010) Immunohistochemical characterization of the rabbit tracheal cartilages. Journal of Biomedical Science and Engineering, 3, 1007-1013. doi: 10.4236/jbise.2010.310131.
nullWemer, R. , Detamore, M. and Weatherly, R. (2010) Immunohistochemical characterization of the rabbit tracheal cartilages. Journal of Biomedical Science and Engineering, 3, 1007-1013. doi: 10.4236/jbise.2010.310131.
References
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[1] Tan, Q., Steiner, R., Hoerstrup, S.P and Weder, W. (2006) Tissue-engineered trachea: History, problems and the future. Eur J Cardiothorac Surg, 30, 782-786. [2] Kojima, K. and Vacanti, C.A. (2004) Generation of a tissue-engineered tracheal equivalent. Biotechnol Appl Biochem, 39, 257-262. [3] Bucheler, M. and Haisch, A. (2003) Tissue engineering in otorhinolaryngology. DNA Cell Biol, 22, 549-564. [4] Grillo, H.C. (2002) Tracheal replacement: a critical review. Ann Thorac Surg, 73, 1995-2004. [5] Grillo, H.C. (2003) Tracheal replacement. J Thorac Cardiovasc Surg, 125, 975. [6] Grillo, H.C. (2005) Bioengineered airway tissue. J Thorac Cardiovasc Surg, 129, 1208. [7] Birchall, M. and Macchiarini, P. (2008) Airway transplantation: a debate worth having? Transplantation, 85, 1075-1080. [8] Doss, A.E., Dunn, S.S., Kucera, K.A., Clemson, L.A. and Zwischenberger, J.B. (2007) Tracheal replacements: Part 2. ASAIO J, 53, 631-639. [9] Kucera, K.A., Doss, A.E., Dunn, S.S., Clemson L.A. and Zwischenberger J.B. (2007) Tracheal replacements: Part 1. ASAIO J, 53, 497-505. [10] Macchiarini, P., Jungebluth, P., Go, T., Asnaghi, M.A., Rees, L.E., Cogan, T.A., Dodson, A., Martorell, J., Bellini, S., Parnigotto, P.P., Dickinson, S.C., Hollander, A.P., Mantero, S., Conconi, M.T. and Birchall, M.A. (2008) Clinical transplantation of a tissue-engineered airway. Lancet. [11] Steger, V., Hampel, M., Trick, I., Muller, M. and Walles, T. (2008) Clinical tracheal replacement: transplantation, bioprostheses and artificial grafts. Expert Rev Med Devices, 5, 605-612. [12] Asnaghi, M.A., Jungebluth, P., Raimondi, M.T., Dickinson, S.C., Rees, L.E., Go, T., Cogan, T.A., Dodson, A., Parnigotto, P.P., Hollander, A.P., Birchall, M.A., Conconi, M.T., Macchiarini, P. and Mantero, S. (2009) A double-chamber rotating bioreactor for the development of tissue-engineered hollow organs: from concept to clinical trial. Biomaterials, 30, 5260-5269. [13] Jungebluth, P., Go, T., Asnaghi, A., Bellini, S., Martorell, J., Calore, C., Urbani, L., Ostertag, H., Mantero, S., Conconi, M.T. and Macchiarini, P. (2009) Structural and morphologic evaluation of a novel detergent-enzymatic tissue-engineered tracheal tubular matrix. J Thorac Cardiovasc Surg, 138, 586-593. [14] Grimmer, J.F., Gunnlaugsson, C.B., Alsberg, E., Murphy, H.S., Kong, H.J., Mooney, D. J. and Weatherly, R.A. (2004) Tracheal reconstruction using tissue-engineered cartilage. Arch Otolaryngol Head Neck Surg, 130, 1191- 1196. [15] Robey, T.C., Eiselt, P.M., Murphy, H.S., Mooney, D.J. and Weatherly, R.A. (2000) Biodegradable external tracheal stents and their use in a rabbit tracheal reconstruction model. Laryngoscope, 110, 1936-1942. [16] Robey, T.C., Valimaa, T., Murphy, H.S., Tormala, P., Mooney, D.J. and Weatherly, R.A. (2000) Use of internal bioabsorbable PLGA “finger-type” stents in a rabbit tracheal reconstruction model. 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[22] Gray, S.D., Titze, I.R., Chan, R. and Hammond, T.H, (1999) Vocal fold proteoglycans and their influence on biomechanics. Laryngoscope, 109, 845-854. [23] Hammond, T.H., Zhou, R., Hammond, E.H., Pawlak, A. and Gray, S.D. (1997) The intermediate layer: a morphologic study of the elastin and hyaluronic acid constituents of normal human vocal folds. Journal of Voice, 11, 59-66. [24] Pawlak, A.S., Hammond, T., Hammond, E. and Gray, S.D. (1996) Immunocytochemical study of proteoglycans in vocal folds. Ann Otol Rhinol Laryngol, 105, 6-11. [25] Chan, R.W., Fu, M., Young, L. and Tirunagari, N. (2007) Relative contributions of collagen and elastin to elasticity of the vocal fold under tension. Ann Biomed Eng, 35, 1471-1483. [26] Melo, E.C., Lemos, M., Aragao, X.F.J., Sennes, L.U., Nascimento, S.P.H. and Tsuji, D.H. (2003) Distribution of collagen in the lamina propria of the human vocal fold. Laryngoscope, 113, 2187-2191. [27] Sivasankar, M. and Ivanisevic, A. (2007) Atomic force microscopy investigation of vocal fold collagen. Laryngoscope, 117, 1876-1881. [28] Tateya, T., Tateya, I. and Bless, D.M.(2006) Collagen subtypes in human vocal folds. Ann Otol Rhinol Laryngol, 115, 469-476. [29] Tateya, T., Tateya, I. and Bless, D.M. (2007) Immuno-scanning electron microscopy of collagen types I and III in human vocal fold lamina propria. Ann Otol Rhinol Laryngol, 116, 156-159. [30] Sakae, F.A,, Imamura, R., Sennes, L.U., Mauad, T., Saldiva, P.H. and Tsuji, D.H. (2008) Disarrangement of collagen fibers in Reinke’s edema. Laryngoscope, 118, 1500-1503. [31] Sato, K., Hirano, M. and Nakashima, T. (2003) 3D structure of the macula flava in the human vocal fold. Acta Otolaryngol, 123, 269-273. [32] Sato, K., Hirano, M. and Nakashima, T. (2002) Age- related changes of collagenous fibers in the human vocal fold mucosa. Ann Otol Rhinol Laryngol, 111, 15- 20. [33] Sato, K. and Hirano, M. 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