Abstract
1. Chitin is known to promote skin wound healing. In this study, chitin, prepared from Zuwai crab shell, was used as a bridge between the proximal and distal stumps of cut hypoglossal nerves in shrews. We compared the effects of chitin on the regeneration of transected right hypoglossal nerve axons, with those of porcine dermis, bovine dermal aterocollagen, and autologous nerve bundles.
2. To assess the survival of neurones, the size of neuronal cell body, and number of motoneurones were determined in the absence of any bridged material and in the presence of porcine dermis, bovine dermal aterocollagen, chitin, or autologous nerve bundles as a bridge.
3. Our results revealed a significantly better outcome in chitin and autologous nerve bridged groups; the size of neuronal cell body and number of hypoglossal neurones were higher than in the other groups. Chitin also enhanced the regeneration of neurones; the number of horseradish peroxide positive neurones indicative of repaired axonal processes was significantly higher in chitin and autologous nerve-bridged groups than in other groups.
4. Our results demonstrated that the use of chitin sheet or autograft successfully prevented the death of severed neurones and promoted the regeneration of the lesioned nerve. Although the mechanisms underlying the effects of chitin are still unknown, chitin seems to be a potentially useful biocompatible material for nerve repair and regeneration.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Ando, M., and Ando, Y. (1983). Clinical use of lyophilized porcine dermal skin for wound cover. Nagoya Med. J. 28:1–13.
Austin, P. R., Brine, C. J., Castle, J. E., and Zikakis, J. P. (1981). Chitin: New facets of research. Science 212:749–753.
Baba, N., Koji, T., Itoh, M., and Mizuno, A. (1999). Reciprocal changes in the expression of Bcl-2 and Bax in hypoglossal nucleus after axotomy in adult rats: Possible involvement in the induction of neuronal cell death. Brain Res. 827:122–129.
Brine, C. J., Sandford, P. A., and Zikakis, J. P. (1991). Clinical application of chitin artificial skin (Beschitin W), The Proceedings of the 5th International Conference on Chitin and Chitosan held in Princeton, NJ, pp. 17–20.
Britis, P. A., Canning, D. R., and Silver. J. (1992). Chondroitin sulfate as a regulator of neuronal patterning in the retina. Science 255:733–736.
Challacombe, J. F., and Elam, J. F. (1997). Chondroitin 4-sulfate stimulates regeneration of goldfish retinal axons. Exp. Neurol. 143:10–17.
Dinarello, C. A. (1988). Biology of interleukin 1. FASEB J. 2:108–115.
Gillardon, F., Klimaschewski, L., Wickert, H., Krajewski S., Reed, J. C., Zimmermann, M. (1996). Expression pattern of candidate cell death effector proteins Bax, Bcl-2, Bcl-X, and c-Jun in sensory and motor neurons following sciatic nerve transection in the rat. Brain Res. 739:244–250.
Gillardon, F., Wickert, H., Krajewski, S., and Zimmermann, M. (1995). Up-regulation of bax and down-regulation of bcl-2 is associated with kainate-induced apoptosis in mouse brain. Neurosci. Lett. 192:85–88.
Hara, H., Iwai, T., Niwa, M., Uematsu, T., Yoshimi, T., Tanaka, T., and Mori, H. (1996). Immunohistochemical detection of Bax and Bcl-2 proteins in gerbil hipocampus following transient forebrain ischemia. Brain Res. 711:249–253.
Hascall, V. C., and Hascall, G. K. (1981). Proteoglycans. In Hay, E. D., (ed.), Cell Biology of Extracellular matrix, Plenum, New York, pp. 39–63.
Ide, C., Tohyama, K., Yokota, R., Nitatori, T., and Onodera, S. (1983). Schwann cell basal lamina and nerve regeneration. Brain Res. 288:61–75.
Kishimoto, S., and Tamaki, K. (1987). Immunohistochemical and histochemical observations in the process of burn wound healing in guinea pig skin under chitin membrane dressing. Acta Dermatol. 82:471–479.
Kurata, S., Itami, S., Teraishi, H., and Takayasu, S. (1994). Successful transplantation of cultured human outer root sheath cells as epithelium. Ann. Plast. Surg. 33:290–294.
Leibovich, S. J., and Ross, R. (1975). The role of the macrophage in wound repair. Am. J. Pathol. 78:71–99.
Letourneau, P. C., Condic, M. L., and Snow, D. M. (1994). Interaction of developing neurons with the extracellular matrix. J. Neurosci. 14:915–916.
Lindholm, D., Heumann, R., Meyer, M., and Thoenen, H. (1987). Interleukin-1 regulates synthesis of nerve growth factor in non-neuronal cells of rat sciatic nerve. Nature 330:658–659.
Machinami, R., Kifune, H., Kawaide, A., and Tsurutani, R. (1991). A histological study of the fate of chitin suture material after intramuscular suturing. Med. Sci. Res. 19:391–392.
Nadim, W., Anderson, P. N., and Turmaine, M. (1990). The role of Schwann cells and basal lamina tubes in the regeneration of axons through long lengths of freeze-killed nerve grafts. Neuropathol. Appl. Neurobiol. 16:411–421.
Nakajima, M., Atsumi, K., Kifune, K., Miura, K., and Kanamaru, H. (1986). Chitin is an effective material for sutures. Jpn. J. Surg. 16:418–424.
Nishimura, K., Nishimura, S., Nishi, N., Saiki, I., Tokura, S., and Azuma I. (1984). Immunological activity of chitin and its derivatives. Vaccine 2:93–99.
Okuno, T., Nomura, Y., Hara, M., Tsurutani, R., and Kifune, K. (1991). Application of beschitin to the inner ear, Transplants and implants in otology. Proceedings of the Second International Symposium on Transplants and Implants in Otology. Matsuyama, Ehime, Japan, pp. 189–192.
Prudden, J. F., Migel, P., Hanson, P., Friedrich, L., and Balassa, L. (1970). The discovery of a potent pure chemical wound-healing accelerator. Am. J. Surg. 119:560–564.
Sinder, W. D., and Thander, S. (1989). Target dependence of hypoglossal motor neurons during development in maturity. J. Comp. Neurol. 279:489–498.
Snow, D. M., Watanabe, M., Letourneau, P. C., and Silver, L. (1991). A chondroitin sulfate proteoglycan may influence the direction of retinal ganglion cell outgrowth. Development 113:1473–1485.
Streit, P. and Reubi, J. C. (1977). A new and sensitive staining method for axonally transported horseradish peroxidase (HRP) in the pigeon visual system. Brain Res. 126:530–537.
Taniuchi, M., Clark, H. B., and Johnson, E. M., (1986). Induction of nerve growth factor receptor in Schwann cells after axotomy. Proc. Natl. Acad. Sci. 83:4094–4098.
Tokuhisa, M., and Tominaga, N. (1994). Effects of chitin on the production of chemical mediators by peripheral monocytes. Jpn. J. Oral Biol. 36:591–597.
Tokuhisa, T., Tominaga, N., Nakamura, M., Fujiki, T., Kuga, Y., Mizuno, A., and Umemoto T. (1996). The effects of products from macrophages cultured on chitin membrane on human gingival fibroblastlike cell. Jpn. J. Oral. Maxillofac. Surg. 42:56–64.
Vekrellis, K., McCathy, M. J., Watson, A., Whitfield, J., Rubin, L. L., and Ham, J. (1997). Bax promotes neuronal cell death and is downregulated during the development of the nervous system. Development 124:1239–1249.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Itoh, MI., Izumi, SI., Uemura, M. et al. Prevention of Death of Axotomized Hypoglossal Neurones and Promotion of Regeneration by Chitin Grafting. Cell Mol Neurobiol 20, 529–540 (2000). https://doi.org/10.1023/A:1007055626632
Issue Date:
DOI: https://doi.org/10.1023/A:1007055626632