ISOLATION, CHARACTERIZATION AND IMMUNOCHEMICAL STUDIES ON FIBROUS PROTEINS FROM COWRY SHELL (CYPRAEA MONETA, LINNAEUS)

Background: Biomaterials are non-drug substances used to treat, enhance or replace functions of body tissues or organs. Natural sources of biomaterials have recently become the focus of several research activities. Cowry shell constitutes one of the most promising natural sources of biomaterials because of its chemical stability, biodegradability and biocompatibility in the body. However, its applications may be limited due to immunogenic and toxic responses that may occur following implantation, hence this study. Materials and Methods: Crude fibrous protein extracted with citrate buffer from pulverised cowry shells (Cypraea moneta (L)), was resolved into two components (CSP1 and CSP2) by gel filtration. Immunological studies were performed with antisera obtained from rabbits by double immunodiffusion and immunoelectrophoresis techniques. Mice treated with the proteins were observed for signs of toxicity and their liver, kidney, lungs and spleen were processed histologically. Results: The native molecular weight of CSP1 and CSP2 determined by gel filtration were 91kDa and 33kDa respectively. CSP1 and CSP2 displayed single bands on SDS-PAGE with subunit molecular weight values of 19kDa and 19.5kDa respectively. Antisera obtained from rabbits immunised with the crude citrate buffer extracts precipitated the antigen in double immunodiffusion tests. Histopathological examinations revealed a dose-dependent damaging effect of the shell proteins on liver, kidney, lung and spleen tissues of the treated mice. Conclusion: This study showed that cowry shells contain fibrous proteins which are immunogenic and toxic in mice at relatively high concentrations, causing visible organ damage without concurrent physical manifestations.

[1]  L. E. Umoru,et al.  Natural Products: A Minefield of Biomaterials , 2012 .

[2]  F. B. Diniz,et al.  Chitosan effect on dental enamel de-remineralization: an in vitro evaluation. , 2010, Journal of dentistry.

[3]  A. Halim,et al.  In vitro biocompatibility of chitosan porous skin regenerating templates (PSRTs) using primary human skin keratinocytes. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.

[4]  J. Jansen,et al.  In vitro and in vivo evaluation on the bioactivity of ZnO containing nano-hydroxyapatite/chitosan cement. , 2009, Journal of biomedical materials research. Part A.

[5]  V. Parameswaran,et al.  Potential use of chitosan nanoparticles for oral delivery of DNA vaccine in Asian sea bass (Lates calcarifer) to protect from Vibrio (Listonella) anguillarum. , 2008, Fish & shellfish immunology.

[6]  O. Oloyede Chemical Constituents of Cowry (Cyparica samplomoneta) , 2008 .

[7]  C. Kim,et al.  Development of Chitosan Dermal Scaffold and its Characterization , 2007 .

[8]  A. Il’ina,et al.  Effect of the molecular weight of chitosan on its antiviral activity in plants , 2006, Applied Biochemistry and Microbiology.

[9]  P. Layrolle,et al.  Caspartin and Calprismin, Two Proteins of the Shell Calcitic Prisms of the Mediterranean Fan Mussel Pinna nobilis* , 2005, Journal of Biological Chemistry.

[10]  H. Tang,et al.  Conchiolin-protein in aragonite shells of mollusks , 1996 .

[11]  H. Baden,et al.  The fibrous proteins of stratum corneum. , 1976, The Journal of investigative dermatology.

[12]  S. Weiner,et al.  Characterization of 80-million-year-old mollusk shell proteins. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[14]  R. Barsacchi,et al.  Molecular charcteristics of chicken kidney arginase. , 1975, The Biochemical journal.

[15]  Lloyd H. Michael,et al.  The Guide for the Care and Use of Laboratory Animals. , 2016, ILAR journal.

[16]  Vikas Kumar,et al.  ACUTE AND SUBCHRONIC TOXICITY STUDY OF CALCIUM BASED AYURVEDIC 'BHASMAS' AND A 'PISHTI' PREPARED FROM MARINE ANIMALS , 2010 .

[17]  J. Carnés,et al.  Selective hypersensitivity to boiled razor shell. , 2007, Journal of investigational allergology & clinical immunology.

[18]  E. Jurkiewicz-Karnkowska Some aspects in nitrogen, carbon and calcium accumulation in molluscs from the Zegrzynski Reservoir ecosystem , 2005 .