[Home ] [Archive]   [ فارسی ]  
:: Main :: About :: Current Issue :: Archive :: Search :: Submit :: Contact ::
:: Volume 20, Issue 4 (Iranian South Medical Journal 2017) ::
Iran South Med J 2017, 20(4): 349-361 Back to browse issues page
Extraction of Chondroitin Sulfate From Cartilage Sturgeon, Stellate Sturgeon (Acipenser stellatus) and its Inductive Effect on Human Fibroblast Proliferation
Saead Delshad1, Katayon Karimzadeh 2, Ali Mostafaie3
1- Young Researchers and Elite Club, Lahijan Branch, Islamic Azad University, Lahijan, Iran
Department of Marine Biology, School of Science, Lahijan Branch, Islamic Azad university, Lahijan, Iran
2- Department of Marine Biology, School of Science, Lahijan Branch, Islamic Azad university, Lahijan, Iran , karimzadehkathy@yahoo.co.uk
3- Department of Immunology, Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Abstract:   (3193 Views)
Background: Chondroitin sulfates (CS) is one of the essential glycosaminoglycan’s which is found in sturgeon fishes and exists in the cartilage part of these fishes. In addition to food and cosmetic industries, these compounds have significant medical and clinical applications, mainly in the treatment of arthritis and wound healing.
Materials and Methods: In this study, the chondroitin sulfate extraction was performed using pepsin and trypsin enzyme digestions in 12 and 18 hours intervals followed by cationic salt, cetyl pyridinium chloride (CPC) treatment. To investigate the effect of Chondroitin sulfate on induction of proliferation of fibroblasts isolated from human skin, MTT assay method was used, which was applied at different incubation times (24, 48 and 72 h). To determine the proper time and enzymatic digestion, after temperature treatment of samples using pepsin and trypsin enzymes, the SDS-PAGE electrophoresis was used. Moreover, the FT-IR analysis was performed to characterize the type of CS.
Results: The maximum yield of enzymatic digestion was observed for trypsin digestion at 18h, according to electrophoresis patterns. The amount of CS yield was estimated at 4.76% in this situation. FT-IR analysis revealed that the CS belongs to the C4S type. Chondroitin sulfates extracting from cartilage showed the concentration-dependent effect on fibroblast cell proliferation. This dose-response effect showed 167% increase in proliferation at 10 µg/ml dose of CS compared to the control in fibroblast cells.
Conclusion: Extracted CS has a positive stimulatory effect on fibroblast cells proliferation in a dose-dependent manner. So it can be used for fibroblast prolioferation induction for wound healing and repairing damaged tissues.
Keywords: Cell proliferation, Cartilage, Chondroitin sulfate, Acipenser stellatus
Full-Text [PDF 1097 kb]   (798 Downloads)    
Type of Study: Original | Subject: General
Received: 2017/08/27 | Accepted: 2017/08/27 | Published: 2017/08/27
1. Roden L. Structure and metabolism of connective tissue proteoglycans. The biochemistry of glycoproteins and proteoglycans. In: Lennarz WJ, editors. Plenum Press: New York, 1980, 267-371.
2. Beaty N, Mello RT. Extracellular mammalian polysaccharides: Glycosaminoglycans and proteoglycans. J Chromatogr B Biomed Sci Appl 1987; 418: 187-222. [DOI:10.1016/0378-4347(87)80009-9]
3. Nakano T, Dixon WT, Ozimek L. Proteoglycan (Glycosaminoglycans/ Mucopolysaccharides). Biopolymers. In: Steinbüchel A, editors. Wiley-VCH Verlag GmbH, Weinheim: Germany, 2002, 575-604.
4. Vynios DH, Karamanos NK, Tsiganos CP. Advances in analysis of glycosaminoglycans: its application for the assessment of physiological states of connective tissues. J Chromatogr B Analyt Technol Biomed Life Sci 2002; 781(1-2): 21-38. [DOI:10.1016/S1570-0232(02)00498-1]
5. Nakano T, Betti M, Pietrasik Z. Extraction, isolation and analysis of chondroitin sulfate glycosaminoglycans. Recent Pat Food Nutr Agric 2010; 2(1): 61-74. [DOI:10.2174/1876142911002010061]
6. Deepa SS, Kalayanamitra K, Ito Y, et al. Novel sulfated octa- and decasaccharides from squid cartilage chondroitin sulfate-E: sequencing and their application for determination of the epitope structure of monoclonal antibody MO-225. Biochemistry 2007; 46(9): 2453-65. [DOI:10.1021/bi602374m]
7. Garnjanagoonchorn W, Wongekalak L, Engkagul A. Determination of chondroitin sulfate from different sources of cartilage. Chem Eng Process: Process Intensification 2007; 46(5): 465-71. [DOI:10.1016/j.cep.2006.05.019]
8. Zhang F, Xie J, Linhardt RJ. Isolation and structural characterization of glycosaminoglycans from heads of red salmon (Oncorhynchus nerka). Jacobs J Biotechnol Bioeng 2014; 19(1): 002.
9. Lamari FN, Karamanos NK. Structure of chondroitin sulfate. Chondroitin sulfate: Structure, role and pharmacological activity, advances in pharmacology. In: Volpi N, editor. Academic Press: San Diego, 2006, 33-48.
10. Volpi N. Quality of different chondroitin sulfate preparations in relation to their therapeutic activity. J Pharm Pharmacol 2009; 61(10): 1271-80. [DOI:10.1211/jpp.61.10.0002]
11. Tat SK, Pelletier JP, Mineau F, et al. Variable effects of 3 different chondroitin sulfate compounds on human osteoarthritic cartilage/chondrocytes: relevance of purity and production process. J Rheumatol 2010; 37(3): 656-64. [DOI:10.3899/jrheum.090696]
12. Capila I, Linhardt RJ. Heparin-protein interactions. Angew Chem Int Ed Engl 2002; 41(3): 391-412. https://doi.org/10.1002/1521-3773(20020201)41:3<390::AID-ANIE390>3.0.CO;2-B [DOI:10.1002/1521-3773(20020201)41:33.0.CO;2-B]
13. Yamada S, Sugahara K. Potential therapeutic application of chondroitin sulfate/dermatan sulfate. Curr Drug Discov Technol 2008; 5(4): 289-301. [DOI:10.2174/157016308786733564]
14. Kuhn MA, Smith PD, Hill DP, et al. In vitro fibroblast populated collagen lattices are not good models of in vivo clinical wound healing. Wound Repair Regen 2000; 8(4): 270-6. [DOI:10.1046/j.1524-475x.2000.00270.x]
15. Vazquez JA, Rodriguez-Amado I, Ignacia Montemayor MI, et al. Chondroitin sulfate, hyaluronic acid and chitin/chitosan production using marine waste sources: characteristics, applications and eco-friendly processes: a review. Mar Drugs 2013; 11(3): 747-74. [DOI:10.3390/md11030747]
16. Luo XM, Fosmire GJ, Leach RM. Chicken keel cartilage as a source of chondroitin sulfate. Poult Sci 2002; 81(7): 1086-9. [DOI:10.1093/ps/81.7.1086]
17. Axelsson I, Heinegard D. Characterization of chondroitin sulfate-rich proteoglycans from bovine corneal stroma. Exp Eye Res 1980; 31(1): 57-66. [DOI:10.1016/0014-4835(80)90090-1]
18. Mourao PA. Perspective on the Use of sulfated polysaccharides from marine organisms as a source of new antithrombotic drugs. Mar Drugs 2015; 13(5): 2770-84. [DOI:10.3390/md13052770]
19. Seno N, Meyer K. Comparative biochemistry of skin the mucopolysaccharides of shark skin. Biochim Biophys Acta 1963; 78: 258-64. [DOI:10.1016/0006-3002(63)91636-6]
20. Vieira RP, Mourao PA. Occurrence of a unique fucosebranched chondroitin sulfate in the body wall of a sea cucumber. J Biol Chem 1988; 263(34): 18176-83.
21. Kinoshita-Toyoda A, Yamada S, Haslam SM, et al. Structural determination of five novel tetrasaccharides containing 3-O-sulfated D-glucuronic acid and two rare oligosaccharides containing a β-D-glucose branch isolated from squid cartilage chondroitin sulfate E. Biochemistry 2004; 43(34): 11063-74. [DOI:10.1021/bi049622d]
22. Maccari F, Ferrarini F, Volpi N. Structural characterization of chondroitin sulfate from sturgeon bone. Carbohydr Res 2010; 345(11): 1575-80. [DOI:10.1016/j.carres.2010.05.016]
23. Williot P, Sabeau L, Gessner J, et al. Sturgeon farming in Western Europe: recent developments and perspectives. Aquat Living Resour 2001; 14(6): 367-74. [DOI:10.1016/S0990-7440(01)01136-6]
24. Norouzi M, Pourkazemi M, Fatemi M. Application of microsatellite markers to study the genetic structure of stellate sturgeon populations (Acipenser stellatus Pallas, 1771) in the south Caspian Sea. Iran J Fish Sci 2009; 8(1): 73-84.
25. Im AR, Sim R, Park JS, et al. Isolation and characterization of chondroitin sulfates from the by-products of marine organisms. Food Sci Biotechnol 2009; 18(4): 872-7.
26. Im AR, Park Y, Kim YS. Isolation and characterization of chondroitin sulfates from sturgeon (Acipenser sinensis) and their effects on growth of fibroblasts. Biol Pharm Bull 2010; 33(8): 1268-73. [DOI:10.1248/bpb.33.1268]
27. Laemmli UK. Cleavage of structural proteins during assembly of head of bacteriophage-T4. Nature 1970; 227(5259): 680-5. [DOI:10.1038/227680a0]
28. Mostafaei, A. Theoretical and practical guide of gel electrophoresis of proteins. 2nd ed. Yadavaran: Tehran, 2003, 19-34. (Persian)
29. Rodén L, Baker JR, Cifonelli JA, et al. Isolation and characterization of connective tissue polysaccharides. Method enzymol 1972; 28: 73-140.‌ [DOI:10.1016/0076-6879(72)28009-0]
30. Taniguchi N. Isolation and analysis of glycosaminoglycans. glycosaminoglycans and proteoglycans in physiological and pathological processes of body systems. In: Varma RS, Varma R, editors. Karger: Basel, 1982; 20-40.
31. Silva LCF. Isolation and purification of chondroitin sulfate. chondroitin sulfate: Structure, role and pharmacological activity, advances in pharmacology. In: Volpi N, editors. Academic Press: San Diego, 2006, 21-31.
32. Nakano T, Nakano K, Sim JS. Extraction of glycosaminoglycan peptide from bovine nasal cartilage with 0.1M sodium acetate. J Agric Food Chem 1998; 46(20: 772-8.
33. Yang H, Liu S, Cai H, et al. Chondroitin sulfate as a molecular portal that preferentially mediates the apoptotic killing of tumor cells by penetratin-directed mitochondria-disrupting peptides. J Biol Chem 2010; 285(33): 25666-76. [DOI:10.1074/jbc.M109.089417]
34. Foot M, Mulholland M. Classification of chondroitin sulfate A, chondroitin sulfate C, glucosamine hydrochloride and glucosamine 6 sulfate. J pharm Biomed Anal 2005; 38(3): 397-407. [DOI:10.1016/j.jpba.2005.01.026]
35. Sugahara K, Masuda M, Harada T, et al. Structural studies on sulfated oligosaccharides derived from the carbohydrate-protein linkage region of chondroitin sulfate proteoglycans of whale cartilage. Eur J Biochem 1991; 202(3): 805-11. [DOI:10.1111/j.1432-1033.1991.tb16436.x]
36. Syrokou A, Tzanakakist G, Tsegenidis T, et al. Effects of glycosaminoglycans on proliferation of epithelial and fibroblast human malignant mesothelioma cells: a structure-function relationship. Cell Prolif 1999; 32(2-3): 85-99. [DOI:10.1046/j.1365-2184.1999.32230085.x]
37. Vazirizadeh A, Naderi-Manesh H, Bargahi A, et al . Impacts of Persian Gulf blackfin stonefish crude venom on the haematological factors and serum enzymes levels of laboratory rat, ISMJ 2014; 17(4): 723-732
38. Marques J, Vilanova E, Mourão PA, et al. Marine organism sulfated polysaccharides exhibiting significant antimalarial activity and inhibition of red blood cell invasion by Plasmodium. Sci Rep 2016; 6: 24368. [DOI:10.1038/srep24368]
Send email to the article author

Add your comments about this article
Your username or Email:


XML   Persian Abstract   Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Delshad S, karimzadeh K, Mostafaie A. Extraction of Chondroitin Sulfate From Cartilage Sturgeon, Stellate Sturgeon (Acipenser stellatus) and its Inductive Effect on Human Fibroblast Proliferation . Iran South Med J. 2017; 20 (4) :349-361
URL: http://ismj.bpums.ac.ir/article-1-887-en.html

Volume 20, Issue 4 (Iranian South Medical Journal 2017) Back to browse issues page
دانشگاه علوم پزشکی بوشهر، طب جنوب ISMJ

Iranian South Medical Journal is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License which allows users to read,
copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly

Copyright © 2017, Iranian South Medical Journal| All Rights Reserved

Persian site map - English site map - Created in 0.06 seconds with 32 queries by YEKTAWEB 4137