Volume 20, Issue 4 (Iranian South Medical Journal 2017)
Iran South Med J 2017, 20(4): 349-361 |
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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
URL: http://ismj.bpums.ac.ir/article-1-887-en.html
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
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 ,
3- Department of Immunology, Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
Abstract: (6991 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.
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.
Type of Study: Original |
Subject:
General
Received: 2016/10/25 | Accepted: 2017/02/21 | Published: 2017/08/27
Received: 2016/10/25 | Accepted: 2017/02/21 | Published: 2017/08/27
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