A Dermal Gel Made of Rutilus Kutum Skin Collagen-Chitosan for Deep Burn Healing

International Journal of Peptide Research and Therapeutics volume 27, pages317–328(2021)Cite this article

Abstract

Natural compounds extracted from marine organisms consisting of biological active materials like collagen provide a major source of compound with medical applications including wound dressing particularly in co-treatment with other natural or synthetic materials to quick the wound healing process. The current study investigates the potential of Rutilus kutum skin-extracted collagen in combination with chitosan [collagen-chitosan (Col-CH) gel] for in vivo second-degree burn wound healing in rats following finding of the most effective collagen-chitosan ratio. Collagen and chitosan were extracted from R. Kutum skin and shell waste of shrimp, respectively, using acid–base method. SDS-PAGE was performed to determine subunit composition of collagen. After amino acid analysis using HPLC, different ratios of collagen and chitosan mixture (Col-CH, 1:3, 1:1 and 3:1) were prepared to fine the most effective of Col-CH gel on burn wound healing and compare to animals treated with silver sulfadiazine ointment. Wound surface area was analyzed using ImageJ software on days 15 and 30. H&E staining was performed to show the epithelialization, collagen deposition, fibroblast cell accumulation and vessel formation on days 15 and 30. The results from H&E were scored and quantified by pathologist. SDS-PAGE technique revealed that extracted-collagen was type 1 composed of two α (α1 and α2) chains. HPLC showed that Glycine was the most abundant amino acid which included one-third of total amino acid in the skin. The wound surface measurement showed a significant reduction in wound size in animals treated with Col-CH (1:1) compared to silver sulfadiazine treated group at day 25. Histopathological findings represented high score in epithelialization, vessel formation and a decrease in inflammatory cells infiltration in Col-CH (1:1) treated group on days 15 and 30. These data suggest collagen in combination with chitosan with 1:1 ratio can significantly restore burn wound compared to silver sulfadiazine ointment.

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References

  1. Alemdaroğlu C et al (2006) An investigation on burn wound healing in rats with chitosan gel formulation containing epidermal growth factor. Burns 32(3):319–327

    Article  Google Scholar 

  2. AlizadehNodeh M, Moradi Z, Nourani MR (2014) Isolation and purification of collagen from the skin of black pomfret (Parastromateus niger) for tissue engineering purpose. J Appl Tissue Eng 1(1):18–21

    Google Scholar 

  3. Basha S et al (2011) Effects of fish scales extracted collagen biocasings on cutaneous wound healing in dogs. Res J Pharm, Biol Chem Sci 2(2):36–49

    Google Scholar 

  4. Bidlingmeyer BA, Tarvin TL, Cohen SA (1986) Amino acid analysis of submicrogram hydrolyzate samples. Methods in protein sequence analysis. Humana Press, Totowa, pp 229–245

    Google Scholar 

  5. Chen R-N et al (2006) Development of N, O-(carboxymethyl) chitosan/collagen matrixes as a wound dressing. Biomacromol 7(4):1058–1064

    CAS  Article  Google Scholar 

  6. Cui F et al (2011) Development of chitosan-collagen hydrogel incorporated with lysostaphin (CCHL) burn dressing with anti-methicillin-resistant Staphylococcus aureus and promotion wound healing properties. Drug Deliv 18(3):173–180

    CAS  Article  Google Scholar 

  7. Duan R et al (2009) Properties of collagen from skin, scale and bone of carp (Cyprinus carpio). Food Chem 112(3):702–706

    CAS  Article  Google Scholar 

  8. Gharagheshlagh SN, Nourani MR, Jamili S, Javad M (2018) Isolation and Characterization of acid-soluble Collagen from the Skin of Scomberomorus guttatus of Persian Gulf. J of Applied Tissue Engineering 5(3)

  9. Gonzalez AC et al (2016) Wound healing-a literature review. An Bras Dermatol 91(5):614–620

    Article  Google Scholar 

  10. Hema GS, Shyni K, Mathew S, Anandan R, Ninan G, Lakshmanan PT (2013) A simple method for isolation of fish skin collagen-Biochemical characterization of skin collagen extracted from albacore tuna (Thunnus alalunga), dog shark (Scoliodon sorrakowah) and rohu (Labeo rohita). Ann Biol Res 4(1):271-278

    CAS  Google Scholar 

  11. Hu Z et al (2017) Marine collagen peptides from the skin of Nile Tilapia (Oreochromis niloticus): characterization and wound healing evaluation. Marine Drugs 15(4):102

    Article  Google Scholar 

  12. Intaraprasita S et al (2012) Efficacy evaluation of the fibroblast-seeded collagen/chitosan scaffold on application in skin tissue engineering. ScienceAsia 38:268–277

    Article  Google Scholar 

  13. Jongjareonrak A et al (2005) Isolation and characterisation of acid and pepsin-solubilised collagens from the skin of Brownstripe red snapper (Lutjanus vitta). Food Chem 93(3):475–484

    CAS  Article  Google Scholar 

  14. Kadler KE et al (1996) Collagen fibril formation. Biochem J 316(1):1–11

    CAS  Article  Google Scholar 

  15. Kirichenko A et al (2013) Morphological study of burn wound healing with the use of collagen-chitosan wound dressing. Bull Exp Biol Med 154(5):692–696

    CAS  Article  Google Scholar 

  16. Kumar NS, Nazeer R, Jaiganesh R (2012) Wound healing properties of collagen from the bone of two marine fishes. Int J Pept Res Ther 18(3):185–192

    Article  Google Scholar 

  17. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680

    CAS  Article  Google Scholar 

  18. Landén NX, Li D, Ståhle M (2016) Transition from inflammation to proliferation: a critical step during wound healing. Cell Mol Life Sci 73(20):3861–3885

    Article  Google Scholar 

  19. Latifi N, Karimi H (2017) Correlation of occurrence of infection in burn patients. Ann Burns Fire Disasters 30(3):172

    Google Scholar 

  20. Liu H, Li D, Guo S (2007) Studies on collagen from the skin of channel catfish (Ictalurus punctaus). Food Chem 101(2):621–625

    CAS  Article  Google Scholar 

  21. Ma L et al (2003) Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering. Biomaterials 24(26):4833–4841

    CAS  Article  Google Scholar 

  22. Milan PB et al (2020) Decellularization and preservation of human skin: A platform for tissue engineering and reconstructive surgery. Methods 171:62-67

    Article  Google Scholar 

  23. Muyonga J, Cole C, Duodu K (2004) Characterisation of acid soluble collagen from skins of young and adult Nile perch (Lates niloticus). Food Chem 85(1):81–89

    CAS  Article  Google Scholar 

  24. Naderi Gharagheshlagh S et al (2019) Isolation and characterization of acid-soluble collagen from the skin of Rutilus Frisii Kutum (Kamensky) of the Caspian Sea. Iran J Fish Sci. https://doi.org/10.22092/ijfs.2019.118957

    Article  Google Scholar 

  25. Nascimento EG et al (2009) Evaluation of chitosan gel with 1% silver sulfadiazine as an alternative for burn wound treatment in rats. Acta Cir Bras 24(6):460–465

    Article  Google Scholar 

  26. Ogawa M et al (2004) Biochemical properties of bone and scale collagens isolated from the subtropical fish black drum (Pogonia cromis) and sheepshead seabream (Archosargus probatocephalus). Food Chem 88(4):495–501

    CAS  Article  Google Scholar 

  27. Rahman MS et al (2019) Characterization of burn wound healing gel prepared from human amniotic membrane and Aloe vera extract. BMC Complement Altern Med 19:115

    Article  Google Scholar 

  28. Rane MM, Mengi SA (2003) Comparative effect of oral administration and topical application of alcoholic extract of Terminalia arjuna bark on incision and excision wounds in rats. Fitoterapia 74(6):553–558

    Article  Google Scholar 

  29. Senaratne L, Park P-J, Kim S-K (2006) Isolation and characterization of collagen from brown backed toadfish (Lagocephalus gloveri) skin. Biores Technol 97(2):191–197

    CAS  Article  Google Scholar 

  30. Shahidi F (1994) Seafood processing by-products. Seafoods: chemistry, processing technology and quality. Springer, New York, pp 320–334

    Google Scholar 

  31. Shen X-R et al (2017) Beneficial effects of a novel shark-skin collagen dressing for the promotion of seawater immersion wound healing. Mil Med Res 4(1):33

    PubMed  PubMed Central  Google Scholar 

  32. Singh P et al (2011) Isolation and characterisation of collagen extracted from the skin of striped catfish (Pangasianodon hypophthalmus). Food Chem 124(1):97–105

    CAS  Article  Google Scholar 

  33. Tahvildari K, Mojahedi H (2016) Investigation of poly aromatic hydrocarbons adsorption using chitosan and its synthetic derivatives. Trends Life Sci 5:2319–5037

    Google Scholar 

  34. Tylingo R et al (2016) Isolation and characterization of acid soluble collagen from the skin of african catfish (Clarias gariepinus), salmon (Salmo salar) and baltic cod (Gadus morhua). J Biotechnol Biomater 6(2):1–6

    Article  Google Scholar 

  35. Ueno H et al (2001) Evaluation effects of chitosan for the extracellular matrix production by fibroblasts and the growth factors production by macrophages. Biomaterials 22(15):2125–2130

    CAS  Article  Google Scholar 

  36. Wang L et al (2007) Isolation and characterization of collagen from the skin of deep-sea redfish (Sebastes mentella). J Food Sci 72(8):E450–E455

    CAS  Article  Google Scholar 

  37. Wang W et al (2008) Acceleration of diabetic wound healing with chitosan-crosslinked collagen sponge containing recombinant human acidic fibroblast growth factor in healing-impaired STZ diabetic rats. Life Sci 82(3–4):190–204

    CAS  Article  Google Scholar 

  38. Ward A (1958) The British gelatine and glue research association. Nature 182(4633):439–440

    Article  Google Scholar 

  39. Xuehui Z, Yuquan C, Hunian Z (2007) Causes of burn-out of articulated phase breaker cable and its prevention measures [J]. Electr Railw 4:32–35

    Google Scholar 

  40. Yan M et al (2008) Characterization of acid-soluble collagen from the skin of walleye pollock (Theragra chalcogramma). Food Chem 107(4):1581–1586

    CAS  Article  Google Scholar 

  41. Yates CC et al (2007) The effect of multifunctional polymer-based gels on wound healing in full thickness bacteria-contaminated mouse skin wound models. Biomaterials 28(27):3977–3986

    CAS  Article  Google Scholar 

  42. Zhang J et al (2009) Characterisation of acid-soluble collagen from skin of silver carp (Hypophthalmichthys molitrix). Food Chem 116(1):318–322

    CAS  Article  Google Scholar 

Download references

Funding

This study was funded by Iran University of Medical Science (Grant Number 93-04-29-25335-1).

Author information

Author notes

  1. Mohammad Javad Fatemi

    Present address: Burn Research Center, Shahid Motahari Hospital, Shahid Yasemi Street, Valiasr Street, Tehran, Iran

Affiliations

  1. Department of Natural Resources and Environment, Science and Research Branch, Islamic Azad University, Tehran, Iran

    Soheila Naderi Gharehgheshlagh

  2. Plastic & Reconstructive Surgery Department, Burn Research Center, Iran University of Medical Sciences, Tehran, Iran

    Mohammad Javad Fatemi

  3. Department of Pharmacology and Razi Drug Research Center, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Ali Mohammad Sharifi

  4. Division of Genomics, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Mohammad Reza Nourani

  5. Iranian Fisheries Science Research Institute, Agricultural Research, Education and Extension Organization, Tehran, Iran

    Shahla Jamili

  6. Medicine, Quran and Hadith Research Center & Department of Community Medicine, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Mohsen Saberi

  7. Burn Research Center, Iran University of Medical Sciences, Tehran, Iran

    Soheila Naderi Gharehgheshlagh

  8. Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran

    Naser Amini & Fatemeh Ganji

  9. Department of Tissue Engineering & Regenerative Medicine, Iran University of Medical Sciences, Tehran, Iran

    Fatemeh Ganji

  10. Department of Plastic and Re-constructive Surgery, Hazrat Fatemeh Hospital, Iran University of Medical Sciences, Tehran, Iran

    Mohammad Javad Fatemi

  11. Institutes of Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran

    Naser Amini

Authors
  1. Soheila Naderi Gharehgheshlagh
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  2. Mohammad Javad Fatemi
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  3. Shahla Jamili
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  4. Mohammad Reza Nourani
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  5. Ali Mohammad Sharifi
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  7. Naser Amini
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  8. Fatemeh Ganji
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Correspondence to Mohammad Javad Fatemi.

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Naderi Gharehgheshlagh, S., Fatemi, M.J., Jamili, S. et al. A Dermal Gel Made of Rutilus Kutum Skin Collagen-Chitosan for Deep Burn Healing. Int J Pept Res Ther 27, 317–328 (2021). https://doi.org/10.1007/s10989-020-10082-y

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Keywords

  • Fish collagen
  • Skin
  • Rutilus kutum
  • Chitosan
  • Burn
  • Healing