Daneshvar Medicine

Daneshvar Medicine

Evaluation of collagen scaffold synthetized by enzymatic method and its efficacy for loading neonatal fibroblast cells

Document Type : Original Article

Authors
Roqieh Shalbafian 1
Sona Zare 2
Rahim Ahmadi 3
Sanaz Sahraee 4
Maryam Hassan Nasab 5
1 Department of Biology, Faculty of Basic Sciences, Hamedan Branch, Islamic Azad University, Hamedan, Iran
2 Skin and Stem Cell Research Center, Tehran University of Medical Sciences, Tehran, Iran
3 Department of Biology, Avicenna International College, Budapest, Hungary
4 Department of Biology, Faculty of Basic Sciences, East Tehran Branch, Islamic Azad University, Tehran, Iran
5 Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran
10.22070/daneshmed.2022.15373.1140
Abstract
Background and Objective: Collagen scaffolds are one of the suitable scaffolds for stem cell loading, but the viability of fibroblast cells loaded on collagen scaffolds is one of the important issues. Therefore, the present study investigated the viability of fibroblasts loaded on collagen scaffold.
Materials and Methods: In this in vitro experimental study, human skin samples were obtained from neonates during circumcision surgery and bovine collagen was prepared. Fibroblast cells were counted using a hemocytometer after isolation from tissue fragments. Cell viability was evaluated using flow cytometry. Vimentin marker were used to identify fibroblast cells. Fibroblast cells were cultured on hydrogels. MTT method was used to evaluate the cytotoxic effect of hydrogels on cells. Data were analyzed using independent t-test.
Results: The viability of fibroblast cells isolated from the foreskin was 89% on the first day after isolation. The results of immunocytochemical examination confirmed the identity of fibroblast cells. MTT results showed that collagen hydrogel had no significant toxicity on loaded fibroblasts. The results of fluorescent microscopy showed that the viability of cells loaded on hydrogel was 96%.
Conclusion: The results of this study showed that collagen scaffolds are suitable for loading fibroblast cells isolated from the foreskin and can be used in tissue engineering and transfer of fibroblast cells to damaged areas.
Keywords
Foreskin
Fibroblast cells
Collagen scaffold
Viability
  1. Lode A, Meyer M, Brüggemeier S, Paul B, Baltzer H, Schröpfer M, et al. Additive manufacturing of collagen scaffolds by three-dimensional plotting of highly viscous dispersions. Biofabrication 2016; 8(1):015015.
  2. Khayyatan F, Emami Sh, Mehrjerdi Nz, Baharvand H. A simple and efficient method to improve mechanical properties of collagen scaffolds by UV irradiation. Science and Technology 2011;23(5):371-8.
  3. Silvipriya KS, Kumar KK, Bhat AR, Kumar BD, John A, Lakshmanan P. Collagen: Animal sources and biomedical application. Journal of Applied Pharmaceutical Science 2015;5(3):123-7.
  4. Lin YL, Chen CP, Lo CM, Wang HS. Stiffness‐controlled three‐dimensional collagen scaffolds for differentiation of human Wharton's jelly mesenchymal stem cells into cardiac progenitor cells. Journal of Biomedical Materials Research 2016;104(9):2234-42.
  5. Chashmpoosh M, Mohammadi A, Abdovis N, Ghasempoor H, Kheirollah A. Isolation and culture of mouse newborn skin fibroblast cells in a simple and inexpensive method. Pars Journal of Medical Sciences 2016;14:34-42.
  6. LeBleu VS, Neilson EG. Origin and functional heterogeneity of fibroblasts. The Federation of American Societies for Experimental Biology Journal 2020;34(3):3519-36.
  7. Langevin HM, Cornbrooks CJ, Taatjes DJ. Fibroblasts form a body wide cellular network. Histochemistry and Cell Biology 2004 ;122(1):7-15.
  8. Kumar G, Tison CK, Chatterjee K, Pine PS, McDaniel JH, Salit ML, et al. The determination of stem cell fate by 3D scaffold structures through the control of cell shape. Biomaterials 2011;32(35):9188-96.
  9. Hashemi SS, Rajabi S, Mahmoudi R, Ghanbari A, Jafari Barmak M. The Investigation of Proliferation of Fibroblasts on Chitosan Scaffold in the Presence of Hyaluronic Acid. Armaghane danesh 2018;23(2):134-45.
  10. Raghunath J, Salacinski HJ, Sales KM, Butler PE, Seifalian AM. Advancing cartilage tissue engineering: the application of stem cell technology. Current Opinion in Biotechnology 2005;16(5):503-9.
  11. Angele P, Johnstone B, Kujat R, Zellner J, Nerlich M, Goldberg V, et al. Stem cell based tissue engineering for meniscus repair. Journal of Biomedical Materials Research 2008 ;85(2):445-55.
  12. Rana D, Zreiqat H, Benkirane‐Jessel N, Ramakrishna S, Ramalingam M. Development of decellularized scaffolds for stem cell‐driven tissue engineering. Journal of Tissue Engineering and Regenerative Medicine 2017;11(4):942-65
  13. Mahmoudi‐Rad M, Abolhasani E, Moravvej H, Mahmoudi‐Rad N, Y. Acellular amniotic membrane: an appropriate scaffold for fibroblast proliferation. Clinical and Experimental Dermatology 2013;38(6):646-51.
  14. Vazifeshiran N, Soleimani M, Mortazavi Y, Kaviani S, Aboualghasemi H, Nikogoftar M. 3D culture of umblical cord blood-drived CD34+cells on a DBM scaffold coated by cord blood-drived unrestricted somatic stem cells (USSC). Modares Journal of Medical Sciences: Pathobiology 2010; 13: 63-81.
  15. Kobayashi K, Suzuki T, Nomoto Y, Tada Y, Miyake M, Hazama A, et al. A tissue-engineered trachea derived from a framed collagen scaffold, gingival fibroblasts and adipose-derived stem cells. Biomaterials 2010;31(18):4855.
  16. Mohammadi M, Shokrgozar MA, Mofid R. Culture of human gingival fibroblasts on a biodegradable scaffold and evaluation of its effect on attached gingiva: A randomized, controlled pilot study. Journal of Periodontology 2007;78(10):1897.
  17. Sheehy EJ, Lemoine M, Clarke D, Gonzalez Vazquez A, O’Brien FJ. The incorporation of marine coral microparticles into collagen-based scaffolds promotes osteogenesis of human mesenchymal stromal cells via calcium ion signalling. Marine Drugs 2020;18(2):74.
  18. Butler DL, Gooch C, Kinneberg KR, Boivin GP, Galloway MT, Nirmalanandhan VS, et al. The use of mesenchymal stem cells in collagen-based scaffolds for tissue-engineered repair of tendons. Nature Protocols 2010;5(5):849-63.
  19. Desimone MF, Hélary C, Rietveld IB, Bataille I, Mosser G, Giraud-Guille MM, et al. Silica–collagen bionanocomposites as three-dimensional scaffolds for fibroblast immobilization. Acta Biomaterialia 2010;6(10):3998-4004.
  20. Chen W, Shi C, Yi S, Chen B, Zhang W, Fang Z, et al. Bladder regeneration by collagen scaffolds with collagen binding human basic fibroblast growth factor. The Journal of Urology 2010;183(6):2432-9.
  21. Helary C, Ovtracht L. Coulomb B, Godeau G, Giraud-Guille MM. Dense fibrillar collagen matrices: A model to study myofibroblast behavior during wound healing. Biomaterials 2006; 27(25): 4443-52.
  22. Breen EC. Mechanical strain increases type I collagen expression in pulmonary fibroblasts in vitro. Journal of Applied Physiology 2000 1;88(1):203-9.
  23. González-Masís J, Cubero-Sesin JM, Corrales-Ureña YR, González-Camacho S, Mora-Ugalde N, Baizán-Rojas M, et al. Increased fibroblast metabolic activity of collagen scaffolds via the addition of propolis nanoparticles. Materials 2020;13(14):3118.
  24. Masci VL, Taddei AR, Gambellini G, Giorgi F, Fausto AM. Ultrastructural investigation on fibroblast interaction with collagen scaffold. Journal of Biomedical Materials Research 2016;104(1):272-82.
  25. Helary C, Bataille I, Abed A, Illoul C, Anglo A, Louedec L, et al. Concentrated collagen hydrogels as dermal substitutes. Biomaterials 2010;31(3):481-90.
  26. Sahraei SS, Kalhor N, Sheykhhasan M. Application of scaffolds in cartilage tissue engineering: a review paper. Razi Journal of Medical Sciences 2019;26(8):42-55.
  27. Ishaug-Riley SL, Crane GM, Gurlek A, Miller MJ, Yasko AW, Yaszemski MJ, et al. Ectopic bone formation by marrow stromal osteoblast transplantation using poly (DL-lactic-co-glycolic acid) foams implanted into the rat mesentery. Journal of Biomedical Materials Research 1997; 36: 1-8.
  28. Shalak R, Fox C. Preface. In: Shalak R, Fox C, editors. Tissue Engineering. New York: Alan R Liss 1998: 26-9.
  29. Lan W, Xu M, Zhang X, Zhao L, Huang D, Wei X, et al. Biomimetic polyvinyl alcohol/type II collagen hydrogels for cartilage tissue engineering. Journal of Biomaterials Science 2020;31(9):1179-98.
  30. Dong C, Lv Y. Application of collagen scaffold in tissue engineering: recent advances and new perspectives. Polymers 2016;8(2):42.
  31. Ding L, Sun H, Su J, Lin N, Péault B, Song T, et al. Transplantation of bone marrow mesenchymal stem cells on collagen scaffolds for the functional regeneration of injured rat uterus. Biomaterials 2014;35(18):4888-900.
  32. Zhao W, Chen B, Li X, Lin H, Sun W, Zhao Y, et al. Vascularization and cellularization of collagen scaffolds incorporated with two different collagen‐targeting human basic fibroblast growth factors. Journal of Biomedical Materials Research 2007;82(3):630-6.
  33. Tedder ME, Liao J, Weed B, Stabler C, Zhang H, Simionescu A, et al. Stabilized collagen scaffolds for heart valve tissue engineering. Tissue Engineering 2009;15(6):1257-68.
  34. Zhao W, Chen B, Li X, Lin H, Sun W, Zhao Y, et al. Vascularization and cellularization of collagen scaffolds incorporated with two different collagen‐targeting human basic fibroblast growth factors. Journal of Biomedical Materials Research 2007 ;82(3):630-6.
  35. Nöth U, Rackwitz L, Heymer A, Weber M, Baumann B, Steinert A, et al. Chondrogenic differentiation of human mesenchymal stem cells in collagen type I hydrogels. Journal of Biomedical Materials Research 2007;83(3):626-35.
  36. Oh SA, Lee HY, Lee JH, Kim TH, Jang JH, Kim HW, et al. Collagen three-dimensional hydrogel matrix carrying basic fibroblast growth factor for the cultivation of mesenchymal stem cells and osteogenic differentiation. Tissue Engineering 2012;18(9-10):1087-100.
  37. Helary C, Zarka M, Giraud‐Guille MM. Fibroblasts within concentrated collagen hydrogels favour chronic skin wound healing. Journal of Tissue Engineering and Regenerative Medicine 2012;6(3):225-37.
  38. Manjari MS, Aaron KP, Muralidharan C, Rose C. Highly biocompatible novel polyphenol cross-linked collagen scaffold for potential tissue engineering applications. Reactive and Functional Polymers 2020; 153:104630.
Daneshvar Medicine
Volume 30, Issue 2 - Serial Number 159
July and August 2022
Pages 1-11