An Effective Method for Decellularization of Human Foreskin: Implications for Skin Regeneration in Small Wounds

Document Type : Original Article

Authors

1 Cancer and Immunology Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

2 Department of Molecular Medicine, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran

3 Department of Applied Mathematics, University of Waterloo, Waterloo, ON, Canada

Abstract

Objective: Acellular matrices of different allogeneic or xenogeneic origins are widely used as structural scaffolds in regenerative medicine. The main goal of this research was to optimize a method for decellularization of foreskin for skin
regeneration in small wounds.
Materials and Methods: In this experimental study, the dermal layers of foreskin were divided into two sections and subjected to two different decellularization methods: the sodium dodecyl sulfate method (SDS-M), and our optimized
foreskin decellularization method (OFD-M). A combination of non-ionic detergents and SDS were used to decellularize the foreskin in OFD-M. The histological, morphological, and biomechanical properties of both methods were compared. In addition, human umbilical cord mesenchymal stem cells (hucMSCs) were isolated, and the biocompatibility and
recellularization of both scaffolds by hucMSC were subsequently determined.
Results: We observed that OFD-M is an appropriate approach for successful removal of cellular components from the foreskin tissue, without physical disturbance to the acellular matrix. In comparison to SDS-M, this new bioscaffold possesses a fine network containing a high amount of collagen fibers and glycosaminoglycans (GAG) (P≤0.03), is biocompatible and harmless for hucMSC (viability 91.7%), and exhibits a relatively high tensile strength.
Conclusion: We found that the extracellular matrix (ECM) structural integrity, the main ECM components, and the
mechanical properties of the foreskin are well maintained after applying the OFD-M decellularization technique, indicating that the resulting scaffold would be a suitable platform for culturing MSC for skin grafting in small wounds

Keywords


1. Debels H, Hamdi M, Abberton K, Morrison W. Dermal matrices and bioengineered skin substitutes: a critical review of current options. Plast Reconstr Surg Glob Open. 2015; 3(1): e284.
2. Cazzell S, Moyer PM, Samsell B, Dorsch K, McLean J, Moore MA. A prospective, multicenter, single-arm clinical trial for treatment of complex diabetic foot ulcers with deep exposure using acellular dermal matrix. Adv Skin Wound Care. 2019; 32(9): 409-415.
3. Rana D, Zreiqat H, Benkirane-Jessel N, Ramakrishna S, Ramalingam M. Development of decellularized scaffolds for stem celldriven tissue engineering. J Tissue Eng Regen Med. 2017; 11(4): 942-965.
4. Begum T, Farrelly PJ,Craigie RJ. Non-cross-linked porcine acellular dermal matrix (Strattice Tissue Matrix) in pediatric reconstructive surgery. J Pediatr Surg. 2016; 51(3): 461-464.
5. Lai C, Song G, Zhao B, Wang H, Pan B, Guo X, et al. Preparation and characterization of human scar acellular dermal matrix. J Biomater Sci Polym Ed. 2019; 30(9): 769-784.
6. Rozario T, DeSimone DW. The extracellular matrix in development and morphogenesis: a dynamic view. Dev Biol. 2010; 341(1): 126-140.
7. Salbach J, Rachner TD, Rauner M, Hempel U, Anderegg U, Franz S, et al. Regenerative potential of glycosaminoglycans for skin and bone. J Mol Med (Berl). 2012; 90(6): 625-635.
8. Heirani-Tabasi A, Hassanzadeh M, Hemmati-Sadeghi S, Shahriyari M, Raeesolmohaddeseen M. Mesenchymal stem cells; defining the future of regenerative medicine. Journal of Genes and Cells. 2015; 1(2): 34-39.
9. Shojaei F, Rahmati S, Banitalebi Dehkordi M. A review on different methods to increase the efficiency of mesenchymal stem cellbased wound therapy. Wound Repair Regen. 2019; 27(6): 661-671.
10. Azzopardi JI, Blundell R. Umbilical Cord Stem Cells. Stem Cell Discovery. 2018; 8: 1-11.
11. Duscher D, Barrera J, Wong VW, Maan ZN, Whittam AJ, Januszyk M, et al. Stem cells in wound healing: the future of regenerative medicine? A mini-review. Gerontology. 2016; 62(2): 216-225.
12. Purpura V, Bondioli E, Cunningham EJ, De Luca G, Capirossi D, Nigrisoli E, et al. The development of a decellularized extracellular matrix–based biomaterial scaffold derived from human foreskin for the purpose of foreskin reconstruction in circumcised males. J Tissue Eng. 2018; 9: 2041731418812613.
13. Young DA, Ibrahim DO, Hu D, Christman KL. Injectable hydrogel scaffold from decellularized human lipoaspirate. Acta Biomater. 2011; 7(3): 1040-1049.
14. Kajbafzadeh AM, Abbasioun R, Sabetkish S, Sabetkish N, Rahmani P, Tavakkolitabassi K, et al. Future prospects for human tissue engineered urethra transplantation: Decellularization and recellularization-
based urethra regeneration. Ann Biomed Eng. 2017; 45(7): 1795-1806.
15. Alizadeh M, Rezakhani L, Soleimannejad M, Sharifi E, Anjomshoa M, Alizadeh A. Evaluation of vacuum washing in the removal of SDS from decellularized bovine pericardium: method and device description. Heliyon. 2019; 5(8): e02253.
16. Luna LG. Manual of histologic staining methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill; 1968.
17. Martinello T, Pascoli F, Caporale G, Perazzi A, Iacopetti I, Patruno M. Might the Masson trichrome stain be considered a useful method for categorizing experimental tendon lesions. Histol Histopathol. 2015; 30(8): 963-969.
18. Ferdowsi Khosroshahi A, Soleimani Rad J, Kheirjou R, Roshangar B, Rashtbar M, Salehi R, et al. Adipose tissue-derived stem cells upon decellularized ovine small intestine submucosa for tissue regeneration: An optimization and comparison method. J Cell Physiol. 2020; 235(2): 1556-1567.
19. Kajbafzadeh AM, Masoumi A, Hosseini M, Borjian MA, Akbarzadeh A, Mohseni MJ. Sheep colon acellular matrix: immunohistologic, biomechanical, scanning electron microscopic evaluation and collagen quantification. J Biosci Bioeng. 2014; 117(2): 236-241.
20. Qiu YL, Chen X, Hou YL, Hou YJ, Tian SB, Chen YH, et al. Characterization of different biodegradable scaffolds in tissue engineering. Mol Med Rep. 2019; 19(5): 4043-4056.
21. Powers JG, Higham C, Broussard K, Phillips TJ. Wound healing and treating wounds: chronic wound care and management. J Am Acad Dermatol. 2016; 74(4): 607-625.
22. Frisch M, Earp BD. Circumcision of male infants and children as a public health measure in developed countries: a critical assessment of recent evidence. Glob Public Health. 2018; 13(5): 626-641.
23. Shimizu R, Kishi K. Skin graft. Plast Surg Int. 2012; 2012: 563493.
24. Cui H, Chai Y, Yu Y. Progress in developing decellularized bioscaffolds for enhancing skin construction. J Biomed Mater Res A. 2019; 107(8): 1849-1859.
25. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials. 2011; 32(12): 3233-3243.
26. Gilpin A, Yang Y. Decellularization strategies for regenerative medicine: from processing techniques to applications. Biomed Res Int. 2017; 2017: 9831534.
27. Wu Q, Bao J, Zhou Yj, Wang Yj, Du Zg, Shi Yj, et al. Optimizing perfusion-decellularization methods of porcine livers for clinicalscale whole-organ bioengineering. Biomed Res Int. 2015; 2015: 785474.
28. Lynch AP, Ahearne M. Strategies for developing decellularized corneal scaffolds. Exp Eye Res. 2013; 108: 42-47.
29. Luo J, Korossis SA, Wilshaw SP, Jennings LM, Fisher J, Ingham E. Development and characterization of acellular porcine pulmonary valve scaffolds for tissue engineering. Tissue Eng Part A. 2014; 20(21-22): 2963-2974.