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Cited by 114Tissue Engineering and Regenerative MedicineCorneal Surgery and Treatments3D Printing in Biomedical Research

Evaluation of Small Intestine Grafts Decellularization Methods for Corneal Tissue Engineering

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Oliveira, Ana Celeste, Garzón, Ingrid, Ionescu, Ana Maria, Carriel, Victor, Cardona, Juan de la Cruz, González-Andrades, Miguel, Pérez, María del Mar, Alaminos, Miguel +1 more·2013·PLoS ONE
DOI 10.1371/journal.pone.0066538PMC PMC3682956
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Abstract

Advances in the development of cornea substitutes by tissue engineering techniques have focused on the use of decellularized tissue scaffolds. In this work, we evaluated different chemical and physical decellularization methods on small intestine tissues to determine the most appropriate decellularization protocols for corneal applications. Our results revealed that the most efficient decellularization agents were the SDS and triton X-100 detergents, which were able to efficiently remove most cell nuclei and residual DNA. Histological and histochemical analyses revealed that collagen fibers were preserved upon decellularization with triton X-100, NaCl and sonication, whereas reticular fibers were properly preserved by decellularization with UV exposure. Extracellular matrix glycoproteins were preserved after decellularization with SDS, triton X-100 and sonication, whereas proteoglycans were not affected by any of the decellularization protocols. Tissue transparency was significantly higher than control non-decellularized tissues for all protocols, although the best light transmittance results were found in tissues decellularized with SDS and triton X-100. In conclusion, our results suggest that decellularized intestinal grafts could be used as biological scaffolds for cornea tissue engineering. Decellularization with triton X-100 was able to efficiently remove all cells from the tissues while preserving tissue structure and most fibrillar and non-fibrillar extracellular matrix components, suggesting that this specific decellularization agent could be safely used for efficient decellularization of SI tissues for cornea TE applications.

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  1. [1]Langer R, Vacanti JP (1993) Tissue engineering. Science 260: 920– 926. 8493529
  2. [2]Alaminos M, Del Carmen Sanchez-Quevedo M, Munoz-Avila JI, Serrano D, Medialdea S, et al (2006) Construction of a complete rabbit cornea substitute using a fibrin-agarose scaffold. Invest Ophthalmol Vis Sci 47: 3311– 3317. 16877396
  3. [3]Ionescu AM, de la Cruz Cardona J, Gonzalez-Andrades M, Alaminos M, Campos A, et al (2010) UV absorbance of a bioengineered corneal stroma substitute in the 240–400 nm range. Cornea 29: 895– 898 doi: 10.1097/ICO.0b013e3181ca3650 20508508 · DOI
  4. [4]Ionescu AM, Alaminos M, de la Cruz Cardona J, de Dios Garcia-Lopez Duran J, Gonzalez-Andrades M, et al (2011) Investigating a novel nanostructured fibrin-agarose biomaterial for human cornea tissue engineering: rheological properties. J Mech Behav Biomed Mater 4: 1963– 1973 doi: 10.1016/j.jmbbm.2011.06.013 22098895 · DOI
  5. [5]Cardona Jde L, Ionescu AM, Gomez-Sotomayor R, Gonzalez-Andrades M, Campos A, et al (2011) Transparency in a fibrin and fibrin-agarose corneal stroma substitute generated by tissue engineering. Cornea 30: 1428– 1435 doi: 10.1097/ICO.0b013e31821bdfd4 21934491 · DOI
  6. [6]Lin CC, Ritch R, Lin SM, Ni MH, Chang YC, et al (2010) A new fish scale-derived scaffold for corneal regeneration. Eur Cell Mater 19: 50– 57. 20186665
  7. [7]Liang Y, Liu W, Han B, Yang C, Ma Q, et al (2011) Fabrication and characters of a corneal endothelial cells scaffold based on chitosan. J Mater Sci Mater Med 22: 175– 183 doi: 10.1007/s10856-010-4190-6 21107657 · DOI
  8. [8]Sharma S, Mohanty S, Gupta D, Jassal M, Agrawal AK, et al (2011) Cellular response of limbal epithelial cells on electrospun poly-epsilon-caprolactone nanofibrous scaffolds for ocular surface bioengineering: a preliminary in vitro study. Mol Vis 17: 2898– 2910. 22128237 PMC3224839
  9. [9]Mi S, Khutoryanskiy VV, Jones RR, Zhu X, Hamley IW, et al (2011) Photochemical cross-linking of plastically compressed collagen gel produces an optimal scaffold for corneal tissue engineering. J Biomed Mater Res A 99: 1– 8 doi: 10.1002/jbm.a.33152 21732526 · DOI
  10. [10]Wu J, Du Y, Watkins SC, Funderburgh JL, Wagner WR (2012) The engineering of organized human corneal tissue through the spatial guidance of corneal stromal stem cells. Biomaterials 33: 1343– 1352 doi: 10.1016/j.biomaterials.2011.10.055 22078813 PMC3254093 · DOI
  11. [11]Ross EA, Williams MJ, Hamazaki T, Terada N, Clapp WL, et al (2009) Embryonic stem cells proliferate and differentiate when seeded into kidney scaffolds. J Am Soc Nephrol 20: 2338– 2347 doi: 10.1681/ASN.2008111196 19729441 PMC2799178 · DOI
  12. [12]Ott HC, Clippinger B, Conrad C, Schuetz C, Pomerantseva I, et al (2010) Regeneration and orthotopic transplantation of a bioartificial lung. Nat Med 16: 927– 933 doi: 10.1038/nm.2193 20628374 · DOI · Paper page
  13. [13]Gonzalez-Andrades M, de la Cruz Cardona J, Ionescu AM, Campos A, Del Mar Perez M, et al (2011) Generation of bioengineered corneas with decellularized xenografts and human keratocytes. Invest Ophthalmol Vis Sci 52: 215– 222 doi: 10.1167/iovs.09-4773 20739475 · DOI
  14. [14]Barakat O, Abbasi S, Rodriguez G, Rios J, Wood RP, et al (2012) Use of decellularized porcine liver for engineering humanized liver organ. J Surg Res 173: e11– 25 doi: 10.1016/j.jss.2011.09.033 22099595 · DOI · Paper page
  15. [15]Cigliano A, Gandaglia A, Lepedda AJ, Zinellu E, Naso F, et al (2012) Fine structure of glycosaminoglycans from fresh and decellularized porcine cardiac valves and pericardium. Biochem Res Int 2012: 1– 10 doi: 10.1155/2012/979351 PMC3296293 22461983 · DOI
  16. [16]Badylak SF, Taylor D, Uygun K (2011) Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds. Annu Rev Biomed Eng 13: 27– 53 doi: 10.1146/annurev-bioeng-071910-124743 21417722 PMC10887492 · DOI · Paper page
  17. [18]Crapo PM, Gilbert TW, Badylak SF (2011) An overview of tissue and whole organ decellularization processes. Biomaterials 32: 3233– 3243 doi: 10.1016/j.biomaterials.2011.01.057 21296410 PMC3084613 · DOI · Paper page
  18. [19]Keckler SJ, Spilde TL, St Peter SD, Tsao K, Ostlie DJ (2007) Treatment of bronchopleural fistula with small intestinal mucosa and fibrin glue sealant. Ann Thorac Surg 84: 1383– 1386. 17889008
  19. [20]Fiala R, Vidlar A, Vrtal R, Belej K, Student V (2007) Porcine small intestinal submucosa graft for repair of anterior urethral strictures. Eur Urol 51: 1702– 1708. 17306922
  20. [21]Lemos NL, Kamergorodsky G, Faria AL, Ribeiro PA, Auge AP, et al (2009) Small intestinal submucosa patch for extensive vaginal endometriosis resection. J Minim Invasive Gynecol 16: 765– 767 doi: 10.1016/j.jmig.2009.07.007 19896606 · DOI
  21. [22]Badylak SF (2004) Xenogeneic extracellular matrix as a scaffold for tissue reconstruction. Transpl Immunol 12: 367– 377. 15157928
  22. [23]Saghizadeh M, Chwa M, Aoki A, Lin B, Pirouzmanesh A, et al (2001) Altered expression of growth factors and cytokines in keratoconus, bullous keratopathy and diabetic human corneas. Exp Eye Res 73: 179– 189. 11446768
  23. [24]Featherstone HJ, Sansom J, Heinrich CL (2001) The use of porcine small intestinal submucosa in ten cases of feline corneal disease. Vet Ophthalmol 4: 147– 153. 11422997
  24. [25]Bussieres M, Krohne SG, Stiles J, Townsend WM (2004) The use of porcine small intestinal submucosa for the repair of full-thickness corneal defects in dogs, cats and horses. Vet Ophthalmol 7: 352– 359. 15310296
  25. [26]Goulle F (2012) Use of porcine small intestinal submucosa for corneal reconstruction in dogs and cats: 106 cases. J Small Anim Pract 53: 34– 43 doi: 10.1111/j.1748-5827.2011.01149.x 22122191 · DOI
  26. [27]Badylak SF, Freytes DO, Gilbert TW (2009) Extracellular matrix as a biological scaffold material: Structure and function. Acta Biomater 5: 1– 13 doi: 10.1016/j.actbio.2008.09.013 18938117 · DOI · Paper page
  27. [28]Carriel VS, Aneiros-Fernandez J, Arias-Santiago S, Garzon IJ, Alaminos M, et al (2011) A novel histochemical method for a simultaneous staining of melanin and collagen fibers. J Histochem Cytochem 59: 270– 277 doi: 10.1369/0022155410398001 21378281 PMC3201150 · DOI
  28. [29]Badylak SF (2007) The extracellular matrix as a biologic scaffold material. Biomaterials 28: 3587– 3593. 17524477
  29. [30]Cortiella J, Niles J, Cantu A, Brettler A, Pham A, et al (2010) Influence of acellular natural lung matrix on murine embryonic stem cell differentiation and tissue formation. Tissue Eng Part A 16: 2565– 2580 doi: 10.1089/ten.tea.2009.0730 20408765 · DOI
  30. [31]Wu J, Du Y, Mann MM, Yang E, Funderburgh J, et al.. (2013) Bioengineering Organized, Multi-Lamellar Human Corneal Stromal Tissue by Growth Factor Supplementation on Highly Aligned Synthetic Substrates. Tissue Eng Part A. In press. PMC3726016 23557404
  31. [32]Kheir E, Stapleton T, Shaw D, Jin Z, Fisher J, et al (2011) Development and characterization of an acellular porcine cartilage bone matrix for use in tissue engineering. J Biomed Mater Res A 99: 283– 294 doi: 10.1002/jbm.a.33171 21858917 · DOI
  32. [33]Zou Y, Zhang Y (2012) Mechanical evaluation of decellularized porcine thoracic aorta. J Surg Res 175: 359– 368 doi: 10.1016/j.jss.2011.03.070 21571306 PMC3100660 · DOI
  33. [34]Zhang AY, Bates SJ, Morrow E, Pham H, Pham B, et al (2009) Tissue-engineered intrasynovial tendons: optimization of acellularization and seeding. J Rehabil Res Dev 46: 489– 498. 19882484
  34. [35]Luo JC, Chen W, Chen XH, Qin TW, Huang YC, et al (2011) A multi-step method for preparation of porcine small intestinal submucosa (SIS). Biomaterials 32: 706– 713 doi: 10.1016/j.biomaterials.2010.09.017 20933271 · DOI
  35. [36]Kreis T, Vale R (1999) Guidebook to the Extracellular Matrix, Anchor, and Adhesion Proteins. New York: Oxford University Press. 335 p.
  36. [37]Gilbert TW (2012) Strategies for tissue and organ decellularization. J Cell Biochem 113: 2217– 2222 doi: 10.1002/jcb.24130 22415903 · DOI · Paper page
  37. [38]Deeken CR, White AK, Bachman SL, Ramshaw BJ, Cleveland DS, et al (2011) Method of preparing a decellularized porcine tendon using tributyl phosphate. J Biomed Mater Res B Appl Biomater 96: 199– 206 doi: 10.1002/jbm.b.31753 21210498 · DOI
  38. [39]Walter RJ, Matsuda T, Reyes HM, Walter JM, Hanumadass M (1998) Characterization of acellular dermal matrices (ADMs) prepared by two different methods. Burns 24: 104– 113. 9625233
  39. [40]Price AP, England KA, Matson AM, Blazar BR, Panoskaltsis-Mortari A (2010) Development of a decellularized lung bioreactor system for bioengineering the lung: the matrix reloaded. Tissue Eng Part A 16: 2581– 2591 doi: 10.1089/ten.TEA.2009.0659 20297903 PMC2947435 · DOI
  40. [41]Richter K, Nessling M, Lichter P (2007) Experimental evidence for the influence of molecular crowding on nuclear architecture. J Cell Sci 120: 1673– 1680. 17430977
  41. [42]Kim ST, Koh JW (2011) Mechanisms of apoptosis on human lens epithelium after ultraviolet light exposure. Korean J Ophthalmol 25: 196– 201 doi: 10.3341/kjo.2011.25.3.196 21655046 PMC3102824 · DOI
  42. [43]Suh H, Lee WK, Park JC, Cho BK (1999) Evaluation of the degree of cross-linking in UV irradiated porcine valves. Yonsei Med J 40: 159– 165. 10333720
  43. [44]Ushiki T (2002) Collagen fibers, reticular fibers and elastic fibers. A comprehensive understanding from a morphological viewpoint. Arch Histol Cytol 65: 109– 126. 12164335

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Authors
Oliveira, A. C., Garzón, I., Ionescu, A. M., Carriel, V., Cardona, J. D. L. C., González-Andrades, M., Pérez, M. D. M., Alaminos, M., & Campos, A.
Affiliations
Universidad de Granada (ES)
Corresponding author
Alaminos, Miguel
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2013-06-14
Volume
8
Issue
6
Pages
e66538
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0066538
PMC
PMC3682956
OpenAlex
W2003040782
Cited by
114
References
1
Citations by year
2026: 4 · 2025: 7 · 2024: 12 · 2023: 6 · 2022: 5 · 2021: 12 · 2020: 14 · 2019: 11 · 2018: 11 · 2017: 8 · 2016: 8 · 2015: 7 · 2014: 9
FWCI
7.89
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, other-oa)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Ghent University Academic Bibliography (Ghent University) (submittedVersion, other-oa)
  • Institutional Repository of the University of Granada (University of Granada) (submittedVersion, cc-by-nc-nd)
  • LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
Funders
Instituto de Salud Carlos III (PI11/)
Topics
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Corneal Surgery and Treatments
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
Keywords
decellularization, small intestine, corneal tissue engineering, decellularized scaffolds, Triton X-100, sodium dodecyl sulfate, sonication, UV irradiation, extracellular matrix preservation, collagen, reticular fibers, proteoglycans, tissue transparency, corneal substitutes
MeSH terms
Animals, Cornea, Detergents, Extracellular Matrix, Glycoproteins, Intestine, Small, Proteoglycans, Reticulin, Sodium Dodecyl Sulfate, Tissue Transplantation, Octoxynol, Tissue Engineering, Fibrillar Collagens, Mice, Tissue Scaffolds, Optical Phenomena

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