J Cosmet Med 2019; 3(2): 86-93  https://doi.org/10.25056/JCM.2019.3.2.86
Supercritical extraction of decellularized extracellular matrix from porcine adipose tissue as regeneration therapeutics
Seungwon Chung, MS1, Hana Kwon, MS2, Namsoo Peter Kim, PhD1,2
1Department of Metallurgical Materials and Biomedical Engineering, The University of Texas at El Paso, El Paso, TX, United States
2Center for Printing Materials Certification, The University of Texas at El Paso, El Paso, TX, United States
Namsoo Peter Kim
E-mail: nkim@utep.edu
Received: November 29, 2019; Revised: December 19, 2019; Accepted: December 22, 2019; Published online: December 31, 2019.
© Korean Society of Korean Cosmetic Surgery. All rights reserved.

cc This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background: Extracellular matrix (ECM) has been broadly applied and shown great promise in medical applications. ECM products should be used after decellularization and purification. Supercritical carbon dioxide treatment is of particular interest for purifying ECM due to its medical availability and rapid process speed. However, it is not fully researched for treatment of biomaterials for tissue engineering. Therefore, we investigated the optimal conditions of supercritical carbon dioxide processing at different extracting parameters in porcine adipose tissue.
Objective: We aimed to identify the optimal supercritical extracting conditions to produce non-cytotoxic and sterile decellularized extracellular matrix (DE-ECM) for regeneration therapeutics.
Methods: The three-day dual treatment including enzymatic decellularization and supercritical fluid extraction of pork adipose tissue was performed. Two protocols using different extracting parameters were applied to evaluate the influence of extracting pressure and temperature on the extraction yield, DNA concentration, and remaining collagen in product.
Results: Yield rate increased when high temperature or pressure was applied and pre-enzyme treatment had higher yield rate percent than pre-supercritical processing. Nearly 90% DNA was removed from the pre-enzyme sample when extracted at 3.04×107 Pa and 30°C±5°C. The pre-enzyme process had efficient extracting ability at each temperature and pressure and the remaining collagen steadily decreased with increase in extracting pressure and temperature. At the lowest temperature (20°C±5°C) and pressure (1.01×107 Pa), remaining collagen was 75.74%±1.83%. Supercritical extraction technology can produce DE-ECM eliminating DNA content efficiently and the remaining proper collagen amount successfully.
Conclusion: This study evaluated the feasibility of utilizing supercritical extraction technology in bio-materials and was proven to be is successful. Through controlling the extracting pressure and temperature, this technology has a potential for DE-ECM mass production, which can be useful as tissue regeneration therapeutics as well new drug delivery paradigm.
Keywords: enzyme decellularization; extracellular matrix; regeneration therapeutics; supercritical extraction process
References
  1. Badylak SF. Xenogeneic extracellular matrix as a scaffold for tissue reconstruction. Transpl Immunol 2004;12:367-77.
    Pubmed CrossRef
  2. Vorotnikova E, McIntosh D, Dewilde A, Zhang J, Reing JE, Zhang L, et al. Extracellular matrix-derived products modulate endothelial and progenitor cell migration and proliferation in vitro and stimulate regenerative healing in vivo. Matrix Biol 2010;29:690-700.
    Pubmed CrossRef
  3. Guler S, Aslan B, Hosseinian P, Aydin HM. Supercritical carbon dioxide-assisted decellularization of aorta and cornea. Tissue Eng Part C Methods 2017;23:540-7.
    Pubmed CrossRef
  4. Bible E, Dell’Acqua F, Solanky B, Balducci A, Crapo PM, Badylak SF, et al. Non-invasive imaging of transplanted human neural stem cells and ECM scaffold remodeling in the strokedamaged rat brain by (19)F- and diffusion-MRI. Biomaterials 2012;33:2858-71.
    Pubmed KoreaMed CrossRef
  5. Ott HC, Matthiesen TS, Goh SK, Black LD, Kren SM, Netoff TI, et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med 2008;14:213-21.
    Pubmed CrossRef
  6. Song JJ, Ott HC. Organ engineering based on decellularized matrix scaffolds. Trends Mol Med 2011;17:424-32.
    Pubmed CrossRef
  7. Choi YC, Choi JS, Kim BS, Kim JD, Yoon HI, Cho YW. Decellularized extracellular matrix derived from porcine adipose tissue as a xenogeneic biomaterial for tissue engineering. Tissue Eng Part C Methods 2012;18:866-76.
    Pubmed KoreaMed CrossRef
  8. Badylak SF. The extracellular matrix as a scaffold for tissue reconstruction. Semin Cell Dev Biol 2002;13:377-83.
    Pubmed CrossRef
  9. Gholipourmalekabadi M, Mozafari M, Salehi M, Seifalian A, Bandehpour M, Ghanbarian H, et al. Development of a cost-effective and simple protocol for decellularization and preservation of human amniotic membrane as a soft tissue replacement and delivery system for bone marrow stromal cells. Adv Healthc Mater 2015;4:918-26.
    Pubmed CrossRef
  10. Jungebluth P, Go T, Asnaghi A, Bellini S, Martorell J, Calore C, et al. Structural and morphologic evaluation of a novel detergent-enzymatic tissue-engineered tracheal tubular matrix. J Thorac Cardiovasc Surg 2009;138:586-93; discussion 592-3.
    Pubmed CrossRef
  11. Haykal S, Soleas JP, Salna M, Hofer SO, Waddell TK. Evaluation of the structural integrity and extracellular matrix components of tracheal allografts following cyclical decellularization techniques: comparison of three protocols. Tissue Eng Part C Methods 2012;18:614-23.
    Pubmed CrossRef
  12. Keane TJ, Swinehart IT, Badylak SF. Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance. Methods 2015;84:25-34.
    Pubmed CrossRef
  13. Prasertsung I, Kanokpanont S, Bunaprasert T, Thanakit V, Damrongsakkul S. Development of acellular dermis from porcine skin using periodic pressurized technique. J Biomed Mater Res B Appl Biomater 2008;85:210-9.
    Pubmed CrossRef
  14. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials 2011;32:3233-43.
    Pubmed KoreaMed CrossRef
  15. Dong X, Wei X, Yi W, Gu C, Kang X, Liu Y, et al. RGD-modified acellular bovine pericardium as a bioprosthetic scaffold for tissue engineering. J Mater Sci Mater Med 2009;20:2327-36.
    Pubmed CrossRef
  16. Reing JE, Brown BN, Daly KA, Freund JM, Gilbert TW, Hsiong SX, et al. The effects of processing methods upon mechanical and biologic properties of porcine dermal extracellular matrix scaffolds. Biomaterials 2010;31:8626-33.
    Pubmed KoreaMed CrossRef
  17. Meyer SR, Chiu B, Churchill TA, Zhu L, Lakey JR, Ross DB. Comparison of aortic valve allograft decellularization techniques in the rat. J Biomed Mater Res A 2006;79:254-62.
    Pubmed CrossRef
  18. Deeken CR, White AK, Bachman SL, Ramshaw BJ, Cleveland DS, Loy TS, et al. Method of preparing a decellularized porcine tendon using tributyl phosphate. J Biomed Mater Res B Appl Biomater 2011;96:199-206.
    Pubmed CrossRef
  19. Rieder E, Kasimir MT, Silberhumer G, Seebacher G, Wolner E, Simon P, et al. Decellularization protocols of porcine heart valves differ importantly in efficiency of cell removal and susceptibility of the matrix to recellularization with human vascular cells. J Thorac Cardiovasc Surg 2004;127:399-405.
    Pubmed CrossRef
  20. Hopkinson A, Shanmuganathan VA, Yeung AM, Gray T, Lowe J, Dua HS, et al. Optimisation of amniotic membrane (AM) denuding for tissue engineering. Acta Ophthalmol 2008. doi:10.1111/j.1755-3768.2008.5436.x.
    Pubmed CrossRef
  21. Phillips M, Maor E, Rubinsky B. Nonthermal irreversible electroporation for tissue decellularization. J Biomech Eng 2010;132:091003.
    Pubmed CrossRef
  22. Wainwright JM, Czajka CA, Patel UB, Freytes DO, Tobita K, Gilbert TW, et al. Preparation of cardiac extracellular matrix from an intact porcine heart. Tissue Eng Part C Methods 2010;16:525-32.
    Pubmed KoreaMed CrossRef
  23. Uriel S, Huang JJ, Moya ML, Francis ME, Wang R, Chang SY, et al. The role of adipose protein derived hydrogels in adipogenesis. Biomaterials 2008;29:3712-9.
    Pubmed CrossRef
  24. Flynn LE. The use of decellularized adipose tissue to provide an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem cells. Biomaterials 2010;31:4715-24.
    Pubmed CrossRef
  25. Cho D, Chung S, Eo J, Kim NP. Super-critical-CO2 de-ECM process. MRS Adv 2018;3:2391-7.
    CrossRef
  26. Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials 2006;27:3675-83.
    Pubmed CrossRef
  27. Roosens A, Somers P, De Somer F, Carriel V, Van Nooten G, Cornelissen R. Impact of detergent-based decellularization methods on porcine tissues for heart valve engineering. Ann Biomed Eng 2016;44:2827-39.
    Pubmed CrossRef
  28. Bamberger T, Erickson JC, Cooney CL, Kumar SK. Measurement and model prediction of solubilities of pure fatty acids, pure triglycerides, and mixtures of triglycerides in supercritical carbon dioxide. J Chem Eng Data 1988;33:327-33.
    CrossRef
  29. Ge Y, Ni Y, Yan H, Chen Y, Cai T. Optimization of the supercritical fluid extraction of natural vitamin E from wheat germ using response surface methodology. J Food Sci 2002;67:239-43.
    CrossRef
  30. Perrut M, Clavier JY, Poletto M, Reverchon E. Mathematical modeling of sunflower seed extraction by supercritical CO2. Ind Eng Chem Res 1997;36:430-5.
    CrossRef
  31. Lack E, Simándi B. 9.6 - Supercritical fluid extraction and fractionation from solid materials. Ind Chem Libr 2001;9:537-75.
    CrossRef
  32. Marsal A, Celma PJ, Cot JM, Cequier M. Supercritical CO2 extraction as a clean degreasing process in the leather industry. J Supercrit Fluids 2000;16:217-23.
    CrossRef
  33. Singer VL, Jones LJ, Yue ST, Haugland RP. Characterization of PicoGreen reagent and development of a fluorescence-based solution assay for double-stranded DNA quantitation. Anal Biochem 1997;249:228-38.
    Pubmed CrossRef
  34. Wilshaw SP, Kearney JN, Fisher J, Ingham E. Production of an acellular amniotic membrane matrix for use in tissue engineering. Tissue Eng 2006;12:2117-29.
    Pubmed CrossRef
  35. Roy BC, Goto M, Hirose T. Extraction of ginger oil with supercritical carbon dioxide: experiments and modeling. Ind Eng Chem Res 1996;35:607-12.
    CrossRef
  36. Vaquero EM, Beltrán S, Sanz MT. Extraction of fat from pigskin with supercritical carbon dioxide. J Supercrit Fluids 2006;37:142-50.
    CrossRef
  37. Jenkins CL, Raines RT. ChemInform abstract: insights on the conformational stability of collagen. ChemInform 2002;33. doi: 10.1002/chin.200218299.
    CrossRef


This Article


Cited By Articles
  • CrossRef (0)

Services
Social Network Service

e-submission

Archives