BACKGROUND AND AIM OF STUDY: Assessment of decellularization of xenogeneic biological scaffolds for tissue engineering has relied primarily on histological cellularity, though this may not ensure the removal of known xenogeneic antigens such as galactose-alpha1,3-galactose (alpha-gal) and MHC I. METHODS: Bovine pericardium (BP) underwent standard (Std) decellularization consisting of hypotonic lysis and treatment with DNAse/RNAse. In addition to Std decellularization, tissues were treated for 24 h with either 0.5% Triton X-100, 0.5% sodium deoxycholate (SD), 0.1% sodium dodecyl sulfate (SDS), alpha-galactosidase (5 U/ml) or phospholipase (PL) A2 (150 U/ml). Tissues underwent a 96-h washout under gentle agitation at 27 degrees C, and then evaluated by light microscopy for % cellularity, and by immunohistochemistry and Western blot for alpha-gal, bovine MHC I and smooth muscle alpha-actin. RESULTS: Standard treatment of BP resulted in only partial removal histological cellularity and persistence of alpha-gal, MHC I and alpha-actin. Adding SD treatment resulted in apparent acellularity, but persistence of xenogeneic antigens. Only the addition of SDS resulted in complete histological acellularity and removal of xenogeneic antigens. Treatment with alpha-galactosidase selectively removed alpha-gal from BP. CONCLUSION: Histological cellularity is not an adequate end-point for assuring removal of antigenicity from xenogeneic biological scaffolds. However, known xenogeneic antigens can be targeted for removal by novel decellularization treatments such as alpha-galactosidase.
BACKGROUND AND AIM OF STUDY: Assessment of decellularization of xenogeneic biological scaffolds for tissue engineering has relied primarily on histological cellularity, though this may not ensure the removal of known xenogeneic antigens such as galactose-alpha1,3-galactose (alpha-gal) and MHC I. METHODS:Bovine pericardium (BP) underwent standard (Std) decellularization consisting of hypotonic lysis and treatment with DNAse/RNAse. In addition to Std decellularization, tissues were treated for 24 h with either 0.5% Triton X-100, 0.5% sodium deoxycholate (SD), 0.1% sodium dodecyl sulfate (SDS), alpha-galactosidase (5 U/ml) or phospholipase (PL) A2 (150 U/ml). Tissues underwent a 96-h washout under gentle agitation at 27 degrees C, and then evaluated by light microscopy for % cellularity, and by immunohistochemistry and Western blot for alpha-gal, bovine MHC I and smooth muscle alpha-actin. RESULTS: Standard treatment of BP resulted in only partial removal histological cellularity and persistence of alpha-gal, MHC I and alpha-actin. Adding SD treatment resulted in apparent acellularity, but persistence of xenogeneic antigens. Only the addition of SDS resulted in complete histological acellularity and removal of xenogeneic antigens. Treatment with alpha-galactosidase selectively removed alpha-gal from BP. CONCLUSION: Histological cellularity is not an adequate end-point for assuring removal of antigenicity from xenogeneic biological scaffolds. However, known xenogeneic antigens can be targeted for removal by novel decellularization treatments such as alpha-galactosidase.
Authors: Matthew A Soicher; Blaine A Christiansen; Susan M Stover; J Kent Leach; Clare E Yellowley; Leigh G Griffiths; David P Fyhrie Journal: J Biomed Mater Res A Date: 2014-02-26 Impact factor: 4.396
Authors: Piotr Wilczek; Anna Lesiak; Aleksandra Niemiec-Cyganek; Barbara Kubin; Ryszard Slomski; Jerzy Nozynski; Grazyna Wilczek; Aldona Mzyk; Michalina Gramatyka Journal: J Mater Sci Mater Med Date: 2015-01-11 Impact factor: 3.896
Authors: Leigh G Griffiths; Leila Choe; Kelvin H Lee; Kenneth F Reardon; E Christopher Orton Journal: Electrophoresis Date: 2008-11 Impact factor: 3.535
Authors: Susan M Mozzicato; Anubha Tripathi; Jonathon B Posthumus; Thomas A E Platts-Mills; Scott P Commins Journal: J Allergy Clin Immunol Pract Date: 2014-07-25
Authors: Leigh G Griffiths; Leila H Choe; Kenneth F Reardon; Steven W Dow; E Christopher Orton Journal: Biomaterials Date: 2008-06-02 Impact factor: 12.479