TY - JOUR
T1 - Comparative proteomic analyses of human adipose extracellular matrices decellularized using alternative procedures
AU - Thomas-Porch, Caasy
AU - Li, Jie
AU - Zanata, Fabiana
AU - Martin, Elizabeth C.
AU - Pashos, Nicholas
AU - Genemaras, Kaylynn
AU - Poche, J. Nicholas
AU - Totaro, Nicholas P.
AU - Bratton, Melyssa R.
AU - Gaupp, Dina
AU - Frazier, Trivia
AU - Wu, Xiying
AU - Ferreira, Lydia Masako
AU - Tian, Weidong
AU - Wang, Guangdi
AU - Bunnell, Bruce A.
AU - Flynn, Lauren
AU - Hayes, Daniel
AU - Gimble, Jeffrey M.
N1 - Funding Information:
The authors thank Dr. James Wade and his staff and patients (Baton Rouge LA) for their support of and participation in this study. Additionally, the authors thank Dr. David Kaplan and Dr. Rosalyn Abbott at the Tufts University Tissue Engineering Resource Center for providing the HFIP silk scaffolds. JMG, XW, and TF are co-owners of Obatala Sciences, Inc. where TF serves as President and CEO while JMG and XW are co-owners, as well as Chief Scientific Officer and Vice President of Research & Development of LaCell LLC, respectively; FS and TF were LaCell LLC employees at the time of this study. CT-P performed this study as partial fullfillment of her PhD thesis requirements at Tulane University School of Medicine.
Publisher Copyright:
© 2018 Wiley Periodicals, Inc.
PY - 2018/9
Y1 - 2018/9
N2 - Decellularized human adipose tissue has potential clinical utility as a processed biological scaffold for soft tissue cosmesis, grafting, and reconstruction. Adipose tissue decellularization has been accomplished using enzymatic-, detergent-, and/or solvent-based methods. To examine the hypothesis that distinct decellularization processes may yield scaffolds with differing compositions, the current study employed mass spectrometry to compare the proteomes of human adipose-derived matrices generated through three independent methods combining enzymatic-, detergent-, and/or solvent-based steps. In addition to protein content, bioscaffolds were evaluated for deoxyribose nucleic acid depletion, extracellular matrix composition, and physical structure using optical density, histochemical staining, and scanning electron microscopy. Mass spectrometry based proteomic analyses identified 25 proteins (having at least two peptide sequences detected) in the scaffolds generated with an enzymatic approach, 143 with the detergent approach, and 102 with the solvent approach, as compared to 155 detected in unprocessed native human fat. Immunohistochemical detection confirmed the presence of the structural proteins actin, collagen type VI, fibrillin, laminin, and vimentin. Subsequent in vivo analysis of the predominantly enzymatic- and detergent-based decellularized scaffolds following subcutaneous implantation in GFP+ transgenic mice demonstrated that the matrices generated with both approaches supported the ingrowth of host-derived adipocyte progenitors and vasculature in a time dependent manner. Together, these results determine that decellularization methods influence the protein composition of adipose tissue-derived bioscaffolds.
AB - Decellularized human adipose tissue has potential clinical utility as a processed biological scaffold for soft tissue cosmesis, grafting, and reconstruction. Adipose tissue decellularization has been accomplished using enzymatic-, detergent-, and/or solvent-based methods. To examine the hypothesis that distinct decellularization processes may yield scaffolds with differing compositions, the current study employed mass spectrometry to compare the proteomes of human adipose-derived matrices generated through three independent methods combining enzymatic-, detergent-, and/or solvent-based steps. In addition to protein content, bioscaffolds were evaluated for deoxyribose nucleic acid depletion, extracellular matrix composition, and physical structure using optical density, histochemical staining, and scanning electron microscopy. Mass spectrometry based proteomic analyses identified 25 proteins (having at least two peptide sequences detected) in the scaffolds generated with an enzymatic approach, 143 with the detergent approach, and 102 with the solvent approach, as compared to 155 detected in unprocessed native human fat. Immunohistochemical detection confirmed the presence of the structural proteins actin, collagen type VI, fibrillin, laminin, and vimentin. Subsequent in vivo analysis of the predominantly enzymatic- and detergent-based decellularized scaffolds following subcutaneous implantation in GFP+ transgenic mice demonstrated that the matrices generated with both approaches supported the ingrowth of host-derived adipocyte progenitors and vasculature in a time dependent manner. Together, these results determine that decellularization methods influence the protein composition of adipose tissue-derived bioscaffolds.
KW - adipose tissue
KW - bioscaffold
KW - decellularization
KW - extracellular matrix
KW - mass spectrometry proteomics
KW - regenerative medicine
UR - http://www.scopus.com/inward/record.url?scp=85053926520&partnerID=8YFLogxK
U2 - 10.1002/jbm.a.36444
DO - 10.1002/jbm.a.36444
M3 - Article
C2 - 29693792
AN - SCOPUS:85053926520
SN - 0021-9304
VL - 106
SP - 2481
EP - 2493
JO - Journal of Biomedical Materials Research - Part A
JF - Journal of Biomedical Materials Research - Part A
IS - 9
ER -