TY  - JOUR
AB  - Use of decellularized pig lung scaffolds recellularized with human lung cells in xenogeneic transplantation is potentially limited by expression of immunogenic galactosylated cell surface glycoproteins. Use of alpha 1,3 galactosyltransferase knock out (α-gal KO) pigs presents a potential alternative. Comparison of decellularized wild type (WT) vs α-gal KO pig lungs by histologic, immunohistochemical, and mass spectrometric techniques demonstrated no obvious differences in histologic structure was observed but an approximate 25% difference in retention of residual proteins including retention of α-galactosylated epitopes in WT lungs. Comparable robust initial recellularization and subsequent growth and proliferation was observed in wild type (WT) vs α-gal KO pig lungs following compartmental inoculation of human lung bronchial epithelial cells (HBE), human lung fibroblasts (HLF), and human bone marrow-derived mesenchymal stromal cells (MSCS) and of human pulmonary vascular endothelial cells (CBF). These initial sutides support further investigation of decellularized α-Gal KO pig lungs for xenogeneic lung regeneration.
AU  - Platz, J.*
AU  - Bonenfant, N.R.*
AU  - Uhl, F.*
AU  - Coffey, A.J.*
AU  - McKnight, T.*
AU  - Parsons, C.*
AU  - Sokocevic, D.*
AU  - Borg, Z.D.*
AU  - Lam, Y.W.*
AU  - Deng, B.*
AU  - Fields, J.*
AU  - Desarno, M.*
AU  - Loi, R.*
AU  - Hoffman, A.*
AU  - Bianchi, J.*
AU  - Dacken, B.*
AU  - Petersen, T.
AU  - Wagner, D.E.*
AU  - Weiss, D.J.*
C1  - 48829
C2  - 41452
CY  - New Rochelle
SP  - 725-739
TI  - Comparative study to the use of decellularized alpha-Gal KO pig lungs for xenogeneic lung transplantation.
JO  - Tissue Eng. Part C Meth.
VL  - 22
IS  - 8
PB  - Mary Ann Liebert, Inc
PY  - 2016
SN  - 1937-3384
ER  - 

TY  - JOUR
AB  - The development of reliable tissue engineering methods using decellularized cadaveric or donor lungs could potentially provide a new source of lung tissue. The vast majority of current lung decellularization protocols are detergent based and incompletely removed residual detergents may have a deleterious impact on subsequent scaffold recellularization. Detergent removal and quality control measures that rigorously and reliably confirm removal, ideally utilizing nondestructive methods, are thus critical for generating optimal acellular scaffolds suitable for potential clinical translation. Using a modified and optimized version of a methylene blue-based detergent assay, we developed a straightforward, noninvasive method for easily and reliably detecting two of the most commonly utilized anionic detergents, sodium deoxycholate (SDC) and sodium dodecyl sulfate (SDS), in lung decellularization effluents. In parallel studies, we sought to determine the threshold of detergent concentration that was cytotoxic using four different representative human cell types utilized in the study of lung recellularization: human bronchial epithelial cells, human pulmonary vascular endothelial cells (CBF12), human lung fibroblasts, and human mesenchymal stem cells. Notably, different cells have varying thresholds for either SDC or SDS-based detergent-induced cytotoxicity. These studies demonstrate the importance of reliably removing residual detergents and argue that multiple cell lines should be tested in cytocompatibility-based assessments of acellular scaffolds. The detergent detection assay presented here is a useful nondestructive tool for assessing detergent removal in potential decellularization schemes or for use as a potential endpoint in future clinical schemes, generating acellular lungs using anionic detergent-based decellularization protocols.
AU  - Zvarova, B.*
AU  - Uhl, F.E.*
AU  - Uriarte, J.J.*
AU  - Borg, Z.D.*
AU  - Coffey, A.L.*
AU  - Bonenfant, N.R.*
AU  - Weiss, D.J.*
AU  - Wagner, D.E.
C1  - 48768
C2  - 41619
CY  - New Rochelle
SP  - 418-428
TI  - Residual detergent detection method for nondestructive cytocompatibility evaluation of decellularized whole lung scaffolds.
JO  - Tissue Eng. Part C Meth.
VL  - 22
IS  - 5
PB  - Mary Ann Liebert, Inc
PY  - 2016
SN  - 1937-3384
ER  - 

TY  - JOUR
AB  - Scaffolds seeded with multipotent precursor cells were hypothesized to heal critically sized bone defects. However, the success of this concept was limited by low cell survival after transplantation due to a lack of nutrients and oxygen. In vivo prevascularization of scaffolds before cell seeding may improve cell survival, yet the best seeding technique and time point of cell application remain elusive. Thus, the aim of this study was to compare different strategies. Demineralized bone matrix scaffolds were implanted around the saphenous arteriovenous (AV) bundle in nude mice. In vivo seeding was performed 0, 5, or 21 days after implantation using enhanced green fluorescent protein (eGFP)-expressing mesenchymal stem cells (MSCs). Cells were applied either by injection or the repetitive dripping technique. In vitro seeded and subcutaneously implanted scaffolds served as controls. Fourteen days after cell application, the fluorescence intensity of transplanted cells and the extent of newly formed vessels were quantified. We found that the AV flow through model as well as cell application increased vessel formation. In vitro seeding resulted in significantly higher cell numbers than in vivo seeding. With increasing time of prevascularization, the number of cells declined dramatically. In vivo seeding by cell injection was superior to the repetitive dripping protocol. On subcutaneously implanted scaffolds, significantly, more cells were found than on axially perfused scaffolds. We conclude that in vitro seeding is more efficient compared to the two novel in vivo seeding techniques of prevascularized scaffolds. With increasing time of prevascularization, the seeding efficiency for the in vivo methods further decreases, presumably due to the ingrowth of connective tissue. Even though, the presence of MSCs and the longer period of prevascularization enhances vessel formation, this conceivable advantage is limited supposedly by the inferior seeding efficiency.
AU  - Polzer, H.*
AU  - Volkmer, E.*
AU  - Saller, M.
AU  - Prall, W.C.
AU  - Haasters, F.*
AU  - Drosse, I.
AU  - Wilhelmi, A.
AU  - Mutschler, W.*
AU  - Schieker, M.*
C1  - 29132
C2  - 33685
SP  - 11-18
TI  - Comparison of different strategies for in vivo seeding of prevascularized scaffolds.
JO  - Tissue Eng. Part C Meth.
VL  - 20
IS  - 1
PB  - Mary Ann Liebert, Inc
PY  - 2014
SN  - 1937-3384
ER  -