Development of a Model Bladder Extracellular Matrix Combining Disulfide Cross‐Linked Hyaluronan with Decellularized Bladder Tissue

Abstract

[Image: see text] In this work we investigate the feasibility of modifying porcine-derived BAM to include HA with a view to developing a model, artificial extracellular matrix for the study of bladder cell-matrix interactions. HA-DPTH was incorporated into BAM disks and then cross-linked oxidatively to a disulfide containing hydrogel. Disks were seeded with bladder smooth muscle cells (BSMC) and UEC under three culture configurations and incubated for 3, 7, and 14 d. At each time point, matrix contraction was measured, and media supernatants assayed for cell-secreted gelatinase activity. To evaluate cell adherence and organization, triple immunofluorescent labeling of cell nuclei, actin cytoskeleton, and focal contacts was performed. HA-modified BAM exhibited a significant increase in matrix contraction and induced a higher level of cell-secreted gelatinase activity compared to unmodified BAM. Immunofluorescent labeling demonstrated that BSMCs remained adherent to both scaffold types over time. The distribution and organization of the cytoskeleton and focal contacts did not appear to be altered by the presence of HA. Interestingly, cellular infiltration into modified BAM was evident by 7 d and continued beyond 14 d, while BSMCs seeded onto unmodified BAM remained localized to the surface out to 14 d, with minimal infiltration evident only at day 28. These differences in cell infiltration support the gelatinase activity results. Increases in cell migration and matrix proteolysis in the presence of HA may be contributing factors toward BAM remodeling leading to increased matrix contraction with time. The model ECM developed in this work will be utilized for future studies aimed at elucidating the mechanisms controlling key remodeling events associated with bladder repair. Matrix contraction of cell-seeded BAM scaffolds.

Characterization of and host response to tyramine substituted-hyaluronan enriched fascia extracellular matrix

“…To the best of our knowledge, there is no precedent for the investigation of in vivo host response to an ECM impregnated with a HA hydrogel, limiting comparison of our results to previous work. However, in one recent study, Brown et al showed that bladder ECM treated with disulfide cross-linked, thiol-modified HA facilitated and expedited the infiltration of bladder smooth muscle cells into bladder ECM in cell culture [59]. Since disulfide cross-linking is reversible and the incorporated HA could be leached or degraded over time (unlike dityramine cross-linking), this result is consistent with our observation of heightened cellularity in TS-HA treated fascia without cross-linking.…”

Controlled processing of a full-sized porcine liver to a decellularized matrix in 24 h

“…The maintenance of GAGs is substantial as they are important in morphogenesis, cell shape changes and motility (26). Contrary to collagen, GAGs levels in decellularized livers were elevated about 5-and 8-fold, respectively, compared to native livers.…”

Life in 3D is never flat: 3D models to optimise drug delivery

“…In addition variables arise depending upon the tissue used, the species from which the tissue/organ is isolated from and the terminal sterilisation procedures used [43]. Such decellularized tissues and organs have been used to culture cells in vitro to better understand disease pathogenesis as well as drug delivery [44,45].…”