Characterization of Porcine Urinary Bladder Matrix Hydrogels from Sodium Dodecyl Sulfate Decellularization Method

Abstract

Urinary bladder matrix (UBM) is one of the most studied extracellular matrixes (ECM) in the tissue engineering field. Although almost all of the UBM hydrogels were prepared by using peracetic acid (PAA), recent studies indicated that PAA was not a trustworthy way to decellularize UBM. A stronger detergent, such as sodium dodecyl sulfate (SDS), may help tackle this issue; however, its effects on the hydrogels’ characteristics remain unknown. Therefore, the objective of this study was to develop a more reliable protocol to decellularize UBM, using SDS, and to compare the characteristics of hydrogels obtained from this method to the widely employed technique, using PAA. The results indicated that SDS was superior to PAA in decellularization efficacy. Different decellularization methods led to dissimilar gelation kinetics; however, the methods did not affect other hydrogel characteristics in terms of biochemical composition, surface morphology and rheological properties. The SDS-treated hydrogels possessed excellent cytocompatibility in vitro. These results showed that the SDS decellularization method could offer a more stable and safer way to obtain acellular UBM, due to reducing immunogenicity. The hydrogels prepared from this technique had comparable characteristics as those from PAA and could be a potential candidate as a scaffold for tissue remodeling.

Evaluating the Effect of Tissue Selection on the Characteristics of Extracellular Matrix Hydrogels from Decellularized Porcine Bladders

“…The two major classes of biomolecules within ECM materials, i.e., sGAG and collagen were accessed quantitatively after decellularization. The results of the UBM were consistent with our previous study [9] with respect to the collagen content (0.48 ± 0.07 compared to 0.49 ± 0.01) and the sGAG content (1.39 ± 0.28 compared to 1.5 ± 0.05, respectively). Interestingly, there was no difference between the soluble collagen and the sGAG amount in UBM, sECM and WB.…”

“…The efficacy of the decellularization process was confirmed on the basis of dsDNA quantification, DAPI staining, HE staining, DNA fragment size according to the procedures presented in our previously published study [9].…”

“…UBM was used as a control to sECM and WB in this study. The tissues were decellularized using our previously published method [9]. In brief, three to five native UBM, sECM and WB were lyophilized, ground into pieces.…”

“…In our previous study [9], we established a reliable protocol to decellularize UBM using sodium dodecyl sulfate (SDS), and successfully prepared UBM hydrogels. At that point, we also questioned the usage of the rest of the bladder after delamination (i.e., the sECM part), and tried to address this problem.…”

Incorporating a structural extracellular matrix gradient into a porcine urinary bladder matrix‐based hydrogel dermal scaffold

“…Our modified nUBM isolation protocol allows for clean mechanical delamination of nUBM from the overlying muscle tissue, as seen by the absence of muscle tissue and lack of visible nuclear material during histological analysis. SDS‐based decellularization has previously been shown to extract cellular material more effectively from UBM when compared to common peracetic acid‐based methods, and subsequent rinsing with serum‐supplemented culture medium further aids DNA removal due to serum nuclease activity 50,51 . DNA in dUBM was well below the commonly accepted threshold of 50 ng DNA/mg dry tissue and comparable to DNA levels in previously‐reported decellularized ECM materials, indicating effective DNA removal from the UBM 48,49,52‐54 .…”