Evaluation of vacuum washing in the removal of SDS from decellularized bovine pericardium: method and device description

Abstract

The aim of this study was to present a new method for removing Sodium dodecyl sulfate (SDS) detergent from decellularized bovine pericardium using vacuum. Materials and Methods: The cows' pericardia were collected and decellularized. The samples were incubated with SDS1% for 48 h at 40 C. To perform vacuum washing (VW: negative pressure was used to wash and remove detergents), every decellularized tissue was cut in 75mm diameter and fixed via a stainless-steel ring with 60mm diameter in the center of filtration Buchner Funnel which was connected to glass filtration flask The system was connected to a vacuum pump by a hose, and a negative pressure of-100 mmHg was applied for 15 min. Then, the samples were shaken and washed at 40-rpm in 100 ml of distilled water for 45 min. This process was repeated for samples of each group (6 times for sample VW6h, 12 times for sample VW12h, and 24 times for sample VW24h). At the end of every cycle, the effluent was collected to take a sample for SDS measurement. The normal washing (NW) group containing distilled water (NWd) and PBS (Phosphate buffered saline) (NWp) were used to wash and remove detergents. SDS measurements, MTT Assay, histological and tensile test, to compare two methods were used. Results: The highest SDS in the effluent was in groups VW12h and VW24h (P 0.001) and the lowest residual SDS in scaffold was in two groups of VW12h and VW24h (P 0.001). MTT assay showed that cell survival in the VW12h and VW24h groups was higher than other groups and there' was no significant difference between cell survival in the VW12h and VW24h groups. Histological study showed destruction of tissue in the VW24h group. The results of the tensile test were shown that the native group had the highest module and the lowest amount was the VW24h sample which was reported with P 0.001 significance for all groups. Conclusion: VW12h can be used as an effective method for SDS removal from decellularized pericardium which morphologically demonstrated a good structure in ECM.

Evaluating the effects of vacuum on the microstructure and biocompatibility of bovine decellularized pericardium

“…It may be due to reduced residual DNA content in the decellularized tissue. According to our previous study, vacuum can reduce residual SDS content in decellularized tissues (Alizadeh et al, 2019).…”

“…For vacuum, we used an apparatus that was provided in our previous study. It consisted of Buchner funnel filtration device (65 mm diameter), glass filtration flask (250 cc), vacuum pump (Millipore Model WPG222050), and silicon hose (Alizadeh et al, 2019). For applying vacuum, each decellularized tissues were cut in 75 mm diameter and were attached to a stainless‐steel ring of 60 mm diameter, and then were placed in the funnel.…”

“…They found that vacuum rapidly generates decellularized scaffolds that are comparable with scaffolds previously decellularized via normal methods. Also, the effect of vacuum on detergent removal from decellularized tissue was evaluated in our previous study (Alizadeh et al, 2019). But the effects of vacuum on different parameters of decellularized tissues such as microstructure and biocompatibility had not been studied yet.…”

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

“…It has been reported in previous studies on decellularized corneas [ 2 ], UBM [ 17 ], SIS [ 25 ] and osteochondral plugs [ 29 ] that the presence of SDS residue within ECM biomaterials has a negative impact on cellular ingrowth due to the its cellular toxicity [ 30 ]. SDS debris inside of the decellularized tissue is difficult to be disposed of, as it binds strongly to the structure of the tissues [ 31 ]. To tackle this situation, the decellularized tissues in many studies were intensively washed with DI water or PBS for at least 48 h [ 31 ].…”

“…SDS debris inside of the decellularized tissue is difficult to be disposed of, as it binds strongly to the structure of the tissues [ 31 ]. To tackle this situation, the decellularized tissues in many studies were intensively washed with DI water or PBS for at least 48 h [ 31 ]. Washing the tissues with Triton X-100 after SDS treatment was also shown to effectively lower the SDS concentration in the materials, thus enhancing the cytocompatibility of the decellularized porcine liver [ 32 ], lung [ 33 ], heart [ 34 ] and kidney [ 35 ].…”