The impact of detergents on the tissue decellularization process: A ToF-SIMS study

Abstract

Biologic scaffolds are derived from mammalian tissues, which must be decellularized to remove cellular antigens that would otherwise incite an adverse immune response. Although widely used clinically, the optimum balance between cell removal and the disruption of matrix architecture and surface ligand landscape remains a considerable challenge. Here we describe the use of time of flight secondary ion mass spectroscopy (ToF-SIMS) to provide sensitive, molecular specific, localized analysis of detergent decellularized biologic scaffolds. We detected residual detergent fragments, specifically from Triton X-100, sodium deoxycholate and sodium dodecyl sulphate (SDS) in decellularized scaffolds; increased SDS concentrations from 0.1% to 1.0% increased both the intensity of SDS fragments and adverse cell outcomes. We also identified cellular remnants, by detecting phosphate and phosphocholine ions in PAA and CHAPS decellularized scaffolds. The present study demonstrates ToF-SIMS is not only a powerful tool for characterization of biologic scaffold surface molecular functionality, but also enables sensitive assessment of decellularization efficacy.

Structural assessments in decellularized extracellular matrix of porcine semilunar heart valves: Evaluation of cell niches

“…The balance between cell removal and ECM conservation is the holy grail of the decellularization process . The harsh process of cell removes by detergent agents potentially causes impact in remodeling once implanted . Cytotoxicity and reduced mechanical resistance are common results in harsh decellularization processes which set detergent concentration only with cell removal focus .…”

Applications of decellularized extracellular matrix in bone and cartilage tissue engineering

“…Zwitterionic detergents such as 3‐([3‐cholamidopropyl] dimethylammonio)‐1‐propanesulfonate (CHAPS) have a net zero charge and show characteristics of both ionic and nonionic detergents . Although zwitterionic detergents result in less denaturing of proteins compared to ionic detergents, they also tend to remove less cellular material than ionic detergents …”

“…15 Although zwitterionic detergents result in less denaturing of proteins compared to ionic detergents, they also tend to remove less cellular material than ionic detergents. 41,42…”

Cellular and Molecular Probing of Intact Transparent Human Organs

“…Furthermore, once the micelles travel into the tissue and interact with the bilayer of lipid, the facial hydrophobic side of CHAPS reclines on the bilayer surface with a larger area rather than being embedded into the lipid as headto-tail detergents. These different interaction behaviors of the detergents suggest that detectable fragments of SDS and Triton X-100 exist after washing while no residual of CHAPS would be left inside of the tissue [30][31][32] (Fig. 1D,E).…”