Decellularization with triton X-100 provides a suitable model for human kidney bioengineering using human mesenchymal stem cells

Shahraki, Samira; Bideskan, Alireza Ebrahimzadeh; Aslzare, Mohammad; Tavakkoli, Mahmoud; Bahrami, Ahmad Reza; Hosseinian, Sara; Matin, Maryam Moghaddam; Rad, Abolfazl Khajavi

doi:10.1016/j.lfs.2021.120167

Cited by 14 publications

(7 citation statements)

References 48 publications

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“…The benchmarks, like the amount of DNA, found to be less than 50 ng dsDNA/mg ECM dry weight ( Figure 2A ), and a complete absence of nuclei, were largely achieved ( Figure 2B ) ( Crapo et al, 2011 ; Gilbert et al, 2006 ). These data indicate a good nuclear removal efficiency, in the case of Triton X 100, already described in the literature ( Spina et al, 2003 ; Wang et al, 2017 ; Shahraki et al, 2022 ) and validated by the decrease in biomaterial thickness, after both treatments ( Supplementary Figure S6D ). The combination of non-ionic detergents, such as Triton X 100 or Tergitol 15 S 9, with bile acid adjuvants, seems to be successful, as it can give excellent results in terms of the amount of final DNA even at low detergent concentrations ( Oliveira et al, 2013 ; Paniagua Gutierrez et al, 2015 ).…”

Section: Discussionsupporting

confidence: 84%

Small intestinal submucosa-derived extracellular matrix as a heterotopic scaffold for cardiovascular applications

Palmosi

Tolomeo

Cirillo

et al. 2022

Front. Bioeng. Biotechnol.

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Structural cardiac lesions are often surgically repaired using prosthetic patches, which can be biological or synthetic. In the current clinical scenario, biological patches derived from the decellularization of a xenogeneic scaffold are gaining more interest as they maintain the natural architecture of the extracellular matrix (ECM) after the removal of the native cells and remnants. Once implanted in the host, these patches can induce tissue regeneration and repair, encouraging angiogenesis, migration, proliferation, and host cell differentiation. Lastly, decellularized xenogeneic patches undergo cell repopulation, thus reducing host immuno-mediated response against the graft and preventing device failure. Porcine small intestinal submucosa (pSIS) showed such properties in alternative clinical scenarios. Specifically, the US FDA approved its use in humans for urogenital procedures such as hernia repair, cystoplasties, ureteral reconstructions, stress incontinence, Peyronie’s disease, penile chordee, and even urethral reconstruction for hypospadias and strictures. In addition, it has also been successfully used for skeletal muscle tissue reconstruction in young patients. However, for cardiovascular applications, the results are controversial. In this study, we aimed to validate our decellularization protocol for SIS, which is based on the use of Tergitol 15 S 9, by comparing it to our previous and efficient method (Triton X 100), which is not more available in the market. For both treatments, we evaluated the preservation of the ECM ultrastructure, biomechanical features, biocompatibility, and final bioinductive capabilities. The overall analysis shows that the SIS tissue is macroscopically distinguishable into two regions, one smooth and one wrinkle, equivalent to the ultrastructure and biochemical and proteomic profile. Furthermore, Tergitol 15 S 9 treatment does not modify tissue biomechanics, resulting in comparable to the native one and confirming the superior preservation of the collagen fibers. In summary, the present study showed that the SIS decellularized with Tergitol 15 S 9 guarantees higher performances, compared to the Triton X 100 method, in all the explored fields and for both SIS regions: smooth and wrinkle.

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Section: Discussionsupporting

confidence: 84%

Small intestinal submucosa-derived extracellular matrix as a heterotopic scaffold for cardiovascular applications

Palmosi

Tolomeo

Cirillo

et al. 2022

Front. Bioeng. Biotechnol.

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“…Therefore, the significant reduction in GAG content in this study can be attributed to the combination of both SDS and SDC in our CHEM protocol. On the other hand, Triton X-100 was reported in several studies to better preserve the components of the ECM [16,18,28,38].…”

Section: Discussionmentioning

confidence: 99%

Decellularization of precision-cut kidney slices—application of physical and chemical methods

et al. 2023

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The extracellular matrix (ECM) obtained by decellularization provides scaffolds with the natural complex architecture and biochemical composition of the target organ. Whole kidney decellularization by perfusion uses the vasculature to remove cells leaving a scaffold that can be recellularized with patient-specific cells. However, decellularization and recellularization are highly complex processes that require intensive optimization of various parameters. In pursuit of this, a huge number of animals must be sacrificed. Therefore, we used precision-cut kidney slices (PCKS) as a source for natural scaffolds, which were decellularized by immersion in chemical reagents allowing the examination of more parameters with less animals. However, chemical reagents have a damaging effect on the structure and components of the ECM. Therefore, this study aimed at investigating the effects of physical treatment methods on the effectiveness of PCKS decellularization by immersion in chemical reagents (CHEM). PCKS were treated physically before or during immersion in chemicals (CHEM) with high hydrostatic pressure (HHP), freezing-thawing cycles (FTC) or in an ultrasonic bath system (UBS). Biochemical and DNA quantification as well as structural evaluation with conventional histology and scanning electron microscopy (SEM) were performed. Compared to decellularization by CHEM alone, FTC treatment prior to CHEM was the most effective in reducing DNA while also preserving glycosaminoglycan content (GAG). Moreover, while UBS resulted in a comparable reduction of DNA, it was the least effective in retaining GAGs. In contrast, despite the pretreatment with HHP with pressures up to 200 MPa, it was the least effective in DNA removal. Histological scoring showed that HHP scaffolds received the best score followed by UBS, FTC and CHEM scaffolds. However further analysis with SEM demonstrated a higher deterioration of the ultrastructure in UBS scaffolds. Altogether, pretreatment with FTC prior to CHEM resulted in a better balance between DNA removal and structural preservation.

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“…Triton™ X-100, a commonly used nonionic detergent, is frequently employed for chemical decellularization. It disrupts cellular lipid-lipid and lipid-protein interactions while preserving protein-protein interactions, leading to cell separation and the release of cytoplasmic materials through cell lysis [15] . It is typically used in various concentrations ranging from 0.1% to 5%, either on its own or in combination with trypsin/EDTA or sodium dodecyl sulfate (SDS) [ 16 , 17 ].…”

Section: Decellularization Of Tissue Ecmmentioning

confidence: 99%

Rise of tissue- and species-specific 3D bioprinting based on decellularized extracellular matrix-derived bioinks and bioresins

Elomaa,

Almalla,

Keshi

et al. 2023

Biomaterials and Biosystems

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Decellularization with triton X-100 provides a suitable model for human kidney bioengineering using human mesenchymal stem cells

Cited by 14 publications

References 48 publications

Small intestinal submucosa-derived extracellular matrix as a heterotopic scaffold for cardiovascular applications

Small intestinal submucosa-derived extracellular matrix as a heterotopic scaffold for cardiovascular applications

Decellularization of precision-cut kidney slices—application of physical and chemical methods

Rise of tissue- and species-specific 3D bioprinting based on decellularized extracellular matrix-derived bioinks and bioresins

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