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Comparison of two decellularization methods of human kidney for the derivation of natural Scaffolds: Regenerative Medicine
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Abolfazl Khajavi Rad   , Samira Shahraki , Maryam Moghaddam Matin , Alireza Ebrahimzadeh Bideskan , Mohammad Asl Zare , Ahmad Reza Bahrami , Sara Hosseinian , Zahra Samadi Noshahr , Behrouz Mohammadi |
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Abstract: (263 Views) |
Introduction: Various diseases can lead to end stage renal disease (ESRD) that is a progressive loss of kidney function with a high rate of morbidity and mortality. Natural biological scaffolds that are prepared by removing cellular components from the donor organs using a decellularization process have many advantages compared with synthetic scaffold. Applying natural kidney scaffolds beside regenerative medicine and tissue engineering techniques are promising to achieve functional kidney.
Methods and Materials: In the current work two decellularization protocols were compared on human kidney sections (cortex, medulla and cortex-medulla). In protocol 1 detergents including 1% Triton X-100 (1A) and 1% SDS (1B) followed by DNase I were used. In the second protocol 0.5% SDS (2A) and 1% SDS (2B) were used. The efficiency of these methods was assessed by using hematoxylin and eosin (H&E), 4', 6 diamidino-2-phenylindole (DAPI), DNA quantification and immunohistochemistry (IHC) staining.
Result: H&E and DAPI staining results revealed a successful elimination of cell nuclei in all sections of protocols 1 and 2. The histological examinations revealed a better extracellular matrix preservation in triton-treated scaffolds (1A) as compared with the SDS-treated scaffolds (1B) at all times protocols. In addition, DNA quantification showed triton X-100 was more efficient as compared with other protocols for removing DNA in kidney sections. Furthermore, IHC staining showed the expression of collagen IV and laminin was preserved during decellularization with triton X-100 on days 5 and 10. In addition, IHC staining showed human leukocyte antigen (HLA) was completely removed in the cortex and medulla of human triton-treated scaffolds on days 5 and 10.
Conclusion: the results of this study showed that triton X-100 was a more effective detergent for decellularization of human kidney compared with SDS.
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| Keywords: Human kidney, Extracellular matrix, Decellularization, Triton, SDS |
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Full-Text [PDF 259 kb]
(35 Downloads)
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Type of Study: Research |
Subject:
General
Received: 2023/12/25 | Accepted: 2023/08/23 | Published: 2023/08/23
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