Surfactant-free preparation of an ostrich carotid artery scaffold using liquefied dimethyl ether and DNase

Kanda, Hideki; Ando, Daichi; Oya, Kento; Diono, Wahyu; Goto, Motonobu

doi:10.1016/j.arabjc.2021.103280

Cited by 5 publications

(15 citation statements)

References 40 publications

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“…Kanda et al demonstrated that DME under subcritical temperatures, followed by DNA fragmentation with DNase, can effectively replace SDS in decellularization protocols for porcine aorta and ostrich carotid artery. 94 , 95 In addition to reducing structural damage to the ECM, DME may also reduce the immunogenic response properties. Urea is a powerful solubilizing agent with a high affinity for antigen removal.…”

Section: Decellularization Techniquesmentioning

confidence: 99%

Decellularization for the retention of tissue niches

Moffat

Jin

2022

J Tissue Eng

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Decellularization of natural tissues to produce extracellular matrix is a promising method for three-dimensional scaffolding and for understanding microenvironment of the tissue of interest. Due to the lack of a universal standard protocol for tissue decellularization, recent investigations seek to develop novel methods for whole or partial organ decellularization capable of supporting cell differentiation and implantation towards appropriate tissue regeneration. This review provides a comprehensive and updated perspective on the most recent advances in decellularization strategies for a variety of organs and tissues, highlighting techniques of chemical, physical, biological, enzymatic, or combinative-based methods to remove cellular contents from tissues. In addition, the review presents modernized approaches for improving standard decellularization protocols for numerous organ types.

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Section: Decellularization Techniquesmentioning

confidence: 99%

Decellularization for the retention of tissue niches

Moffat

Jin

2022

J Tissue Eng

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“…As a new approach for decellularization without SDS, a method for removing lipids from tissues using liquefied dimethyl ether (DME) as the solvent instead of SDS was proposed in 2021. DNA degradation by DNase and washing after lipid removal allow decellularization of the porcine aorta [5] and ostrich carotid arteries [21]. This method fulfilled all three conditions considered prerequisites for successful decellularization.…”

Section: Introductionmentioning

confidence: 86%

“…HPLC-grade methanol, chloroform, and water were purchased from FujiFilm Wako Pure Chemical Corporation (Osaka, Japan). The reagents and equipment not listed here are the same as those used in previous studies [5,21].…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, when used as an extraction solvent, it must be pressurized above the saturated vapor pressure. Its low boiling point also means that it leaves little residue in the extract or the extraction residue [5,21]. The European Union permits its use as an extraction medium in a number of cases [39].…”

Section: Introductionmentioning

confidence: 99%

“…These C=N bonds are degraded by aqueous DNase treatment after lipid extraction with liquefied DME [5]; however, the elasticity of the decellularized tissue in the porcine aorta is reduced [4]. In the case of the ostrich carotid arteries, temporary C=N bonds were not formed [21]. On the other hand, this C=N bond was not formed in the ostrich carotid artery, and the elasticity of the ostrich carotid artery was not reduced at all [42].…”

Section: Introductionmentioning

confidence: 99%

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Surfactant-Free Decellularization of Porcine Auricular Cartilage Using Liquefied Dimethyl Ether and DNase

et al. 2023

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The most common decellularization method involves lipid removal using surfactant sodium dodecyl sulfate (SDS) and DNA fragmentation using DNase, and is associated with residual SDS. We previously proposed a decellularization method for the porcine aorta and ostrich carotid artery using liquefied dimethyl ether (DME), which is free from the concerns associated with SDS residues, instead of SDS. In this study, the DME + DNase method was tested on crushed porcine auricular cartilage tissues. Unlike with the porcine aorta and the ostrich carotid artery, it is important to degas the porcine auricular cartilage using an aspirator before DNA fragmentation. Although approximately 90% of the lipids were removed using this method, approximately 2/3 of the water was removed, resulting in a temporary Schiff base reaction. The amount of residual DNA in the tissue was approximately 27 ng/mg dry weight, which is lower than the regulatory value of 50 ng/mg dry weight. Hematoxylin and eosin staining confirmed that cell nuclei were removed from the tissue. Residual DNA fragment length assessment by electrophoresis confirmed that the residual DNA was fragmented to less than 100 bp, which was lower than the regulatory limit of 200 bp. By contrast, in the uncrushed sample, only the surface was decellularized. Thus, although limited to a sample size of approximately 1 mm, liquefied DME can be used to decellularize porcine auricular cartilage. Thus, liquefied DME, with its low persistence and high lipid removal capacity, is an effective alternative to SDS.

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