Trypsin as enhancement in cyclical tracheal decellularization: Morphological and biophysical characterization

Giraldo-Gómez, David M.; León-Mancilla, Benjamín; Prado-Audelo, María Luisa Del; Sotres-Vega, Avelina; Villalba-Caloca, Jaime; Garciadiego-Cázares, David; Piña-Barba, María C.

doi:10.1016/j.msec.2015.10.094

Cited by 52 publications

(37 citation statements)

References 16 publications

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“…SEM showed that the three dimensional hierrarchial arrangement of the matrix was maintained at day 14 for protocol B. Protocol A was not ideal since it not only necessitated 35 days to decellularize the cartilage but in doing so it obliterated the ECM’s three dimensional (3D) structure. Surface characterization of cartilage post decellularization with SEM has also been reported by Utomo 15 as well as Gomez 18 and is an effective method of ECM analysis to asses for gross changes in the scaffold’s structure.…”

Section: Discussionmentioning

confidence: 74%

“…Trypsin was used in protocol B as it has shown to accelerate decellularization in the case of hyaline cartilage of the trachea as reported by Gomez et al . 18 .…”

Section: Discussionmentioning

confidence: 99%

“…The aim of this study was to identify a protocol that could optimize the decellularization of human auricular cartilage. The technique was based on the success of previous methods applied in the case of tracheal 17 , 18 and ear decellularization 15 . Our methodology consisted of applying three different decellularization protocols to human ear cartilage consisting of both physical and chemical methods.…”

Section: Introductionmentioning

confidence: 99%

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Optimising the decellularization of human elastic cartilage with trypsin for future use in ear reconstruction

Rahman

Griffin

Naik

et al. 2018

Sci Rep

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Decellularized scaffolds can induce chondrogenic differentiation of stem cells. This study compares different methods to optimise the decellularization of auricular cartilage. The process consisted of an initial 12 hour dry freeze thaw which froze the cartilage specimens in an empty tube at −20 °C. Samples were allowed to thaw at room temperature followed by submersion in phosphate buffer solution in which they were frozen at −20 °C for a 12 hour period. They were then allowed to thaw at room temperature as before. Protocol A subsequently involved subjecting specimens to both deoxyribonuclease and sodium deoxycholate. Protocol B and C were adaptations of this using 0.25% trypsin (7 cycles) and a 0.5 molar solution of ethylenediaminetetraacetic acid (3 hours for each cycle) respectively as additional steps. Trypsin accelerated the decellularization process with a reduction in DNA content from 55.4 ng/μL (native) to 17.3 ng/μL (P-value < 0.05) after 14 days. Protocol B showed a faster reduction in DNA content when compared with protocol A. In comparison to protocol C after 14 days, trypsin also showed greater decellularization with a mean difference of 11.7 ng/μL (P-value < 0.05). Histological analysis with H&E and DAPI confirmed depletion of cells at 14 days with trypsin.

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Section: Discussionmentioning

confidence: 74%

“…Trypsin was used in protocol B as it has shown to accelerate decellularization in the case of hyaline cartilage of the trachea as reported by Gomez et al . 18 .…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Optimising the decellularization of human elastic cartilage with trypsin for future use in ear reconstruction

Rahman

Griffin

Naik

et al. 2018

Sci Rep

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“…Collagen fibers, on the other hand, deformed, a result which can be detrimental to the heart valve's structural function. In porcine trachea decellularization, trypsin was supplemented to the SD/DNase I in a five-cycle protocol [71]. Each cycle involved the addition of 1% trypsin (3 hr at 4°C), 4% SD (4 hr at room temperature), and DNase I (3 hr at room temperature) with wash cycles in between each new solution.…”

Section: Decellularization Strategiesmentioning

confidence: 99%

Decellularization Strategies for Regenerative Medicine: From Processing Techniques to Applications

Gilpin

Yang

2017

BioMed Research International

597

578

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As the gap between donors and patients in need of an organ transplant continues to widen, research in regenerative medicine seeks to provide alternative strategies for treatment. One of the most promising techniques for tissue and organ regeneration is decellularization, in which the extracellular matrix (ECM) is isolated from its native cells and genetic material in order to produce a natural scaffold. The ECM, which ideally retains its inherent structural, biochemical, and biomechanical cues, can then be recellularized to produce a functional tissue or organ. While decellularization can be accomplished using chemical and enzymatic, physical, or combinative methods, each strategy has both benefits and drawbacks. The focus of this review is to compare the advantages and disadvantages of these methods in terms of their ability to retain desired ECM characteristics for particular tissues and organs. Additionally, a few applications of constructs engineered using decellularized cell sheets, tissues, and whole organs are discussed.

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“…In this context, decellularized lung tissue has been tested as an experimental alternative to transplantation. Perfusion of detergents such as SDS and Triton X-100 is a technique useful to decellularize mouse and rat cadaveric lung tissue, thereby preserving the vascular and airway structures of the tissue [141][142][143]. Due to the complexity of lung tissue and the need for instant functionality of used implants, a lung's dECM requires recellularization with epithelial and endothelial cells via cell infusion and bioreactors for ex vivo generation, maturation, and maintenance of the so-called bioartificial lungs.…”

Section: Tissues Related To Respiratory Systemmentioning

confidence: 99%

Tissue-Specific Decellularization Methods: Rationale and Strategies to Achieve Regenerative Compounds

Mendibil

Ruiz-Hernández

Retegi-Carrión

et al. 2020

IJMS

203

187

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The extracellular matrix (ECM) is a complex network with multiple functions, including specific functions during tissue regeneration. Precisely, the properties of the ECM have been thoroughly used in tissue engineering and regenerative medicine research, aiming to restore the function of damaged or dysfunctional tissues. Tissue decellularization is gaining momentum as a technique to obtain potentially implantable decellularized extracellular matrix (dECM) with well-preserved key components. Interestingly, the tissue-specific dECM is becoming a feasible option to carry out regenerative medicine research, with multiple advantages compared to other approaches. This review provides an overview of the most common methods used to obtain the dECM and summarizes the strategies adopted to decellularize specific tissues, aiming to provide a helpful guide for future research development.

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Trypsin as enhancement in cyclical tracheal decellularization: Morphological and biophysical characterization

Cited by 52 publications

References 16 publications

Optimising the decellularization of human elastic cartilage with trypsin for future use in ear reconstruction

Optimising the decellularization of human elastic cartilage with trypsin for future use in ear reconstruction

Decellularization Strategies for Regenerative Medicine: From Processing Techniques to Applications

Tissue-Specific Decellularization Methods: Rationale and Strategies to Achieve Regenerative Compounds

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