Mechanical characterisation of unidirectional and cross-directional multilayered urinary bladder matrix (UBM) scaffolds

Callanan, Anthony; Davis, Niall F.; Walsh, Michael T.; McGloughlin, Timothy M.

doi:10.1016/j.medengphy.2012.06.022

Cited by 17 publications

(17 citation statements)

References 26 publications

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“…SDS treatment resulted in connective tissue with collagen bands slightly disorganized in some areas and arranged compactly in others, while the Triton X-100 protocol retained the natural connective tissue structure with lengthwise and homogeneous collagen bands. The Masson's trichrome staining, used to detected collagens (Callanan et al 2012), corroborated that the material observed in UBM decellularized with SDS (Fig. 2C) and Triton X-100 (Fig.…”

Section: Resultssupporting

confidence: 75%

“…Also note the absence of nuclei. Rosario et al 2008;Gilbert et al 2009;Narayanan et al 2009;Callanan et al 2012). Some authors maintain that the absence of nuclei on an H&E stain or by other IF methods alone is enough to confirm tissue decellularization (Gilbert 2012;Sawkins et al 2013); however, we believe that due to the subjectivity of this method and the necessary training of technicians, this cannot be the only method for evaluation.…”

Section: Discussionmentioning

confidence: 89%

“…Tissue engineering is a growing field with huge potential in tissue regeneration and repair, providing solutions for conditions in which the native tissue is diseased, damaged, or lost (Sarikaya et al 2002;Lee et al 2004;Rose et al 2009;Lu et al 2011a, b;Selim et al 2011;Callanan et al 2012;Teodori et al 2014). To reconstruct a new tissue, a triad of components is required: cells, biomaterials to be used as scaffolds, and growth factors (Ahn et al 2007;Narayanan et al 2009); the biomaterials play a central role because most mammalian cell types are anchorage dependent (Khang et al 2006;Liu et al 2011;Lu et al 2011a, b;Liu, et al 2012).…”

Section: Introductionmentioning

confidence: 99%

“…Decellularization involves a combination of physical, chemical, and enzymatic treatments (Bolland et al 2007;Badylak et al 2009;Lu et al 2011a, b;Callanan et al 2012). It is important to assess whether the selected decellularization procedure removes cellular components completely because many commonly used methods appear to be insufficient for complete cell removal and result in the retention of some DNA ).…”

Section: Introductionmentioning

confidence: 99%

“…The complete removal of the DNA from donor ECM is a critical step to prevent a potential inflammatory response following its implantation Gilbert et al 2009). Currently, the methods to evaluate the presence of DNA in ECM include spectrophotometry (Rosario et al 2008;Narayanan et al 2009), histological stains (Bolland et al 2007;Chun et al 2007;Rosario et al 2008;Gilbert et al 2009;Callanan et al 2012), and immunohistochemical methods (Woods and Gratzer 2005). The disadvantages of these methods include subjectivity and a lack of specificity.…”

Section: Introductionmentioning

confidence: 99%

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Quantification of DNA in urinary porcine bladder matrix using the ACTB gene

Silva-Benítez

Soto-Sáinz

Pozos‐Guillén

et al. 2015

In Vitro Cell.Dev.Biol.-Animal

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Extracellular matrix (ECM) is a rich network of proteins and proteoglycans that has proved to be very useful in tissue regeneration. Porcine ECM has been proposed as a biological scaffold, and urinary bladder matrix (UBM) has demonstrated superior biological properties; however, its use in human treatment requires ensuring that it is DNA free. Several protocols have been used for decellularization and to demonstrate the absence of DNA, but until now, a porcine housekeeping gene for quantifying DNA by real-time quantitative PCR (qPCR) has been limiting. The aim of this study was to propose a protocol to quantify the DNA content of decellularized UBM by qPCR for the beta-actin gene (ACTB). A total of 20 porcine bladders were used, and each bladder was divided into three pieces: one as a control and the others decellularized with either SDS or Triton X-100 detergent. The presence of DNA was assessed by histology, spectrophotometry, conventional PCR, and qPCR for the ACTB. Histological analysis demonstrated the absence of nuclei using both protocols. Spectrophotometrical evaluation resulted in DNA concentrations of 1561.4 ± 357.1 and 1211.9 ± 635.2 ng of DNA/mg dry weight after the SDS and Triton X-100 protocols, respectively. DNA was not detected in any protocol by conventional PCR. In contrast, using qPCR, we found 3.9 ± 2.8 ng of DNA/mg dry weight in the Triton X-100 protocol. Therefore, the use of qPCR is a reliable method to quantify residual DNA content after decellularization procedures.

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Section: Resultssupporting

confidence: 75%

Section: Discussionmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Quantification of DNA in urinary porcine bladder matrix using the ACTB gene

Silva-Benítez

Soto-Sáinz

Pozos‐Guillén

et al. 2015

In Vitro Cell.Dev.Biol.-Animal

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Influence of aorta extracellular matrix in electrospun polycaprolactone scaffolds

Reid

Callanan

2019

J of Applied Polymer Sci

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Cardiovascular disease is the leading cause of mortality worldwide. Therefore, new research strategies for the treatment of cardiovascular disease are required. Previously, extracellular matrices (ECMs) have been used alongside polymers to generate hybrid bioscaffolds. Herein, we propose combining aortic ECMs with a polycaprolactone electrospun scaffold and biomechanically evaluating the scaffolds. We electrospun three scaffolds with varying ECM concentrations and found that increasing the ECM concentration leads to decreased stiffness at low strains, increased elasticity at high strain, reduction in failure strain, and an increase in yield strength. We also noted a decrease in water droplet contact angle with the increasing ECM concentration. Furthermore, we found that all three scaffolds were capable of maintaining human umbilical vein endothelial cell attachment and survival. These findings show the wide spectrum of mechanical properties that can be achieved through the addition of different concentrations of ECM into the fibers. © 2019 The Authors. Journal of Applied Polymer Science published by Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 48181.

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On the potential of hydrated storage for naturally derived ECMs and associated effects on mechanical and cellular performance

et al. 2013

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Tissue engineered acellular vascular grafts are an emerging concept in the development of vascular prostheses for the minimally invasive treatment of cardiovascular diseases. Extracellular matrix (ECM) scaffolds, such as small intestinal submucosa (SIS) and urinary bladder matrix (UBM), offer many advantages over currently available synthetic devices. However, storage of such biomaterials can unduly influence the scaffold properties. This study evaluated the effects of up to 16 weeks hydrated storage on the mechanical and cellular performance of stented and unstented tubular scaffolds. This study aimed to demonstrate the viability, mechanical integrity, and bioactive potential of xenogeneic ECMs as potential off-the-shelf vascular prosthetic devices. Rehydrated ECM samples versus the lyophilized controls showed an increase in UTS and stiffness. The mechanical strength of all samples evaluated was above the average reported aortic tissue failure strength and more compliant than current synthetic materials employed. Post-storage cellular bioactivity investigations indicated that both ECM scaffolds tested were unaffected by increased hydrated storage duration when compared with the controls. Overall, the results indicate that the biomechanical and biologic properties of ECMs are not negatively affected by long-term hydrated storage. Therefore, with further investigations, naturally derived ECM materials may offer potential as an off-the-shelf therapeutic treatment of cardiovascular diseases.

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Mechanical characterisation of unidirectional and cross-directional multilayered urinary bladder matrix (UBM) scaffolds

Cited by 17 publications

References 26 publications

Quantification of DNA in urinary porcine bladder matrix using the ACTB gene

Quantification of DNA in urinary porcine bladder matrix using the ACTB gene

Influence of aorta extracellular matrix in electrospun polycaprolactone scaffolds

On the potential of hydrated storage for naturally derived ECMs and associated effects on mechanical and cellular performance

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