Novel Sodium Deoxycholate-Based Chemical Decellularization Method for Peripheral Nerve

McCrary, Michaela W.; Vaughn, Natalie; Hlavac, Nora; Song, Young Hye; Wachs, Rebecca A.; Schmidt, Christine E.

doi:10.1089/ten.tec.2019.0135

Cited by 44 publications

(57 citation statements)

References 49 publications

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“…Ionic detergents are subdivided into cationic and anionic solutions. Among them, sodium dodecyl sulfate (SdS) and sodium deoxycholate are representative examples for the removal of cellular debris from tissues (39). SdS is commonly used in the removal of nuclear remnants and cytoplasmic proteins, while sodium deoxycholate proved to be superior for solubilizing cytoplasmic and nuclear membranes.…”

Section: Methods Of Decellularization Treatmentsmentioning

confidence: 99%

Extracellular matrix grafts: From preparation to application (Review)

Jiang

Han

et al. 2020

Int J Mol Med

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Recently, the increasing emergency of traffic accidents and the unsatisfactory outcome of surgical intervention are driving research to seek a novel technology to repair traumatic soft tissue injury. From this perspective, decellularized matrix grafts (ECM-G) including natural ECM materials, and their prepared hydrogels and bioscaffolds, have emerged as possible alternatives for tissue engineering and regenerative medicine. Over the past decades, several physical and chemical decellularization methods have been used extensively to deal with different tissues/organs in an attempt to carefully remove cellular antigens while maintaining the non-immunogenic ECM components. It is anticipated that when the decellularized biomaterials are seeded with cells in vitro or incorporated into irregularly shaped defects in vivo , they can provide the appropriate biomechanical and biochemical conditions for directing cell behavior and tissue remodeling. The aim of this review is to first summarize the characteristics of ECM-G and describe their major decellularization methods from different sources, followed by analysis of how the bioactive factors and undesired residual cellular compositions influence the biologic function and host tissue response following implantation. Lastly, we also provide an overview of the in vivo application of ECM-G in facilitating tissue repair and remodeling.

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Section: Methods Of Decellularization Treatmentsmentioning

confidence: 99%

Extracellular matrix grafts: From preparation to application (Review)

Jiang

Han

et al. 2020

Int J Mol Med

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“…A diffusion method of chemical decellularization, adapted from previously published methods (30,31), was employed. Embryonic hearts were placed in 2 mL low retention tubes (Fisher Scientific 02-681-321), submerged with detergent and/or buffer, and placed on a rotator at 15 rpm (Thermo Fisher) for the following durations.…”

Section: Decellularization Processmentioning

confidence: 99%

“…Analysis of decellularized tissue, rather than unprocessed tissue, allows for better proteomic detection of ECM proteins, since highly abundant cellular proteins (e.g., myosin, actin) can mask the signal of generally lesser-abundant ECM proteins (36,37). Hudson et al (30) and McCrary et al (31)] utilizing anionic detergent sodium deoxycholate, zwitterionic detergents sulfobetaine-10 and−16, and enzyme deoxyribonuclease (DNase). Immunostaining was used to confirm removal of intracellular content and preservation of extracellular matrix proteins.…”

Section: Decellularization Creates Ecm-rich Hearts For Proteomic Assementioning

confidence: 99%

Extracellular Matrix Disparities in an Nkx2-5 Mutant Mouse Model of Congenital Heart Disease

Bousalis

Lacko

Hlavac

et al. 2020

Front. Cardiovasc. Med.

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Congenital heart disease (CHD) affects almost one percent of all live births. Despite diagnostic and surgical reparative advances, the causes and mechanisms of CHD are still primarily unknown. The extracellular matrix plays a large role in cell communication, function, and differentiation, and therefore likely plays a role in disease development and pathophysiology. Cell adhesion and gap junction proteins, such as integrins and connexins, are also essential to cellular communication and behavior, and could interact directly (integrins) or indirectly (connexins) with the extracellular matrix. In this work, we explore disparities in the expression and spatial patterning of extracellular matrix, adhesion, and gap junction proteins between wild type and Nkx2-5 +/R52G mutant mice. Decellularization and proteomic analysis, Western blotting, histology, immunostaining, and mechanical assessment of embryonic and neonatal wild type and Nkx2-5 mutant mouse hearts were performed. An increased abundance of collagen IV, fibronectin, and integrin β-1 was found in Nkx2-5 mutant neonatal mouse hearts, as well as increased expression of connexin 43 in embryonic mutant hearts. Furthermore, a ventricular noncompaction phenotype was observed in both embryonic and neonatal mutant hearts, as well as spatial disorganization of ECM proteins collagen IV and laminin in mutant hearts. Characterizing such properties in a mutant mouse model provides valuable information that can be applied to better understanding the mechanisms of congenital heart disease.

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“…As shown in Figure 8d, the collagen content was not statistically different across different groups. To determine what ECM proteins are present in the tissues after decellularization, proteomic analysis can be performed 45 , e.g., mass spectrometry and Western blots. Finally, tissue weights at different steps of the Campto+SB10 method were measured as an indicator of cell removal.…”

Section: Quantitative Assessment Of Decellularizationmentioning

confidence: 99%

Development of novel apoptosis-assisted lung tissue decellularization methods

Song

Maynes

Hlavac

et al. 2020

Preprint

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Decellularized tissues hold great potential for both regenerative medicine and disease modeling applications.The acellular extracellular matrix (ECM)-enriched scaffolds can be recellularized with patient-derived cells prior to transplantation, or digested to create thermally-gelling hydrogels for 3D cell culture. Current methods of decellularization clear cellular components using detergents, which can result in loss of ECM proteins and tissue architectural integrity. Recently, an alternative approach utilizing apoptosis to decellularize excised murine sciatic nerves resulted in superior ECM preservation, cell removal, and immune tolerance in vivo. However, this apoptosis-assisted decellularization approach has not been optimized for other tissues with a more complex geometry, such as lungs. To this end, we developed an apoptosis-assisted lung tissue decellularization method using a combination of camptothecin and sulfobetaine-10 (SB-10) to induce apoptosis and facilitate gentle and effective removal of cell debris, respectively. Importantly, combination of the two agents resulted in superior cell removal and ECM preservation compared to either of the treatments alone, presumably because of pulmonary surfactants. In addition, our method was superior in cell removal compared to a previously established detergentbased decellularization protocol. Furthermore, thermally-gelling lung ECM proteins supported high viability of rat lung epithelial cells for up to 2 weeks in culture. This work demonstrates that apoptosis-based lung tissue decellularization is a superior technique that warrants further utilization for both regenerative medicine and disease modeling applications. IntroductionAdvances in tissue engineering have led to broad use of naturally-derived biomaterials such as collagen, fibrin, hyaluronan, and decellularized tissue extracellular matrix (ECM) scaffolds, for regenerative medicine as well as for ex vivo disease modeling 1-4 . Naturally-derived biomaterials offer unique advantages in both applications because they provide natural cell attachment sites and are biocompatible, biodegradable, and cause little to no immune response post-implantation 2 . Additionally, natural biomaterial scaffolds allow assessment of cell behavior in a physiologically relevant manner as two-dimensional (2D) substrate coatings and as threedimensional (3D) cell culture scaffolds 1 . Studies have collectively demonstrated the importance of ECM-derived cues in cell response in healthy and diseased states, further highlighting the importance of environmental relevance when understanding cell behavior and translating the results to clinical applications.Among different naturally-derived biomaterials, decellularized tissues and organs offer unique advantages in generating acellular scaffolds with high retention of the majority of tissue-specific ECM components while removing immunogenic cellular debris 5 . As such, decellularized tissues have been widely used to repair damaged tissues and replace lost or non-functional tissues/organs...

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Novel Sodium Deoxycholate-Based Chemical Decellularization Method for Peripheral Nerve

Cited by 44 publications

References 49 publications

Extracellular matrix grafts: From preparation to application (Review)

Extracellular matrix grafts: From preparation to application (Review)

Extracellular Matrix Disparities in an Nkx2-5 Mutant Mouse Model of Congenital Heart Disease

Development of novel apoptosis-assisted lung tissue decellularization methods

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