Protocols for the Preparation and Characterization of Decellularized Tissue and Organ Scaffolds for Tissue Engineering

Hsieh, Dar-Jen; Periasamy, Sivakumar; Yen, Ko-Chung; Yeh, Yi-Chun; Chen, Yun‐Ju; Wang, Hung-Chou; Tarng, Yih-Wen

doi:10.2144/btn-2020-0141

Cited by 17 publications

(14 citation statements)

References 15 publications

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“…Detergent decellularization is known to reduce the number of valuable growth factors that are vital for the recellularization of tissues. Moreover, residual surfactants and chemicals often cause cytotoxicity [15] inducing adverse effects in the recellularization of tissue and organ scaffolds (Table 3) [13,14,16].…”

Section: Tissues and Organsmentioning

confidence: 99%

Section: Adipose Tissuementioning

confidence: 99%

“…We produce tissue and organ scaffold using SCCO 2 extraction technology, such as liver, kidney, heart, pancreas, artery, skin, bone, cartilage, and cornea [16]. Table 5 listed our works on the various tissue and organ scaffolds extracted by SCCO 2 technology for tissue engineering applications [16]. The ultimate goal of TERM is to use the tissues and organs produced by SCCO 2 from the porcine or bovine to regenerate the human tissues and organs (Figure 7).…”

Section: Adipose Tissuementioning

confidence: 99%

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Supercritical Carbon Dioxide Facilitated Collagen Scaffold Production for Tissue Engineering

Periasamy¹,

Hsieh²

2022

Collagen Biomaterials

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The rise of tissue engineering and regenerative medicine (TERM) is a developing field that focuses on the advancement of alternative therapies for tissue and organ restoration. Collagen scaffold biomaterials play a vital role as a scaffold to promote cell growth and differentiation to promote the repair and regenerate the tissue lesion. The goal of this chapter will be to evaluate the role of supercritical carbon dioxide extraction technology in the production of collagen scaffold biomaterials from various tissues and organs and relate it to the traditional decellularization techniques in the production of collagen biomaterials for TERM. Therefore, we will study the collagen scaffold biomaterials produced using supercritical carbon dioxide extraction technology and their characteristics, such as chemical-physical properties, toxicity, biocompatibility, in vitro and in vivo bioactivity that could affect the interaction with cells and living system, relative to traditional decellularization technique-mediated collagen scaffolds. Furthermore, the chapter will focus on supercritical carbon dioxide extraction technology for the production of collagen scaffolds biomaterial appropriate for TERM.

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Section: Tissues and Organsmentioning

confidence: 99%

Section: Adipose Tissuementioning

confidence: 99%

Section: Adipose Tissuementioning

confidence: 99%

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Supercritical Carbon Dioxide Facilitated Collagen Scaffold Production for Tissue Engineering

Periasamy¹,

Hsieh²

2022

Collagen Biomaterials

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“…Although such decellularized ECM-based bioinks may be the best choice for a bioprinted tissue equivalent, to dissolve the tissue-derived ECM it has to be proteolytically digested, which in turn will destroy many ECM-anchored signaling molecules. In one recent paper, this problem was circumvented by milling and filtering ECM to µm-sized particles [ 12 ], which can be printed and are expected to represent more ‘native’ ECM than e.g. pepsin-digested decellularized matrix.…”

Section: Bioinks As the Crucial Components For Cell Survival Differentiation And Structural Integritymentioning

confidence: 99%

3D bioprinting: novel approaches for engineering complex human tissue equivalents and drug testing

Hagenbuchner

Nothdurfter

Ausserlechner

2021

Essays in Biochemistry

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Conventional approaches in drug development involve testing on 2D-cultured mammalian cells, followed by experiments in rodents. Although this is the common strategy, it has significant drawbacks: in 2D cell culture with human cells, the cultivation at normoxic conditions on a plastic or glass surface is an artificial situation that significantly changes energy metabolism, shape and intracellular signaling, which in turn directly affects drug response. On the other hand, rodents as the most frequently used animal models have evolutionarily separated from primates about 100 million years ago, with significant differences in physiology, which frequently leads to results not reproducible in humans. As an alternative, spheroid technology and micro-organoids have evolved in the last decade to provide 3D context for cells similar to native tissue. However, organoids used for drug testing are usually just in the 50–100 micrometers range and thereby too small to mimic micro-environmental tissue conditions such as limited nutrient and oxygen availability. An attractive alternative offers 3D bioprinting as this allows fabrication of human tissue equivalents from scratch with hollow structures for perfusion and strict spatiotemporal control over the deposition of cells and extracellular matrix proteins. Thereby, tissue surrogates with defined geometry are fabricated that offer unique opportunities in exploring cellular cross-talk, mechanobiology and morphogenesis. These tissue-equivalents are also very attractive tools in drug testing, as bioprinting enables standardized production, parallelization, and application-tailored design of human tissue, of human disease models and patient-specific tissue avatars. This review, therefore, summarizes recent advances in 3D bioprinting technology and its application for drug screening.

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“…While this method provides an initial quantitative estimate of DNA content, the tissue is conventionally classified as either successfully decellularized or not, depending on whether it passes the gold standard set by Crapo et al in 2011 [8]: tissues are considered decellularized if their DNA content is below 50 ng per mg of dry tissue. In addition, cell nuclei can be visualized with a histological stain such as Hematoxylin and Eosin (H&E) [9][10][11] or by staining the DNA using 4',6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) or Hoechst 33342 [10,[12][13][14]. Optical microscopy images are then used to confirm the lack of visible nuclei when compared to the native tissue.…”

Section: Introductionmentioning

confidence: 99%

Image-Based Method to Quantify Decellularization of Tissue Sections

Narciso

Otero

Navajas

et al. 2021

IJMS

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Tissue decellularization is typically assessed through absorbance-based DNA quantification after tissue digestion. This method has several disadvantages, namely its destructive nature and inadequacy in experimental situations where tissue is scarce. Here, we present an image processing algorithm for quantitative analysis of DNA content in (de)cellularized tissues as a faster, simpler and more comprehensive alternative. Our method uses local entropy measurements of a phase contrast image to create a mask, which is then applied to corresponding nuclei labelled (UV) images to extract average fluorescence intensities as an estimate of DNA content. The method can be used on native or decellularized tissue to quantify DNA content, thus allowing quantitative assessment of decellularization procedures. We confirm that our new method yields results in line with those obtained using the standard DNA quantification method and that it is successful for both lung and heart tissues. We are also able to accurately obtain a timeline of decreasing DNA content with increased incubation time with a decellularizing agent. Finally, the identified masks can also be applied to additional fluorescence images of immunostained proteins such as collagen or elastin, thus allowing further image-based tissue characterization.

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Protocols for the Preparation and Characterization of Decellularized Tissue and Organ Scaffolds for Tissue Engineering

Cited by 17 publications

References 15 publications

Supercritical Carbon Dioxide Facilitated Collagen Scaffold Production for Tissue Engineering

Supercritical Carbon Dioxide Facilitated Collagen Scaffold Production for Tissue Engineering

3D bioprinting: novel approaches for engineering complex human tissue equivalents and drug testing

Image-Based Method to Quantify Decellularization of Tissue Sections

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