<i>In vitro</i>evaluation of surface biological properties of decellularized aorta for cardiovascular use

Kobayashi, Mako; Ohara, Masako; Hashimoto, Yoshihide; Nakamura, Naoko; Fujisato, Toshiya; Kimura, Tsuyoshi; Kishida, Akio

doi:10.1039/d0tb01830a

Cited by 11 publications

(8 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In one study, HHP-decellularized liver powder was implanted in a rat model of acute myocardial infarction, inducing cell migration and neovascularization, thus resulting in the suppression of myocardial necrosis [ 16 ]. The role of ECM microstructure in the good in vivo performance of HHP-decellularized tissues has been previously investigated [ 17 , 18 , 19 , 20 , 21 ]; however, the mechanism involving the bioactive components is still unknown. In this study, to evaluate whether MBVs are present in decellularized tissues, regardless of decellularization method, MBVs were extracted, and their presence was identified within HHP-decellularized tissues.…”

Section: Introductionmentioning

confidence: 99%

Extraction and Biological Evaluation of Matrix-Bound Nanovesicles (MBVs) from High-Hydrostatic Pressure-Decellularized Tissues

Kobayashi

Ishida

Hashimoto

et al. 2022

IJMS

Self Cite

View full text Add to dashboard Cite

Decellularized tissues are widely used as promising materials in tissue engineering and regenerative medicine. Research on the microstructure and components of the extracellular matrix (ECM) was conducted to improve the current understanding of decellularized tissue functionality. The presence of matrix-bound nanovesicles (MBVs) embedded within the ECM was recently reported. Results of a previous experimental investigation revealed that decellularized tissues prepared using high hydrostatic pressure (HHP) exhibited good in vivo performance. In the current study, according to the hypothesis that MBVs are one of the functional components in HHP-decellularized tissue, we investigated the extraction of MBVs and the associated effects on vascular endothelial cells. Using nanoparticle tracking assay (NTA), transmission electron microscopy (TEM), and RNA analysis, nanosized (100–300 nm) and membranous particles containing small RNA were detected in MBVs derived from HHP-decellularized small intestinal submucosa (SIS), urinary bladder matrix (UBM), and liver. To evaluate the effect on the growth of vascular endothelial cells, which are important in the tissue regeneration process, isolated SIS-derived MBVs were exposed to vascular endothelial cells to induce cell proliferation. These results indicate that MBVs can be extracted from HHP-decellularized tissues and may play a significant role in tissue remodeling.

show abstract

Section: Introductionmentioning

confidence: 99%

Extraction and Biological Evaluation of Matrix-Bound Nanovesicles (MBVs) from High-Hydrostatic Pressure-Decellularized Tissues

Kobayashi

Ishida

Hashimoto

et al. 2022

IJMS

Self Cite

View full text Add to dashboard Cite

show abstract

“…A well-preserved tissue structure demonstrates structural strength and tolerance to high shearing force 23 . HHP-based decellularization is a procedure involving high hydrostatic pressure for cellular antigen annihilation, and is associated with less damage to the tissue structure 13,24,25 . Ex vivo recellularization was performed with the aim of ex vivo tissue reconstruction prior to in vivo implantation for restoring vascular function, such as prevention of clot formation.…”

Section: Discussionmentioning

confidence: 99%

A novel approach for the endothelialization of xenogeneic decellularized vascular tissues by human cells utilizing surface modification and dynamic culture

Kobayashi

Murata

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Decellularized xenogeneic vascular grafts can be used in revascularization surgeries. We have developed decellularization methods using high hydrostatic pressure (HHP), which preserves the extracellular structure. Here, we attempted ex vivo endothelialization of HHP-decellularized xenogeneic tissues using human endothelial cells (ECs) to prevent clot formation against human blood. Slices of porcine aortic endothelium were decellularized using HHP and coated with gelatin. Human umbilical vein ECs were directly seeded and cultured under dynamic flow or static conditions for 14 days. Dynamic flow cultures tend to demonstrate higher cell coverage. We then coated the tissues with the E8 fragment of human laminin-411 (hL411), which has high affinity for ECs, and found that Dynamic/hL411showed high area coverage, almost reaching 100% (Dynamic/Gelatin vs Dynamic/hL411; 58.7 ± 11.4 vs 97.5 ± 1.9%, P = 0.0017). Immunostaining revealed sufficient endothelial cell coverage as a single cell layer in Dynamic/hL411. A clot formation assay using human whole blood showed low clot formation in Dynamic/hL411, almost similar to that in the negative control, polytetrafluoroethylene. Surface modification of HHP-decellularized xenogeneic endothelial tissues combined with dynamic culture achieved sufficient ex vivo endothelialization along with prevention of clot formation, indicating their potential for clinical use as vascular grafts in the future.

show abstract

“…Fresh porcine blood was obtained from a local slaughterhouse and stored at 4°C until use. Blood coagulation tests were performed according to a previously described protocol [ 10 ]. Briefly, whole blood containing 0.324% citric acid was prepared with a tenth of calcium chloride (CaCl 2 ) for the coagulation of blood for 15 min on glass (C022221, Matsunami Glass Ind.,Ltd.).…”

Section: Methodsmentioning

confidence: 99%

Effect of luminal surface structure of decellularized aorta on thrombus formation and cell behavior

et al. 2021

Self Cite

View full text Add to dashboard Cite

Due to an increasing number of cardiovascular diseases, artificial heart valves and blood vessels have been developed. Although cardiovascular applications using decellularized tissue have been studied, the mechanisms of their functionality remain unknown. To determine the important factors for preparing decellularized cardiovascular prostheses that show good in vivo performance, the effects of the luminal surface structure of the decellularized aorta on thrombus formation and cell behavior were investigated. Various luminal surface structures of a decellularized aorta were prepared by heating, drying, and peeling. The luminal surface structure and collagen denaturation were evaluated by immunohistological staining, collagen hybridizing peptide (CHP) staining, and scanning electron microscopy (SEM) analysis. To evaluate the effects of luminal surface structure of decellularized aorta on thrombus formation and cell behavior, blood clotting tests and recellularization of endothelial cells and smooth muscle cells were performed. The results of the blood clotting test showed that the closer the luminal surface structure is to the native aorta, the higher the anti-coagulant property. The results of the cell seeding test suggest that vascular cells recognize the luminal surface structure and regulate adhesion, proliferation, and functional expression accordingly. These results provide important factors for preparing decellularized cardiovascular prostheses and will lead to future developments in decellularized cardiovascular applications.

show abstract

In vitroevaluation of surface biological properties of decellularized aorta for cardiovascular use

Abstract: The aim of this study was to determine an in vitro evaluation method that could directly predict in vivo performance of decellularized tissues for cardiovascular use. We hypothesized that key...

Cited by 11 publications

References 68 publications

Extraction and Biological Evaluation of Matrix-Bound Nanovesicles (MBVs) from High-Hydrostatic Pressure-Decellularized Tissues

Extraction and Biological Evaluation of Matrix-Bound Nanovesicles (MBVs) from High-Hydrostatic Pressure-Decellularized Tissues

A novel approach for the endothelialization of xenogeneic decellularized vascular tissues by human cells utilizing surface modification and dynamic culture

Effect of luminal surface structure of decellularized aorta on thrombus formation and cell behavior

Contact Info

Product

Resources

About