Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure.

Daiki Zemmyo,Shogo Miyata,Masashi Yamamoto

doi:10.3390/mi11111008

Abstract

Decellularized tissues are promising materials that mainly consist of extracellular matrices (ECMs) obtained by removing all cells from organs and tissues. High hydrostatic pressure (HHP) has been used for decellularization to remove cells physically from organs or tissues rather than by chemical methods. However, ultrahigh pressure induces denaturation of the ECM structure. In this study, we examined the effects of cyclic HHP at low and high pressures on the cell membrane structure to establish a novel decellularization method that enables decellularization without the denaturation of the ECM. A decellularization device using cyclic HHP (maximum pressure: 250 MPa, cycle number: 5) was developed. NB1RGB cell suspension was injected into a plastic bag to be subjected to cyclic HHP. After applying cyclic HHP, the amount of DNA inside the cells and the morphological changes of the cells were evaluated. As a result, the amount of DNA inside the cells decreased after the cyclic HHP compared to the static HHP. In addition, cyclic HHP was suggested to promote the destruction of the cell and nuclear membrane. In conclusion, it was revealed that the cell structure could be denatured and destroyed by cyclic HHP at a lower level than that of previous approaches.

Highlights

Tissue engineering is a discipline that aims to regenerate tissues and organs using living cells and scaffold materials
Bioabsorbable polymers are used as raw materials for scaffold materials, and these polymers are classified into two types: natural polymers and bioabsorbable synthetic polymers
scanning electron microscopy (SEM) observations were performed to evaluate the cell microstructure subjected to High hydrostatic pressure (HHP) (Figure 4b)

Summary

Introduction

Tissue engineering is a discipline that aims to regenerate tissues and organs using living cells and scaffold materials. Organs and tissues are composed of three elements: cells, scaffold materials, and physiologically active substances [1]. Bioabsorbable polymers are used as raw materials for scaffold materials, and these polymers are classified into two types: natural polymers and bioabsorbable synthetic polymers. They are used in hydrogel form as well as porous materials [2,3]. Synthetic polymers grater mechanical strength, but do not interact as favorably as biologically derived scaffolds [11,12]. In recent years, decellularized tissues obtained by removing cells from tissues obtained from humans or animals have been used as scaffold materials [13,14]

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