Abstract
Decellularized tissues are considered superior scaffolds for cell cultures, preserving the microstructure of native tissues and delivering many kinds of cytokines. High hydrostatic pressure (HHP) treatment could remove cells physically from biological tissues rather than chemical methods. However, there are some risks of inducing destruction or denaturation of extracellular matrices (ECMs) at an ultrahigh level of HHP. Therefore, efficient decellularization using moderate HHP is required to remove almost all cells simultaneously to suppress tissue damage. In this study, we proposed a novel decellularization method using a moderate HHP with supercooling pretreatment. To validate the decellularization method, a supercooling device was developed to incubate human dermal fibroblasts or collagen gels in a supercooled state. The cell suspension and collagen gels were subjected to 100, 150, and 200 MPa of HHP after supercooling pretreatment, respectively. After applying HHP, the viability and morphology of the cells and the collagen network structure of the gels were evaluated. The viability of cells decreased dramatically after HHP application with supercooling pretreatment, whereas the microstructures of collagen gels were preserved and cell adhesivity was retained after HHP application. In conclusion, it was revealed that supercooling pretreatment promoted the denaturation of the cell membrane to improve the efficacy of decellularization using static application of moderate HHP. Furthermore, it was demonstrated that the HHP with supercooling pretreatment did not degenerate and damage the microstructure in collagen gels.
Highlights
Tissue engineering is a therapeutic approach that aims to regenerate tissues and organs using living cells and three-dimensional scaffolds
We reported that the decellularization effect was realized by cyclic application of moderate hHP [31]. in our previous study, the cell removal efficiency of our cyclic hHP application method was insufficient for clinical applications
The number of viable cells significantly decreased in the experimental group that was subjected to hHP at 150 MPa supercooling at −4 °C in the case of no high hydrostatic pressure (HHP) treatment (Figure 4)
Summary
Tissue engineering is a therapeutic approach that aims to regenerate tissues and organs using living cells and three-dimensional scaffolds. Tissue engineering methods reconstruct organs and biological tissues from cells, scaffold materials, and cytokines [1]. Cells adhere to scaffold materials to promote matrix synthesis and tissue reconstruction. Bioabsorbable polymers are usually used as scaffold materials, and they are classified into natural polymers and bioabsorbable synthetic polymers. Natural polymers are superior in cell proliferation and extracellular matrix (ECM) production because they contain cell adhesion molecules, whereas their mechanical strength is not sufficient as a scaffold material for three-dimensional cell cultures. Synthetic polymers have superior mechanical strength and are useful for controlling their geometric shape, whereas these kinds of polymers do not activate cells as biologically derived scaffolds [9,10]. Neither of these polymers can replicate the complex structures of organs and living tissues
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
