Pre-Loading of Cells via Vapor Sublimation and the Deposition Polymerization Process with a 3D Porous Scaffold for Cell Cultures.

Abstract

In this study, we fabricate a three-dimensional (3D) porous poly-p-xylylene scaffold via a preloading technique and tailor it for cell culture. The fabrication process utilizes vapor sublimation and deposition polymerization, which exploits an ice template for sublimation and subsequent deposition of poly-p-xylylene under lower pressure and room temperature conditions. During this process, living cells are incorporated within a protective oil-in-water emulsion system, which facilitates high cell viability, and this construction forms a poly-p-xylylene scaffold with multiscale pores in the scaffold architecture that can be maintained for a tested time frame of 21 days in the current study. This reported fabrication method addresses inherent limitations of traditional methods, such as restricted biocompatibility, the need for modification procedures to achieve adequate porosity, and postseeding/loading of cells. By facilitating precise control over both micro- and nanostructures, the approach simultaneously preloads and accommodates multiple cell types and/or the necessary bioactive factors in the water solution and becomes an ice template. Finally, a single vapor phase fabrication step can lead to the construction of devised multifunctional scaffolds. The resulting scaffolds exhibit high porosity, featuring interconnected pores for cell migration and nutrient diffusion. Furthermore, controlled nanoroughness and microporosity promote cell attachment and enhance cell-cell and cell-matrix interactions, which are critical for tissue integration. Various types of cell cultures alongside diverse lineages of differentiations, including adipogenic, osteogenic, and neurogenic lineages, were examined in this study. Finally, the creation of anisotropic directional scaffolds that mimic native tissue architecture and promote cell attachment is particularly relevant for applications such as dental tissue regeneration and vascularization. Overall, the presented methodology represents a significant advancement in scaffold fabrication technology with considerable potential for versatility in regenerative medicine and complex tissue regeneration.