Artificial decellularized extracellular matrix improves the regenerative capacity of adipose tissue derived stem cells on 3D printed polycaprolactone scaffolds

Abstract

Ideal tissue engineering frameworks should be both an optimal biological microenvironment and a shape and stability providing framework. In this study we tried to combine the advantages of cell-derived artificial extracellular matrix (ECM) with those of 3D printed polycaprolactone (PCL) scaffolds. In Part A, both chondrogenic and osteogenic ECMs were produced by human adipose derived stem cells (hASCs) on 3D-printed PCL scaffolds and then decellularized to create cell free functionalized PCL scaffolds, named acPCL and aoPCL respectively. The decellularization resulted in a significant reduction of the DNA content as well as the removal of nuclei while the ECM was largely preserved. In Part B the bioactivation and the effect of the ac/aoPCL scaffolds on the proliferation, differentiation, and gene expression of hASCs was investigated. The ac/aoPCL scaffolds were found to be non-toxic and allow good adhesion, but do not affect proliferation. In the in vitro investigation of cartilage regeneration, biochemical analysis showed that acPCL scaffolds have an additional effect on chondrogenic differentiation as gene expression analysis showed markers of cartilage hypertrophy. The aoPCL showed a large influence on the differentiation of hASCs. In control medium they were able to stimulate hASCs to produce calcium alone and all genes relevant investigated for osteogenesis were significantly higher expressed on aoPCL than on unmodified PCL. Therefore, we believe that ac/aoPCL scaffolds have a high potential to improve regenerative capacity of unmodified PCL scaffolds and should be further investigated.

Comparative study of dedifferentiated fat cell and adipose‐derived stromal cell sheets for periodontal tissue regeneration: In vivo and in vitro evidence

“…Several approaches have been adopted to enhance the osteogenic potential of ADSCs. For example, decellularized trabecular bone (Shridhar et al, 2019) and decellularized ECM from osteogenic‐induced ADSCs (Blum et al, 2021) have been reported to enhance the osteogenic differentiation of ADSCs. Synthetic materials such as β‐tricalcium phosphate/collagen‐I fibre scaffolds (Wang et al, 2021), hydrogel scaffolds (Kim et al, 2020), and PCL‐based nanofibrous composite scaffolds (Mansour et al, 2022) have also been employed to stimulate osteogenic differentiation of ADSCs.…”

Modified Histopathological Protocol for Poly-ɛ-Caprolactone Scaffolds Preserving Their Trabecular, Honeycomb-like Structure

“…Despite the considerable developments in imaging techniques allowing for comprehensive in vivo and postmortem scaffold evaluation, and even the creation of 3D virtual models [ 29 , 30 ], little attention has been paid to the refinement and updating of histopathological techniques, which remain the basic tool for assessments of scaffold morphology, biocompatibility, and interactions with the surrounding cells. The standard techniques used today only allow for evaluations of selected representative fragments, where the scaffold is often cleared, melted, or swollen, and it is not possible to obtain the full cross-section view with the preserved scaffold’s honeycomb-like structure [ 31 , 32 ].…”

Evaluation of the Usability of a Low-Cost 3D Printer in a Tissue Engineering Approach for External Ear Reconstruction

“…Subsequently, the self-made PCL-scaffold was compared to a commercially available PCL-scaffold, which was successfully used for chondrogenic and osteogenic tissue engineering in an earlier study [28]. In vitro cultivation with human ASCs revealed comparable cell viability, distribution, and proliferation capacity on both priorly sterilized PCL-scaffolds.…”