Mechanosensitive Osteogenesis on Native Cellulose Scaffolds for Bone Tissue Engineering

Abstract

Accurate physical representation of the tissue microenvironment is essential for implant development. In this study, we applied cyclic hydrostatic pressure (HP) to mimic the effect of cyclic hydrostatic pressure (HP) in a bone-like environment, using a custom-made, remote controlled bioreactor. In recent years, plant-derived cellulosic biomaterials have become a popular way to create scaffolds for a variety of tissue engineering applications. Moreover, such scaffolds possess similar physical properties (porosity, stiffness) that resemble bone tissues and have been explored as potential biomaterials for tissue engineering applications. Here, plant-derived cellulose scaffolds (derived from apple hypanthium tissue) were seeded with MC3T3-E1 pre-osteoblast cells. After 1 week of proliferation, cell-seeded scaffolds were exposed to HP up to 270 KPa at a frequency of 1Hz, once per day, for up to 2 weeks. Scaffolds were incubated in osteogenic inducing media or regular culture media. The effect of cyclic hydrostatic pressure combined with osteogenic inducing media on cell-seeded scaffolds resulted in an increase of differentiated cells. This corresponded with an upregulation of alkaline phosphatase activity and scaffold mineralization. The results reveal that in vitro, the mechanosensitive pathways which regulate osteogenesis appear to be functional on novel plant-derived cellulosic biomaterials.

Wood-Derived Vascular Patches Loaded With Rapamycin Inhibit Neointimal Hyperplasia

“…Recently, plant-derived scaffolds have been used in cell culture and show promising applications ( Contessi Negrini et al, 2020 ; Gershlak et al, 2017 ) and continue to attract attention ( Latour and Pelling 2022 ; Ianovici et al, 2022 ). Plant-based scaffolds can be used as a vascular patch and a drug delivery system ( Xie et al, 2021 ; Bai et al, 2021a ).…”

Translating Material Science into Bone Regenerative Medicine Applications: State-of-The Art Methods and Protocols

“…Different cell lines also find an approved use in the field: the most used is the mouse calvarial preosteoblast MC3T3-E1 cell line [ 52 , 54 , 62 , 63 , 71 , 78 , 80 , 82 , 85 , 86 , 87 , 90 , 114 ]. Other reported cell lines are: human osteosarcoma SaOS-2 cells [ 56 , 64 , 88 ], the mouse osteoblastic KUSA-A1 cell line [ 84 ], the human osteoblast NHOst-Osteoblasts OGM cell line [ 66 ], and the human osteosarcoma MG63 osteoblast-like cells [ 58 , 95 , 97 ].…”

“…The analysis of alkaline phosphatase (ALPL) activity is one of the most common tests used to evaluate the osteogenic process and numerous easy-to-use and highly sensitive assays to measure ALPL activity are available on the market [ 14 , 53 , 54 , 55 , 58 , 61 , 65 , 66 , 67 , 68 , 70 , 71 , 73 , 76 , 79 , 80 , 83 , 85 , 86 , 88 , 90 , 91 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 107 , 114 , 115 ].…”

“…During osteogenic differentiation, calcium deposition can be measured by Alizarin Red S staining [ 14 , 53 , 54 , 55 , 57 , 61 , 64 , 65 , 67 , 68 , 69 , 71 , 72 , 76 , 83 , 84 , 85 , 86 , 87 , 90 , 91 , 93 , 94 , 96 , 99 , 114 ]. The direct visualization and analysis of the Alizarin Red stained deposits provides a qualitative measure of the mineralized matrix deposition, but it is possible only on transparent supports and not on thick opaque materials like ceramics and graphene composites.…”

“…The direct visualization and analysis of the Alizarin Red stained deposits provides a qualitative measure of the mineralized matrix deposition, but it is possible only on transparent supports and not on thick opaque materials like ceramics and graphene composites. However, it is possible to quantify the amount of staining and compare different conditions by dissolving the Alizarin Red from the stained monolayer and obtaining a measure of the absorbance of each sample [ 14 , 53 , 57 , 61 , 64 , 65 , 72 , 86 , 91 , 96 , 114 , 116 ].…”