Abstract
A three-dimensional (3D) culture system that closely replicates the in vivo microenvironment of calcifying osteoid is essential for in vitro cultivation of bone-like material. In this regard, the 3D cellulose constructs of plants may well serve as scaffolds to promote growth and differentiation of osteoblasts in culture. Our aim in this study was to generate bone-like tissue by seeding pluripotent stem cells (hiPSCs), stimulated to differentiate as osteoblasts in culture, onto the decellularised scaffolds of various plants. We then assessed expression levels of pertinent cellular markers and degrees of calcium-specific staining to gauge technical success. Apple scaffolding bearing regular pores of 300 μm seemed to provide the best construct. The bone-like tissue thus generated was implantable in a rat calvarial defect model where if helped form calcified tissue. Depending on the regularity and sizing of scaffold pores, this approach readily facilitates production of mineralized bone.
Highlights
It is well established that three-dimensional (3D), rather than two-dimensional (2D), culture offers a microenvironment closer to in vivo conditions, better enabling in vitro development of organoids
Once seeded with hiPSCs, only cells cultivated in apple scaffolding survived, as confirmed by Cell Counting Kit-8 (CCK-8) assay after 96 h (Fig. 2C) and by scanning electron microscopy (Fig. 2D)
Viable hiPSCs were confirmed within apple scaffolding under phase contrast microscopy and in haematoxylin and eosin (H&E)-stained histological preparations (Fig. 2E)
Summary
It is well established that three-dimensional (3D), rather than two-dimensional (2D), culture offers a microenvironment closer to in vivo conditions, better enabling in vitro development of organoids. The range of inherent properties is an important aspect of any scaffolding biomaterial, which should be biocompatible, manipulated, and structurally sound, providing proper mechanical support and bioactivity. To this end, various natural or synthetic materials have been utilised as biomaterials in scaffold development. A cellulose-based scaffold has been applied for culturing and osteoblastic differentiation of human mesenchymal stem cells[11]. These successes prompted our use of this natural, readily available, and handled scaffolding in the 3D engineering of bone. We explored the prospective clinical merits, using resultant bone organoid as in vivo implants
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