Abstract
Skeletal reconstruction is necessary in cases of bone defects created by tumors, trauma, and abnormalities. Regeneration of bone defects remains a critical problem, and current approaches are based on biocompatible scaffolds. Spheroids represent a simple 3D system since no supporting material is required for cell growth. Different techniques are used to generate spheroids, such as hanging drop, low-attachment plates, and magnetic nanoparticles. The idea of using magnetic nanoparticles is to cross-link through cell membrane overnight to create complex 3D cellular spheroid by using magnets to guide the cellular response. Herein, the current study aimed to achieve 3D human fetal osteoblast (hFOB) spheroid under magnetic levitation. Formation of 3D spheroid culture under magnetic levitation was evaluated by cell viability at 3, 7, and 14 days. Morphology of the 3D hFOB spheroid was analyzed by SEM and fluorescence microscopy and the differentiation towards mineralized lineage by ALP assay, qPCR, and alizarin red staining. The cell viability indicated that the 3D hFOB spheroid still viable after 14 days of culture. ALP assay, qPCR analysis expression of Col1, ALP, and Itg-β1 molecules, and calcium deposition with alizarin red showed a high level of bioactivity of the 3D hFOB spheroid. SEM images allowed the morphological analysis of the 3D microtissue-like spheroid with the presence of matrix deposition. These results indicate that magnetic levitation culture enables 3D stable osteoblast spheroids and could be a promising strategy for engineering application in the 3D construct in surgery regeneration of mineralized tissue.
Highlights
Bone defects resulting from trauma, disease, surgery, or congenital malformations are a significant health problem worldwide
This study presents a set of experiments to propose a consolidated and straightforward methodology based on magnetic levitation to develop a threedimensional (3D) spheroid from human fetal osteoblasts culture
The present results indicate that 3D human fetal osteoblast (hFOB) spheroids developed by magnetic levitation systems can be evaluated for biocompatibility and show that long-term culture is feasible
Summary
Bone defects resulting from trauma, disease, surgery, or congenital malformations are a significant health problem worldwide. The problems associated with transplanted grafts have raised interest in bone tissue engineering in searching and improving the design of three-dimensional (3D) cellular constructs This growing interest in culturing adherent cells for developing a 3D construct is one of the most important issues for tissue engineering. It opens a new research area in 3D cell culture techniques because there is a Traditionally, 2D cell culture systems are a standard method for proliferating cells in which the cells grow as a monolayer and do not fully reflect tissues’ physiology due to some limitations in mimic in vivo multicellular conditions. Spheroids cultures represent a simple 3D system, where no scaffold or supporting material is required for 3D cell growth and has significant advantages as facilitate cell-cell and cell-matrix interaction networks, provide a similar physicochemical environment, secrete cytokines, chemokines, and angiogenic factors and maintain intrinsic phenotypic properties identical to the in vivo, and tissue morphology and function is different from that of cells in the monolayer culture system (Griffith and Swartz, 2006; Fennema et al, 2013)
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