Abstract
BackgroundBiomaterials are widely used to regenerate or substitute bone tissue. In order to evaluate their potential use for clinical applications, these need to be tested and evaluated in vitro with cell culture models. Frequently, immortalized osteoblastic cell lines are used in these studies. However, their uncontrolled proliferation rate, phenotypic changes or aberrations in mitotic processes limits their use in long-term investigations. Recently, we described a new pluripotent-like subpopulation of dental pulp stem cells derived from the third molars (DPPSC) that shows genetic stability and shares some pluripotent characteristics with embryonic stem cells. In this study we aim to describe the use of DPPSC to test biomaterials, since we believe that the biomaterial cues will be more critical in order to enhance the differentiation of pluripotent stem cells.MethodsThe capacity of DPPSC to differentiate into osteogenic lineage was compared with human sarcoma osteogenic cell line (SAOS-2). Collagen and titanium were used to assess the cell behavior in commonly used biomaterials. The analyses were performed by flow cytometry, alkaline phosphatase and mineralization stains, RT-PCR, immunohistochemistry, scanning electron microscopy, Western blot and enzymatic activity. Moreover, the genetic stability was evaluated and compared before and after differentiation by short-comparative genomic hybridization (sCGH).ResultsDPPSC showed excellent differentiation into osteogenic lineages expressing bone-related markers similar to SAOS-2. When cells were cultured on biomaterials, DPPSC showed higher initial adhesion levels. Nevertheless, their osteogenic differentiation showed similar trend among both cell types. Interestingly, only DPPSC maintained a normal chromosomal dosage before and after differentiation on 2D monolayer and on biomaterials.ConclusionsTaken together, these results promote the use of DPPSC as a new pluripotent-like cell model to evaluate the biocompatibility and the differentiation capacity of biomaterials used in bone regeneration.
Highlights
Biomaterials are widely used to regenerate or substitute bone tissue
Characterization of dental pulp pluripotent-like stem cells dental pulp pluripotentlike stem cells (DPPSC) were isolated as a new subpopulation of dental pulp stem cells (DPSC) with pluripotent-like characteristics
In order to summarize the genetic pattern during the osteogenic differentiation of DPPSC, a schematic representation with the different stages was performed (Fig. 2d)
Summary
Biomaterials are widely used to regenerate or substitute bone tissue. In order to evaluate their potential use for clinical applications, these need to be tested and evaluated in vitro with cell culture models. For bone related biomaterials, most studies have examined their osteogenic potential using immature osteoblasts, immortalized cell lines or mesenchymal stem cells among others Primary cells, such as lineage-specific osteoblasts, can be isolated and cultivated relatively ; they have a limited lifespan [3]. Immortalized cell lines, such as the human sarcoma osteogenic cell line (SAOS-2) have been frequently used in applied biology since they are from human origin while providing unlimited number of cells [2, 4] These cell lines, due to their cancer origin, usually possess phenotype changes between passages, aberrations in mitotic processes and lack of growth inhibition, which limits their use in long-term investigations [5]. There is a need to find a cell type with genetic stability and stemness characteristics to be used to evaluate biomaterials in cell therapy applications
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