Abstract
The growth of bone morphogenetic protein 7 (BMP-7) has been applied for tissue regeneration due to its osteoinductive properties. The aim of this research is to analyze the enhancing effects of BMP-7 on the osteogenic differentiation and mineralization of human bone marrow-derived stem cells cultured on the bovine bone particle. After the stem cells were loaded onto the bone graft material, their morphology was observed on day 7. Viability assays based on the application of fluorescent stains were used for qualitative analyses. Alkaline phosphatase activity assays and Alizarin red staining were used for the assessment of osteogenic differentiation on days 7 and 14. Next-generation mRNA sequencing was applied to evaluate global gene expression. Gene ontology and pathway analysis was used to propose the underlying mechanism. Fibroblast-like morphology was attained with the stem cells. The cells were shown to be firmly attached to the bone particle. Most of the stem cells produced an intense green fluorescence. The relative cellular viability assay values for BMP-7 groups at 0, 10, and 100 ng/mL on day 7 were 0.295 ± 0.003, 0.250 ± 0.002, and 0.240 ± 0.003, respectively (p < 0.05). Alkaline phosphatase activity was significantly higher in BMP-7 groups at concentration of 100 ng/mL compared to the control on days 7 and 14 (p < 0.05). The results of the mineralization assay showed significantly higher values for BMP-7 groups at 100 ng/mL concentration when compared with the control (p < 0.05). The expression of RUNX2 was increased with application of BMP-7 and mitogen-activated protein kinase pathway was associated with the target genes. Overall, this study shows that in vitro application of BMP-7 increases alkaline phosphorylase activity and mineralization of stem cells culture on deproteinized bovine bone mineral. The study suggests that combining stem cells with osteoinductive growth factors with scaffolds can have synergy effects on osteogenic differentiation.
Highlights
The purpose of this study is to evaluate the bone regeneration ability through the simultaneous application of bone morphogenetic protein 7 (BMP-7) and stem cells cultured on deproteinized bovine bone
The number of cells presented on deproteinized bovine bone mineral particles seemed to decrease with addition of bone morphogenetic proteins (BMPs)-7 (Figure 1)
50 and 100 ng/mL of BMP-7 induced proliferation of stem cells derived from periodontal ligament, contrary to this study [25] and this may be due to the differences in culture condition, passage and origin of the stem cells [26]
Summary
In the first half of the 21st century, the number of people over 65 years of age dramatically increased [1]. There might be a demographic shift within the elderly population in the coming decades in which disease incidences and mortality increase [2]. Most adults suffer from periodontal disease, which is the second most-seen oral disease along with dental caries [3]. Periodontal disease is a major cause of tooth loss in the elderly population [4]. The economic burden and quality of life for the elderly tend to drop significantly due to the need for immediate and continuous treatment of
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