Effect of Boron on Osteogenic Differentiation of Human Bone Marrow Stromal Cells

Abstract

Bone marrow stromal cells (BMSCs) have been well established as an ideal source of cell-based therapy for bone tissue engineering applications. Boron (B) is a notable trace element in humans; so far, the effects of boron on the osteogenic differentiation of BMSCs have not been reported. The aim of this study was to evaluate the effects of boron (0, 1, 10,100, and 1,000 ng/ml) on osteogenic differentiation of human BMSCs. In this study, BMSCs proliferation was analyzed by cell counting kit-8 (CCK8) assay, and cell osteogenic differentiation was evaluated by alkaline phosphatase (ALP) activity assay, Von Kossa staining, and real-time PCR. The results indicated that the proliferation of BMSCs was no different from the control group when added with B at the concentration of 1, 10, and 100 ng/ml respectively (P > 0.05); in contrast, 1,000 ng/ml B inhibited the proliferation of BMSCs at days 4, 7, and 14 (P < 0.05). By ALP staining, we discovered that BMSCs treated with 10 and 100 ng/ml B presented a higher ALP activity compared with control (P < 0.05). By real-time PCR, we detected the messenger RNA expression of ALP, osteocalcin, collagen type I, and bone morphogenetic proteins 7 were also increased in 10 and 100 ng/ml B treatment groups (P < 0.05). The calcium depositions were increased in 1 and 10 ng/ml B treatment groups (P < 0.05). Taken all together, it was the first time to report that B could increase osteogenic effect by stimulating osteogenic differentiation-related marker gene synthesis during the proliferation and differentiation phase in human BMSCs and could be a promising approach for enhancing osteogenic capacity of cell-based construction in bone tissue engineering.

Boron containing poly-(lactide-co-glycolide) (PLGA) scaffolds for bone tissue engineering

“…Several in vivo studies have proved that boron takes an important place in calcium and bone metabolism [36]. We and two other groups have shown that boric acid and its salts have increased mineralization, bone related protein expression, and osteogenic differentiation of various precursor cells [24][25][26]. In line with these studies, the results revealed that despite there were no detectable structural differences due to NaB addition, boron incorporation within PLGA scaffolds significantly increased the number of attached stem cells and calcium mineralization levels.…”

“…Collagen type I is an early marker of bone healing and is strictly necessary for osteoblast maturation, osteocalcin is a late marker of regeneration that is involved in the mineralization process and VEGF is the main marker of neo-vascularization [41]. Previous in vitro studies have proved that boron treatment has increased odontogenesis/ osteogenesis-related gene expression levels including collagen type I, osteonectin, osteocalcin and alkaline phosphatase in progenitor and stem cells [24][25][26]. In consistence with these reports and a study claiming high VEGF expression trough TNF-α signaling pathway after boron administration [42], we found that boron containing scaffold treatment resulted in high prevalence and density of VEGF along with osteocalcin and collagen type I compared to control groups.…”

“…Three independent studies have demonstrated the possible molecular mechanisms of boron on bone mineralization and growth. Boron has significantly increased the level of mineralization and bone associated protein expression in osteoblasts [24], bone marrow stromal cells [25] and dental stem cells [26]. Therefore, application of boron compounds in non-toxic doses for bone tissue engineering to provide calcium mineralization would be of very fundamental importance for clinical applications.…”

Enhanced hydrophilicity and in vitro bioactivity of porous TiO2 film through the incorporation of boron

“…They found that B could be a promising element to induce osteogenesis by regulating mRNA expression of extracellular matrix proteins (COL I, OPN, BSP and OCN) and protein expression related to bone morphogenesis (BMP-4,-6 and -7). The effects of B on the osteogenetic differentiation of human bone marrow stromal cells (BMSCs) were investigated by Ying et al [19]. They reported that B could increase osteogenic effect by stimulating osteogenic differentiationrelated marker gene synthesis (ALP, COL I, OCN and BMP-7).…”

Dose-dependent Effect of Boric Acid on Myogenic Differentiation of Human Adipose-derived Stem Cells (hADSCs)

“…Along with understanding molecular pathway, emergence of this unique era allows scientist to develop new drugs and supplements for tissue regeneration and fetal development rapidly and effectively. In this line, the most prominent and well-observed effect of boron on bone and teeth growth has been elucidated using in vitro approaches in which adult progenitor and mesenchymal stem cells has been differentiated into osteogenic cell lineage in the presence of boron [11][12][13]. Discovering positive contributions of such additives in in vitro would direct scientist to develop biocompatible and bioinductive scaffold system to be used in regenerative medicine.…”