188 The Effect of Vitamin B12 on the Differentiation of Adipose-Derived Stem Cells into Osteoblasts
T. A. Bane A , J. C. Bertels A , K. M. Polkoff A , M. Rubessa A B and M. B. Wheeler A BA Department of Animal Sciences, University of Illinois, Urbana-Champaign, IL, USA;
B Carl R. Woese Institute of Genomic Biology, University of Illinois, Urbana-Champaign, IL, USA
Reproduction, Fertility and Development 30(1) 234-234 https://doi.org/10.1071/RDv30n1Ab188
Published: 4 December 2017
Abstract
Large bone defects present a tremendous challenge to the treating surgeon. Tissue engineering using scaffolds of various sizes and shapes that contain stem cells and other osteoinductive molecules offer a potential solution to this difficult problem. The aim of this project was to evaluate if the osteogenic medium infused with vitamin B12 influences the differentiation of adipose-derived stem cells (ASC) into osteoblasts. Vitamin B12 has been shown to have a stimulatory effect on osteoclastogenesis in vitro (Vaes et al. 2009 Calcified Tissue Int. 84, 413-422). Our hypothesis was that the presence of vitamin B12 in the osteogenic medium would positively influence the number of osteoblastic nodules formed. Swine ASC were isolated as described (Monaco et al. 2009 Open Tissue Eng. Regen. Med. J. 2, 20-33). The ASC were divided in 8 different treatments: 8 concentrations of vitamin B12 in the osteogenic medium (0.1, 0.2, 1, 2, 10, and 20 μM) plus 2 control treatments (osteogenic medium without vitamin B12 and a negative control, DMEM). The medium was changed twice a week for 4 weeks. The experiment was replicated 6 times. At the end of the culture period, cells were stained with Alizarin Red and Von Kossa stains. In each well, we counted the nodules and then divided them in 2 categories: formed and forming nodules. Data was analysed using the generalized linear model (GLM) procedure in SPSS (IBM/SPSS, Armonk, NY, USA). Bonferroni’s post hoc test was used to perform statistical multiple comparison. The α-level was set at 0.01. The results showed that the concentration of 20 μM vitamin B12 was detrimental for nodule formation. Table 1 illustrates the number of formed and forming nodules in addition to their standard deviation. There was no positive effect on nodule formation when different concentrations of vitamin B12 were added to the osteogenic medium. More experiments need to be conducted to determine if vitamin B12 can act synergistically with other vitamins to produce a significant role in ASC differentiation into osteoblasts. This preliminary experiment is the first step towards the analysis of the behaviour of ASC on scaffolds with vitamin B12 incorporated into their matrix.
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