Journal of

Background and Aim: Bone sialoprotein (BSP) is a specific marker of osteoblastic differentiation. In this research, the effect of Zoledronic Acid on BSP expression and methylation status during osteoblastic differentiation of mesenchymal stem cells (MSCs) was evaluated.

Materials and Methods: In this experimental study, MSCs were isolated from human bone marrow. For osteogenic differentiation, hMSCs were pulse treated with zoledronic acid, and were incubated in osteogenic differentiation medium for 3 weeks. The DNA and RNA were extracted after the first, second and third weeks of culture and also from undifferentiated MSCs. After Sodium bisulfate (SBS) treatment, gene specific methylation analysis for BSP was carried out using Methylation Specific PCR technique.

Results: BSP expression was observed in osteoblastic differentiated cells whereas it was not seen in MSCs. MSP showed that BSP was unmethylated during osteoblastic differentiation.

Conclusion: BSP was expressed from the first week of differentiation. This confirms that zoledronic acid accelerates osteoblastic differentiation. Unmethylation status of BSP indicates that zoledronic acid does not have any effect on BSP methylation status. Other genetic or epigenetic mechanisms may control BSP expression during osteoblastic differentiation induction by zoledronic acid.

Background and Aim: The use of probiotics has been surveyed in different studies. For example, many researches have been conducted on immunity, cancer and embryonic cells proliferation. Mesenchymal stem cells (MSCs) have a rather slower growth and a more limited number of passages. In this research, the effect of supernatant of Lactobacillus acidophilus (LS) as one of the most commonly used probiotics on the proliferation of mesenchymal stem cells has been studied with the aim of increasing their proliferation for the treatment of patients in need of transplantation using mesenchymal stem cells.

Materials and Methods: The MSCs separated from rats’ bone marrow were led to bone and fat levels in the second passage, and their mesenchymal state was confirmed. The cells were then treated by the supernatant of Lactobacillus acidophilus (LS) and MRS broth medium, which have been separated in advance. The number of cells was examined by MTT test equated with the standard.

Results: A curve was drawn for the cells’ growth in different amounts and the number of cells was obtained by converting the results of MTT test to a standard curve. For statistical analyses, SPSS and ANOVA were employed.

Conclusion: The findings show the double effect of LS on the weak effect of MRS broth medium. The use of supernatant of Lactobacillus acidophilus (LS) can be a practical and economical method for increasing the proliferation of MSCs isolated from bone marrow.