Abstract
Purpose: Application of Mummy material for treatment of different diseases such as bone fracture, cutaneous wounds and joint inflammation has been advised since hundred years ago in Persian traditional medicine. Due to the claims of indigenous people and advice of traditional medicine for application of this material in healing of bone fractures, this study has been designed to evaluate whether Mummy material can promote the differentiation of mesenchymal stem cells into osteoblasts and enhance the expression of bone specific genes and proteins.Methods: Adipose derived stem cells (ASCs) at fourth cell passage were divided into control, osteogenesis group (received osteogenic medium), Mummy group (received Mummy at concentration of 500 µg/ml). ASCs in the fourth group were treated with both osteogenic medium and Mummy (500µg/ml). Cells in all groups were harvested on days 7, 14 and 21 days for further evaluation through Real time RT-PCR, Von kossa staining, Immunocytochemistry and flowcytometery.Results: Treatment of ASCs with Mummy at concentration of 500µg/ml promotes the expression level of Osteocalcin, RUNX-2 and β1-integrin genes in different time points but that of the Osterix did not changed. Furthermore the expression of Osteocalcin protein enhanced significantly in ASCs treated with Mummy detected by Immunocytochemistry and flowcytometery technique compared to the control groups. The results of this study also showed that treatment of ASCs with Mummy resulted in formation of mineral deposits which was evaluated by Von Kossa staining method.Conclusion: Obtained data from this study reveals that Mummy is a potent enhancer for differentiation of ASCs into osteoblasts in in vitro system, probably through increasing the level of bone specific genes and proteins.
Highlights
Bone is a specialized type of connective tissue which is composed of two different cells named as osteoblasts and osteoclasts.[1]
Effects of Mummy on gene expression in Adipose derived stem cells (ASCs) differentiated into osteoblasts The effect of Mummy on differentiation of ASCs to osteoblasts were evaluated through expression of osteocalcin, RUNX-2 and osterix genes, β1-integrin gene expression was evaluated to examine whether Mummy can enhance the cell-cell and cell-matrix adhesion which is crucial for cell survival
After 14 days, the expression level of this transcription factor increased significantly in ASCs induced with osteogenic medium and cells co-treated with osteogenic medium and Mummy( 500μg/ml) compared to the control cells (P
Summary
Bone is a specialized type of connective tissue which is composed of two different cells named as osteoblasts and osteoclasts.[1] Coordinated balance between activities of these cells is responsible for maintenance of bone homeostasis and remodeling.[2] Synthesis of organic components of bone matrix is attributed to osteoblasts, while bone resorption occurs through activity of osteoclasts.[3] Disruption of this precisely regulated balance can result in bone related diseases as osteoporosis which is defined as bone mass reduction.[4] The prevalence of bone disorders is increasing, in part because of growing of aging population. On the other hand bone defects can occur due to trauma, surgery, tumor metastasis and metabolic diseases, so finding efficient therapeutic methods for bone defects is of great importance. These cells can be harvested from different sources such as bone marrow, umbilical cord Wharton's jelly, muscular and
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