Eyvazi Maryam, Farahzadi Raheleh, Karimian Fathi Nahid, Karimipour Mohammad, Soleimani Rad Jafar, Montaseri Azadeh
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2018 Aug;8(3):457-464. doi: 10.15171/apb.2018.053. Epub 2018 Aug 29.
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. 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. 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. 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.
早在百年前,波斯传统医学就已建议将木乃伊材料用于治疗诸如骨折、皮肤创伤和关节炎症等不同疾病。鉴于当地居民的说法以及传统医学关于该材料在骨折愈合中应用的建议,本研究旨在评估木乃伊材料是否能够促进间充质干细胞向成骨细胞分化,并增强骨特异性基因和蛋白质的表达。将处于第四代细胞传代的脂肪来源干细胞(ASC)分为对照组、成骨组(接受成骨培养基)、木乃伊组(接受浓度为500μg/ml的木乃伊材料)。第四组的ASC同时用成骨培养基和木乃伊材料(500μg/ml)处理。在第7天、14天和21天收集所有组的细胞,通过实时逆转录聚合酶链反应、冯·科萨染色、免疫细胞化学和流式细胞术进行进一步评估。用浓度为500μg/ml的木乃伊材料处理ASC可在不同时间点促进骨钙素、RUNX-2和β1整合素基因的表达水平,但osterix的表达未发生变化。此外,与对照组相比,通过免疫细胞化学和流式细胞术检测发现,用木乃伊材料处理的ASC中骨钙素蛋白的表达显著增强。本研究结果还表明,用木乃伊材料处理ASC会导致矿化沉积的形成,这通过冯·科萨染色法进行评估。本研究获得的数据表明,在体外系统中,木乃伊材料可能通过提高骨特异性基因和蛋白质的水平,成为ASC向成骨细胞分化的有效增强剂。