Zaminy Arash, Ragerdi Kashani Iraj, Barbarestani Mohammad, Hedayatpour Azim, Mahmoudi Reza, Farzaneh Nejad Ahmadreza
Department of Anatomy, School of Medicine, Medical Sciences/University of Tehran, Tehran, Iran.
Dept. of Anatomy, School of Medicine, Yasouj University of Medical Sciences, Yasouj, Iran.
Iran Biomed J. 2008 Jul;12(3):133-41.
Adipose-derived stem cells (ADSC) could be an appealing alternative to bone marrow stem cells (BMSC) for engineering cell-based osteoinductive grafts. Meanwhile, prior studies have demonstrated that melatonin can stimulate osteogenic differentiation. Therefore, we assayed and compared the melatonin effect on osteogenic differentiation of BMSC with that of ADSC.
Mesenchymal stem cells (MSC) were isolated from the bone marrow and fat of adult rats. Both cell types were cultured in osteogenic medium in the absence and presence of melatonin at physiological concentrations (20-200 pg/ml). After 4 weeks, the expression of osteocalcin gene was analyzed by reverse transcription-PCR, alkaline phosphatase (ALP) activity was assayed and alizarin red S and von Kossa staining were done. Cell viability and apoptosis were also assayed by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, a tetrazole (MTT) and flow cytometry, respectively.
The osteoblastic differentiation of ADSC as demonstrated by ALP activity was less than that of BMSC. The amount of matrix mineralization has shown by alizarin red S and von Kossa staining also showed statistical differences between the two MSC. The incidence of apoptotic cells was higher among ADSC than BMSC. The flow cytometry proves that cell growth reduction is due to a decrease in the number of the cells entering the S phase of the cell cycle. MTT assay indicated that viable cells were fewer among ADSC than BMSC in control groups.
The results of the study suggest that BMSC have greater osteogenic potential than ADSC and that melatonin promotes osteogenic differentiation to BMSC, but has a negative effect on ADSC osteogenic differentiation.
脂肪来源干细胞（ADSC）可能是骨髓干细胞（BMSC）用于构建基于细胞的骨诱导移植物的一种有吸引力的替代选择。同时，先前的研究表明褪黑素可以刺激成骨分化。因此，我们测定并比较了褪黑素对BMSC和成骨分化的影响。
从成年大鼠的骨髓和脂肪中分离间充质干细胞（MSC）。两种细胞类型均在无和有生理浓度（20 - 200 pg/ml）褪黑素的成骨培养基中培养。4周后，通过逆转录 - PCR分析骨钙素基因的表达，测定碱性磷酸酶（ALP）活性，并进行茜素红S和冯·科萨染色。还分别通过3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐（一种四氮唑，MTT）和流式细胞术测定细胞活力和凋亡。
ALP活性表明ADSC的成骨细胞分化低于BMSC。茜素红S和冯·科萨染色显示的基质矿化量在两种MSC之间也存在统计学差异。ADSC中凋亡细胞的发生率高于BMSC。流式细胞术证明细胞生长减少是由于进入细胞周期S期的细胞数量减少。MTT分析表明，对照组中ADSC中的活细胞比BMSC中的少。
研究结果表明，BMSC比ADSC具有更大的成骨潜力，褪黑素促进BMSC的成骨分化，但对ADSC的成骨分化有负面影响。
