Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Mol Biol Rep. 2020 May;47(5):3663-3674. doi: 10.1007/s11033-020-05461-z. Epub 2020 Apr 25.
Bone regeneration is a significant and crucial health issue worldwide. Tissue bioengineering has shown itself to be the best substitute for common clinical treatment of bone loss. The suitable cell source is human endometrial stem cells (hEnSCs) which have several suitable characteristics for this approach. Since sex steroid hormones are involved in expansion and conservation of the skeleton, the effect of two sex steroid hormones known as estrogen (17-β estradiol) and progesterone on osteogenic differentiation of hEnSCs were examined. For this purpose, hEnSCs were treated with 17-β estradiol and progesterone separately (1 × 10 M) and simultaneously (1 × 10 M). Osteogenic differentiation tests including measurement of total mineral calcium content, Alizarin Red staining, the quantitative expression levels of some osteogenic markers by Real-time RT-PCR, and immunofluorescence staining were performed at 7 and 14 days of differentiation. To exhibit the morphology of the cells in osteogenic and culture medium, the hEnSCs were stained with Acridine Orange (AO) solution. In this research, MTT assay and AO staining revealed progesterone and 17-β estradiol increase the proliferation of hEnSCs in a dose-dependent manner. Furthermore, the results of calcium content analysis, Real-time RT-PCR assay, and all tests of differentiation staining have shown that 17-β estradiol and progesterone cannot induce hEnSCs' osteogenic differentiation. In conclusion, it is indicated that 17-β estradiol and progesterone do not have positive effects on hEnSCs' osteogenic differentiation in vitro.
骨再生是全球范围内一个重要且关键的健康问题。组织生物工程已被证明是治疗骨丢失的常用临床方法的最佳替代方法。合适的细胞来源是人类子宫内膜干细胞（hEnSCs），其具有几个适合这种方法的特性。由于性激素参与骨骼的扩张和维持，因此研究了两种已知的性激素雌激素（17-β雌二醇）和孕激素对 hEnSCs 成骨分化的影响。为此，分别用 17-β雌二醇和孕激素（1×10 -5 M）以及同时用这两种激素（1×10 -5 M）处理 hEnSCs。在分化的第 7 天和第 14 天进行包括总钙含量测定、茜素红染色、实时 RT-PCR 测定一些成骨标志物的定量表达水平以及免疫荧光染色的成骨分化试验。为了展示成骨和培养基中细胞的形态，用吖啶橙（AO）溶液对 hEnSCs 进行染色。在这项研究中，MTT 分析和 AO 染色显示孕激素和 17-β雌二醇以剂量依赖的方式增加 hEnSCs 的增殖。此外，钙含量分析、实时 RT-PCR 分析以及所有分化染色试验的结果表明，17-β雌二醇和孕激素不能诱导 hEnSCs 的成骨分化。总之，结果表明 17-β雌二醇和孕激素对 hEnSCs 的体外成骨分化没有积极作用。
