Department of Cellular and Molecular Biology, Faculty of Biology, Science and Research Branch of Islamic Azad University, Tehran, Iran.
Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.
Mol Biol Rep. 2023 Dec;50(12):10037-10045. doi: 10.1007/s11033-023-08877-5. Epub 2023 Oct 30.
Bioactive polysaccharides are a promising way for bone disease prevention with high efficiency. Schizophyllan (SPG) is a polysaccharide derived from a species of fungus with anticancer, antitumor, and anti-inflammatory effects. In the present study, for the first time, the cell proliferation, osteogenic markers, mineral deposition, and osteogenic gene expression of human adipose tissue-derived mesenchymal stem cells (hADMSCs) grown on SPG were evaluated by in vitro assays.
The cytotoxicity of SPG was measured using the MTT assay and acridine orange staining. Differentiation of hADMSCs was assessed using alkaline phosphatase (ALP) activity test, cellular calcium content assay, and mineralized matrix staining. To this end, Alizarin red S, von Kossa staining, and the expression of bone-specific markers, including ALP, Runx2, and osteonectin, were used by real-time RT-PCR over a 2-week period. According to the results, SPG at 10 µg/ml concentration was determined as the optimal dosage for differentiation studies. The results of osteogenic differentiation tests showed that compared to the control groups in vitro, SPG enhanced the osteogenic markers and mineralization as well as upregulation of the expression of bone specific genes in differentiated hADMSCs during differentiation.
The results revealed that SPG could be applied as effective factor for osteogenic differentiation in the future. The current study provides insights into the hADMSC-based treatment and introduces promising therapeutic material for individuals who suffer from bone defects and injuries.
具有高效性的生物活性多糖是预防骨疾病的一种有前途的方法。裂褶菌多糖(SPG)是一种从真菌中提取的多糖,具有抗癌、抗肿瘤和抗炎作用。在本研究中,首次通过体外试验评估了生长在 SPG 上的人脂肪组织来源间充质干细胞(hADMSCs)的细胞增殖、成骨标志物、矿物质沉积和成骨基因表达。
通过 MTT 测定法和吖啶橙染色测定 SPG 的细胞毒性。通过碱性磷酸酶(ALP)活性试验、细胞钙含量测定和矿化基质染色评估 hADMSCs 的分化。为此,在 2 周的时间内通过实时 RT-PCR 检测了骨特异性标志物,包括 ALP、Runx2 和骨粘连蛋白的表达。结果表明,在 10μg/ml 浓度下,SPG 被确定为分化研究的最佳剂量。成骨分化试验的结果表明,与体外对照组相比,SPG 增强了成骨标志物和矿物质沉积,并上调了分化后的 hADMSCs 中骨特异性基因的表达。
结果表明,SPG 将来可作为成骨分化的有效因子。本研究为基于 hADMSC 的治疗提供了新的见解,并为患有骨缺损和损伤的个体引入了有前途的治疗材料。