Study of the effect of phytoextract based on rice husk and germ film, green tea on the functional properties of cells involved in bone tissue metabolism

N. Sitnikova, N. Bondarenko, A. O. Kushnarenko
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Abstract

Osteoporosis is a disease associated with a decrease of bone density and quality. Bone tissue is especially sensitive to changes in trace element content in the body. Deficiency of macro- and microelements (silicon, calcium, manganese, etc.) can lead to the development of osteoporosis. The aim of the research was to study the effect of NaBiKat phytoextract on the viability and functional activity of fibroblasts, mesenchymal stem cells (MSCs), osteoblasts and osteoclasts, involved in bone metabolism in osteoporosis. Material and methods. An aqueous solution of the NaBiKat phytocomplex consisting of rice husks and germ film, green tea was used in the work; its content of basic chemical elements was measured using atomic immersion spectral analysis. The cytotoxicity of the phytoextract was evaluated on fibroblasts, MSCs, and osteoblasts of humans using the MTT test. The activity of mitochondria in cells was assessed by staining with a fluorescent dye TMRM. The production of intracellular proteins in osteoblasts and osteoclasts was determined using flow cytometry. Results and discussion. In an aqueous solution of phytoextract, a large amount of sodium, potassium, silicon, manganese, magnesium and a small amount of calcium, iron are found. The phytoextract did not have a cytotoxic effect on fibroblasts and stimulated proliferation of MSCs and osteoblasts up to 112–128 % and up to 130 %, respectively. At maximum concentration, the phytocomplex reduced osteoblast proliferation. Phytocomplex enhanced mitochondria activity in MSCs and osteoblasts, reduced in fibroblasts and osteoclasts. Under its influence, an increase in RANKL synthesis was observed in osteoblasts, the amount of type 1 collagen did not change, and cathepsin K production decreased in osteoclasts, while it did not significantly affect MMP-9 content. Conclusions. The results obtained indicate that an aqueous phytoextract based on rice husk and germ film, green tea leaves is safe for MSCs, fibroblasts, osteoblasts, and osteoclasts in humans and is able to stimulate the functional activity of osteoblasts, as well as reduce the activity of osteoclasts due to the high content of macro- and microelements.
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基于稻壳和胚芽膜、绿茶的植物提取物对参与骨组织代谢的细胞功能特性的影响研究
骨质疏松症是一种与骨密度和骨质量下降有关的疾病。骨组织对体内微量元素含量的变化特别敏感。缺乏宏量和微量元素(硅、钙、锰等)会导致骨质疏松症的发生。本研究旨在研究 NaBiKat 植物提取物对参与骨质疏松症骨代谢的成纤维细胞、间充质干细胞(MSCs)、成骨细胞和破骨细胞的活力和功能活性的影响。材料和方法工作中使用了由稻壳和胚芽膜、绿茶组成的 NaBiKat 植物复合物水溶液;使用原子浸透光谱分析法测定了其中基本化学元素的含量。使用 MTT 试验评估了植物提取物对成纤维细胞、间充质干细胞和成骨细胞的细胞毒性。用荧光染料 TMRM 染色评估了细胞中线粒体的活性。使用流式细胞仪测定成骨细胞和破骨细胞中细胞内蛋白质的生成。结果与讨论植物提取物的水溶液中含有大量的钠、钾、硅、锰、镁和少量的钙、铁。植物提取物对成纤维细胞没有细胞毒性作用,对间叶干细胞和成骨细胞的增殖刺激分别高达 112-128 % 和 130 %。在最高浓度下,植物复合物会减少成骨细胞的增殖。植物复合体增强了间充质干细胞和成骨细胞的线粒体活性,降低了成纤维细胞和破骨细胞的线粒体活性。在它的影响下,成骨细胞中 RANKL 的合成增加,1 型胶原蛋白的数量没有变化,破骨细胞中 cathepsin K 的产生减少,而它对 MMP-9 的含量没有显著影响。结论研究结果表明,基于稻壳和胚芽膜、绿茶叶的水性植物提取物对人体间充质干细胞、成纤维细胞、成骨细胞和破骨细胞是安全的,并且由于含有大量的宏量和微量元素,能够刺激成骨细胞的功能活性,同时降低破骨细胞的活性。
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