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首页> 外文期刊>Biopreservation and biobanking >A short-term preservation of human cultured periosteal sheets, osteogenic grafting materials, using a commercial preservation solution containing epigallocatechin-3-gallate (Theliokeep~?) under hypothermic conditions
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A short-term preservation of human cultured periosteal sheets, osteogenic grafting materials, using a commercial preservation solution containing epigallocatechin-3-gallate (Theliokeep~?) under hypothermic conditions

机译：在低温条件下，使用含有表没食子儿茶素-3-没食子酸酯（Theliokeep〜？）的商业保存溶液，对人类培养的骨膜片，成骨移植材料进行短期保存

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In the past decade, it has increasingly been reported that epigallocatechin-3-gallate (EGCG), a major catechin derivative extracted from Green tea, has various bioactivities, including a cell-protective action on mammalian cells and tissues. In this study, we have tested a commercial preservation solution containing EGCG (Theliokeep~?) in both two- and three-dimensional cultures of human periosteal sheets, which have been used as an osteogenic grafting material for periodontal regenerative therapy. When periosteal sheets were 3D-cultured on collagen mesh, cell viability was maintained for 2 days using the hypothermic EGCG preservation solution. Replenishment of EGCG solution with 2-day intervals prevented the time-dependent decline in cell viability at 3 days and later. As observed in nonpreserved control cultures, most cells were positive for proliferating cell-nuclear antigen (PCNA) in the cultures preserved at 4°C in the EGCG solution, whereas PCNA-negative cells were increased in the cultures preserved at 4°C in the MesenPRO medium. In periosteal sheets 2D-cultured in plastic dishes, the EGCG solution occasionally was associated with vacuole formation in the cytoplasm, but cells could again expand in the culture medium at 37°C. As observed in the nonpreserved periosteal sheets control, the osteogenic induction upregulated alkaline phosphatase in those cells and tissues preserved in the EGCG solution. The EGCG solution protected cells from the cold shock-induced membrane phospholipid peroxidation. Our data suggest that the EGCG solution acts as an antioxidant to protect periosteal cells from cold shock and preserves cells under chilled conditions. The limited period of preservation time could be expanded by repeating replenishment of the EGCG solution or by optimizing the formula to be more favorable for human periosteal sheets without sacrificing cell viability. This methodology of preserving human cultured periosteal sheets with EGCG would be expected to support and spread the clinical use of regenerative therapy with autologous periosteal sheets.
机译：在过去的十年中，越来越多地报道了从绿茶中提取的主要儿茶素衍生物表没食子儿茶素-3-没食子酸酯（EGCG）具有多种生物活性，包括对哺乳动物细胞和组织的细胞保护作用。在这项研究中，我们已经在人骨膜片的二维和三维培养物中测试了包含EGCG（Theliokeep？）的商业保存溶液，该溶液已用作牙周再生治疗的成骨移植材料。当在胶原网上对骨膜片进行3D培养时，使用低温EGCG保存溶液将细胞活力维持2天。间隔2天补充EGCG溶液可防止3天及以后细胞活力随时间的下降。如在未保存的对照培养物中观察到的，大多数细胞在EGCG溶液中于4°C保存的培养物中增殖细胞核抗原（PCNA）呈阳性，而在4°C于EGCG溶液中保存的培养物中PCNA阴性细胞增加。 MesenPRO媒体。在塑料培养皿中二维培养的骨膜片中，EGCG溶液有时与细胞质中的液泡形成有关，但细胞可以在37°C的培养基中再次扩增。如在未保存的骨膜片对照中所观察到的，成骨诱导上调了在EGCG溶液中保存的那些细胞和组织中的碱性磷酸酶。 EGCG溶液可保护细胞免受冷冲击诱导的膜磷脂过氧化作用。我们的数据表明，EGCG溶液可作为抗氧化剂来保护骨膜细胞免受冷冲击，并在寒冷条件下保存细胞。可以通过重复补充EGCG溶液或通过优化配方使其更适合人体骨膜片而又不牺牲细胞生存力来延长有限的保存时间。这种用EGCG保存人类培养的骨膜片的方法学有望支持和推广自体骨膜片再生疗法的临床应用。

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