Igawa Kazunari, Ohara Naoko, Kawakubo Atsushi, Sugimoto Kouji, Yanagiguchi Kajiro, Ikeda Takeshi, Yamada Shizuka, Hayashi Yoshihiko
Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
Department of Conservative Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan.
Biomed Res Int. 2014;2014:485867. doi: 10.1155/2014/485867. Epub 2014 Feb 11.
D-Glucosamine is a useful medicament in various fields of medicine and dentistry. With respect to stability of the cell membrane, it has been reported that bradykinin-induced nociceptive responses are significantly suppressed by the direct application of D-glucosamine. Electroporation is usually used to effectively introduce foreign genes into tissue culture cells. Buffers for electroporation with or without D-glucosamine are used in experiments of transfection vectors. This is the first study to indirectly observe the stability and protection of the osteoblast membrane against both electric stress and gene uptake (the proton sponge hypothesis: osmotic rupture during endosomes prior to fusion with lysosomes) in electroporation with D-glucosamine application. The transfection efficiency was evaluated as the fluorescence intensity of the transfected green fluorescent protein (GFP) in the cultured cells (osteoblasts; NOS-1 cells). The transfection efficiency increased over 30% in the electroporation samples treated with D-glucosamine-supplemented buffer after one day. The membrane absorption of D-glucosamine is the primary mechanism of membrane stress induced by electric stress. This new function of D-glucosamine is useful and meaningful for developing more effective transformation procedures.
D-葡萄糖胺是医学和牙科各个领域中一种有用的药物。关于细胞膜的稳定性，据报道，直接应用D-葡萄糖胺可显著抑制缓激肽诱导的伤害性反应。电穿孔通常用于有效地将外源基因导入组织培养细胞。在转染载体实验中使用含或不含D-葡萄糖胺的电穿孔缓冲液。这是第一项间接观察在应用D-葡萄糖胺的电穿孔过程中，成骨细胞膜对电应激和基因摄取（质子海绵假说：内体在与溶酶体融合之前的渗透破裂）的稳定性和保护作用的研究。转染效率通过培养细胞（成骨细胞；NOS-1细胞）中转染的绿色荧光蛋白（GFP）的荧光强度来评估。一天后，用补充了D-葡萄糖胺的缓冲液处理的电穿孔样品中转染效率提高了30%以上。D-葡萄糖胺的膜吸收是电应激诱导膜应激的主要机制。D-葡萄糖胺的这一新功能对于开发更有效的转化程序是有用且有意义的。
