Abstract

The effect of an aqueous solution of hydrated C60-fullerene (HyFn) on the growth and “stationary phase aging” (accumulation of “age-related” changes in cultured cells during the slowing down of their proliferation within a single passage and the subsequent “aging” in the stationary phase of growth) of transformed B11-dii FAF28 Chinese hamster cells was studied. The final calculated concentration of HyFn in the growth medium was 10−19 M. A paradoxical result contrasting the available data on the absence of HyFn cytotoxicity at higher concentrations was obtained in our experiments: namely, HyFn decelerated cell proliferation (estimated by the growth of mass culture, as well as by the efficiency of colony formation) and accelerated the “stationary phase aging” of the cell culture. Moreover, repeated addition of an aqueous solution of HyFn (to the final calculated concentration of 10−19 M) to the cells that had already reached the stationary phase of growth caused a rapid (within no more than 24 h) death of a significant part of the cell population. The observed effect of HyFn at ultralow concentration is supposed to arise from the alterations in the properties of the water surrounding the fullerene molecule: namely, water becomes a donor and acceptor of electrons and regulates redox processes (especially those involving oxygen) in aqueous systems. This effect of HyFn at an ultralow concentration may be specific for transformed cells, and, therefore, experiments on normal fibroblasts with limited mitotic potential are planned as a continuation of the present study. It is also possible that the reported antiaging effect of HyFn in experimental animals is due to its anticancer, immunostimulatory, antiviral, and antibacterial properties manifested only at the whole-organism level.

Highlights

  • Ôóëëåðåíû, íàèáîëåå òèïè÷íûì ïðåäñòàâèòåëåì êîòîðûõ ÿâëÿåòñÿ C60-ôóëëåðåí, îáëàäàþò øèðîêèì ñïåêòðîì áèîëîãè÷åñêîãî äåéñòâèÿ

  • Ôëàêîíû ïîìåùàëè íà 4—5 ñóò â òåðìîñòàò (37°Ñ), ïîñëå ÷åãî êëåòêè ñíèìàëè ñ ïîâåðõíîñòè ðîñòà ñìåñüþ ðàñòâîðîâ âåðñåíà è òðèïñèíà è ïîäñ÷èòûâàëè èõ êîëè÷åñòâî ñ ïîìîùüþ êàìåð Ãîðÿåâà (5—6 ôëàêîíîâ íà òî÷êó, 4 êàìåðû íà ôëàêîí)

  • The observed features of HyFn at ultra-low concentration are determined by some special properties of the water surrounding C60, namely, its ability to serve as an electron donor and acceptor regulating redox processes in aqueous systems, especially those in which oxygen is involved

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Summary

Ìàòåðèàëû è ìåòîäû

Èñõîäíûé âîäíûé ðàñòâîð ÃÔ (C60FWS) ñ êîíöåíòðàöèåé 14,4 ìã/ìë (20 ìêÌ) ïðèîáðåòàëè ó ðàçðàáîò÷èêà è èçãîòîâèòåëÿ Ôëàêîíû ïîìåùàëè íà 4—5 ñóò â òåðìîñòàò (37°Ñ), ïîñëå ÷åãî êëåòêè ñíèìàëè ñ ïîâåðõíîñòè ðîñòà ñìåñüþ ðàñòâîðîâ âåðñåíà è òðèïñèíà è ïîäñ÷èòûâàëè èõ êîëè÷åñòâî ñ ïîìîùüþ êàìåð Ãîðÿåâà (5—6 ôëàêîíîâ íà òî÷êó, 4 êàìåðû íà ôëàêîí). Ïîñëå ýòîãî ÷åðåç îïðåäåëåííûå ïðîìåæóòêè âðåìåíè èçâëåêàëè èç òåðìîñòàòà 4 ôëàêîíà, êëåòêè ñíèìàëè ñ ïîâåðõíîñòè ðîñòà ñìåñüþ âåðñåíà è òðèïñèíà, ñóñïåíäèðîâàëè â ðîñòîâîé ñðåäå áåç ÑÊÐÑ è îöåíèâàëè êîëè÷åñòâî êëåòîê â êàæäîì ôëàêîíå ñ ïîìîùüþ 4 êàìåð Ãîðÿåâà. ×åðåç 7 ñóò ïîñëå ïîñåâà â ïîëîâèíó îñòàâøèõñÿ ôëàêîíîâ äîáàâëÿëè ïî 15 ìêë ðàñòâîðà ÃÔ äî êîíå÷íîé ðàñ÷åòíîé êîíöåíòðàöèè 10–19 Ì, à â äðóãóþ ïîëîâèíó — ïî 15 ìêë äèñòèëëèðîâàííîé âîäû, ïîñëå ÷åãî ïðîäîëæàëè êóëüòèâèðîâàíèå êëåòîê, ïåðèîäè÷åñêè èçâëåêàÿ èç òåðìîñòàòà ïî 3 êîíòðîëüíûõ è 3 îïûòíûõ ôëàêîíà, â êîòîðûõ îöåíèâàëè êîëè÷åñòâî êëåòîê ïî âûøåîïèñàííîé ñõåìå. Ìàòåìàòè÷åñêèå ðàñ÷åòû è ñòàòèñòè÷åñêóþ îáðàáîòêó ðåçóëüòàòîâ ïðîèçâîäèëè ñ ïîìîùüþ ïðîãðàìì Microsoft Excel 2007 è SigmaPlot 12

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