ABSTRACTS FROM: The Prolongation of the Lifespan of Rats by Repeated Oral Administration of [60]Fullerene Dec 12, 2016

Link to article: https://www.researchgate.net/publication/224004891_The_prolongation_of_the_lifespan_of_rats_by_repeated_oral_administration_of_60fullerene

the metabolic fate and the in vivo chronic effects of C60itself still remain unknown. fullerene is active only in soluble form. the extremely slow dissolution of C60in biological media prevents  controlling accurately the active fraction .

 the chronic toxicity of C60has never been investigated to our knowledge.
C60is soluble in lipid droplets inside living cells as well as in fats in general [2. Moreover, C60can freely cross membrane barriers. C60interactions with living systems as well as its toxicity should be determined using solubleforms.

While C60solubility in vegetable oils  solutions should be quite appropriate for studying its chronic toxicity at reiterated doses.  the in vivo behaviour of soluble forms of C60 was unknown, we determined the in vivo fate of C60dissolved in olive oil before studying its chronic effects at reiterated doses.

Virgin olive oil is obtained from a Chemlali Boughrara cultivar from Tunisia planted in the Sahel area. C60(purity 99.98%) was obtained from SES ResearchCorporation (USA) and used without further purification. Fifty mg of C60were dissolved in 10 ml of olive oil by stirring for 2 weeks at ambient temperature in the dark. The resulting mixture was centrifuged at 5.000 g for1 h and the supernatant was filtered through a Millipore filter with 0.25mm porosity.

 Chronic toxicity and effects of C60on survival of rats

The rats were housed three per cage and acclimated for 14 days, before dosing.Three groups of 6 rats (10 months old, weighing 465  31 g) were administered daily for one week, then weekly until the end of the second month and then every two weeks until the end of the 7th month, by gavages with 1 ml of water or olive oil orC60dissolved in olive oil (0.8 mg/ml), respectively.

The rats were weighed before each dosing. Routine observations following official recommendations [27] were made on all animals inside and outside the cageonce a day throughout the study for signs of departure from normal activity,morbidity and mortality.

Sixty rats randomly divided into 10 groups of 6 rats were pre-treated daily for 7days by oral gavages (og groups) or by i.p. injection (ip groups). Groups A (GAog andGAip), received 1 ml of water. Groups B and C (GBog, GCog and GBip, GCip) were pre-treated with 1 ml of olive oil while groups D and E (GDog, GEog and GDip, GEip)were pre-treated with 1 ml of C60-olive oil.

The resulting C60-olive oil solution is purple and contains0.80  0.02 mg/ml (n ¼ 6) The stability of both oily and control solutions stored at ambient temperature and in the dark was checked monthly during 48months. No change was recorded under our chromatographic conditions  C60is well distributed in tissues.

 Chronic toxicity and effects of C60on lifespan of rats

Fig. 3 shows the animal survival and growth. After five months of treatment (M15) one rat treated with water only exhibited some palpable tumours in the abdomen region. Due to the rapid development of tumours (about 4 cm of diameter) this rat died at M17. As rats are known to be sensitive to gavages, we decided to stop the treatment for all rats and to observe their behaviour and overall survival.

All remaining animals survived with no apparent sign of behavioural trouble until M25(Fig. 3a). At the end of M25 the animals of the control groups showed signs of ulcerative dermatitis with ageing while C60-treated animals remained normal. As the growths of all surviving animals showed no significant difference until M30(Fig. 3b) indicating that the treatment did not alter their food intake, we continued observing their survival.

At M38 all water-treated control rats were dead (Fig. 3a). This agrees with the expected lifespan of this animal species that is thirty to thirty six months. At this time 67% of olive-oil-treated rats and 100% of C60-treated rats were still alive.

 The estimated median lifespan (EML) for the C60-treated rats was 42months while the EMLs for control rats and olive oil-treated ratswere 22 and 26 months, respectively. These are increases of 18 and 90% for the olive-oil and C60-treated rats, respectively, as compared to controls.

olive oil extends the lifespan of rats with respect to water with a probability of 0.99 while C60-olive oil extends the lifespan of C60 treated rats with a probability of 0.999 and 0.995 with respect to water and olive oil treatments, respectively.

. C60-olive oil solution preparation

The C60-olive oil solution used in this study can be considered as free of C60aggregates because its colour is purple that is characteristic of C60solutions AND it is freely and instantaneously soluble in toluene.

The stability of C60-olive oil solution determined under our experimental conditions agrees with recently published results showing that the addition of [60]fullerene significantly hampers the peroxide formation thus increasing the stability of the tested oils . The results of this pharmacokinetic study show for the first timethat C60is absorbed by the gastro-intestinal tract

The elimination process follows a non-urinary route because unmodified C60was not detected in urine samples C60is mainly eliminated through the bile ducts  C60reacts inside the liver cells with vitamin A. As C60and some of its derivatives mainly accumulate in the livers and spleens we also studied the accumulation of C60in these organs after 7 successive days of administration.

 C60molecules are eliminated from the organs in a few hours after both oral and i.p. administrations. the presence of significant amounts in the brain 24 hours after both oral and i.p. administrations under our experimental conditions confirms that solubilized C60can cross the bloodebrain barrier

Chronic toxicity and effects of C60on survival of treated rats

C60has no acute or sub-acute toxicity inrodents  it can act as an antioxidant, we investigated its chronic toxicity concomitantly with its effects on the survival of rats. Ten-month old male rats (M10) were chosen instead of young ratsas officially recommended, in order to avoid possible compensatory effects that can occur during early development.   we treated the rats daily only during 7 days and weekly during the first two months, then every two weeks until one control rat died.

Our results show that while olive oil treatment can lead to anincrease of 18% of lifespan of treated rats, C60-olive oil can increaseit up to 90%, as compared to controls.  at M38 all C60-treated rats were still alive.  C60should be the most efficient ever material for extending lifespan.  C60itself is a powerful antioxidant

 This is the first investigation of the in vivo chronic effects ofa soluble form of C60. C60is a powerful antioxidant .C60can scavenge large numbersof free radicals. C60isa powerful liver-protective agent. C60is ableto scavenge in vitro a large number of radicals per molecule  and be this property can be involved in the mechanism of protection against toxicity,The results obtained for antioxidant enzyme activities also confirm the antioxidant effect of C60against toxicity.

the protective effect involves a free-radical scavenging mechanism. dissolved C60appears hundred times more active than when it is in suspension. In fact the action of soluble C60is immediate while that of suspended C60is delayed because it has to be dissolved to act. In all cases, based on C60-livercontent (Table 2), these results prove that this fullerene is active atthe nano molar level. However, the involved mechanism remains to be established.

For the time being the hypothesis of free-radical scavenging remains possible. C60could act as a free-radicalscavenger as it has been widely demonstrated in vitro, but up to now no resulting C60adduct has been observed in vivo. 


The effect of pristine C60on lifespan emphasizes the absence of chronic toxicity. In the present case, the treatment was stopped when a control rat died at M17, which proves that the effects of the C60treatment are long-lasting as the estimated median lifespan for C60-treated rats is 42 months. It can be thought that a longer treatment could have generated even longer lifespans. Anyway, this work should open the road towards the developmentof the considerable potential of C60in the biomedical field,including cancer therapy, neurodegenerative disorders and ageing.Furthermore, in the field of ageing, as C60can be administered orally and as it is now produced in tons,


Published on May 19, 2018

A great interview with Chris Burres, owner and founder of SES Research on making C60, differences in C60 and how they are not created or treated equally, and common questions, concerns and confusion answered and explained. This IS the video to watch on C60!

FOR INFO ON THE MOLECULE C60: MUST LISTEN:PART 1 C60 Olive Oil Revealed with Chris Burres/SES Research INTERVIEW 51 MINS MP3

MUST LISTEN:PART 2 C60 Olive Oil Revealed with Chris Burres/SES Research INTERVIEW 15 MINS MP3


MUST LISTEN 33min interview with Mr. Geska who made and provided the C60 used in the original C60 toxicity rat study  http://jokebook.com/mp3/33min-Interview%20-Bob%20Greska-made-orig-C60-used-in-orig-rat-study.mp3

Mr. Geska explain how the C60 molecule heals:

1. The C60 molecule neutralize toxins and frees up cellular energy.

2. Toxins lack an electron and they receives one and are neutralized when making contact with electron donor C60.

3. The more cellular energy is available the more the body is healed.

4. C60 is the closest to the "fountain of youth" that has been found.