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TOXICITY AND BIOCOMPATIBILITY STUDIES.

Evidence of safety in toxicity testing is one of the first hurdles any fullerene material must clear before environmental and biological exposureIt should be a first and foremost concern for anyone considering using or giving fullerene material to a pet.  Although, in a general sense, pure pristine fullerene molecules are considered to be “biocompatible”, however the purchasing of these molecules poses somewhat of a crapshoot.  The potential for poor quality or contaminated/modified fullerene molecule material is an ever-present risk. 

Our Grafex®Super [C-60] fullerene molecules has undergone numerous university and independent lab tests that have all shown excellent biocompatibilityOver a number of years these tests have involved different batches of Grafex®Super [C-60] being administered at different time frames, with different concentrations and environments. All with excellent results. The proprietary production process of Grafex®Super [C-60] fullerene molecules results in a consistent, spherical multi-layered carbon material with uniform structures that does not require “cleaning or harvesting” and thereby avoids potential contamination

Toxicity and life extension considerations of the Grafex®Super [C-60] fullerene carbon material would be expected to hold, at a minimum, a strong concordance to its forerunner the Regular C60 Fullerene molecule coming with 60 carbon atoms

The previous Regular C60 fullerene toxicity and life extension results appear to be mirrored by the Grafex®Super [C-60] carbon material, as a result of the subcutaneous injection and follow on ablation of human cancer cells in a 10 mice study. This testing was conducted over a three-year period at the University of New York, at Stony Brook, by Dr. Mark Desantis (see first Mice Study).

The results of this action prompted an almost three-year follow up study by a company-performed informal internal ingestion study, on field mice. During this research study of approximately 3 years, nearly seven generations of the  “control group micepassed away. During that timeframe, all of the mice fed the Grafex® Super (C-60) carbon continued in extraordinary youthful vitality as well as surprising extended longevity

This study was conducted to determine potential longevity, variable dosing volume reactions, toxicity, teratogenicity, as well as cancer occurrence/reoccurance. (see Second  mice study).

The Grafex®Super [C-60]carbon material has undergone at least five known formal in vitro and invivo studies following various established protocols from a broad assortment of sophisticated laboratories across the nation, as well as over 100 formal tests on animals:

Below is a list of some of the facilities that have conducted formal and controlled research laboratory testing’s on the Grafex® Super [C-60] fullerene molecule.

Our desire is to foster strategic cooperative relationships to facilitate additional research and product development.

Grafex® Super [C-60] fullerenes are the highest quality/purity (99.99%) nano carbon fullerene material on the market today. We are able to produce and can make it available for any level of research needs. At this time the Grafex material is being offered in reasonable requested quantities, only to University and Certified Research Facility researchers, in the U.S.

  

A 75% discount will be given to those research facilities that agrees to share the results of their testing with Graphitic Nano Onions LLC

If you have an interest, please provide us with your contact information, as well as your volume requirements and a sales representative will contact you within 24 hrs to complete your order. 

 

 

 

 

This research includes:

®2013Desantis, et al., In-situ Nanocarbon (Grafex®Super [C-60] assisted Microwave treatment (NAMT), Hyperthermia Cancer Treatment Journal of Vascular and Interventional Radiology Volume 24, Issue 4, Supplement, April 2013, Pages S116–S117

Research was performed by Dr. Mark Desantis, an interventional radiologist at the University of New York, Stony Brook Medical Center, Stony Brook NY to evaluate use of spherical nanocarbon Grafex®Super [C-60] fullerene molecules injected into known Human Prostatic carcinoma to increase absorption of microwave energy, specifically into tumor cells. Over 90% of the mice responded to the treatment without toxic effects of the retained nanocarbon within the dermis or skin. This study was selected to evaluate if Grafex®Super [C-60] and microwave energy could be used as a primary process due to a demonstrated range of energy absorption capabilities superior to other non-metallic materials. The study went on to note that there “is no observed toxic side effects in mice from this two and one half year study.”(see mice New York Power Pt.)

2014 – Abstract of a Poster ‘MR Imaging during “Thermal Ablation of Breast Tumor” presented at Society of Nuclear Medicine and Molecular Imaging 2014 Annual Meeting St. Louis, MO June 7-11, 2014 by Dr. Desantis and his team at Stony Brook University Medical Center, Stony Brook, NY.

This breast cancer tumor study was performed by the authors of the previous 2013 prostate cancer ablation study, which revealed that “Spherical nanocarbon (Grafex® Super [C-60]) appears to provides a nontoxic method of aiding in thermally ablating the tumors utilizing MRI safe microwave antenna, while allowing for imaging during the treatment. 

2013- Wheeler, et al., Mechanical & cell culture properties of elastin-like polypeptide, collagen, bioglass, and carbon nanosphere composites. Ann Biomed Eng. 2013 Oct;41(10):2042-55 (Ole  Miss –Jackson, MS)

Grafex® was –added to ELP collagen hydrogels to enhance tissue engineering in guided bone regeneration techniques.  Such composite membranes must possess good osteoconductive and biocompatibility with various cell types including pre-osteoblasts.  This study found that not only did “Grafex® contribute to the nearly doubling of the average tensile strength and elastic modulus compared to the other ELP-collagen hydrogels;” that it also had “no adverse impact on cell viability“, DNA content, ALP activity, and OCN production compared to other ELP-collagen hydrogels tested, suggesting “good biocompatibility”.

(UAMS–Little Rock, AR)

2016 – 2019 – Ongoing Research at the University of Arkansas, Cancer Research Center,

A multi-part study to test Grafex®Super [C-60] fullerenes in combination with radiotherapy and microwave treatment for localized therapies for different cancers is being conducted at the University of Arkansas Cancer Research Center. As part of this study a cell viability assay was performed. Cells were seeded in 96 well plate at a concentration of 1000 cells/well and all conditions were run in triplicate. The result of a standard viability assay run with exposure of tumor and normal endothelial cells mixture to a range of 0.1-100 ug/ml of Grafex® Super [C-60], demonstrated little or no toxicity of the carbon on these cells

Overall the results of the viability study showed that Grafex® Super [C-60] had “no significant cytotoxicity” up to a concentration of 100 ppm. These concentrations also do not appear to adversely affect spheroid formation.  In two cell migration studies, we found limited to no effect of the Grafex®Super [C-60] on either brain tumor or breast tumor cell motility alone or when combined with radiation. Consistent with other reports in the literature, “the Grafex®Super [C-60] at ~1 ppm seems to provide some protection from radiation.”   Based on this testing, Grafex®Super [C-60] appears to be “non-toxic” and does not perturb cellular functions to a significant degree, suggesting “good biocompatibility”.

The study also used a 3D cell culture model to study Grafex® Super [C-60’s] possible influence on cell to cell adhesion and growth The results demonstrated that the presence of carbon in the growth medium seemed to stimulate the growth of multicellular spheroids (mixed tumor and endothelial cells, 4T1 and 2H11).   There was no statistically significant difference in response to concentration, however the highest concentration of Grafex® Super [C-60] tested tended to be the largest spheroid size further confirming the biocompatibility of the Grafex® Super C-60 material.

Based on their previous assessment of the low potential for toxicity and excellent biocompatibility of the Grafex®Super [C-60] material, the University of Arkansas has begun several in vivo research projects that include ongoing toxicity research, investigating distribution, metabolism, and clearance research into imaging capabilities and treatment options, in cancer models, using 100 mice. 

The cancer studies include an imaging study of Grafex®Super [C-60] fullerenes with solid tumors (4T1 breast tumor model) to determine the imaging capabilities for bio distribution and tumor targeting as well as real-time treatment optimization using advanced non-invasive imaging platforms; as well as using Grafex®Super [C-60] for microwave phototherapy ablation for tumor destruction and control. 

 

 

(ITG – Angleton, TX)

2016- Radiolabeling and Cell Toxicity of Grafex™Super [C-60] Report prepared by IsoTherapeutics Group, LLC of Angleton, TX.

The results of this study included two types of experiments that were performed by IsoTherapeutics Group on Grafex® Super [C-60] material. Radioactive labeling strategies for the particles were explored and several strategies to radiolabel with I-131 were attempted in the laboratory. In addition, the effect of nanoparticles on cell cultures was studies in order to determine if the particles have toxicity to human fibroblast cells.

 From the radiolabeling experiments we concluded that: Analytical methods using size exclusion chromatography were developed and used to determine the yield of reactions. We recommend pursuing these and other chromatographic methods.

Radiolabeling using a bifunctional chelating agent and a radioactive metal is possible but it would require a significant effort to accomplish.

From the cell toxicity studies we concluded that in this study:

  1. All three Grafex® samples were not toxic to human fibroblast cells.
  2. Grafex® samples are expected to be safe in animals.  

 

2018 – A retrospective investigation of evidence of novel pristine nanocarbon fullerene material toxicity after prolonged inhalation, dermal, and ingestion exposure in humans Bourassa, et al., Einstein Institute, USAT – School of Medicine. 

Dr. Bourassa and colleagues presented their findings after conducting a limited retrospective study on a small number of individuals that have had extensive inhalation, dermal, and oral exposure to a novel multi-walled nanocarbon fullerene-like material called Grafex®Super [C-60].

Dr. Bourassa and colleagues interviewed the subjects as well as assessed their individual serum laboratory results, chest imaging and medical records.

All of the subjects reported varying positive effects from consumption of the Grafex®Super [C-60] material.  These effects ranged from increased energy and endurance, improved mental clarity and cognition, to noticeable reduction in previous joint pain and inflammation.

One subject with prior occupationally acquired lung pathology related to Black Lung disease reported that his ability to climb hills, walk long distances and work 10 to 12 hours days, had improved immensely.

Another subject who had been diagnosed with COPD and had been using Spiriva for over a year, began taking Grafex® Super [C-60] orally. After several months of use he stopped taking the Spiriva because his respiratory function improved enough that he was able to resume ‘singing in his church choir again.That was two years ago. He is still singing.

Another subject, who had been exposed to, and had breathed this material for 12 years,   began a regimen of self-testing with this material, through oral consumption over the past 6 year period. He had previously been diagnosed with severe bilateral degenerative joint disease of the knees, as was confirmed on MRI and X-Ray imaging studies that was presented. Shortly after consuming the Grafex® He was able to resume running activities and tennis without pain of inflammation in his knees.

The same individual had also been diagnosed (via x-rays) as having a left shoulder rotator cup tear and was scheduled for surgery. Because of scheduling problems, he delayed the surgery. Within several months after consuming the Grafex® Super [C-60] material he was pain free and is still lifting weights on a regular basis. That was 8 years ago.  

There were no reported respiratory problems and all subjects reported feeling good to excellent health, as well as having been able to stop taking the prescribed drugs that they had previously been administered.

It was their conclusion that there was no evidence of nanocarbon related pathology associated with prolonged inhalation and dermal exposure, by the oral consumption of this novel nanocarbon fullerene material in lipid suspensions

Additionally, the positive effects from oral consumption of Grafex® Super [c-60], such as improved energy, endurance, reduction of inflammation and arthritic painas well as cognitive improvements and increased well-being, were reported by all subjects.  There was some evidence that the magnitude of effects may correlate with an increase in the material consumption.

Finally, the subjective claims of beneficial effects and absence of negative effects are consistent with the observations reported by researchers in the Baati and Desantis studies and the widespread antidotal claims made by other individuals consuming fullerenes in lipid (oil) suspensions. 

Their conclusion was that future medical research into the mechanism of fullerene absorption, tissue concentrations, metabolism, elimination, and interaction in human cell biology can be approached with greater confidence using this multi-layered fullerene-like material and likely other pure, high quality fullerenes when general precautions are followed including in vitro studies using the fullerenes that have demonstrated excellent  biocompatibility.

They further concluded that although the small population and limitations of this retrospective study are recognized; the length and extent of exposure present a significant opportunity to evaluate the subjects for either evidence of pathology and nanocarbon fullerene toxicity; as well as to indicate the relative degree of safety and biocompatibility of the Grafex® Super [C-60]  multi-layered nanocarbon fullerene-like material, in adult humans.

Over the last 10 years there have been hundreds of human self- experimenters that have utilized the Grafex®Super [C-60] material in various ways. Not one has had any adverse effect to the use of the material to date. To the contrary, everyone has had a variety of positive results, depending on what their particular aliments were. (see personal reviews pg).

Biomedical research, based on classical “concepts of physics and biology,” into applications of fullerene material, has so far, been overwhelmingly positive

April, 2018

Gabriel Gojon Sr., Ph.D., Co-Founder and Chief Technical Officer SulfaGenix Inc.

Dr. Gojon, a past recipient of the prestigious Andres Manuel del Rio award (2007) from the National Chemical Society Mexico, is the Director of Research at NANT and the co-discoverer of SG1002. Dr. Gojon began his career in industry as the Director of Research at Reactimex, SA de CV, in Monterrey, where he was in charge of new product research and development. Throughout his career Dr. Gojon has received numerous recognitions, distinctions and awards.

Here is the answer from Dr. Gabriel Gojon on the results of his first testing of the Grafex® Super [C-60] material; “here you can quote Dr Gojon about the research (preliminary result):

Grafex®Super [C-60] nano onions are at least an order of magnitude (10 times) better at neutralizing free radicals than C60 Fullerenes”        

Dr. Gabriel Gojon Zorrilla PhD Sulfagenix, Inc.

 Friday, August 31, 2018 12:54 PM,

 “Sorry for the delay in reporting findings. We could not disclose some results due to the patent process. What we can report right now is that the Grafex® Nano material (both types we have been provided):

prevents a host of degenerative diseases in rat models, decrease free radical damage due to senescent cells overproducing them (thus having the same final effect as a senolytic drug but with a different mechanism) in rat models.

is a universal organ protector (brain, liver, kidneys, etc.), in rats.

is a possible treatment for eye diseases in rat models.” 

Dr. Gabriel Gojon Zorrilla PhD Sulfagenix, Inc. 

Many of these include nano research labs that bring to bear highly sophisticated equipment and techniques for identification and verifying the properties, purity and structural features of the Grafex®Super [C-60] fullerene material.

Such characterization proceeded during a rapid advancement of the new and growing field of nano technology.

 



Aim:

To develop the potential application of carbon nanomaterials as antioxidants calls for better understanding of how the specific structure affects their antioxidant activity. Materials & methods: Several typical carbon nano materials, including graphene quantum dots and fullerene derivatives were characterized and their radical scavenging activities were evaluated; in addition, the invitro and in vivo “radioprotection experiments were performed.”

Results:

These carbon nanomaterials can “efficiently scavenge” free radicals in a structure-dependent manner. In vitro assays demonstrate that administration of these carbon nanomaterials markedly increases the surviving fraction of cells exposed to ionizing radiation. Moreover, in vivo experiments confirm that their administration can also increase the survival rates of mice exposed to radiation.

Conclusion:

All results confirm that “large, buckyball-shaped fullerenes” show the strongest antioxidant properties and the best radioprotective efficiency. Our work will be useful in guiding the design and optimization of nanomaterials for potential antioxidant and radioprotection bio-applications. Published online: 15 March 2018

Antioxidant activity and radical scavenging (Nakamura S, 2009)  (Chistyakov VA, 2013) (Baati T, 2012) (Petrovic D, 2015) (Satoh & Takayanagi, 2006)       (Syrensky A, 2006), and  NrF2 –induced cellular antioxidant defense (Ye S, 2014) (Ershova E, 2016)

•  Antimicrobial activity through growth inhibition (Aoshima H, 2009) (Taherpour A, 2017)

•   Cancer therapy (Prylutska S B. A., 2011) (Orlova M, 2013) (Petrovic D, 2015) (Satoh & Takayanagi, 2006) (Pacor S, 2015) (Chen Z, 2012) (Mitra A, 2015) (Srivastava I,

2017), anti-neoplastic activity (Lynchack O, 2017), and Caspase-independent autophagy induced cancer cell death  (Q Zhang, 2009) (Orlova M, 2013)

•  Biosensors (Saini J, 2016)

•  Photodynamic therapy (Orlova M, 2013) (Sharma S, 2011) (Syrensky A, 2006)

Anti-virus and anti-HIV activity of fullerenes (Orlova M, 2013)   (Satoh & Takayanagi, 2006) (Marchesan S, 2005), against hepatitis C (Mashino T, 2005), and influenza virus (Shoji M, 2013)

Anti-inflammatory action (Dellinger A, 2015) (Petrovic D, 2015) (Liu Q J. I.,  2013)

Treatment of osteoarthritis and IVD disease  (Liu Q C. Q., 2014) (Satoh & Takayanagi, 2006)

Immune System effects  (Bunz H, 2012) (Petrovic D, 2015)

Neurodegeneration, neuronal protection, and stroke  (Giust D, 2011) (Orlova M, 2013) (Petrovic D, 2015)  (D giust, 2014) (Satoh & Takayanagi, 2006) (Syrensky A, 2006) (Baldrighi M, 2016)

Skin (Satoh & Takayanagi, 2006)

EMF and Radiation protection (Orlova M, 2013)

 Fullerene in drug delivery (Orlova M, 2013) (Prylutska S G. I., 2014) (Petrovic D, 2015) (Pietersz G, 2017) (Tripathi A, 2015) (Mitra A, 2015)

Bio imaging in medicine  (Lalwani G, 2013) (Chen Z, 2012) Ischemia and reperfusion injury  (Syrensky A, 2006)

Antiproliferative and anti-atherogenic effects (Syrensky A, 2006)

Internalization of Carbon Nano-onions by Hippocampal Cells Preserves Neuronal Circuit Function and Recognition Memory. 
Massimo Trusel Michele Baldrighi Roberto Marotta Francesca Gatto Mattia Pesce Marco Frasconi Tiziano Catelani Francesco Papaleo Pier Paolo Pompa Raffaella Tonini*Silvia Giordani* April 2018.