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CNTs Nontoxic in Mice

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STANFORD, Calif., Jan. 31, 2008 -- Carbon nanotubes (CNTs) tracked inside mice for months were found to be nontoxic, research that may give a boost to using the tiny cylinders in biomedical applications.

HongjieDai.jpgCNT cylinders are so small that it takes 50,000 lying side by side to equal the width of one human hair. They have potential as highly accurate ways to administer medicine and other therapeutic agents to patients, but a lack of data about what happens to the tubes after they deliver their payloads has hampered progress. Now some answers have been found by Stanford University researchers, who spent months tracking them in vivo.

Studies in mice already had shown that most nanomaterials tend to accumulate in organs such as the liver and spleen, which was a concern because no one knew how long they could linger. But fears that the tubes might be piling up in vital organs can now be put to rest, said Hongjie Dai, the J. G. Jackson and C. J. Wood Professor of Chemistry at Stanford, whose research team has demonstrated that the nanotubes exit the organs.

Dai and his group found that the CNTs leave the body primarily through the feces, with some by way of the urine. "That's nice to know," he said. "This now proves that they do get out of the system."

The news from the three-month-long study is even better, he said, allaying worries that nanotubes, by simply remaining in the organs for a long time, would prove toxic to the mice.Nanotube1.jpg
Detail of a carbon nanotube, composed of linked hexagonal rings, with a representative molecule of branching polyethylene glycol (PEG) attached. (Images courtesy Hongjie Dai)
"None of the mice died or showed any anomaly in the blood chemistry or in the main organs," said Dai. "They appear very healthy, and they are gaining weight, just like normal mice. There's no obvious toxicity observed."

The lack of toxicity of nanotubes in mice is consistent with a previous pilot study done by Sanjiv Gambhir, a professor of radiology at Stanford, and his research group in collaboration with Dai's group.

"This is the first time anyone has done a systematic circulation and excretion study like this for nanotubes, and data on other nanoparticles is also scarce," Dai said. "The excretion pathway may apply to other nanomaterials and may need to be looked at closely like this also."

Previous research published by Dai's group has demonstrated the potential for using nanotubes in treating cancerous cells and targeting tumors in mice. His group used Raman spectroscopy, a method of applying light from a laser beam that effectively "illuminates" the presence of the target molecules in the organs of the mice.

Being hit with light from the beam causes a detectable change in the state of a molecule's energy. Carbon nanotubes, composed entirely of carbon atoms that are mostly arranged in linked hexagonal rings, give off a strong signal in response to the beam. This allowed the researchers to pinpoint the position of the chosen molecules, as well as ascertain their abundance in the blood or organs.

Previous detection methods that relied on attaching fluorescent labels or spectroscopic tags to the nanotubes yielded unreliable results. The attachments tended to either come loose from the tubes or decay over time spans ranging from a few days to only a few hours -- far too short to reveal the ultimate fate of the nanotubes.

While knowing the CNTs will move through the digestive system at a healthy pace is critical to future practical applications, it is also crucial that the nanotubes not enter the digestive system too soon after being injected; they need to spend enough time in the circulatory system to find their way to their target location.

The key to fine-tuning the CNTs' speed of circulation turns on how the basic, bare-bones floor model is chemically accessorized.
A representation of a carbon nanotube accessorized with a coating of branched PEG.
"You can make the nanotubes circulate a very long time in the blood, if the chemistry is done right," Dai said. The researchers found that coating their CNTs with polyethylene glycol (PEG), a common ingredient in cosmetics, worked best.

They used a form of PEG with three little limbs sprouting off a central trunk. "Those provide better shielding to the nanotube than just a single branch. Therefore, they interact less with the biological molecules around them," Dai said.

The team stuffed the PEG liberally into the linked hexagonal rings that compose the nanotubes, prompting Dai to describe the end result as resembling rolled-up chicken wire with feathers sticking out all over. The PEG coating made the nanotubes highly water soluble, which helped them to stay in the blood instead of being absorbed.

"They circulate in the blood for about 10 hours or so in mice, which seems to be a good length of time," Dai said.

The right chemical coating on nanotubes also can help ease them out of the mouse in a timely fashion, and the three-branched PEG was effective there, too. Dai's earlier research demonstrated that nanotubes have promise for treating cancer with two different approaches. Once they have zeroed in on the target cells, shining light on the nanotubes causes them to generate heat, which can kill cancer cells. The other method is to rig the nanotubes to accumulate at targeted sites, where they can deliver medication from within the tubes.

"(CNTs) seem to be promising for biomedical applications and for potentially treating cancer, either using drugs or using the physical properties," Dai said. "This is the reason we carried out the study of the fate of nanotubes in mice. I think this is really a very fundamental issue."

The research was funded by the Cancer Center for Nanotechnology Excellence, which is funded by the National Institutes of Health and the National Cancer Institute.

The work was scheduled to be published this week in the online early edition of Proceedings of the National Academy of Sciences; Dai is senior author.

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Jan 2008
The technology of generating and harnessing light and other forms of radiant energy whose quantum unit is the photon. The science includes light emission, transmission, deflection, amplification and detection by optical components and instruments, lasers and other light sources, fiber optics, electro-optical instrumentation, related hardware and electronics, and sophisticated systems. The range of applications of photonics extends from energy generation to detection to communications and...
raman spectroscopy
That branch of spectroscopy concerned with Raman spectra and used to provide a means of studying pure rotational, pure vibrational and rotation-vibration energy changes in the ground level of molecules. Raman spectroscopy is dependent on the collision of incident light quanta with the molecule, inducing the molecule to undergo the change.
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