University of Pennsylvania: In the Future Seeing Your Dentist May ‘Go Nano’ ~ Using nanoparticles to break up plaque and prevent cavities

The bacteria that live in dental plaque and contribute to tooth decay often resist traditional antimicrobial treatment, as they can “hide” within a sticky biofilm matrix, a glue-like polymer scaffold.

A new strategy conceived by University of Pennsylvania researchers took a more sophisticated approach. Instead of simply applying an antibiotic to the teeth, they took advantage of the pH-sensitive and enzyme-like properties of iron-containing nanoparticles to catalyze the activity of hydrogen peroxide, a commonly used natural antiseptic. The activated hydrogen peroxide produced free radicals that were able to simultaneously degrade the biofilm matrix and kill the bacteria within, significantly reducing plaque and preventing the tooth decay, or cavities, in an animal model.

“Even using a very low concentration of hydrogen peroxide, the process was incredibly effective at disrupting the biofilm,” said Hyun (Michel) Koo, a professor in the Penn School of Dental Medicine’s Department of Orthodontics and divisions of Pediatric Dentistry and Community Oral Health and the senior author of the study, which was published in the journal Biomaterials. “Adding nanoparticles increased the efficiency of bacterial killing more than 5,000-fold.”

The paper’s lead author was Lizeng Gao, a postdoctoral researcher in Koo’s lab. Coauthors were Yuan Liu, Dongyeop Kim, Yong Li and Geelsu Hwang, all of Koo’s lab, as well as David Cormode, an assistant professor of radiology and bioengineering with appointments in Penn’s Perelman School of Medicine and School of Engineering and Applied Science, and Pratap C. Naha, a postdoctoral fellow in Cormode’s lab.

The work built off a seminal finding by Gao and colleagues, published in 2007 in Nature Nanotechnology, showing that nanoparticles, long believed to be biologically and chemically inert, could in fact possess enzyme-like properties. In that study, Gao showed that an iron oxide nanoparticle behaved similarly to a peroxidase, an enzyme found naturally that catalyzes oxidative reactions, often using hydrogen peroxide.

When Gao joined Koo’s lab in 2013, he proposed using these nanoparticles in an oral setting, as the oxidation of hydrogen peroxide produces free radicals that can kill bacteria.

“When he first presented it to me, I was very skeptical,” Koo said, “because these free radicals can also damage healthy tissue. But then he refuted that and told me this is different because the nanoparticles’ activity is dependent on pH.”

Gao had found that the nanoparticles had no catalytic activity at neutral or near-neutral pH of 6.5 or 7, physiological values typically found in blood or in a healthy mouth. But when pH was acidic, closer to 5, they become highly active and can rapidly produce free radicals.

The scenario was ideal for targeting plaque, which can produce an acidic microenvironment when exposed to sugars.

Gao and Koo reached out to Cormode, who had experience working with iron oxide nanoparticles in a radiological imaging context, to help them synthesize, characterize and test the effectiveness of the nanoparticles, several forms of which are already FDA-approved for imaging in humans.

Beginning with in vitro studies, which involved growing a biofilm containing the cavity-causing bacteria Streptococcus mutans on a tooth-enamel-like surface and then exposing it to sugar, the researchers confirmed that the nanoparticles adhered to the biofilm, were retained even after treatment stopped and could effectively catalyze hydrogen peroxide in acidic conditions.

They also showed that the nanoparticles’ reaction with a 1 percent or less hydrogen peroxide solution was remarkably effective at killing bacteria, wiping out more than 99.9 percent of the S. mutans in the biofilm within five minutes, an efficacy more than 5,000 times greater than using hydrogen peroxide alone. Even more promising, they demonstrated that the treatment regimen, involving a 30-second topical treatment of the nanoparticles followed by a 30-second treatment with hydrogen peroxide, could break down the biofilm matrix components, essentially removing the protective sticky scaffold.

Moving to an animal model, they applied the nanoparticles and hydrogen peroxide topically to the teeth of rats, which can develop tooth decay when infected with S. mutans just as humans do. Twice-a-day, one-minute treatments for three weeks significantly reduced the onset and severity of carious lesions, the clinical term for tooth decay, compared to the control or treatment with hydrogen peroxide alone. The researchers observed no adverse effects on the gum or oral soft tissues from the treatment.

“It’s very promising,” said Koo. “The efficacy and toxicity need to be validated in clinical studies, but I think the potential is there.”

Among the attractive features of the platform is the fact that the components are relatively inexpensive.

“If you look at the amount you would need for a dose, you’re looking at something like 5 milligrams,” Cormode said. “It’s a tiny amount of material, and the nanoparticles are fairly easily synthesize, so we’re talking about a cost of cents per dose.”

In addition, the platform uses a concentration of hydrogen peroxide, 1 percent, which is lower than many currently available tooth-whitening systems that use 3 to 10 percent concentrations, minimizing the chance of negative side effects.

Looking ahead, Gao, Koo, Cormode and colleagues hope to continue refining and improving upon the effectiveness of the nanoparticle platform to fight biofilms.

“We’re studying the role of nanoparticle coatings, composition, size and so forth so we can engineer the particles for even better performance,” Cormode said.

Explore further: Nanoparticles release drugs to reduce tooth decay

More information: Lizeng Gao et al. Intrinsic peroxidase-like activity of ferromagnetic nanoparticles, Nature Nanotechnology (2007). DOI: 10.1038/nnano.2007.260

Lizeng Gao et al. Nanocatalysts promote Streptococcus mutans biofilm matrix degradation and enhance bacterial killing to suppress dental caries in vivo, Biomaterials (2016). DOI: 10.1016/j.biomaterials.2016.05.051

 

 

The “Nano-Tooth Fairy” – Nanoparticles Release Drugs to Reduce Tooth Decay

Therapeutic agents intended to reduce dental plaque and prevent tooth decay are often removed by saliva and the act of swallowing before they can take effect. But a team of researchers has developed a way to keep the drugs from being washed away.

Dental plaque is made up of bacteria enmeshed in a sticky matrix of polymers—a polymeric matrix—that is firmly attached to teeth. The researchers, led by Danielle Benoit at the University of Rochester and Hyun Koo at the University of Pennsylvania’s School of Dental Medicine, found a new way to deliver an antibacterial agent within the plaque, despite the presence of saliva.

Their findings have been published in the journal ACS Nano.

“We had two specific challenges,” said Benoit, an assistant professor of biomedical engineering. “We had to figure out how to deliver the anti-bacterial agent to the teeth and keep it there, and also how to release the agent into the targeted sites.”

To deliver the agent—known as farnesol—to the targeted sites, the researchers created a spherical mass of particles, referred to as a nanoparticle carrier. They constructed the outer layer out of cationic—or positively charged—segments of the polymers. For inside the carrier, they secured the drug with hydrophobic and pH-responsive polymers.

The positively-charged outer layer of the carrier is able to stay in place at the surface of the teeth because the enamel is made up, in part, of HA (hydroxyapatite), which is negatively charged. Just as oppositely charged magnets are attracted to each other, the same is true of the nanoparticles and HA. Because teeth are coated with saliva, the researchers weren’t certain the nanoparticles would adhere. But not only did the particles stay in place, they were also able to bind with the polymeric matrix and stick to dental plaque.

Since the nanoparticles could bind both to saliva-coated teeth and within plaque, Benoit and colleagues used them to carry an anti-bacterial agent to the targeted sites. The researchers then needed to figure out how to effectively release the agent into the plaque.

Farnesol is released from the nanoparticle carriers into the cavity-causing dental plaque. Credit: Michael Osadciw/University of Rochester 

A key trait of the inner carrier material is that it destabilizes at acidic—or low pH—levels, such as 4.5, allowing the drug to escape more rapidly. And that’s exactly what happens to the pH level in plaque when it’s exposed to glucose, sucrose, starch, and other food products that cause tooth decay. In other words, the nanoparticles release the drug when exposed to cavity-causing eating habits—precisely when it is most needed to quickly stop acid-producing bacteria.

The researchers tested the product in rats that were infected with Streptococcus mutans—a microbe that causes tooth decay. “We applied the test solutions to rats’ mouths twice daily for 30 seconds, simulating what a person might do using a mouth rinse morning and night,” said Hyun Koo, a professor in the Department of Orthodontics and co-senior author of the work. “When the drug was administered without the nanoparticle carriers, there was no effect on the number of cavities and only a very small reduction in their severity. But when it was delivered by the nanoparticle carriers, both the number and severity of the cavities were reduced.”

Plaque formation and tooth decay are chronic conditions that need to be monitored through regular visits to the dental office. The researchers hope their results will someday lead to better—and perhaps permanent—treatments for dental plaque and tooth decay, as well as other biofilm-related diseases.