Tag Archives: Benedetto marelli

Silk micro or nanoparticles for tracking seeds

I hadn’t heard of counterfeit seeds before but this March 22, 2023 news item on phys.org describes the problem and announces an approach to prevention,

Average crop yields in Africa are consistently far below expected, and one significant reason is the prevalence of counterfeit seeds whose germination rates are far lower than those of the genuine ones. The World Bank estimates that as much as half of all seeds sold in some African countries are fake, which could help to account for crop production that is far below potential.

There have been many attempts to prevent this counterfeiting through tracking labels, but none have proved effective; among other issues, such labels have been vulnerable to hacking because of the deterministic nature of their encoding systems. But now, a team of MIT [Massachusetts Institute of Technology] researchers has come up with a kind of tiny, biodegradable tag that can be applied directly to the seeds themselves, and that provides a unique randomly created code that cannot be duplicated.

I found this picture helped me to understand how ‘tracking’ these seeds could be useful,

As a way to reduce seed counterfeiting, MIT researchers developed a silk-based tag that, when applied to seeds, provides a unique code that cannot be duplicated. Credits: Credit: Photograph courtesy of the researchers. Edited by Jose-Luis Olivares, MIT

A March 22, 2023 Massachusetts Institute of Technology (MIT) news release by David Chandler (also on EurekAlert), which originated the news item, describes the work in more detail (Note: I’m glad to see David Chandler is getting bylines again even if it’s on the EurekAlert version only),

The new system, which uses minuscule dots of silk-based material, each containing a unique combination of different chemical signatures, is described today in the journal Science Advances in a paper by MIT’s dean of engineering Anantha Chandrakasan, professor of civil and environmental engineering Benedetto Marelli, postdoc Hui Sun, and graduate student Saurav Maji.

The problem of counterfeiting is an enormous one globally, the researchers point out, affecting everything from drugs to luxury goods, and many different systems have been developed to try to combat this. But there has been less attention to the problem in the area of agriculture, even though the consequences can be severe. In sub-Saharan Africa, for example, the World Bank estimates that counterfeit seeds are a significant factor in crop yields that average less than one-fifth of the potential for maize, and less than one-third for rice. 

Marelli explains that a key to the new system is creating a randomly-produced physical object whose exact composition is virtually impossible to duplicate. The labels they create “leverage randomness and uncertainty in the process of application, to generate unique signature features that can be read, and that cannot be replicated,” he says.

What they’re dealing with, Sun adds, “is the very old job of trying, basically, not to get your stuff stolen. And you can try as much as you can, but eventually somebody is always smart enough to figure out how to do it, so nothing is really unbreakable. But the idea is, it’s almost impossible, if not impossible, to replicate it, or it takes so much effort that it’s not worth it anymore.”

The idea of an “unclonable” code was originally developed as a way of protecting the authenticity of computer chips, explains Chandrakasan, who is the Vannevar Bush Professor of Electrical Engineering and Computer Science. “In integrated circuits, individual transistors have slightly different properties coined device variations,” he explains, “and you could then use that variability and combine that variability with higher-level circuits to create a unique ID for the device. And once you have that, then you can use that unique ID as a part of a security protocol. Something like transistor variability is hard to replicate from device to device, so that’s what gives it its uniqueness, versus storing a particular fixed ID.” The concept is based on what are known as physically unclonable functions, or PUFs.

The team decided to try to apply that PUF principle to the problem of fake seeds, and the use of silk proteins was a natural choice because the material is not only harmless to the environment but also classified by the Food and Drug Administration in the “generally recognized as safe” category, so it requires no special approval for use on food products.

“You could coat it on top of seeds,” Maji says, “and if you synthesize silk in a certain way, it will also have natural random variations. So that’s the idea, that every seed or every bag could have a unique signature.”

Developing effective secure system solutions have long been one of Chandrakasan’s specialties, while Marelli has spent many years developing systems for applying silk coatings to a variety of fruits, vegetables, and seeds, so their collaboration was a natural for developing such a silk-based coding system towards enhanced security. 

“The challenge was what type of form factor to give to silk,” Sun says, “so that it can be fabricated very easily.” They developed a simple drop-casting approach that produces tags that are less than one-tenth of an inch in diameter. The second challenge was to develop “a way where we can read the uniqueness, in also a very high throughput and easy way.”

For the unique silk-based codes, Marelli says, “eventually we found a way to add a color to these microparticles [emphasis mine] so that they assemble in random structures.” The resulting unique patterns can be read out not only by a spectrograph or a portable microscope, but even by an ordinary cellphone camera with a macro lens. This image can be processed locally to generate the PUF code and then sent to the cloud and compared with a secure database to ensure the authenticity of the product. “It’s random so that people cannot easily replicate it,” says Sun. “People cannot predict it without measuring it.”

And the number of possible permutations that could result from the way they mix four basic types of colored silk nanoparticles [emphasis mine] is astronomical. “We were able to show that with a minimal amount of silk, we were able to generate 128 random bits of security,” Maji says. “So this gives rise to 2 to the power 128 possible combinations, which is extremely difficult to crack given the computational capabilities of the state-of-the-art computing systems.”

Marelli says that “for us, it’s a good test bed in order to think out-of-the-box, and how we can have a path that somehow is more democratic.” In this case, that means “something that you can literally read with your phone, and you can fabricate by simply drop casting a solution, without using any advanced manufacturing technique, without going in a clean room.”

Some additional work will be needed to make this a practical commercial product, Chandrakasan says. “There will have to be a development for at-scale reading” via smartphones. “So. that’s clearly a future opportunity.” But the principle now shows a clear path to the day when “a farmer could at least, maybe not every seed, but could maybe take some random seeds in a particular batch and verify them,” he says.

I’ve looked at the paper (very quickly) and haven’t spotted any mention of silk nanoparticles. It’s all silk microparticles.

Here’s a link to and a citation for the paper,

Integrating biopolymer design with physical unclonable functions for anticounterfeiting and product traceability in agriculture by Hui Sun, Saurav Maji, Anantha P. Chandrakasan, and Benedetto Marelli. Science Advances 22 Mar 2023 Vol 9, Issue 12 DOI: 10.1126/sciadv.adf1978

This paper is open access.

Food sensor made from of silk microneedles looks like velco

These sensors really do look like velcro,

The Velcro-like food sensor, made from an array of silk microneedles, can pierce through plastic packaging to sample food for signs of spoilage and bacterial contamination. Image: Felice Frankel

A September 9, 2020 news item on Nanowerk announces some research from the Massachusetts Institute (MIT),

MIT engineers have designed a Velcro-like food sensor, made from an array of silk microneedles, that pierces through plastic packaging to sample food for signs of spoilage and bacterial contamination.

The sensor’s microneedles are molded from a solution of edible proteins found in silk cocoons, and are designed to draw fluid into the back of the sensor, which is printed with two types of specialized ink. One of these “bioinks” changes color when in contact with fluid of a certain pH range, indicating that the food has spoiled; the other turns color when it senses contaminating bacteria such as pathogenic E. coli.

A Sept. 9, 2020 MIT news release (also on EurekAlert), which originated the news item, delves further into the research,

The researchers attached the sensor to a fillet of raw fish that they had injected with a solution contaminated with E. coli. After less than a day, they found that the part of the sensor that was printed with bacteria-sensing bioink turned from blue to red — a clear sign that the fish was contaminated. After a few more hours, the pH-sensitive bioink also changed color, signaling that the fish had also spoiled.

The results, published today in the journal Advanced Functional Materials, are a first step toward developing a new colorimetric sensor that can detect signs of food spoilage and contamination.

Such smart food sensors might help head off outbreaks such as the recent salmonella contamination in onions and peaches. They could also prevent consumers from throwing out food that may be past a printed expiration date, but is in fact still consumable.

“There is a lot of food that’s wasted due to lack of proper labeling, and we’re throwing food away without even knowing if it’s spoiled or not,” says Benedetto Marelli, the Paul M. Cook Career Development Assistant Professor in MIT’s Department of Civil and Environmental Engineering. “People also waste a lot of food after outbreaks, because they’re not sure if the food is actually contaminated or not. A technology like this would give confidence to the end user to not waste food.”

Marelli’s co-authors on the paper are Doyoon Kim, Yunteng Cao, Dhanushkodi Mariappan, Michael S. Bono Jr., and A. John Hart.

Silk and printing

The new food sensor is the product of a collaboration between Marelli, whose lab harnesses the properties of silk to develop new technologies, and Hart, whose group develops new manufacturing processes.

Hart recently developed a high-resolution floxography technique, realizing microscopic patterns that can enable low-cost printed electronics and sensors. Meanwhile, Marelli had developed a silk-based microneedle stamp that penetrates and delivers nutrients to plants. In conversation, the researchers wondered whether their technologies could be paired to produce a printed food sensor that monitors food safety.

“Assessing the health of food by just measuring its surface is often not good enough. At some point, Benedetto mentioned his group’s microneedle work with plants, and we realized that we could combine our expertise to make a more effective sensor,” Hart recalls.

The team looked to create a sensor that could pierce through the surface of many types of food. The design they came up with consisted of an array of microneedles made from silk.

“Silk is completely edible, nontoxic, and can be used as a food ingredient, and it’s mechanically robust enough to penetrate through a large spectrum of tissue types, like meat, peaches, and lettuce,” Marelli says.

A deeper detection

To make the new sensor, Kim first made a solution of silk fibroin, a protein extracted from moth cocoons, and poured the solution into a silicone microneedle mold. After drying, he peeled away the resulting array of microneedles, each measuring about 1.6 millimeters long and 600 microns wide — about one-third the diameter of a spaghetti strand.

The team then developed solutions for two kinds of bioink — color-changing printable polymers that can be mixed with other sensing ingredients. In this case, the researchers mixed into one bioink an antibody that is sensitive to a molecule in E. coli. When the antibody comes in contact with that molecule, it changes shape and physically pushes on the surrounding polymer, which in turn changes the way the bioink absorbs light. In this way, the bioink can change color when it senses contaminating bacteria.

The researchers made a bioink containing antibodies sensitive to E. coli, and a second bioink sensitive to pH levels that are associated with spoilage. They printed the bacteria-sensing bioink on the surface of the microneedle array, in the pattern of the letter “E,” next to which they printed the pH-sensitive bioink, as a “C.” Both letters initially appeared blue in color.

Kim then embedded pores within each microneedle to increase the array’s ability to draw up fluid via capillary action. To test the new sensor, he bought several fillets of raw fish from a local grocery store and injected each fillet with a fluid containing either E. coli, Salmonella, or the fluid without any contaminants. He stuck a sensor into each fillet. Then, he waited.

After about 16 hours, the team observed that the “E” turned from blue to red, only in the fillet contaminated with E. coli, indicating that the sensor accurately detected the bacterial antigens. After several more hours, both the “C” and “E” in all samples turned red, indicating that every fillet had spoiled.

The researchers also found their new sensor indicates contamination and spoilage faster than existing sensors that only detect pathogens on the surface of foods.

“There are many cavities and holes in food where pathogens are embedded, and surface sensors cannot detect these,” Kim says. “So we have to plug in a bit deeper to improve the reliability of the detection. Using this piercing technique, we also don’t have to open a package to inspect food quality.”

The team is looking for ways to speed up the microneedles’ absorption of fluid, as well as the bioinks’ sensing of contaminants. Once the design is optimized, they envision the sensor could be used at various stages along the supply chain, from operators in processing plants, who can use the sensors to monitor products before they are shipped out, to consumers who may choose to apply the sensors on certain foods to make sure they are safe to eat.

Here’s a link to and a citation for the paper,

A Microneedle Technology for Sampling and Sensing Bacteria in the Food Supply Chain by Doyoon Kim, Yunteng Cao, Dhanushkodi Mariappan, Michael S. Bono Jr., A. John Hart, Benedetto Marelli. DOI: https://doi.org/10.1002/adfm.202005370 First published: 09 September 2020

This paper is behind a paywall.

Preprogramming silk protein-based materials

A new material based on silk proteins has been developed at Tufts University (US), according to a Dec. 26, 2016 news item on ScienceDaily,

Tufts University engineers have created a new format of solids made from silk protein that can be preprogrammed with biological, chemical, or optical functions, such as mechanical components that change color with strain, deliver drugs, or respond to light, according to a paper published online this week [Dec. 26 -30, 2016] in Proceedings of the National Academy of Sciences (PNAS).

Caption: This image shows examples of engineered 3-D silk constructs. Credit: Silklab, Department of Biomedical Engineering, School of Engineering, Tufts University

A Dec. 26, 2016 Tufts University news release (also on EurekAlert), which originated the news item, describes the research in more detail,

Using a water-based fabrication method based on protein self-assembly, the researchers generated three-dimensional bulk materials out of silk fibroin, the protein that gives silk its durability. Then they manipulated the bulk materials with water-soluble molecules to create multiple solid forms, from the nano- to the micro-scale, that have embedded, pre-designed functions.

For example, the researchers created a surgical pin that changes color as it nears its mechanical limits and is about to fail, functional screws that can be heated on demand in response to infrared light, and a biocompatible component that enables the sustained release of bioactive agents, such as enzymes.

Although more research is needed, additional applications could include new mechanical components for orthopedics that can be embedded with growth factors or enzymes, a surgical screw that changes color as it reaches its torque limits, hardware such as nuts and bolts that sense and report on the environmental conditions of their surroundings, or household goods that can be remolded or reshaped.

Silk’s unique crystalline structure makes it one of nature’s toughest materials. Fibroin, an insoluble protein found in silk, has a remarkable ability to protect other materials while being fully biocompatible and biodegradable.

Here’s a link to and a citation for the paper,

Programming function into mechanical forms by directed assembly of silk bulk materials by Benedetto Marelli, Nereus Patel, Thomas Duggan, Giovanni Perotto, Elijah Shirman, Chunmei Li, David L. Kaplan, and Fiorenzo G. Omenetto. PNAS 10.1073/pnas.1612063114 December 27, 2016

This paper is behind a paywall.

Coat fruit with silk to keep it fresh

A May 6, 2016 news item on ScienceDaily describes a way to keep fruit fresh without refrigeration,

Half of the world’s fruit and vegetable crops are lost during the food supply chain, due mostly to premature deterioration of these perishable foods, according to the Food and Agriculture Organization (FAO) of the United Nations.

Tufts University biomedical engineers have demonstrated that fruits can stay fresh for more than a week without refrigeration if they are coated in an odorless, biocompatible silk solution so thin as to be virtually invisible. The approach is a promising alternative for preservation of delicate foods using a naturally derived material and a water-based manufacturing process.

A May 6, 2016 Tufts University news release (also on EurekAlert), which originated the news item, describes the work,

Silk’s unique crystalline structure makes it one of nature’s toughest materials. Fibroin, an insoluble protein found in silk, has a remarkable ability to stabilize and protect other materials while being fully biocompatible and biodegradable.

For the study, researchers dipped freshly picked strawberries in a solution of 1 percent silk fibroin protein; the coating process was repeated up to four times.  The silk fibroin-coated fruits were then treated for varying amounts of time with water vapor under vacuum (water annealed) to create varying percentages of crystalline beta-sheets in the coating. The longer the exposure, the higher the percentage of beta-sheets and the more robust the fibroin coating. The coating was 27 to 35 microns thick.

The strawberries were then stored at room temperature. Uncoated berries were compared over time with berries dipped in varying numbers of coats of silk that had been annealed for different periods of time. At seven days, the berries coated with the higher beta-sheet silk were still juicy and firm while the uncoated berries were dehydrated and discolored.

Tests showed that the silk coating prolonged the freshness of the fruits by slowing fruit respiration, extending fruit firmness and preventing decay.

“The beta-sheet content of the edible silk fibroin coatings made the strawberries less permeable to carbon dioxide and oxygen. We saw a statistically significant delay in the decay of the fruit,” said senior and corresponding study author Fiorenzo G. Omenetto, Ph.D. Omenetto is the Frank C. Doble Professor in the Department of Biomedical Engineering and also has appointments in the Department of Electrical Engineering and in the Department of Physics in the School of Arts and Sciences.

Similar experiments were performed on bananas, which, unlike strawberries, are able to ripen after they are harvested. The silk coating decreased the bananas’ ripening rate compared with uncoated controls and added firmness to the fruit by preventing softening of the peel.

The thin, odorless silk coating did not affect fruit texture.  Taste was not studied.

“Various therapeutic agents could be easily added to the water-based silk solution used for the coatings, so we could potentially both preserve and add therapeutic function to consumable goods without the need for complex chemistries,” said the study’s first author, Benedetto Marelli, Ph.D., formerly a post-doctoral associate in the Omenetto laboratory and now at MIT.

Here’s a link to and a citation for the paper,

Silk Fibroin as Edible Coating for Perishable Food Preservation by B. Marelli, M. A. Brenckle, D. L. Kaplan & F. G. Omenetto. Scientific Reports 6, Article number: 25263 (2016) doi:10.1038/srep25263 Published online: 06 May 2016

This is an open access paper.

Silk inks containing enzymes, antibiotics, antibodies, nanoparticles, and growth factors

There’s an almost euphoric tone to a June 16, 2015 Tufts University news release (also on EurekAlert) about research which has resulted in the ability to print silk-based inks,

Silk inks containing enzymes, antibiotics, antibodies, nanoparticles and growth factors could turn inkjet printing into a new, more effective tool for therapeutics, regenerative medicine and biosensing, according to new research led by Tufts University  biomedical engineers and published June 16 [2015] in the journal Advanced Materials online in advance of print.

Until now, heat used in the inkjet printing process made using silk a challenge (as it does for cellulose nanomaterials used in 3D printers, noted in my June 17, 2015 posting), from the Tufts news release,

Inkjet printing is one of the most immediate and accessible forms of printing technology currently available, according to the researchers, and ink-jet printing of biomolecules has been previously proposed by scientists. However, the heat-sensitive nature of these unstable compounds means printed materials rapidly lose functionality, limiting their use.

Enter purified silk protein, or fibroin, which offers intrinsic strength and protective properties that make it well-suited for a range of biomedical and optoelectronic applications. This natural polymer is an ideal “cocoon” that can stabilize compounds such as enzymes, antibodies and growth factors while lending itself to many different mechanically robust formats, said Fiorenzo Omenetto, Ph.D., senior author on the paper and associate dean for research and Frank C. Doble Professor of Engineering at Tufts School of Engineering.

“We thought that if we were able to develop an inkjet-printable silk solution, we would have a universal building block to generate multiple functional printed formats that could lead to a wide variety of applications in which inks remain active over time,” he said.

By using this simple approach and starting with the same base material, the research team created and tested a “custom library” of inkjet-printable, functional silk inks doped with a variety of components:

  • Bacterial-sensing polydiacetylenes (PDAs) printed on surgical gloves; the word “contaminated” printed on the glove changed from blue to red after exposure to E. coli
  • Proteins that stimulate bone growth (BMP-2) printed on a plastic dish to test topographical control of directed tissue growth
  • Sodium ampicillin printed on a bacterial culture to test the effectiveness of a topographical distribution of the antibiotic
  • Gold nanoparticles printed on paper, for possible application in photonics and biology (e.g., color engineering, surface plasmon resonance based sensing and bio-imaging)
  • Enzymes printed on paper to test the ability of the ink to entrain small functional biomolecules

The researchers, who included collaborators from the University of Illinois at Urbana-Champaign, foresee wide potential for future investigation and application of this technology.

For example, Omenetto envisions more work on the bio-sensing gloves, which he says could selectively react to different pathological agents. The ability to print antibiotics in topographical patterns could address the need for “smart” bandages, where therapeutics are incorporated and delivered to match a complex injury.

The published research was restricted to one ink cartridge, but the scientists believe it could extend to multi-cartridge printing combining complex functions.

Omenetto and Kaplan are pioneers in the use of silk as an alternative to plastics. Omenetto’s 2011 TED Talk called silk a “new old material” that could have a profound impact in many technical fields.

Here’s a link to and citation for the paper,

Inkjet Printing of Regenerated Silk Fibroin: From Printable Forms to Printable Functions by Hu Tao, Benedetto marelli, Miaomiao Yang, Bo An, Serdar Onses, John A. Rogers, David L. Kaplan, & Fiorenzo G. Omenetto. Advanced Materials DOI: 10.1002/adma.201501425 First published: 16 June 2015

This article is behind a paywall.