Tag Archives: silk fibroin

Grow better organ-like tissues by using silkworms

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A June 6, 2024 news item on ScienceDaily describes a technique, which could lead to better organ-on-a-chip (OOC) systems,

Biomedical engineers at Duke University [North Carolina, US] have developed a silk-based, ultrathin membrane that can be used in organ-on-a-chip models to better mimic the natural environment of cells and tissues within the body. When used in a kidney organ-on-a-chip platform, the membrane helped tissues grow to recreate the functionality of both healthy and diseased kidneys.

By allowing the cells to grow closer together, this new membrane helps researchers to better control the growth and function of the key cells and tissues of any organ, enabling them to more accurately model a wide range of diseases and test therapeutics.

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A June 6, 2024 Duke University news release (also on EurekAlert), which originated the news item, describes the OOC system and the problem these researchers are seeking to solve,

Often no larger than a USB flash drive, organ-on-a-chip (OOC) systems have revolutionized how researchers study the underlying biology of the human body, whether it’s creating dynamic models of tissue structures, studying organ functions or modeling diseases. These platforms are designed to stimulate cell growth and differentiation in a way that best mimics the organ of interest. Researchers can even populate these tools with human stem cells to generate patient-specific organ models for pre-clinical studies.

But as the technology has evolved, problems in the chip’s design have also emerged –– most notably with the materials used to create the membranes that form the support structure for the specialized cells to grow on. These membranes are typically composed of polymers that don’t degrade, creating a permanent barrier between cells and tissues. While the extracellular membranes in human organs are often less than one micron thick, these polymer membranes are anywhere from 30 to 50 microns, hindering communication between cells and limiting cell growth.

“We want to handle the tissues in these chips just like a pathologist would handle biopsy samples or even living tissues from a patient, but this wasn’t possible with the standard polymer membranes because the extra thickness prevented the cells from forming structures that more closely resemble tissues in the human body,” said Samira Musah, an assistant professor of biomedical engineering and medicine at Duke. “We thought, ‘Wouldn’t it be nice if we could get a protein-based material that mimics the structure of these natural membranes and is thin enough for us to slice and study?’”

This question led Musah and George (Xingrui) Mou, a PhD student in Musah’s lab and first author on the paper, to silk fibroin, a protein created by silkworms that can be electronically spun into a membrane. When examined under a microscope, silk fibroin looks like spaghetti or a Jackson Pollock painting. Made out of long, intertwining fibers, the porous material better mimics the structure of the extracellular matrix found in human organs, and it has previously been used to create scaffolds for purposes like wound healing.

“The silk fibroin allowed us to bring the membrane thickness down from 50 microns to five or fewer, which gets us an order of magnitude closer to what you’d see in a living organism,” explained Mao.

To test this new membrane, Musah and Mao applied the material to their kidney chip models. Made out of a clear plastic and roughly the size of a quarter, this OOC platform is meant to resemble a cross section of a human kidney––specifically the glomerular capillary wall, a key structure in the organ made from clusters of blood vessels that is responsible for filtering blood.

Once the membrane was in place, the team added human induced pluripotent stem cell derivatives into the chip. They observed that these cells were able to send signals across the ultrathin membrane, which helped the cells differentiate into glomerular cells, podocytes and vascular endothelial cells. The platform also triggered the development of endothelial fenestrations in the growing tissue, which are holes that allow for the passage of fluid between the cellular layers.

By the end of the test, these different kidney cell types had assembled into a glomerular capillary wall and could efficiently filter molecules by size.

“The new microfluidic chip system’s ability to simulate in vivo-like tissue-tissue interfaces and induce the formation of specialized cells, such as fenestrated endothelium and mature glomerular podocytes from stem cells, holds significant potential for advancing our understanding of human organ development, disease progression, and therapeutic development,” said Musah.

As they continue to optimize their model, Musah and colleagues are hoping to use this technology to better understand the mechanisms behind kidney disease. Despite affecting more than 15 percent of American adults, researchers lack effective models for the disease. Patients are also often not diagnosed until the kidneys have been substantially damaged, and they are often required to undergo dialysis or receive a kidney transplant.

“Using this platform to develop kidney disease models could help us discover new biomarkers of the disease,” said Mao. “This could also be used to help us screen for drug candidates for several kidney disease models. The possibilities are very exciting.”

“This technology has implications for all organ-on-a-chip models,” said Musah. “Our tissues are made up of membranes and interfaces, so you can imagine using this membrane to improve models of other organs, like the brain, liver, and lungs, or other disease states. That’s where the power of our platform really lies.”

This work was supported by a Whitehead Scholarship in Biomedical Research, Chair’s Research Award from the Department of Medicine at Duke University, MEDx Pilot Grant on Biomechanics in Injury or Injury Repair, Burroughs Wellcome Fund PDEP Career Transition Ad Hoc Award, Duke Incubation Fund from the Duke Innovation & Entrepreneurship Initiative, Genetech Research Award, a George M. O’Brien Kidney Center Pilot Grant (P30 DK081943), an NIH [National Institutes of Health] Director’s New Innovator Grant (DP2DK138544).

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

An Ultrathin Membrane Mediates Tissue-Specific Morphogenesis and Barrier Function in a Human Kidney Chip by Xingrui Mou, Jessica Shah, Yasmin Roye, Carolyn Du, Samira Musah. Science Advances. June 4, 2024 Vol 10, Issue 23 DOI: https://doi.org/10.1126/sciadv.adn2689

This paper is open access.

Preprogramming silk protein-based materials

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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.

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

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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.

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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.