REU student profile: Christopher Glover

Christopher Glover is a rising senior in bioengineering at the University of Missouri. He worked this summer as an REU intern in the laboratory of professor Jeff Tza-Huei Wang, who has joint appointments in mechanical engineering, biomedical engineering and oncology. The Research Experience for Undergraduates, hosted by Johns Hopkins Institute for NanoBioTechnology, attracts nearly 800 undergraduate applicants for just 10 research positions.

Christopher Glover

Christopher Glover

Christopher’s project involved a proof-of-concept experiment to test a device used to digitally sort and amplify DNA samples.

The device consists of a silicone chip imprinted with 3,000 tiny wells to contain DNA. A thermoplastic lid covers the top of the chip to keep the DNA in place in the wells. After a segment of DNA is added to the chip, the number of copies of that DNA segment is amplified using a device called a thermal cycler. “The goal is to either get zero or one copy of the DNA segment in each well, which makes the device “digital,” he said.

“We aren’t concerned about the type of DNA we are amplifying but just to see if it will work,” Christopher said. “This could be used for medical screening where a specific allele could be detected within a gene to see if someone is more susceptible to getting a disease,” he said.

Christopher said that working in the Wang lab has helped him learn much more about nanotechnology than he had previously known. His future plans include earning a PhD in biomedical engineering.

For all press inquiries regarding INBT, its faculty and programs, contact Mary Spiro, mspiro@jhu.edu or 410-516-4802.

REU student profile: Rebecca Majewski

DNA, the genetic sequence that tells cells what proteins to manufacture, typically resides inside the nucleus of a cell, but not always. Rebecca Majewski is studying the uptake of DNA into cell nuclei using a different polymer chains. Rebecca is a rising senior in BioMolecular Engineering from the Milwaukee School of Engineering and is working as a summer intern in the Johns Hopkins Institute for Nanobiotechnology’s REU program.

“We are interested in how much of the DNA with the polyplex can get into the nucleus,” she said, but explains that DNA associated outside of the nucleus can cause false higher measurements.

Rebecca Majewski. Photo by Mary Spiro

Rebecca Majewski. Photo by Mary Spiro

Rebecca is washing the cells with the nuclei to get rid of DNA outside the nucleus and then comparing the measurement of uptake of the DNA by the cell versus the measurement of the uptake of DNA by the nucleus.

“We are interested in what DNA gets inserted into the nucleus because that is what is ultimately expressed. It is important to find out how much makes it to the final destination and then is expressed. The goal of this work is to test different polymer chains to see which one actually does the better job of getting the DNA into the nucleus,” she said.

Rebecca works alongside PhD students and postdoctoral fellows in the biomedical engineering lab of Jordan Green lab at the Johns Hopkins School of Medicine. She says she highly values the opportunity for a research experience through INBT’s REU because her undergraduate institution does not train graduate students.

For all press inquiries regarding INBT, its faculty and programs, contact Mary Spiro, mspiro@jhu.edu or 410-516-4802.

REU student profile: Ian Reucroft

Sitting at what looks like a pottery wheeled turned on its side, Ian Reucroft is using a method called electrospinning to create a nano-scale polymer fiber embedded with a drug that encourages nerve growth. The strand is barely visible to the eye, but the resulting fibers resemble spider web.

Ian Reucroft, a rising junior in Biomedical Engineering at Rutgers University, is working in the medical school campus laboratory of Hai-Quan Mao, professor of materials sciences and engineering at Johns Hopkins University. He is part of Johns Hopkins Institute for NanoBioTechnology’s summer REU, or research experience for undergraduates program.

Ian Reucroft in the Mao lab. Photo by Mary Spiro.

Ian Reucroft in the Mao lab. Photo by Mary Spiro.

“We are developing a material to help regrow nerves, either in central or peripheral nervous systems,” said Ian. One method of doing that he explained is to make nanofibers and incorporating a drug into those fibers, drugs that promote neuronic growth or cell survival or various other beneficial qualities. The Mao lab is looking into a relatively new and not well-studied drug called Sunitinib that promotes neuronal survival.

“We make a solution of the component to make the fiber, which is this case is polylactic acid (PLA), and the drug, which I have to dissolve into the solution,” Ian said. Although the drug seems to remain stable in solution, one of the challenges Ian has faced has been improving the distribution of the drug along the fiber.

This is Ian’s first experience with electrospinning but not his first time conducting research. He plans to pursue a PhD in biomedical engineering and remain in academia.

For all press inquiries regarding INBT, its faculty and programs, contact Mary Spiro, mspiro@jhu.edu or 410-516-4802.

Konstantopoulos to present distinguished lecture on tumor cell migration

Biomedical Engineering 8 5 x 11 4-7Biomedical Engineering 8 5 x 11 4-7Biomedical Engineering 8 5 x 11 4-7Professor and Chair of the Department of Chemical and Biomolecular Engineering, Konstantinos Konstantopoulos will present a distinguished lecture for the Department of Biomedical Engineering on Monday, April 7 at 4 p.m. in the Mason Hall Auditorium on the Homewood campus of Johns Hopkins University.  His talk. “Joining Forces with Biology: A Bioengineering Perspective on Tumor Cell Migration,” will reveal some of his laboratory’s current findings on metastasis. The talk is free and open the Johns Hopkins University community. Refreshments follow the lecture.

Here’s the abstract of his talk:

“Understanding the mechanisms of cell migration is a fundamental question in cell, developmental and cancer biology. Unraveling key, physiologically relevant motility mechanisms is also crucial for developing technologies that can control, manipulate, promote or stop cell migration in vivo. Much of what we know about the mechanisms of cell migration stems from in vitro studies using two-dimensional (2D) surfaces. Cell locomotion in 2D is driven by cycles of actin protrusion, integrin-mediated adhesion and myosin-dependent contraction. A major pitfall of 2D assays is that they fail to account for the physical confinement that cells  encounter within the physiological tissue environment. The seminar will challenge the conventional wisdom regarding cell motility mechanisms, and show that migration through physically constricted spaces does not require beta1 integrin dependent adhesion or myosin contractility. Importantly, confined migration persists even when filamentous actin is disrupted. This seminar will also discuss a novel mechanism of confined cell migration based on an osmotic en

Nanotechnology for gene therapy

Editor’s Note: The following is a summary of one of the talks from the 2013 Nano-bio Symposium hosted by Johns Hopkins Institute for NanoBioTechnology held May 17. This summary was written by Randall Meyer, a doctoral candidate in the biomedical engineering and a member of the Cancer Nanotechnology Training Center. Look for other symposium summaries on the INBT blog.

One of the key features of nanotechnology is its wide range of applicability across multiple biological scenarios ranging from gene therapy to immune system modulation. Jordan Green, an assistant professor of Biomedical Engineering at Johns Hopkins University, summarized some of the fascinating applications of nanotechnology on which his laboratory has been working. Green is an INBT affiliated faculty member.

One of the Green lab projects involves the design and implementation of nanoparticle based vectors for delivery of genetic material to the cell. Green demonstrated how these particles could be used to deliver DNA and induce expression of a desired gene, or small interfering RNA (siRNA) to silence the expression of a target gene. These genetic therapeutics are being developed to target a wide variety of retinal diseases and cancers.

Jordan Green (Photo by Marty Katz)

Jordan Green (Photo by Marty Katz)

 

As opposed to viral based vectors for gene therapy, nonviral vectors such as nanoparticles are safer, more flexible in their range of cellular targets, and can carry larger cargoes than viruses, Green explained.

 

Another project in the Green lab involves the development of micro and nano dimensional artificial antigen presenting cells (aAPCs) for cancer immunotherapy. These aAPCs mimic the natural signals that killer T-cells receive when there is an invader (bacteria, virus, cancer cell, etc.) in the body. The Green lab is currently working with these particles to stimulate the immune system to fight melanoma.

 

Green Group

Nanoscale scaffolds spur stem cells to cartilage repair

Scanning electron micrographs showing chondroitin sulfate (CS) and poly(vinyl alcohol)-methacrylate (PVA) nanofibers after electrospinning and processing to render the nanofiber scaffolds water-insoluble. Image by Jeannine Coburn/JHU first appeared in PNAS.

A spun 3-D scaffold of nanofibers did a better job of producing larger quantities of and a more durable type of the cartilage protein than flat, 2-D sheets of fibers did. 

Johns Hopkins tissue engineers have used tiny, artificial fiber scaffolds thousands of times smaller than a human hair to help coax stem cells into developing into cartilage, the shock-absorbing lining of elbows and knees that often wears thin from injury or age.

Reporting online June 4 in the Proceedings of the National Academy of Sciences, investigators say they have produced an important component of cartilage in both laboratory and animal models. While the findings are still years away from use in people, the researchers say the results hold promise for devising new techniques to help the millions who endure joint pain.

“Joint pain affects the quality of life of millions of people. Rather than just patching the problem with short-term fixes, like surgical procedures such as microfracture, we’re building a temporary template that mimics the cartilage cell’s natural environment, and taking advantage of nature’s signals to biologically repair cartilage damage,” says Jennifer Elisseeff, Ph.D., Jules Stein Professor of Ophthalmology and director of the Translational Tissue Engineering Center at the Johns Hopkins University School of Medicine. Elisseeff is also an affiliated faculty member of Johns Hopkins Institute for NanoBioTechnology.

Unlike skin, cartilage can’t repair itself when damaged. For the last decade, Elisseeff’s team has been trying to better understand the development and growth of cartilage cells called chondrocytes, while also trying to build scaffolding that mimics the cartilage cell environment and generates new cartilage tissue. This environment is a three-dimensional mix of protein fibers and gel that provides support to connective tissue throughout the body, as well as physical and biological cues for cells to grow and differentiate.

In the laboratory, the researchers created a nanofiber-based network using a process called electrospinning, which entails shooting a polymer stream onto a charged platform, and added chondroitin sulfate — a compound commonly found in many joint supplements — to serve as a growth trigger. After characterizing the fibers, they made a number of different scaffolds from either spun polymer or spun polymer plus chondroitin. They then used goat bone marrow-derived stem cells (a widely used model) and seeded them in various scaffolds to see how stem cells responded to the material.

Elisseeff and her team watched the cells grow and found that compared to cells growing without scaffold, these cells developed into more voluminous, cartilage-like tissue.

“The nanofibers provided a platform where a larger volume of tissue could be produced,” says Elisseeff, adding that three-dimensional nanofiber scaffolds were more useful than the more common nanofiber sheets for studying cartilage defects in humans.

The investigators then tested their system in an animal model. They implanted the nanofiber scaffolds into damaged cartilage in the knees of rats, and compared the results to damaged cartilage in knees left alone.

They found that the use of the nanofiber scaffolds improved tissue development and repair as measured by the production of collagen, a component of cartilage. The nanofiber scaffolds resulted in greater production of a more durable type of collagen, which is usually lacking in surgically repaired cartilage tissue. In rats, for example, they found that the limbs with damaged cartilage treated with nanofiber scaffolds generated a higher percentage of the more durable collagen (type 2) than those damaged areas that were left untreated.

“Whereas scaffolds are generally pretty good at regenerating cartilage protein components in cartilage repair, there is often a lot of scar tissue-related type 1 collagen produced, which isn’t as strong,” says Elisseeff. “We found that our system generated more type 2 collagen, which ensures that cartilage lasts longer.”

“Creating a nanofiber network that enables us to more equally distribute cells and more closely mirror the actual cartilage extracellular environment are important advances in our work and in the field. These results are very promising,” she says.

Other authors included Jeannine M. Coburn, Matthew Gibson, Sean Monagle and Zachary Patterson, all from The Johns Hopkins University.

From a press release by Audrey Huang.

 

It’s a small world: Micro/nanotechnology in regenerative medicine and cancer

Sageeta Bhatia

Nanotechnology, regenerative medicine and cancer will be the topic of a special biomedical engineering seminar on March 6 at 3 p.m. in the Darner Conference Room, Ross Building, Room G007 at the Johns Hopkins School of Medicine. Speaker Sangeeta Bhatia, MD, PhD, director, of the Laboratory for Multiscale Regenerative Technologies at Massachusetts Institute of Technology will present “It’s a small world: Micro/Nanotechnology in Regenerative Medicine and Cancer. ” She will discuss the role of micro and nanotechnology for mimicking, monitoring and perturbing the tissue microenvironment.

“I will present our work on reconstructing normal liver microenvironments using microtechnology, biomaterials and induced pluripotent stem cells as well as our work on normalizing diseased cancer microenvironments using both inorganic and organic nano materials,” Bhatia noted in an announcement.  Bhatia is a professor of Health Sciences and Technology and professor of Electrical Engineering and Computer Science at MIT.

The talk is hosted by associate professor of Materials Science and Engineering and affiliated faculty member of the Institute for NanoBioTechnology Hai-Quan Mao. The event is free and open to the Johns Hopkins Community. Refreshments will be served.

 

 

Engineered hydrogel helps grow new, scar-free skin

In early testing, this hydrogel, developed by Johns Hopkins researchers, helped improve healing in third-degree burns. Photo by Will Kirk/HomewoodPhoto.jhu.edu

Johns Hopkins researchers have developed a jelly-like material and wound treatment method that, in early experiments on skin damaged by severe burns, appeared to regenerate healthy, scar-free tissue.

In the Dec. 12-16 online Early Edition of Proceedings of the National Academy of Sciences, the researchers reported their promising results from mouse tissue tests. The new treatment has not yet been tested on human patients. But the researchers say the procedure, which promotes the formation of new blood vessels and skin, including hair follicles, could lead to greatly improved healing for injured soldiers, home fire victims and other people with third-degree burns.

The treatment involved a simple wound dressing that included a specially designed hydrogel—a water-based, three-dimensional framework of polymers. This material was developed by researchers at Johns Hopkins’ Whiting School of Engineering, working with clinicians at the Johns Hopkins Bayview Medical Center Burn Center and the Department of Pathology at the university’s School of Medicine.

Third-degree burns typically destroy the top layers of skin down to the muscle. They require complex medical care and leave behind ugly scarring. But in the journal article, the Johns Hopkins team reported that their hydrogel method yielded better results. “This treatment promoted the development of new blood vessels and the regeneration of complex layers of skin, including hair follicles and the glands that produce skin oil,” said Sharon Gerecht, an assistant professor of chemical and biomolecular engineering who was principal investigator on the study.

Guoming Sun, left, a postdoctoral fellow, and Sharon Gerecht, an assistant professor of chemical and biomolecular engineering, helped develop a hydrogel that improved burn healing in early experiments. Photo by Will Kirk/HomewoodPhoto.jhu.edu

Gerecht said the hydrogel could form the basis of an inexpensive burn wound treatment that works better than currently available clinical therapies, adding that it would be easy to manufacture on a large scale. Gerecht suggested that because the hydrogel contains no drugs or biological components to make it work, the Food and Drug Administration would most likely classify it as a device. Further animal testing is planned before trials on human patients begin. But Gerecht said, “It could be approved for clinical use after just a few years of testing.”

John Harmon, a professor of surgery at the Johns Hopkins School of Medicine and director of surgical research at Bayview, described the mouse study results as “absolutely remarkable. We got complete skin regeneration, which never happens in typical burn wound treatment.”

If the treatment succeeds in human patients, it could address a serious form of injury. Harmon, a coauthor of the PNAS journal article, pointed out that 100,000 third-degree burns are treated in U. S. burn centers like Bayview every year. A burn wound dressing using the new hydrogel could have enormous potential for use in applications beyond common burns, including treatment of diabetic patients with foot ulcers, Harmon said.

Guoming Sun, Gerecht’s Maryland Stem Cell Research Postdoctoral Fellow and lead author on the paper, has been working with these hydrogels for the last three years, developing ways to improve the growth of blood vessels, a process called angiogenesis. “Our goal was to induce the growth of functional new blood vessels within the hydrogel to treat wounds and ischemic disease, which reduces blood flow to organs like the heart,” Sun said. “These tests on burn injuries just proved its potential.”

Gerecht says the hydrogel is constructed in such a way that it allows tissue regeneration and blood vessel formation to occur very quickly. “Inflammatory cells are able to easily penetrate and degrade the hydrogel, enabling blood vessels to fill in and support wound healing and the growth of new tissue,” she said. For burns, the faster this process occurs, Gerecht added, the less there is a chance for scarring.

Originally, her team intended to load the gel with stem cells and infuse it with growth factors to trigger and direct the tissue development. Instead, they tested the gel alone. “We were surprised to see such complete regeneration in the absence of any added biological signals,” Gerecht said.

Sun added, “Complete skin regeneration is desired for various wound injuries. With further fine-tuning of these kinds of biomaterial frameworks, we may restore normal skin structures for other injuries such as skin ulcers.”

Gerecht and Harmon say they don’t fully understand how the hydrogel dressing is working. After it is applied, the tissue progresses through the various stages of wound repair, Gerecht said. After 21 days, the gel has been harmlessly absorbed, and the tissue continues to return to the appearance of normal skin.

The hydrogel is mainly made of water with dissolved dextran—a polysaccharide (sugar molecule chains). “It also could be that the physical structure of the hydrogel guides the repair,” Gerecht said. Harmon speculates that the hydrogel may recruit circulating bone marrow stem cells in the bloodstream. Stem cells are special cells that can grow into practically any sort of tissue if provided with the right chemical cue. “It’s possible the gel is somehow signaling the stem cells to become new skin and blood vessels,” Harmon said.

Additional co-authors of the study included Charles Steenbergen, a professor in the Department of Pathology; Karen Fox-Talbot, a senior research specialist from the Johns Hopkins School of Medicine; and physician researchers Xianjie Zhang, Raul Sebastian and Maura Reinblatt from the Department of Surgery and Hendrix Burn and Wound Lab. From the Whiting School’s Department of Chemical and Biomolecular Engineering, other co-authors were doctoral students Yu-I (Tom) Shen and Laura Dickinson, who is a Johns Hopkins Institute for NanoBioTechnology (INBT) National Science Foundation IGERT fellow. Gerecht is an affiliated faculty member of INBT.

The work was funded in part by the Maryland Stem Cell Research Fund Exploratory Grant and Postdoctoral Fellowship and the National Institutes of Health.

The Johns Hopkins Technology Transfer staff has filed a provisional patent application to protect the intellectual property involved in this project.

Related links:

Sharon Gerecht’s Lab

Johns Hopkins Burn Center

Johns Hopkins Institute for NanoBioTechnology

 

Story by Mary Spiro

Engineers put a new ‘twist’ on lab-on-a-chip

Close-up of a cylindrically-shaped microfluidic device with two fluorescent solutions flowing through. Reproduced with permission from Nature Communications.

A leaf works something like a miniature laboratory. While the pores on the leaf surface allow it to channel nutrients in and waste products away from a plant, part of a leaf’s function also lies in its ability to curl and twist. Engineers use polymers to create their own mini-labs, devices called “labs-on-a-chip,” which have numerous applications in science, engineering and medicine. The typical flat, lab on a chip, or microfluidic device, resembles an etched microscopy cover slip with channels and grooves.

But what if you could get that flat lab-on-a-chip to self-assemble into a curve, mimicking the curl, twist or spiral of a leaf? Mustapha Jamal, a PhD student and IGERT fellow from Johns Hopkins Institute for NanoBioTechnology, has created a way to make that so.

Jamal is the lead author on “Differentially photo-crosslinked polymers enable self-assembling microfluidics,” published November 8, 2011 in Nature Communications. Along with principle investigator David Gracias, associate professor of Chemical and Biomolecular Engineering in the Whiting School of Engineering, and fellow graduate student Aasiyeh Zarafshar, Jamal has developed, for the first time, a method for creating three-dimensional lab-on-a-chip devices that can curl and twist.

The process involves shining ultraviolet (UV) light on a film of a substance called SU-8. Film areas closer to the light source become more heavily crosslinked than layers beneath, which on solvent conditioning creates a stress gradient.

Immersing the film in water causes the film to curl. Immersion in organic solvents like acetone causes the film to flatten. The curling and flattening can be reversed. The result, Jamal said, is the “self-assembly of intricate 3D devices that contain microfluidic channels.” This simple method, he added, can “program 2D polymeric (SU-8) films such that they spontaneously and reversibly curve into intricate 3D geometries including cylinders, cubes and corrugated sheets.”

Members of the Gracias lab have previously created curving and folding polymeric films consisting of two different materials. This new method achieves a stress gradient along the thickness of a single substance. “This provides considerable flexibility in the type and extent of curvature that can be created by varying the intensity and direction of exposure to UV light,” Gracias said.

Gracias explained that the method works with current protocols and materials for fabricating flat microfluidic devices. For example, one can design a 2D film with one type of lab-on-a-chip network, and then use their method to shape it into another geometry, also with microfluidic properties.

Fluorescent image of curved, self-assembled microfluidic device. Reproduced with permission from Nature Communications.

“Since our approach is compatible with planar lithography methods, we can also incorporate optical elements such as split ring resonators that have unique optical features. Alternatively, flexible electronic circuits could be incorporated and channels could be used to transport cooling fluids” Gracias said.

Tissue engineering is among the many important applications for 3D microfluidic devices, Gracias said. “Since many hydrogels can be photopolymerized, we can use the methodology of differential cross-linking to create stress gradients in these materials,” Gracias explained. “We plan to create biodegradable, vascularized tissue scaffolds using this approach.”

Link to the journal article here.

Story by Mary Spiro

 

 

Hopkins Imaging Initiative to host first annual conference

The Johns Hopkins University Imaging Initiative will host the first annual Imaging Conference, October 6, 2011 at the Turner Auditorium on the medical campus. The conference features afternoon lectures from various Hopkins faculty followed by a research poster session and happy hour. Anyone interested in imaging is welcome to attend.

Speakers include Elliot McVeigh, director of the Department of Biomedical Engineering; Elliot Fishman, MD, director of diagnostic imaging at body CT at Johns Hopkins Hospital; Jerry Prince, the William B. Kouwenhoven Professor of Electrical and Computer Engineering at the Whiting School of Engineering; Xingde Li, associate professor of biomedical engineering and head of the Laboratory of Biophotonics Imaging and Therapy at the Whiting School; Peter van Zijl, professor of radiology at the school of medicine and director of the F.M. Kirby Research Center for Functional Brain Imaging; and several others to be announced.

Abstracts will be accepted until Sept 6 and conference registration will be accepted until October 1. For complete information about this event and to register, go to http://imaging.jhu.edu/conferences/imaging-conference-2011