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RNA Technology Allows for Fast Vaccine Development

Vaccine syringe


5 July 2016. Engineers at Massachusetts Institute of Technology designed a process using genetic material in nanoscale particles for vaccines that in lab mice protect against dangerous diseases with a single dose. A report from the team in the lab of biology and engineering professor Daniel Anderson appears this week in Proceedings of the National Academy of Sciences (paid subscription required).

Researchers led by postdoctoral fellows Jasdave Chahal and Omar Khan, first authors of the paper, are seeking to overcome the long lead times needed to produce vaccines, as well as provide a more predictable and efficient development process. Many vaccines today use inactivated or weakened microbes, or proteins generated by pathogens that require additional compounds called adjuvants to boost their immune response. While vaccines, like any drugs, need careful testing with human clinical trials, the initial discovery tasks can add many months and years to an already extended process.

Chahal, Khan, and colleagues seek to improve the process for discovering new types of vaccines with messenger RNA as the active ingredients. Messenger RNA is genetic material related and complementary to DNA used by cells to produce the amino acids in proteins. While genetic technologies are available for making vaccines, they sometimes use delivery mechanisms like benign viruses, which while not causing disease, can generate unwanted immune responses on their own.

The MIT team developed a set of techniques for quickly producing vaccines formulated in nanoscale particles containing messenger RNA molecules. These molecules are configured into star-shaped branches called dendrimers that can be concentrated into particles about 150 nanometers in diameter; 1 nanometer equals 1 billionth of a meter. This size is about the same as many viruses, making it possible for the nanoparticles to target the same surface proteins that attract viruses.

The techniques encode messenger RNA molecules to address specific disease-causing proteins that replicate when they reach the target, where they generate antibodies or produce pathogen-fighting T-cells in the immune system. The researchers were able to use these techniques to design and produce quantities of individual vaccines for testing against specific pathogens, each in about 7 days.

The team produced with this process protective vaccines against H1N1 influenza or swine flu, Ebola, and Toxoplasma gondii parasites, single-cell organisms causing toxoplasmosis, a type of food poisoning from undercooked or contaminated meat. Vaccines for H1N1 influenza strains are produced routinely today, while Ebola vaccines are still in clinical trials, and no vaccines are yet available to protect against Toxoplasma gondii.

In lab tests, mice were given single doses of the vaccines in intramuscular injections, with comparable mice not injected. The mice were then given lethal doses of the pathogens. All mice receiving the H1N1 vaccine produced antibodies that protected against the virus, while the unvaccinated mice died within 7 days. For the Ebola virus, the team tested different dosage levels, and found a dose that generated protective T-cells against the virus. As with the H1N1 virus, mice not protected with vaccines died from the Ebola virus within 7 days.

With Toxoplasma gondii, the researchers developed a more complex vaccine that protects against different life stages of the parasite. As with the the other pathogens, mice given the vaccine survived the lethal dose of Toxoplasma gondii, while the unvaccinated mice died with 12 days. The researchers say this was the first demonstration of a protective nanoparticle vaccine against the parasite.

“This nanoformulation approach allows us to make vaccines against new diseases in only 7 days, allowing the potential to deal with sudden outbreaks or make rapid modifications and improvements,” says Anderson in an MIT statement. The university says Chahal and Khan plan to form a company to license and commercialize the technology. They also plan to extend the technology to vaccines that protect against Zika and Lyme disease.

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Independence Day 2016

Pregame parachute jump
4 July 2016. It’s the Fourth of July, a.k.a. Independence Day, in the U.S. today, so Science & Enterprise is taking the day off. We’ll resume regular posting tomorrow.

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Bacterial-Biopolymer Delivery System Designed for Vaccines

Blaine Pfeifer and Charles Jones

Blaine Pfeifer, left, and Charles Jones (Onion Studio, University at Buffalo)

1 July 2016. A biotechnology company and university lab developed and tested in mice a technique for packaging antigens that induce an immune response in vaccines, which combines E. coli bacteria and a biocompatible polymer material. The team from University at Buffalo in New York and spin-off enterprise Abcombi Biosciences published its findings in today’s issue of the journal Science Advances.

Researchers led by Buffalo engineering professor Blaine Pfeifer and Charles Jones, CEO of Abcombi Biosciences Inc., are seeking to improve the delivery of antigens, proteins that induce an immune response, in vaccines. In some cases, vaccines need an extra boost, provided by adjuvants to promote a stronger immune response. Aluminum gels and salts are sometimes used a vaccine adjuvants.

Pfeifer, Jones, and colleagues aim to reduce or eliminate the need for adjuvants by designing a technique for packaging and presenting antigens that already enhances their immune-response capabilities. The team started with cells from benign strains of E. coli bacteria, a sub-type of the bacteria that does not cause disease. Cell walls from E. coli bacteria contain lipopolysaccharides, natural compounds known to stimulate an immune response.

The bacteria cells are encased in a biocompatible material, known as poly (beta amino ester), or PBAE, a polymer tested in nanoparticles for gene and stem cell delivery. The PBAE cage has a positive charge, and the bacteria cell walls have a negative charge, which when combined, form into containers for the vaccine’s antigen payloads. The containers are small enough to be taken in by phagocytes, white blood cells in the immune system that ingest bacteria.

The Buffalo-Abcombi team designed the hybrid containers to be payload-independent, capable of carrying antigens, other proteins, or nucleic acids. As a result, say the authors, the system can pack components of a vaccine and generate antigens for more powerful payloads while being delivered.

For this study, the researchers tested the delivery system in lab mice with antigens designed to protect against pneumococcal disease that includes ear and sinus infections, as well as more serious pneumonia and blood stream infections. When compared to vaccines with conventional adjuvants, mice receiving the hybrid containers provided more antibody production and longer-term protection against pneumococcal infections. In addition, the improved vaccine performance occurs in both injected and nasal formulations. The higher performance of vaccines using hybrid containers also makes it possible to reduce the size of the doses, compared to conventional adjuvants.

Jones received his doctorate in engineering at Buffalo in 2015. He and Pfeifer are among the co-founders of Abcombi Biosciences that began in business in June 2015. The company has exclusive licensing agreements with the university to develop and commercialize their vaccine technology. While located in Buffalo, the company will incubate in Toronto, Canada at JLabs, a life sciences start-up incubator sponsored by Johnson & Johnson.

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Hundreds of U.S. Clinics Found Selling Stem Cell Services

Stem cell clinic map

Map of continental U.S. with locations of stem cell clinics. Blue stars indicate hot-spot concentrations. Click on image to view full size. (Leigh Turner and Paul Knoepfler)

1 July 2016. A comprehensive Internet search finds nearly 600 clinics in the U.S. claiming to offer stem cell treatments, many having dubious scientific or medical validity. The analysis by bioethicist Leigh Turner and stem cell researcher Paul Knoepfler is published in yesterday’s issue of the journal Cell Stem Cell.

Turner at University of Minnesota and Knoepfler at University of California in Davis, sought to better understand the extent of medical treatments marketed in the U.S. that claim to based on therapeutic benefits of stem cells. Recent news reports, including a story just last week in the New York Times, tell about catastrophic health consequences of stem-cell tourism offered by clinics in Russia, China, and South America. Turner and Knoepfler indicate that similar problems may be developing much closer to home, inside the country’s borders.

The authors conducted systematic and comprehensive key-word Internet searches, on terms such as “stem cell treatment” and “stem cell therapy,” combined with text mining and content analyses of Web sites. Their searches revealed 351 businesses offering stem cell medical services through 570 clinics. Nearly 4 in 10 of these clinics (38%) are located in either Florida or California, with 18 clinics alone in Beverly Hills and another 12 in Los Angeles. Other concentrations of clinics are in New York City, San Antonio and Austin in Texas, and Scottsdale and Phoenix in Arizona.

Many businesses with stem cell services offer treatments for a wide range of conditions, but the two leading applications are for orthopedic and pain disorders. Many clinics offer anti-aging and cosmetic applications, including face lifts and breast augmentation based on stem cells. However, the researchers also found therapies for more serious disorders including spinal cord injuries, immunological, cardiac, pulmonary, ophthalmologic, and urological diseases.

In addition, the researchers found a number of marketing pitches for stem cell treatments aimed at parents and guardians of children with inherited disorders, and caregivers of individuals with neurodegenerative conditions, such as dementia. The team found 33 claims for treatments of muscular dystrophy, as well as 9 therapies for autism and cerebral palsy. Another 27 enterprises offer treatments for Alzheimer’s disease.

Most of the businesses offering stem cell services use autologous stem cells, those taken from the patient. About 6 in 10 companies derive stem cells from adipose or fat-storing tissue, while nearly half (48%) take stem cells from bone marrow. About 1 in 5 enterprises advertised allogenic stem cells, those from external sources, mainly amniotic materials or fluids, as well as placenta tissue, and umbilical cords.

Turner and Knoepfler say a legal or regulatory analysis of these services is outside the scope of their paper, but they conclude that their results raise concerns about safety and efficacy of these treatments as well as the ethics of raising false hopes and charging high prices for therapies with dubious marketing claims. The authors note FDA issued draft guidelines for regulating products based on stem cells in October 2015, and cited news reports the agency is preparing to crack down on clinics marketing phony cures.

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Protein Biotech Lands $51.5M in First Venture Round

Finance, calculator

(stevepb, Pixabay)

30 June 2016. A new spin-off biotechnology enterprise from Harvard University developing therapies that block signaling proteins causing disease, is raising $51.5 million its first venture funding round. This first-round financing for Morphic Therapeutic in Waltham, Massachusetts is led by the venture capital arms of pharmaceutical companies GlaxoSmithKline and Pfizer.

Morphic Therapeutic creates small molecule, or low molecular weight, treatments that block the activity of integrins, a class of proteins in humans and other animals that attach cell skeletons to the extracellular matrix, the network of molecules providing structural and biochemical support for cells. Integrins provide signaling pathways going into the cell from outside, and out of the cell from inside, acting as receptors for binding molecules affecting cell activities, as well as other proteins. Many biological processes function normally with integrins, but when integrins send aberrant signals, a number of diseases can result.

Timothy Springer, an immunologist and biophysicist at Harvard Medical School and Boston Children’s Hospital, has studied integrins since the 1980s. His lab’s research led to early treatments — administered with injections — already approved to address integrins associated with a number of diseases: multiple sclerosis, ulcerative colitis, Crohn’s disease, plaque psoriasis, acute coronary syndrome, and complications during procedures implanting a stent to open arteries.

Springer founded Morphic Therapeutic to bring to market more recent research with small molecule compounds that could lead to oral drugs rather than injections. The company says that technology licensed from Springer’s lab at Harvard is designed to overcome challenges that up to now prevented development of small-molecule oral drugs blocking signaling activity of integrin targets in diseased tissue. Morphic expects to design treatments for immunological diseases, fibrotic and vascular disorders, and neoplastic conditions, those causing abnormal growth of tissue from rapid or uncontrolled cell division.

The company’s first financing round is lead by SR One, the venture capital arm of GlaxoSmithKline and Pfizer Venture investments. Joining the round are AbbVie Ventures, another pharmaceutical venture group, as well as Omega Ventures. Polaris Partners, ShangPharma Investment Group, Schrödinger Inc., and Springer himself were original seed investors. The $51.5 million raised is expected to support research and development of multiple drug candidates into clinical trials.

Original investor Schrödinger Inc., a company making software for computational biology, is also collaborating with Morphic on design and discovery of small molecule drug candidates.

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Disclosure: The author owns shares in Pfizer.

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Radiation Enhances Nanoparticle Cancer Treatments

Cancer in headline


30 June 2016. A strategy targeting vulnerable proteins on tumors with chemotherapy delivered in nanoscale particles and preceded with shots of radiation was shown in lab mice to boost treatment effectiveness in a range of cancers. A team from the lab of Daniel Heller at Memorial Sloan Kettering Cancer Center in New York published its findings in yesterday’s issue of the journal Science Translational Medicine (paid subscription required).

Heller’s lab studies the use of nanoscale drug delivery technologies — 1 nanometer equals 1 billionth of a meter — for treating metastatic cancer that spreads from original cancer sites and is responsible for 90 percent of cancer deaths. Delivering cancer drugs in nanoscale pieces makes it possible to target tumors directly, preventing drugs from building up and harming healthy tissue, a major problem encountered with many current cancer treatments.

Delivering drugs as nanoparticles, however, encounters problems with extravasation, or leakage into tissue surrounding the tumor site. To overcome this problem, Heller and colleagues designed a technique that aims nanoparticles with chemotherapy drugs directly at a protein called P-selectin found in blood platelets and cells lining blood vessels, as well as expressed on some metastatic tumor cells, including lung, ovarian, breast, and liver. The nanoparticles are made of fucoidan, a natural carbohydrate material derived from seaweed, and known to attract and bind to P-selectin.

The researchers tested fucoidan nanoparticles delivering chemotherapy drugs paclitaxel and doxorubicin with lab mice induced with melanoma, an advanced and aggressive form of skin cancer, and breast cancer. Results were compared to similar tumors on mice treated with nanoparticles made with dextran sulfate, a sodium salt compound used frequently as a stabilizer, and chemotherapy drugs delivered in free form. The results show mice receiving the fucoidan nanoparticles had greater tumor reduction and longer survival than mice receiving dextran sulfate nanoparticles or chemotherapy in free form.

While P-selectin is found on a number tumor cells, not all tumors express this protein target. The researchers also tested radiation as a way to make tumors that do not express P-selectin on their surface more vulnerable to fucoidan nanoparticles. The team grafted on the hind limbs of lab mice a form of lung cancer without P-selectin, and subjected one of the limbs to X-ray doses. The results show the tumors on the limb receiving X-rays started expressing P-selectin in about 4 hours, and increased over 24 hours.

In addition, the team found the tumor on the limb not receiving X-rays also expressing P-selectin proteins about 24 hours after the radiation, a phenomenon known as abscopal effect. Tests of fucoidan nanoparticle treatments in lab mice with tumors receiving X-rays show more chemotherapy drugs delivered to the tumor sites, as well as less tumor growth, and in some cases complete tumor regression.

The authors say this process still needs further refinement, particularly when dealing with radiation that can be toxic in some cases. Nonetheless, Heller and first author Yosi Shamay filed a patent for the targeted nanoparticle technology.

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Moderna, Merck Partner on Personal Cancer Vaccines

Human T-cell lymphocyte

Scanning electron micrograph of a human T-cell lymphocyte (National Institute of Allergy and Infectious Diseases, NIH)

29 June 2016. Biotechnology company Moderna Therapeutics and drug maker Merck are developing therapeutic vaccines using messenger RNA that address unique genetic patterns in patients’ tumors. The deal is expected to bring Moderna an immediate payment of $200 million, and the chance to earn further payments to develop cancer treatments combining Moderna’s technology with an approved targeted antibody therapy for cancer made by Merck.

Moderna Therapeutics in Cambridge, Massachusetts, develops medications that use genetic material to produce therapeutic proteins in the body, with a technology based on research licensed from Harvard University and MIT. That technology harnesses messenger RNA, a nucleic acid related to DNA used by cells to produce the amino acids in proteins for carrying out bodily functions. Moderna designs what it calls modified messenger RNA to produce proteins that act like drugs as treatments for diseases, creating antibodies able to cut the time and expense for therapeutic proteins over current genetic engineering methods.

Merck in Kenilworth, New Jersey developed pembrolizumab, a targeted antibody marketed under the brand name Keytruda that harnesses the immune system to fight tumors. Keytruda is in a class of drugs called checkpoint inhibitors that limit the actions of tumor cells to block the immune system. In this case, Keytruda stops receptor proteins on the surface of tumor cells from blocking the activation of T-cells in the immune system to attack tumors. Keytruda is already approved by Food and Drug Administration to treat melanoma, an advanced and metastatic form of skin cancer, and non–small cell lung cancer.

Under the agreement, the companies will develop therapeutic cancer vaccines using Moderna’s messenger RNA technology to target neoantigens, unique sets of mutations expressed in cancer patients’ tumors. The vaccines will be designed to induce immune responses specifically targeted to those mutations. The companies believe this personalized activation of immune responses will work well with Keytruda and other checkpoint inhibitors.

The deal calls for Merck to make an immediate payment of $200 million for Moderna to design and evaluate messenger RNA treatments with Keytruda. Moderna will be responsible for all initial research and development through proof-of-concept. The initial payment will also cover building a small-batch manufacturing facility on a Moderna site outside Boston meeting Good Manufacturing Practice, or GMP, standards for pharmaceuticals, designed to deliver personalized vaccines.

Following proof-of-concept tests, Moderna and Merck may move ahead on further development of personalized cancer vaccine therapies. The deal calls for the companies to share costs and profits equally in a worldwide collaboration, with Moderna having an option to co-promote products from the partnership in the U.S. Under the agreement, Merck will make another payment of an undisclosed amount to Moderna, if the companies go ahead with second stage.

Merck and Moderna are already collaborating on development of antiviral vaccines and passive immunity therapies, those induced with antibodies delivered from outside the body. That deal, begun in January 2015, runs for 3 years with an optional 1-year extension.

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More Device Approvals, Higher Risks in Europe than U.S.

EKG graphic

(PublicDomainPictures, Pixabay)

29 June 2016. An analysis of regulatory approvals and outcomes shows medical devices are reviewed and approved faster in Europe than the U.S., but also face more safety issues later on. A team from Kings College London and Harvard University published its findings yesterday in the journal BMJ.

Researchers led by Aaron Kesselheim, a bioethicist at Harvard Medical School and professor of medicine at Brigham and Women’s Hospital in Boston, sought to better understand the impact of different regulatory approaches for medical devices between Europe and the U.S. In Europe, medical devices can receive Conformité Européenne or CE certification from the European Union if developers can show the devices work as intended and are likely to be safe. Clinical trials are required only on some high-risk systems. In the U.S., the Food and Drug Administration requires high risk devices to demonstrate safety and effectiveness, usually in clinical trials, before approval for marketing.

As a result, medical devices are generally approved faster in Europe and before the U.S. Regulators in both jurisdictions, say the authors, need to balance faster access to new devices against the public safety in setting rules for their review. In 2012, however, FDA issued a report listing 12 devices, such as breast implants and stents to repair aneurysms, that were approved in Europe but later found to harm patients. In this study, the team aimed to provide more systematic evidence of the impact on safety from the different approaches to regulation, which up to now has been limited by a lack of public registries of devices in Europe — FDA has such a registry — decentralized nature of regulation in Europe, and confidentiality of information provided to regulators.

Kesselheim and colleagues reviewed public announcements, through news releases and reports such as financial filings, of medical devices receiving CE certification from 2005 through 2010. Their review yielded a collection of 309 devices designed to treat cardiovascular, neurologic, and orthopedic conditions. The team also reviewed FDA registries and records for these same devices, as well as public records of clinical trials testing the devices for safety and effectiveness.

The researchers rated each device a “major innovation” if it represented the first of a new class of device, introduced new technology, made new claims of safety and effectiveness, or was intended for a new patient population. Devices not meeting any of these criteria were considered “other changes.” In addition, the team reviewed public databases and records in the U.S., U.K., and Germany for product recalls, safety notices, or software upgrades to fix deficiencies.

The Harvard-Kings College team found nearly 8 in 10 of the devices receiving CE marks (79%) were for cardiovascular conditions with the remainder split about evenly between neurologic and orthopedic devices, and about a quarter (24%) rated as major innovations. About two-thirds of the devices approved in Europe (67%) were also approved by FDA.

The researchers report as of January 2016, about a quarter of the devices (24%) receiving CE certification were recalled or the subject of a safety alert, such as an automated system to assist heart pumping that shut down and stopped without warning. Of the recalls and alerts, nearly twice the number occurred among devices approved first in Europe (27%) than those approved first by FDA (14%). Using statistical models of risk over time, devices approved first in Europe had a 3 to 4.5 times greater chance of recall or safety alerts than first approved in the U.S.

In addition, only about half (49%) of devices considered major innovations reported published clinical trials supporting their approval. Reports of these trials were published a median of about 3 years (37 months) following regulatory approval. Devices approved in the U.S. using FDA’s pre-market approval pathway for higher-risk systems were more likely to have published clinical trials than devices using other processes.

The authors recommend caution in overhauling regulatory processes to increase the speed of reviews, but also greater transparency for regulatory decisions in Europe, beginning with making public the current European database of devices. Kesselheim and colleagues also note that there is not necessarily a trade-off between regulation and market success. They cite a recent study showing coronary stents evaluated in clinical trials were more likely to be commercially successful than those that were not tested. Thus higher regulatory standards could help improve product sales.

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Animal-Free Leather Company Gains $40M in Venture Funds

Cowboy boots

(skeeze, Pixabay)

28 June 2016. A company making leather for consumer goods with tissue engineering and gene-editing from living cells is raising $40 million in its second venture funding round. The enterprise, Modern Meadow Inc., in Brooklyn, New York, says it now raised $53.5 million since its founding in 2011.

Modern Meadow produces leather without environmentally harmful methods of raising animals and tanning hides. The company applies principles of cell engineering developed in the labs of its scientific founder Gabor Forgacs at University of Missouri and Clarkson University in Potsdam, New York. Its process starts with cells for producing collagen, an abundant protein providing substance to connective tissue such as bones, tendons, and skin, including the skin or hides of animals.

The basic collagen cells are then genetically modified to produce a leather-like material with specified properties, such as strength or suppleness, then the cells are cultured to proliferate and produce complex collagen molecules in sufficient quantities. The collagen molecules are assembled into nanoscale fibers that connect into networks that assemble further into raw three-dimensional structures resembling animal skins.

Because the engineered hide is produced in a lab, the traditional tanning process using chemicals to remove hair, flesh, and fat is largely eliminated. Only a final finishing process to preserve the engineered leather and provide suppleness and surface qualities is required. Modern Meadow says its processes reduce waste by about 80 percent, including fewer inputs of land, water, energy, and chemicals.

The new funding round is led by Horizons Ventures and Iconiq Capital with participation by ARTIS Ventures, Temasek, Breakout Ventures, Red Swan Ventures, Collaborative Fund and Tony Fadell. As reported in Science & Enterprise, Modern Meadow was an early recipient of seed funding from Breakout Labs, a program of the Thiel Foundation, founded by entrepreneur and investor Peter Thiel, co-founder of PayPal and early venture backer of Facebook.

The company plans to use the new financing to convert from a research and development to a manufacturing enterprise, taking its first products to the marketplace. Andras Forgacs, co-founder and CEO — and son of Gabor Forgacs — says in a company statement that the $100 billion leather market “is subject to fluctuations in availability, quality, price, and growing demand. At Modern Meadow, we’re re-imagining this millennia-old material to create revolutionary new features without harming animals or the environment.”

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3-D Printing Technique Devised for Replacement Cartilage

Knee X-ray

(Akha, Wikimedia Commons)

28 June 2016. Biomedical engineers developed a process for directly printing replacement cartilage without the scaffolds needed in previous tissue engineering techniques. The team at Pennsylvania State University, led by engineering professor Ibrahim Ozbolat, describe their process in yesterday’s issue of the journal Scientific Reports.

Ozbolat and colleagues, from Penn State and University of Iowa where he was previously on the faculty, are seeking to simplify production of replacement cartilage tissue for people with osteoarthritis, or wear and tear on joints, and other cartilage damage. Once damaged, cartilage tissue does not grow back on its own. In addition, replacement cartilage not only needs to provide physical and structural support, but also the biological and cell signaling functions of original tissue.

Current tissue engineering methods for replacement cartilage require first creating a scaffold for the new tissue, usually from hydrogel, a material made up largely of water with polymer chains that provides a framework. Cartilage cells are then seeded on the hydrogel framework, where they proliferate and grow. “Hydrogels don’t allow cells to grow as normal,” says Ozbolat in a university statement. “The hydrogel confines the cells and doesn’t allow them to communicate as they do in native tissues.” Because of these limitations, adds Ozbolat, scaffold-grown tissue often lacks the needed mechanical properties and is susceptible to toxins from degrading hydrogel.

The researchers designed a process that produces new cartilage tissue directly with strands of cells that can be extruded through a 3-D printer. The cells are first cultured for 10 days inside thin tubes made of alginate, a biocompatible extract of algae used in wound healing, drug delivery, and tissue engineering. The cells adhere into strands that are easily removed from the alginate. The strands are thin enough to fit through a 3-D printer, with a specially-designed nozzle, and fabricated into tissue patches. The patches are then cultured further in nutrients where they self-assemble into replacement cartilage.

“We can manufacture the strands in any length we want,” notes Ozbolat. “Because there is no scaffolding, the process of printing the cartilage is scalable, so the patches can be made bigger as well.”

The Penn State team demonstrated the process in proof-of-concept tests with cartilage cells from cattle. The tests show the printed cartilage has biochemical and some mechanical properties similar to natural cartilage. The researchers also attached the printed cartilage to a cattle bone model. While superior to scaffold-grown cartilage, however, the replacement cartilage did not fully integrate with the bone, like original cartilage. The authors recommend a bio-compatible glue to improve adhesion.

Ozbolat tells more about 3-D printing of cartilage in the following video.

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