Bioprinter patent for Extra Cellular Matrix
Uhohinc
[0036] "Bioprinting," as used herein, generally refers to the deposition of living cells, as well as other components (e.g., a flowable ECM; synthetic matrices) onto a surface using standard or modified printing technology, e.g., ink jet printing technology. Basic methods of depositing cells onto surfaces, and of bioprinting cells, including cells in combination with hydrogels, are described in Warren et al. U.S. Pat. No. 6,986,739, Boland et al. U.S. Pat. No. 7,051,654, Yoo et al. US 2009/0208466 and Xu et al. US 2009/0208577, the disclosures of each of which are incorporated by reference herein their entirety. Additionally, bioprinters suitable for the methods described herein are commercially available, e.g., the 3D-Bioplotter® from Envisiontec GmbH (Gladbeck, Germany); and the NovoGen MMX Bioprinter® from Organovo (San Diego, Calif.).
[0037] The bioprinter used in the methods described herein may include mechanisms and/or software that enables control of the temperature, humidity, shear force, speed of printing, and/or firing frequency, by modifications of, e.g., the printer driver software and/or the physical makeup of the printer. In certain embodiments, the bioprinter software and/or hardware preferably may be constructed and/or set to maintain a cell temperature of about 37° C. during printing.
[0038] In certain embodiments, the inkjet printing device may include a two-dimensional or three-dimensional printer. In certain embodiments, the bioprinter comprises a DC solenoid inkjet valve, one or more reservoir for containing one or more types of cells, e.g., cells in a flowable composition, and/or ECM (e.g., a flowable ECM) prior to printing, e.g., connected to the inkjet valve. The bioprinter may have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more reservoirs, e.g., one for each cell type or each ECM used to construct the tissues and organs described herein. The cells may be delivered from the reservoir to the inkjet valve by air pressure, mechanical pressure, or by other means. Typically, the bioprinter, e.g., the print heads in the bioprinter, is/are computer-controlled such that the one or more cell types, and said ECM, are deposited in a predetermined pattern. Said predetermined pattern can be a pattern that recreates or recapitulates the natural arrangement of said one or more types of cells in an organ or tissue from which the cells are derived or obtained, or a pattern that is different from the natural arrangement of said one or more types of cells.
[0039] In certain embodiments, the bioprinter used in the methods provided herein may be a thermal bubble inkjet printer, see, e.g., Niklasen et al. U.S. Pat. No. 6,537,567, or a piezoelectric crystal vibration print head, e.g., using frequencies up to 30 kHz and power sources ranging from 12 to 100 Watts. Bioprinter print head nozzles, in some embodiments, are each independently between 0.05 and 200 micrometers in diameter, or between 0.5 and 100 micrometers in diameter, or between 10 and 70 micrometers in diameter, or between 20 and 60 micrometers in diameter. In further embodiments, the nozzles are each independently about 40 or 50 micrometers in diameter. Multiple nozzles with the same or different diameters may be used. In some embodiments the nozzles have a circular opening; in other embodiments, other suitable shapes may be used, e.g., oval, square, rectangle, etc., without departing from the spirit of the invention.
[0040] In certain embodiments, the bioprinter used in accordance with the methods described herein comprises a plurality of print heads and/or a plurality of print jets, wherein said plurality of print heads or print jets may, in certain embodiments, be separately controllable. In certain embodiments, each of said print heads or print jets operates independently from the remaining said print heads or print jets. In certain embodiments, at least one of said plurality of print heads or plurality of print jets may print compositions comprising cells, and at least one of said plurality of print heads or plurality of print jets may print compositions comprising ECM. In certain embodiments, at least one of said plurality of print heads or plurality of print jets may print compositions comprising cells, at least one of said plurality of print heads or plurality of print jets may print compositions comprising ECM, and at least one of said plurality of print heads or plurality of print jets may print compositions comprising an additional component.
[0041] In certain embodiments, one or more print heads or print jets of a bioprinter used in accordance with the methods described herein may be modified so that it is suitable for printing on certain surface. For example, a print head or print jet may be modified by attaching it to a certain surgical instrument, e.g., a laparoscope. In accordance with such embodiments, the surgical instrument may be fitted with one or more other components that aid in the printing procedure, e.g., a camera.
[0042] In certain embodiments, an anatomical image of the tissue or organ to be printed on may be constructed using software, e.g., a computer-aided design (CAD) software program. In accordance with such embodiments, programs can be generated that allow for three-dimensional printing on a three-dimensional surface that is representative of the structure of the tissue or organ to be printed on. For example, if it is desired to print on a bone, an anatomical image of the bone may be constructed and a program may be generated that directs the printer heads of the bioprinter to rotate around the three-dimensional bone inside the subject surface during printing.
[0043] In certain embodiments, the surface of the tissue or organ to be printed on is scanned in or on said the subject so as to form a surface map, and said surface map is used to guide the depositing of the cells, ECM, and/or any additional components to be printed. Such scanning may comprise, without limitation, the use of a laser, electron beam, magnetic resonance imaging, microwave, or computed tomography. The scan of said surface may comprise a resolution of least 1000, 100, 10, 1, or 0.1 microns.
[0044] In certain embodiments, the methods of bioprinting provided herein comprise the delivery/deposition of individual droplets of cells (e.g., compositions comprising single cells or compositions comprising multiple cells) and flowable extracellular matrix (ECM) on a surface in or on a subject.
[0045] In certain embodiments, the methods of bioprinting provided herein comprise the deposition of a single cell type and flowable ECM on a surface in or on a subject. Exemplary cell types that can be used in accordance with such methods are provided in Section 4.1.1, below. ECM, including flowable ECM, is described in Section 4.1.3, below.
[0046] In other embodiments, the methods of bioprinting provided herein comprise the deposition of multiple (e.g., two, three, four, five or more) cell types and flowable ECM on a surface in or on a subject. In a specific embodiment, the multiple cell types are deposited as part of the same composition, i.e., the source of the cells is a single composition that comprises the multiple cell types. In another specific embodiment, the multiple cell types are deposited as part of different compositions, i.e., the source of the cells are distinct compositions that comprise the multiple cell types. In another specific embodiment, a portion of the multiple cell types are deposited as part of one composition (e.g., two or more cell types are in a single composition) and another portion of the multiple types are deposited as a different composition (e.g., one or more cell types are in a single composition). Exemplary cell types that can be used in accordance with such methods are provided in Section 4.1.2, below.
[0047] In a specific embodiment, the cells to be deposited and the flowable ECM are deposited on a surface in or on a subject together (e.g., simultaneously) as part of the same composition. In another specific embodiment, the cells to be deposited and the flowable ECM are deposited on a surface in or on a subject together as part of different compositions. In another specific embodiment, the cells to be deposited and the flowable ECM are deposited on a surface in or on a subject separately (e.g., at different times).
Read more: http://www.faqs.org/patents/app/20150246072#ixzz3klmkvbDV
Uhohinc
By Serenitie Wang and Katie Hunt, CNN
Updated 1:01 AM ET, Tue January 10, 2017
Scientists have successfully grafted 3-D printed blood vessels onto rhesus monkeys.
Scientists have successfully grafted 3-D printed blood vessels onto rhesus monkeys.
Story highlights
Chinese scientists have implanted 3-D printed blood vessels into monkeys
The new vessels can merge with the monkeys' own organs to become functional
Beijing, China (CNN)A major breakthrough in 3-D printed medicine could bring hope to nearly 1.8 billion patients with cardiovascular disease.
Chinese scientists working for Sichuan Revotek have successfully 3-D printed blood vessels and implanted them in rhesus monkeys, the company said.
It is a major step on the road to mass printing human organs for transplants.
The company is "the first to have maintained the viability of the cells with the 3-D printing technology," said James Kang, chief scientist and CEO of Sichuan Revotek.
Producing 'bio-ink'
The key to the experiment's success was the biological material developed by Sichuan Revotek, known as bio-ink, which is made from stem cells derived from the fat tissue of monkeys.
Stem cells have the ability to grow into any cells within the body, and the use of the monkey's own stem cells means the vessels won't be rejected by its immune systems once implanted.
Patented as "Biosynsphere," the bio-ink consists of stem cells in a micro-environment of growth factors and nutrients that can stimulate the cells to grow into the types of cells needed to form a functioning blood vessel.
The use of stem cells acquired from fat tissue is also safer than the usual source -- embryos, Kang said.
Scientists develop new 3D organ printer
Scientists develop new 3D organ printer 03:20
How it works
The technology could one day be used in humans.
The technology could one day be used in humans.
During the surgery, Kang's team replaced a 2-centimeter segment of abdominal artery with a 3-D printed blood vessel in 30 rhesus monkeys.
Within five days of implantation, the stem cells were able to grow into the various types of cells needed to make a functioning blood vessel, including endothelial cells and smooth muscle cells.
One month later, the grafted vessels had completely merged into the monkey's own artery and functioned "exactly the same" as the monkey's original vessel, the company said.
The printed vessels have been shown to support vascular functions, such as aiding blood flow and transporting nutrients throughout the body as the bio-ink helps them fully develop into living vessels, Kang said.
"And the bio-ink's capacity to develop collagen, a necessity for the tissue to mold into different shapes, is the first of its kind," he said.
Other experts agree the work is groundbreaking.
How a 3-D-printer changed a 4-year-old's heart and life
How a 3-D-printed heart changed a girl's life
"This is an important breakthrough in the field," said Alex Lee, an assistant professor at the Chinese University of Hong Kong, where his team has been using 3-D printing to prototype heart models to personalize cardiac surgeries for patients.
However, Lee said it would require years to observe the long-term effect of these vessels and cautioned that they "may become blocked again years later."
Kang said the company was applying for regulatory approval to test the process on humans.
Last year, Sichuan Revotek created the world's first blood vessel bio-printer, which is said to be able to produce living tissue and organs. The privately owned company is one of a number of tech start-ups in China's rising innovation hub in the Sichuan capital of Chengdu, offering great hope for the field to progress.
Sichuan Revotek has also produced the world's first 3D blood vessel bio-printer.
Sichuan Revotek has also produced the world's first 3D blood vessel bio-printer.
The long road to 3-D organs
Last year saw an explosion of attempts to produce 3-D printed biological structures.
Russian scientists transplanted a 3-D printed thyroid gland into a lab mouse in November and expect to print human organs in 15 years. French cosmetic giant L'Oreal announced that it is developing 3-D printed skin tissues for product testing. A New York start-up, EpiBone, is attempting to 3-D print customized bone grafts.
However, these advances have aroused some ethical concerns, and analysts say they're likely to spark major debate leading to regulation of the technology.
But Lee sees 3-D printed vessels as a real possibility for humans and expects to see them "tested in clinical trials in a couple of years." Other simple structures could also become a reality, she predicts.
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"What we can print right now are cardiac tissue patches and small-sized blood vessels," he said, adding that they are parts "that have simple, hollow structures."
But the parts making up a human heart, such as valves, have more complex shapes and tissues, so they will be more technologically challenging for the field of 3-D printing.
The printing of living organs, such as a whole heart, still has "a long way to go to become a clinical reality," said Lee.
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Human heart bioprinted, functional.
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> https://www.dailymail.co.uk/sciencetech/article-6955367/German-scientists-create-human-organs.html
Can we heal wounds by printing skin?
Published Monday 11 March 2019 By Monica Beyer Fact checked by Jasmin Collier
Printing layers of skin to help treat chronic wounds or burns may be on the horizon, thanks to a newly developed mobile skin bioprinting system.
Skin bioprinter being used
The skin bioprinter at work.
Image credit: WFIRM
Scientists at the Wake Forest Institute for Regenerative Medicine (WFIRM) in Winston-Salem, NC, have created a bioprinter that uses a person's own skin cells to create layers of new skin and apply them directly to the wound.
A new paper, which now appears in the journal Scientific Reports, details the development of this new technology.
The procedure involves harvesting major skin cells called dermal fibroblasts and epidermal keratinocytes from a biopsy of a person's normal skin tissue.
The scientists expanded the cells and mixed them into a hydrogel. They then placed them into the bioprinter, which scans the person's wound, feeds the data into the software, and tells the device where to place the printed layers of skin.
The resulting material is uniquely printed to match the exact areas in a person's wound where it is needed without the need for a donor skin graft.
This technique can replicate the natural function of skin and accelerate how skin naturally forms, which means that healing can take less time and has fewer risks.
"The unique aspect of this technology is the mobility of the system and the ability to provide on-site management of extensive wounds by scanning and measuring them in order to deposit the cells directly where they are needed to create skin."
Lead study author Sean Murphy, Ph.D.
Chronic wounds are difficult to treat
A wound normally takes up to 4–6 weeks to heal, depending on severity and size. However, when it does not heal within this timeframe, doctors consider it a chronic wound. Many factors can cause a chronic wound, including diabetes and poor nutrition.
There are many ways that healthcare providers attempt to heal a chronic wound. Wound dressing is an important part of the care and healing of wounds, but these vary in price and effectiveness.
Scientists design 'smart' wound healing technique
Scientists design 'smart' wound healing technique
A recent study looked into innovative ways of encouraging natural wound healing.
READ NOW
Skin grafts are another option for both chronic wounds and burns that encompass a large surface area of tissue. Skin grafts can either come from another area of the person's body, called autologous sources, or in the form of skin substitutes.
Bioprinted skin layers may be the next revolution in wound healing. Dr. James Yoo, Ph.D., who led the research team, explains that bioprinted layers of skin help people start their healing process much sooner.
He also mentions that other methods of treating and closing wounds do not really help create skin, as this method would.
The researchers note that skin grafts are common, but they can have multiple disadvantages. For example, autologous grafts can be limited due to a shortage of healthy tissue. Also, donor skin grafts can bring a risk of tissue rejection.
The new bioprinter helps ameliorate these risks because it helps skin form outward from the center of the wound, which only happens when a person's own cells are used in its creation.
'No significant difference'
The findings are encouraging, but our in-
Uhohinc
Recent and rapid progression in three-dimensional (3D) printing techniques has revolutionized conventional therapies in medicine; 3D printed constructs are gradually being recognized as common substitutes for the replacement of skin wounds. As gel-inks, large numbers of natural and synthetic (e.g., collagen and polyurethane, respectively) substances were used to be printed into different shapes and sizes for managing both acute and chronic skin wounds. The resultant 3D printed scaffolds not only provide physical support but also act as supporting niches for improving immunomodulation and vascularization and subsequent accelerated wound healing. Recently, the use of thermosensitive and pH-responsive gels has made it possible to prepare 3D printed constructs with the ability to facilitate in situ crosslinking within the biopolymer and with native wound edge tissue as well as to fill the exact shape of wound damage. In this chapter, we aim to introduce the current state of 3D printable gel-inks utilized for skin wound treatment and illustrate future prospects in this amazing area of science.