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Nanobots the Artificial Blood: Design, Manufacturing, and Performance of Respirocytes and Clottocyte



HBOCs from expired human blood or fresh bovine blood have to undergo numerous modifications to make them safe and effective oxygen carriers. The RBCs are first lysed to release their hemoglobin, and then the stroma is removed by a variety of methods, including centrifugation, filtration and chemical extraction. The stroma-free hemoglobin is then purified and undergoes modifications to cross-link, polymerize or conjugate it to other compounds. Without these modifications, the oxygen affinity of the stroma-free hemoglobin is too great to facilitate oxygen release in the tissues. Also, when it is outside the RBC, hemoglobin rapidly dissociates into 32 kDa αβ dimers and 16 kDa α or β monomers, both of which are rapidly filtered in the kidney and can precipitate in the loop of Henle, resulting in severe renal toxicity. The dimer haem iron is oxidized more easily than in the tetramer, leading to molecules unable to bind oxygen. Moreover, the formation of ferric ions triggers a cascade of reactions that generate reactive oxygen species and reactive nitrogen species, the molecular basis of oxidative damage.


The field of small scale robotics is developing rapidly, in large part due to the potential for these machines to operate with precision on the cellular and sub-cellular level. The smallest such constructs, synthetic molecular machines1,2, can consist of fewer than 20 atoms3, however there are a significant number of challenges yet to overcome before they are of any practical use4. On the other hand, microorganisms use biomolecular machines to perform myriad useful tasks. For example, in low Reynolds number fluidic environments, many microorganisms propel themselves via biomolecular motors which utilize viscous drag. To accomplish this, these organisms often employ some form of nonreciprocal motion5. One example of this motion is the oblique beating of flagella and cilia, which protrude from prokaryotes and eukaryotes. This method of swimming has been mimicked in an artificial swimmer composed of streptavidin coated magnetic microparticle chains linked together through biotinylated DNAs, attached to an erythrocyte, and actuated using an oscillating field6. Similarly, other swimmers that use flexible metallic nanowires have also been reported7,8. However, in nature there is another example of nonreciprocal motion, namely the rotation of helical flagella used by bacteria9. In particular, this method of propulsion has been extensively investigated for robotic swimmers, and the design and actuation of biomimetic micro- and nanoscale artificial flagella has been well established10,11,12,13. Furthermore, potential biological and biomedical applications, including micro transportation13, drug delivery14, in vitro cell manipulation15,16, and in vivo imaging17 have been demonstrated using such devices. While these small scale helical shaped devices have proven to be effective swimmers, many limitations in both their fabrication and actuation ability are still prevalent. In terms of fabrication, most of the reported nano sized helical swimmers have been created using top-down fabrication methods, such as shadow-growth10 or direct laser writing13, that require specialized equipment and often involve complex fabrication steps. Also, these fabrication methods have been traditionally limited to just inorganic (i.e. metals and metalloids) or photoactive polymer materials18,19,20,21. With respect to actuation, for the nanoswimmers reported thus far, surface coatings have been used to augment additional functionality14,22.




nanobots the artificial blood pdf



The ability to rapidly and reversibly convert between different superhelical forms allows the flagellum to balance external forces that would otherwise cause it to yield or buckle, and lose its rigid form during swimming28. Thus far, it has been shown that polymorphic transformations can be induced though environmental changes in pH and ionic strength29,30,31, temperature24,32, addition of organic solvents33,34, by applying electric fields35, or mechanical force24,36. The ability of flagella to undergo polymorphic transformation when exposed to a variety of different stimuli gives flagella the potential to be used as both nanoscale sensors and mechanical transducers. These capabilities are not present in previously reported artificial helical nanoswimmers, however there has been a recent report of a sub-millimeter scale soft reconfigurable helical swimmer37. Also, in a previous work we developed a microscale swimmer using bacterial flagella, however we were unable steer these devices nor visualize flagella during swimming38. Notably, many microorganisms, especially those responsible for infection, utilize the ability to change morphology to effectively infiltrate and bypass physical biological barriers present with host systems. Robotic nanoswimmers that utilize flagella, which have a programmable morphology, could potently mimic this mechanism to navigate complex biological microenvironments. Thus, in an effort to harness the properties of bacterial flagella for both swimming and sensing, we report on the fabrication, visualization, and actuation of shape reconfigurable nanorobotic swimmers.


Since 2017, six CAR T-cell therapies have been approved by the Food and Drug Administration (FDA). All are approved for the treatment of blood cancers, including lymphomas, some forms of leukemia, and, most recently, multiple myeloma.


As part of their immune-related duties, T cells release cytokines, chemical messengers that help stimulate and direct the immune response. In the case of CRS, the infused T cells flood the bloodstream with cytokines, causing serious side effects, including dangerously high fevers and precipitous drops in blood pressure. In some cases, severe CRS can be fatal.


Although CD19 and BCMA are the only antigens for which there are FDA-approved CAR T-cell therapies, CAR T-cell therapies have been developed that target other antigens commonly found in blood cancers, including therapies that target multiple antigens at one time.


A blood substitute (also called artificial blood or blood surrogate) is a substance used to mimic and fulfill some functions of biological blood. It aims to provide an alternative to blood transfusion, which is transferring blood or blood-based products from one person into another. Thus far, there are no well-accepted oxygen-carrying blood substitutes, which is the typical objective of a red blood cell transfusion; however, there are widely available non-blood volume expanders for cases where only volume restoration is required. These are helping doctors and surgeons avoid the risks of disease transmission and immune suppression, address the chronic blood donor shortage, and address the concerns of Jehovah's Witnesses and others who have religious objections to receiving transfused blood.


The main categories of "oxygen-carrying" blood substitutes being pursued are hemoglobin-based oxygen carriers (HBOC) and perfluorocarbon emulsions.[1] Oxygen therapeutics are in clinical trials in the U.S. and Europe, and Hemopure is available in South Africa.


After William Harvey discovered blood pathways in 1616, many people tried to use fluids such as beer, urine, milk, and non-human animal blood as blood substitute.[2] Sir Christopher Wren suggested wine and opium as blood substitute.[3]


At the beginning of the 20th century, the development of modern transfusion medicine initiated through the work of Landsteiner and co-authors opened the possibility to understanding the general principle of blood group serology.[4] Simultaneously, significant progress was made in the fields of heart and circulation physiology as well as in the understanding of the mechanism of oxygen transport and tissue oxygenation.[5][6]


Restrictions in applied transfusion medicine, especially in disaster situations such as World War II, laid the grounds for accelerated research in the field of blood substitutes.[7] Early attempts and optimism in developing blood substitutes were very quickly confronted with significant side effects, which could not be promptly eliminated due to the level of knowledge and technology available at that time. The emergence of HIV in the 1980s renewed impetus for development of infection-safe blood substitutes.[3] Public concern about the safety of the blood supply was raised further by mad cow disease.[3][8] The continuous decline of blood donation combined with the increased demand for blood transfusion (increased ageing of population, increased incidence of invasive diagnostic, chemotherapy and extensive surgical interventions, terror attacks, international military conflicts) and positive estimation of investors in biotechnology branch made for a positive environment for further development of blood substitutes.[8]


Efforts to develop blood substitutes have been driven by a desire to replace blood transfusion in emergency situations, in places where infectious disease is endemic and the risk of contaminated blood products is high, where refrigeration to preserve blood may be lacking, and where it might not be possible or convenient to find blood type matches.[9]


The first approved oxygen-carrying blood substitute was a perfluorocarbon-based product called Fluosol-DA-20, manufactured by Green Cross of Japan. It was approved by the Food and Drug Administration (FDA) in 1989. Because of limited success, complexity of use and side effects, it was withdrawn in 1994. However, Fluosol-DA remains the only oxygen therapeutic ever fully approved by the FDA. As of 2017 no hemoglobin-based product had been approved.[9]


Perfluorochemicals are not water soluble and will not mix with blood, therefore emulsions must be made by dispersing small drops of PFC in water. This liquid is then mixed with antibiotics, vitamins, nutrients and salts, producing a mixture that contains about 80 different components, and performs many of the vital functions of natural blood. PFC particles are about .mw-parser-output .sfracwhite-space:nowrap.mw-parser-output .sfrac.tion,.mw-parser-output .sfrac .tiondisplay:inline-block;vertical-align:-0.5em;font-size:85%;text-align:center.mw-parser-output .sfrac .num,.mw-parser-output .sfrac .dendisplay:block;line-height:1em;margin:0 0.1em.mw-parser-output .sfrac .denborder-top:1px solid.mw-parser-output .sr-onlyborder:0;clip:rect(0,0,0,0);height:1px;margin:-1px;overflow:hidden;padding:0;position:absolute;width:1px1/40 the size of the diameter of a red blood cell (RBC). This small size can enable PFC particles to traverse capillaries through which no RBCs are flowing. In theory this can benefit damaged, blood-starved tissue, which conventional red cells cannot reach. PFC solutions can carry oxygen so well that mammals, including humans, can survive breathing liquid PFC solution, called liquid breathing.[citation needed] 2ff7e9595c


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