What is Colossus skin made of?

Colossus' impenetrable skin is not a unique superpower in the superhero universe. By being referred to as “organic steel” skin, the reader is led to believe that his mutant skin is made up of steel, an alloy with iron as the primary element. Roughly 96% of the human body is carbon, hydrogen, oxygen, and nitrogen.

journals.physiology.org - The physiology of impenetrable skin: Colossus of the X-Men

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INTRODUCTION

The fictitious superhero worlds of Marvel and DC are filled with innumerate characters possessing extraordinary superpowers. Some characters, such as Iron Man (63), Superman, and Captain Marvel, can fly, whereas Sue Storm of the Fantastic Four has the power of invisibility (25, 28). Other characters, such as the super-soldiers Captain America (9) and the Winter Soldier (86), have been genetically modified, whereas Hawkeye of the Avengers has advanced eyesight similar to birds of prey due to genetic inheritance and evolution (26, 28). Given the range of superpowers, it is a difficult task to select the greatest superpower. Many people would contend that the greatest superpower is impenetrability, which is possessed by a number of characters via technological and biological means. From a technological viewpoint, Iron Man, Black Panther, and War Machine all have the power of impenetrability, thanks to wearable exoskeleton suits. Conversely, characters such as Superman, Wonder Woman, and Luke Cage have the power of impenetrability due to their natural or modified genetics. For example, in the 2016 film, Batman v Superman: Dawn of Justice (5), bullets and projectiles are shown to bounce off Superman’s skin without any noticeable discomfort for the Kryptonian. It is important to stress that Superman is an alien; thus his DNA more than likely differs significantly from that of human DNA and is the key to his powers. This difference in DNA also applies to Wonder Woman, who is an Amazonian god. However, Luke Cage of the Defenders acquires an impenetrable skin after experimental genetic treatment, and thus he did not naturally develop his ability. The impenetrable skin of Superman and Luke Cage is continually “active,” meaning that they are unable to “turn on or off” their power as desired. This has led to health treatment issues for Luke Cage, particularly when he has required medical attention, as hypodermic needles cannot breach his skin. However, one superhero character with the ability to “turn on or off” his power of impenetrable skin is Colossus of the X-Men. The current crop of superhero films can ascribe their success and popularity to the pioneering X-Men films, such as the 2000 film X-Men (101). The X-Men are a group of superheroes, often referred to as mutants, with a wide array of individual powers and abilities that can be attributed to the presence of genetic mutations associated with the X-Gene. The abilities portrayed by each character are dictated by how the X-Gene has been integrated in his DNA. In the case of Colossus, he can create an “organic steel” layer on his skin that facilitates an impenetrable layer around his body. In addition, he gains superhuman strength when in his steel form. In previous studies, I have outlined how superheroes can be used in the classroom to supplement lessons on the physiology of the eye (26) and learning objectives in a physics syllabus (28). In this article, I will demonstrate how Colossus can be used to address a diverse range of physiological concepts and motivate multidisciplinary overlap in the classroom. Colossus is the primary focus since, unlike Superman, Wonder Woman, or Luke Cage, Colossus’ skin undergoes a significant biological change when he changes to his steel form. In addition, I reflect on the use of Colossus in the promotion of responsible innovation, particularly in relation to technologies that could artificially augment the physiological capabilities of a person. Similar to my previous studies (26−28), a pedagogical approach for using Colossus in the classroom is also presented. Colossus: The Superhero Colossus was created by Len Wein and David Cockrum, and first appeared in Giant-Size X-Men no. 1 in 1975 (95). The character’s real name is Piotr Rasputin, a Russian farmer born and reared in the former Soviet Union near Lake Baikal in Siberia. Similar to a number of characters in Marvel literature, Colossus has the ability to change his external appearance. His genetic mutation allows him to create an “organic steel” layer, which acts as an impenetrable external shell. In the Hollywood superhero films, Colossus has been depicted in two different ways. In films such as X-Men 2 (99), X-Men: The Last Stand (102), and X-Men: Days of Future Past (100), Colossus is shown changing from his human form to his steel form. However, in the films Deadpool (15) and Deadpool 2 (14), a rebooted version of the character is always in the “organic steel” form. Here, I focus on the physiology of the earlier cinematic depiction of Colossus, where the character can be seen changing from a human form to his mutant form and back again. Nonetheless, recent appearances of the character in the Deadpool films will also be referenced. Colossus Skin The key to Colossus’ power of impenetrability is a change in the structure of his skin. A key question that must be addressed is how much of Colossus’ skin changes to steel during a transformation process. Before answering this question, the structure and function of human skin is considered. Structure of human skin. Skin, which is part of the integumentary system and the largest organ in the human body, performs a number of key physiological functions, such as preventing water or pathogens from entering the body, and in the thermoregulation of body temperature via responses such as vasodilatation, vasoconstriction, and sweating. Thus skin is highly relevant for the homeostasis, or the stable biological state, of a person. Skin consists of two primary layers known as the epidermis and dermis (Fig. 1). The epidermis is the outermost layer of the skin with an average thickness of ~60 μm (21). The top part of the epidermis is known as the stratum corneum, which acts as the final barrier between the human body and the outside world and consists of several layers of densely interlocked corneocytes or dead keratinized cells (54). These cells are constantly subject to shedding in a process known as desquamation. As the corneocytes are shed, they are replaced by cells originating in the stratum basale, a part of the epidermis located below the stratum corneum. Below the epidermis lies the dermis, which is composed of connective tissue that separates the epidermis from the hypodermis or subcutaneous tissue. The dermis also contains blood vessels, sweat glands, nerves, and hair follicles. Finally, the hypodermis or subcutaneous tissue attaches the skin layers to the bone and muscles. In addition, the hypodermis contains a rich network of blood vessels, fats, and hair follicle roots. Fig. 1.Illustration of the layers of skin: epidermis and dermis. The dermis connects the epidermis with the hypodermis or subcutaneous tissue below. [From Fitzgerald (25) with permission.] Download figureDownload PowerPoint In the laboratory, noninvasive medical imaging can be used to image microvascular flow in the dermis (59, 60, 68). For example, linearly polarized light can resolve red blood cell (RBC) concentration after occlusion and demonstrate the effects of vasodilatation (60). Optical coherence tomography (OCT) is an noninvasive imaging method that is the optical equivalent of ultrasound that can produce three-dimensional (3D) images of biological samples, such as retinal cross-sectional imaging (41, 73) and has been extensively applied to visualize the layers of skin (23, 35, 58, 102a). A number of OCT variants have been devised, such as time domain OCT (41), swept source OCT (12), spectral domain OCT (6), and multiple-reference OCT (MR-OCT) (19, 58). MR-OCT, which is a method that can quickly create cross-sectional images of a tissue sample over a large depth, could lead to the development of low-cost, compact OCT devices built from off-the-shelf components for use in medical institutes and research laboratories. Figure 2 shows an in vivo image of a fingertip using an MR-OCT device. There are a number of key features highlighted in the image. First, both the epidermis and dermis layers and the interface between the layers are evident. Second, the same variations in the interface between the epidermis and dermis layers are seen in the stratum corneum of the epidermis. Finally, the image resolves eccrine sweat ducts, which serve a key function in thermoregulation. Fig. 2.Multiple-reference optical coherence tomography (MR-OCT) image of the skin layers in a fingertip. Indicated on the image are the epidermis, dermis, and stratum corneum. (Image provided by Paul M. McNamara. Similar image is available in Ref. 58.) Download figureDownload PowerPoint Hindrance of thermoregulation. The structure of skin shown in Fig. 1 and imaged using MR-OCT in Fig. 2 is representative of Colossus’ skin when in his human form. However, when Colossus is in his steel form, his body’s ability to thermoregulate may be severely hampered. Thermoregulation is a key physiological process in the human body (43a, 44, 91, 92) and is related to the maintenance of a body core temperature of ~37°C. While the body is constantly regulating its core temperature, this process becomes noticeable during exercise due to an increase in metabolic heat production (98). The process of heat loss can be affected by numerous factors, including age, individual fitness, chronic diseases, the presence of clothing, and ambient conditions (43a). To maintain physiological functionality, the body must be able to regulate core body temperature, which can be influenced by both internal and external factors. An expression for heat storage including these factors is Heat storage = metabolism − external work − evaporation ± radiation ± conduction ± convection In an idealized case, the heat gain in a person would equal heat loss, such that the heat storage would be zero. Of principle interest for Colossus’ thermoregulation are metabolism and evaporation. When fighting against villains alongside the X-Men (100) or with Deadpool (14), Colossus’ metabolism and working muscles will produce large amounts of heat. As a mutant, it can be speculated that Colossus’ metabolism and method of food intake differ from a human. However, in Deadpool (15), Colossus is shown eating cereal for breakfast and professing the importance of a good breakfast to Negasonic Teenage Warhead, a colleague in the X-Men. Therefore, it is assumed that Colossus requires sustenance just like any other person, and he is subject to the same metabolic processes to fuel muscle during physical exertion. Evaporation refers to the process by which water is vaporized at the skin surface to promote heat loss from the body. The process of evaporation is influenced by the surface area exposed to the environment, ambient air conditions such as humidity and temperature, and the magnitude of convective air currents near the body. Thermoregulatory responses associated with heat loss from exercise, such as cutaneous vasodilatation and sweat production, take place in the epidermis and dermis layers (29, 91). In vasodilatation, the blood vessels widen to allow more blood to be brought closer to the surface of the skin, a process controlled by the autonomic nervous system (91). Coupled with vasodilatation is sweat production and its subsequent evaporation from the surface of the skin. Eccrine sweat glands and ducts secrete fluids onto the surface of the skin. These glands are distributed throughout the body, with the number of eccrine sweat glands in the range 1.6–4 million, depending on the surface area of the body (81, 91). An eccrine sweat gland consists of a secretory coil located in the lower dermis that leads to a duct in the epidermis. Examples of sweat ducts are shown in the MR-OCT image of the epidermis and dermis in Fig. 2. When in human form, Colossus’ body exhibits the usual skin physiological responses, such as thermoregulation via cutaneous vasodilatation and sweating. However, when in his steel form, these processes of thermoregulation will be adversely affected, given that the sweat ducts, and possibly the sweat glands, are covered or masked by a steel-like material. It is conceivable that Colossus’ metabolic heat production may internally self-adjust without the need for traditional thermoregulation responses. Heat loss via convection may compensate for the disruption of sweat production and associated evaporation. Convection is the transfer of body heat to the surrounding gas or liquid. As air molecules interact with a warm skin surface, the molecules become warmer. The process of convection could be further promoted by the increase in thermal conductivity of Colossus’ skin. The thermal conductivity of steel varies from 12 to 45 W·m−1·K−1, while the epidermis and dermis have a thermal conductivity of ~0.3 W·m−1·K−1 (13). Heat produced by the metabolic process could be brought close to the “organic steel” skin via vasodilatation, and the increased thermal conductivity of this skin could lead to increased heat transfer to the surface of the skin. For Colossus, convective transfer to the environment on its own may not be sufficient to regulate heat storage. Colossus’ steel skin acts as an impenetrable barrier between his body and the surrounding environment. Hence, it can be classified as a biological armor similar to osteoderms, which is a natural form of chain mail-like armor composed of bone plates located within the dermis of species such as the armadillo lizard (Ouroborus cataphractus) and giant girdled lizard (Smaug giganteus) (8, 43). Recently it has been demonstrated that this armor is not only important for protection, but also actively involved in thermoregulation. The osteoderms in armadillo lizards have been shown to be better insulators and have a lower thermal conductivity than those in the giant girdled lizard. Although the armadillo lizard has better thermoregulation responses than the giant girdled lizard, the increased thermal insulation of their osteoderms leads to a decrease in the strength of their armor (8). For the giant girdled lizard, the strength of their osteoderms might be greater, but the thermal insulation is lower compared with the armadillo lizard. Evidently, there is a trade-off between the effectiveness of thermoregulation responses and the strength in the osteoderms of these species. It is improbable that Colossus would accept a compromise on the strength of his “organic steel” skin just to retain a viable thermoregulation process. His impenetrable skin has proven to be a formidable defense against villains, such as Angel Dust and the Juggernaut. However, surrounding his body with a steel alloy does not conform with the expectations of thermoregulation responses. In the final battle of X-Men: The Last Stand (102), Colossus and the other X-Men fight Magneto and his Brotherhood of Mutants on Alcatraz Island. Over the course of the battle, Colossus is shown running, battling other mutants and even throwing Wolverine, whose skeleton has been bonded to adamantium, a fictional metal with a high density and whose mass is more than likely in excess of 150 kg. Inevitably these exertions will lead to increased heat storage in Colossus’ body. Given that his “organic steel” skin negatively affects traditional thermoregulation responses, hyperpyrexia can occur, where his core temperature could exceed 42°C, a temperature beyond which normal physiological functioning is adversely affected. For instance, a temperature of 42°C or above can result in the cessation of normal biological processes, such as DNA synthesis, and lead to arterial thrombosis, vomiting, and eventually death. Ironically, rather than protecting his well-being, his mutation could actually kill him. Evidently, this has not been the case, and Colossus has been shown to function normally when in his mutant form. This suggests that his skin does not consist of steel or an associated alloy. Before outlining a realistic candidate material for his impenetrable skin, I will demonstrate, using numerical and physiological analysis, that it is inconceivable that his impenetrable skin is composed of steel. Debunking “organic steel” skin. When comic book writers formulate a new character, their main priority is the narrative, and they are not strictly bound by scientific laws. Colossus’ impenetrable skin is not a unique superpower in the superhero universe. By being referred to as “organic steel” skin, the reader is led to believe that his mutant skin is made up of steel, an alloy with iron as the primary element. Roughly 96% of the human body is carbon, hydrogen, oxygen, and nitrogen. On the other hand, iron accounts for just 0.00006% (22). To demonstrate the improbability of Colossus’ skin consisting of steel, Colossus’ mass increase when he changes to his steel appearance is estimated. For ease of calculation, a number of assumptions are outlined. First, it is assumed that when Colossus changes, only the epidermis and dermis layers of his skin change. The hypodermis layer remains unchanged and retains biological functionality. Figure 2 shows an MR-OCT image of the epidermis and the dermis for a fingertip. According to the scale in the figure, the epidermis and dermis layers in the image have a combined thickness of 1 mm. The combined thickness of these layers depends on the location in the body. For instance, the skin thickness at an eyelid is ~0.5 mm, whereas the thickness is 4.0 mm at the heel. On average, the total thickness of the epidermis and dermis is 2 mm, and it is assumed that this thickness changes to steel. Second, the epidermis and dermis layers have a combined mass of 6.3 kg, where Colossus’ total mass is assumed to be 100 kg and skin mass is 1/16th of his total mass (53), whereas the skin area covering his body is ~2 m2 (105). Hence, with a skin thickness of 2 mm, the volume of skin that changes V skin = 0.004 m3. The third assumption relates to the density of “organic steel” skin. Steel alloys have a density varying from 7,750 to 8,050 kg/m3, depending on the other elements added to the alloy, such as carbon, chromium, and nickel. For Colossus, his steel skin is assumed to have a density ρ colossus = 8,000 kg/m3. In comparison, human skin has a density slightly higher than water (ρ skin = 1,050 kg/m3) due in part to the presence of biological components, such as nutrients, hairs, vessels, and nerves in the epidermis and dermis layers. Using the above assumptions, the change in Colossus’ mass when he transforms to his steel profile can be calculated. After transformation, and given that ρ colossus = 8,000 kg/m3 and V skin = 0.004 m3, his skin, more precisely the epidermis and dermis layers, have a mass of 32 kg. This equates to an overall mass increase of 25.7 kg, as his skin has a mass of 6.3 kg before transformation. Therefore, every time Colossus changes into his “organic steel” form his mass increases by 25.7 kg. For Colossus’ body to support this additional mass, he would require a dense skeletal structure and a highly defined muscular form. Both characteristics are evident from the portrayal of the character in the films, where Colossus is depicted as muscular and athletic in both his human and mutant forms. A mass increase of 25.7 kg should not pose any problems for Colossus. In fact, this mass increase is moderate compared with the mass of equipment that soldiers carry in modern combat. For instance, Australian soldiers on military operations between 2001 and 2010 carried a mean absolute load of 47.7 kg, with some artillery personnel carrying in excess of 70 kg (70). Colossus can have no complaints about his mass increase, given the loads carried by these soldiers. Colossus’ transformation does pose a problem in terms of the conversation of mass and energy. Mass-energy equivalence theory states that mass m of an object is directly proportional to the energy stored in the object E as summarized by E = mc2, where c = 299,792,458 m/s, the speed of light. For Colossus to change into his steel form and increase his mass by 25.7 kg, he requires E change = 2.313 × 1018 J from the Universe, which is a significant number. The energy requirement per day for an adult male is ~1.0 × 107 J (75), while the maximum fuel energy for an Airbus A380 is roughly 1.1 × 1013 J, both of which are many orders of magnitude lower than E change . Indeed Colossus’ energy requirements are even larger than the energy released during the eruption of the volcano Krakatoa in 1883, which is estimated to have been 8 × 1017 J! Like Colossus, the Hulk is another superhero character for which mass/energy conversion would be a critical issue. When Bruce Banner transforms into the Hulk, he markedly increases both his mass and height. In comparison to Colossus, Banner’s energy requirements are likely to be larger, given that Banner needs to create a vast and defined muscular appearance, which Colossus already has in his human form. Thus, when Banner becomes the Hulk, it is likely that, at the very least, his mass doubles, meaning that his energy requirements for the transformation dwarf Colossus’ needs and the energy released by Krakotoa. This energy calculation is based on the idealized view that the transformation process does not dissipate any energy. During the transformation process, energy will inevitably be used by the cells in biochemical processes to enable the transformation and lead to the production of waste thermal energy. A portion of this thermal energy may be transmitted via conduction to the surrounding tissue, such as the dermis skin layer. Thereafter, the energy could make its way to the epidermis and then be transmitted to the clothing and external environment via convection. Transfer of thermal energy in this manner can only take place in regions of the skin not yet covered by the steel layer. In addition, some energy will be lost in the form of sound as the steel “clicks” into place on his body. These energy losses make it impossible for Colossus to return the exact energy he “borrowed” from the Universe to create his steel skin layer. Therefore, the “organic steel” transformation process is not very energy efficient. Of greater concern though is the energy required for the transformation, which is 3.033 × 1018 J and effectively impossible to attain. However, there is an alternative material option for Colossus that takes advantage of the elemental composition of the human body. That material is graphene. Feasible Colossus Skin: Graphene What is graphene? The human body is predominantly made from nitrogen, oxygen, hydrogen, and carbon. Of these elements, carbon could hold the key for an impenetrable skin for Colossus comprising graphene. In nature, carbon is allotropic, meaning that atoms can be arranged in a number of different ways leading to differing material properties. For instance, in diamonds, carbon atoms are arranged in a diamond cubic crystal structure that directly influences the high strength and hardness of diamonds. On the other hand, graphite is composed of layered sheets where carbon atoms in each layer are arranged as a honeycomb lattice. Unlike diamond, graphite is a brittle material. A single honeycomb lattice layer, which is one carbon atom thick, is known as graphene (25, 32) and is illustrated in Fig. 3. Fig. 3.Illustration of the honeycomb arrangement of carbon atoms in a graphene lattice. Each disk represents a carbon atom. Download figureDownload PowerPoint

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For decades, the possibility of extracting a two-dimensional (2D) graphene layer from graphite was proposed, but a viable technique was lacking. However, in 2004, a technique for the exfoliation of graphene from bulk graphite was developed (66). Often referred to as the “Scotch-tape” technique, it involves the repeated stripping of layers from graphite using an adhesive surface such as Scotch tape (32, 66). A similar technique was subsequently developed, where graphitic layers rubbed against a solid surface leads to the deposition of flakes of graphite on the solid surface (67). Optical microscopy of these flakes on SiO 2 substrates can then be used to identify 2D graphene crystals. In recent years, graphene has generated huge interest in the scientific community due to its unique mechanical (1) and electrical properties (32). Despite being a million times thinner than paper, graphene is stronger than diamond and more conductive than copper. Graphene beams have been shown to be capable of supporting very large loads that can be millions of times the mass of the beams, thus demonstrating graphene’s impressive stiffness (7). In addition, graphene is extremely flexible and offers the potential for chemical modification (32, 33). Andre Geim and Kostya Novoselov were jointly awarded the 2010 Nobel Prize in Physics for their work on graphene (62). As of yet, though, commercial success has eluded graphene-based applications (71). Nonetheless, graphene has been used in the development of field-effect transistors (31) and flexible optically transparent films for tabletlike devices (2). In biological applications, graphene has been used in the development of wearable patches for sweat-based diabetes monitoring and microneedle-based drug delivery (50). Nanosheets of graphene oxide (GO) have been used in the photothermal treatment of bacterial and fungal wound healing in mice, whereby the GO sheets were mediated using a laser, thus leading to faster healing of wounds and demonstrating graphene’s biocompatibility (79a). Of particular significance for Colossus’ power, bio-inspired materials (82, 83) and nanomaterials based on graphene (51, 96) are actively being considered for state-of-the-art armors with advanced ballistic protection capabilities. In one study, silica μ-bullets with velocities close to 1 km/s were fired at various material targets (51). Layers of graphene were shown to outperform traditional protective materials such as steel and Kevlar, due to its higher strength and stiffness, and ability to delocalize energy from the region of impact. The protective capability and biocompatibility of graphene certainly provides a viable alternative to the steel skin of Colossus portrayed in the comic books and films. Protein folding and protein self-assembly. The potential formation of any protective bio-graphene layers in the dermis of Colossus, or any person for that matter, is critically dependent on the body’s ability to build biological aggregates. Fortunately, the human body facilitates the construction of complex aggregates via protein self-assembly, where proteins can assemble into structures ranging in size from nanometers to micrometers (57, 85, 94, 97). Protein self-assembly is highly important for many physiological processes in the human body. For instance, type I collagen, which is the most abundant protein in the human body, self-assembles into fibril structures that are, in turn, used in the formation of various tissues, such as tendons (38, 84). The triskelion-shaped clathrin protein is capable of self-assembling into polyhedral coats or cages near membranes. Clathrin cages are used in endocytosis, the process of transporting molecules into a biological cell. Notably, the cages have a regular lattice-like appearance (77). Another form of protein self-assembly takes place in a virus through the construction of a viral capsid, which is a coat that contains and protects the genome of a virus (57, 69). Multiprotein self-assembly processes are fundamentally dependent on the accurate folding of a protein into its native structure; otherwise, the protein will not function as intended (17). It has been demonstrated that the failure of proteins to fold into their correct native structure or to remain correctly folded for too long can influence the development of a number of diseases (16, 57). For example, it is now widely accepted that the self-assembly of amyloid-β peptides into fibrils and other biological structures is associated with the development of Alzheimer’s disease (61, 74). A peptide is a generic chain of amino acids, while an amyloid-β peptide is a specific peptide chain consisting of ~40 amino acids. Deposition of amyloid-β aggregates leads to the formation of senile plaques, neurofibrillary tangles, and neuronal cell loss, which are viewed as key pathological indicators for the disease (61, 76). Recently, protein self-assembly has inspired research in nanotechnology, in particular functional nanostructures that can self-assemble via controlled protein-protein interactions (36). There is also considerable interest in the field of 2D self-assembled protein arrays or membranes for applications in medical diagnosis and treatments. In a recent study, protein crystals with specific structures were shown to self-assemble to form 2D square lattices (88). Single- or double-point mutations of the C 4 -symmetric homotetramer RhuA facilitated the assembly of the proteins into 2D lattice structures via various intermolecular interactions. Interestingly, one variant of RhuA produced dynamic 2D lattices that, when stretched, become thicker in the direction perpendicular to the applied force. In the natural world, there are examples of regular or lattice-like structures within dermal layers (56, 93). Transmission electron microscopy images of the skin of the panther chameleon (Furcifer pardalis), a lizard found in Madagascar that is capable of color change for camouflage and social interaction with other chameleons, revealed the presence of two layers of iridophore cells containing guanine crystals (93). The upper layer, known as the superficial (S-) iridophore layer, consists of close-packed guanine crystals, each with a diameter of ~128 nm arranged in a triangular lattice. Differences in the refractive index of guanine and cytoplasm allows the S-layer to respond in a manner similar to a photonic crystal. When the chameleon undergoes a color change, the distance between adjacent crystals increases, while the size of the crystals remains constant. The process of color change takes roughly 2−3 min to complete, and it is completely reversible. Below the S-iridophores lies the deep (D-) iridophores layer, which is composed of a layer of bricklike disorganized guanine crystals. The D-iridophores are not associated with color change. Instead, they may help control the degree of absorption of sunlight, and hence aid in the thermal regulatory process of the chameleon. This discovery of regular lattices in the dermal layers of chameleons has already inspired the development of flexible photonic structures that change color when subject to deformation (49, 104). Colossus: forming a bio-graphene layer. The color-changing lattice layers in the dermal layers of the panther chameleon is supportive of the proposal that Colossus’ impenetrable power is based on a bio-graphene honeycomb lattice layer. Here, I outline a process for the creation of a bio-graphene layer in Colossus’ dermis that facilitates impenetrable skin. First, expression of the bio-graphene gene in dermal cells is initiated by an external source, such as a hormone that interacts with a receptor protein in the cell that, in turn, moves to the nucleus with the instruction for gene expression. This leads to the transcription of the bio-graphene messenger RNA, which is transported to the ribosomes. Construction of the primary protein structure then commences as the necessary proteinogenic amino acids are delivered to the ribosomes by transfer RNAs. It may be the case that nonstandard amino acids, which are amino acids that are not traditionally associated with the construction of proteins, are also required. Second, when the ribosome has completed construction of the primary structure of the protein, the native structure of the protein is formed via folding of the protein. The folding process is expedited by chaperones (17), which are proteins that guide the folding process and increase efficiency of the process by reducing the likelihood of misfolding and aggregate formation. Misfolded proteins would impede the accurate self-assembly of the bio-graphene layer. Once the proteins are in their native structure, the self-assembly process commences. Chaperone proteins may be involved in the self-assembly process to ensure that the bio-graphene proteins are properly orientated relative to neighboring proteins. This self-assembly process may be similar to the self-assembly of the 2D RhuA protein lattices (88). Recall that graphene is a single carbon atom thick with the atoms arranged in a honeycomb lattice. Therefore, to mimic the properties of graphene, the self-assembled 2D lattice must consist of carbon-atom-thick layer with structural support beneath the lattice. The key would be the formation of a pseudo-2D lattice that mimics the material properties of graphene, in particular its strength, stiffness, and flexibility. This lattice would be the ultimate protein superstructure, given that it would have a size that is of the order of meters as opposed to micrometers for the known largest protein superstructures (57, 94). Finally, the bio-graphene layer would then migrate to the interface between the epidermis and dermis (as depicted in the MR-OCT image of Fig. 2). When Colossus no longer requires the bio-graphene layer, the layer can be shed in a manner similar to the desquamation of corneocytes from the stratum corneum. A summary of the key steps in the production of the bio-graphene layer is given in Fig. 4. Fig. 4.Summary of the key steps in the production of a bio-graphene layer in Colossus’ body. 2D, two-dimensional. Download figureDownload PowerPoint While this process enables the generation of a bio-graphene layer, there is an issue with the timescale of the transformation. In the films and comic books, the timescale for any transformation process is of the order of seconds. However, the timescale for production of proteins from DNA in the human body can be of the order of minutes. To overcome this timescale issue, Colossus’ body could continually produce the bio-graphene proteins in the same manner that the body continually produces other proteins such as collagen and hemoglobin. The bio-graphene proteins are then added to Colossus’ blood and transported throughout the body along with the RBCs, white blood cells, other proteins, nutrients, platelets, and clotting factors. The addition of bio-graphene proteins can also affect blood viscosity, more specifically the plasma viscosity. Blood consists of blood cells, such as erythrocytes and leukocytes, dispersed in plasma. While blood is a non-Newtonian fluid in that its viscosity decreases with increasing shear strain, plasma is a Newtonian fluid such that its viscosity is invariant to changes in shear. Blood viscosity depends on a number of factors, such as hematocrit, RBC deformability, and plasma viscosity, while plasma viscosity is partially dependent on the type and number of proteins that it contains (45). For instance, the addition of lipoproteins has been shown to increase plasma viscosity (79), while a recent clinical study suggests that the level of plasma viscosity may be associated with cardiovascular disease (72). Thus the introduction of bio-graphene proteins will likely lead to an increase in plasma viscosity and hence an increase in blood viscosity. Speculating further, Colossus could have a greater risk of cardiovascular disease due to the presence of bio-graphene proteins in comparison to anyone without the ability to produce this protein. When Colossus requires the bio-graphene layer, biological cells in the dermis produce bio-graphene cytokines, which are proteins involved in cell signaling, such as in wound healing (37). As the blood moves through vessels in the dermis, the bio-graphene proteins in the blood interact with the cytokines, which signal the proteins to start the assembly of the honeycomb lattice. During the assembly process, blood viscosity may also increase locally. This process does not rely on a potentially time-consuming protein production and folding step. Nonetheless, the timescale for the formation of the bio-graphene lattice may still be 1 or 2 min, which is a similar timescale for the S-iridophore lattice of panther chameleons to deform and initiate a color change (93). In addition, there are issues with the construction of a biological graphene layer. Large-scale 2D structures such as graphene are susceptible to thermal fluctuations that can force 2D planes to develop 3D structures, which are much more stable (32). While an underlying protein scaffold may offer structural support for such a structure, thermal fluctuations at the molecular scale could certainly disrupt the development and prevalence of a 2D bio-graphene layer. Graphene solves physiological problems. In the Colossus Skin section above, I discussed how an “organic steel” layer could negatively affect Colossus’ thermoregulation processes and potentially lead to his death. Conversely, if Colossus’ impenetrable skin were composed of a bio-graphene protein lattice layer, a number of physiological problems could be addressed. First and foremost, Colossus’ ability to thermoregulate his core temperature via sweating would not be affected by the presence of a bio-graphene layer. Recent research has demonstrated that graphene can be employed in ultrathin membranes for water desalination purposes (78, 87). In one study, nanometer-sized pores were fabricated in a membrane consisting of a single layer of graphene. This membrane displayed both rapid water flux and a salt rejection rate of ∼100% (87). These findings could pave the way for the future use of graphene-based membranes in water treatment techniques. For Colossus, the ability for a bio-graphene layer to act as a permeable membrane would prove invaluable toward maintaining his natural thermoregulatory processes. The bio-graphene layer would not cover the sweat ducts, and, as sweat is produced by the sweat glands, the layer would allow for the transportation of fluids to the stratum corneum. Of greater significance is graphene’s ability to prevent the motion of ions such as Na+ and Cl−. During exercise, a person loses both water and key electrolytes, such as Na+ and Cl−, which can negatively impact performance (3). Initial losses of Na+ and Cl− due to sweating are balanced by reabsorption. However, with increased sweating rate, Na+ and Cl− losses become more evident as reabsorption methods for Na+ and Cl− are unable to balance losses. Thus a bio-graphene layer around Colossus’ body could help his body in the reabsorption of key electrolytes, maintaining athletic performance and allowing Colossus to battle villains for extended periods. A second physiological problem that graphene would solve is in relation to Colossus’ eyesight. When in human form, Colossus’ eyes function as normal. However, when covered by steel, his eyes are depicted as being completely covered in both the comic books and films. If his eyes were coated in steel, intuitively one would expect that this would seriously affect the functionality of the eye. If light cannot be focused onto the retina and interact with the photoreceptor cells, then Colossus would not be able to see. Nevertheless, it is clear from both the comic books and the films that Colossus can see when in his mutant form. A bio-graphene layer covering Colossus’ body, including his eyes, could address this inconsistency. One of the unique properties of graphene is its transparency, which would not inhibit light from interacting with the lens of the eye. Graphene is actively been considered for advanced smart contact lenses due to its advanced electrical and mechanical properties as well as its biocompatibility (11, 52). This lens reduces exposure to strong electromagnetic waves and prevents dehydration of the eye. Graphene absorbs the electromagnetic energy and dissipates it as thermal energy, thus protecting the interior of the eye (52). If Colossus’ impenetrable skin is composed of a bio-graphene layer, this will also lead to a major change in his appearance. Given graphene’s transparency, when in his mutant form, the bio-graphene layer would be indistinguishable from his stratum corneum. Graphene films produced via chemical vapor deposition have been shown to be 97.4% optically transparent (2). Nevertheless, the color of graphene and GO layers have been shown to depend on the type of substrate below the layers and the thickness of the layer (42). For instance, graphene layers with a thickness of ~50 nm deposited on SiO 2 /Si layer have a silverlike color, depending on the illumination level. This color would roughly match the color of Colossus in the comic books and films. One major issue though is the SiO 2 /Si layer, given that the average human body is composed of just 0.001 kg of silicon. This does not discount that another material substrate could lead to the same color of the graphene layers. However, for the moment, it can be assumed that, if Colossus can generate a bio-graphene layer around his body, then it is a transparent layer. A final physiological problem that a bio-graphene layer could address is the flexibility portrayed by Colossus’ body during a battle with the Juggernaut in the 2018 film Deadpool 2 (14). During the fight, the Juggernaut blocks a punch from Colossus and instantly bends Colossus’ radiocarpal joint into an extremely unnatural position. Colossus’ shriek is an indicator that it is distinctly painful. However, later in the fight, Colossus’ hand and wrist have returned to their normal configuration, and there is no suggestion of discomfort for Colossus. This suggests that his skin, and bones for that matter, are exceptional flexible, while resisting permanent damage or fracture. Graphene has a high material strength and is exceptionally flexible (32). The flexibility of a bio-graphene layer at the interface between the epidermis and dermis would explain the ability for Colossus’ skin to elastically respond to deformation. This scene with the Juggernaut does suggest that other parts of Colossus’ body may be made of bio-graphene. However, for brevity, the focus here is on graphene in the skin. The physiological problems addressed by bio-graphene are summarized in Fig. 5. Fig. 5.Physiological problems with steel as a skin for Colossus that are solved or addressed with a bio-graphene-impenetrable skin. EM, electromagnetic. Download figureDownload PowerPoint Human Enhancement and Ethics Colossus and his impenetrable power could be used to motivate real-life beneficial technological developments and prompt ethical discussions in relation to human enhancement and ethics. Graphene-based materials are already being explored for more beneficial applications. In current technological developments, bio-inspired composite materials containing carbon nanotubes or graphene are being considered as new material options for the protection of spacecraft against the impact of meteoroids and space debris (82), which can of course serve to maintain the habitable environment for astronauts. It can be envisaged that these materials would also be utilized in structures and transport vehicles on Earth, such as cars, trains, and buildings. The biocompatibility of graphene makes it a highly suitable material for medical treatments. GO has been used for the development of treatments that could lead to the faster healing of wounds (79a). In addition, GO could be used for the delivery of osteoinductive proteins and stem cell recruitment agents for bone regeneration in the vicinity of titanium implants (48). Research is also ongoing in relation to the integration of graphene in advanced biomedical sensors and regulators for pharmaceutical dosages (50). While these studies are unlikely to have been inspired by Colossus, the character and his power could stimulate new research on protective materials, medical treatments, and advancements in biocompatible implants. A critical issue with regards to the use of graphene-based medical treatments is whether or not a person is enhanced by the treatment. Human enhancement can be defined as “biomedical treatments to improve human capacities, performance, dispositions, and well-being beyond the traditional scope of therapeutic medicine” (34). This definition can be applicable to cognitive enhancement, for which there has been debate on appropriate governance issues (80). However, the use of graphene for wound-healing, bone regeneration, or controllers for the administration of medicines leads to the enhancement of key physiological responses in the human body. In the case of bone regeneration, GO serves to promote bone regeneration by transporting osteoinductive proteins and stem cells to the region of interest. While the GO primarily serves to hasten the integration of implants in a person’s body, such treatments might have unexpected advantageous side effects for the person. For example, some of the GO could be transported to other skeletal areas of the body. If there is a bone fracture or break, the associated osteoinductive proteins and stem cells could speed up the bone regeneration process. In effect, the person would heal faster and over a timescale that is shorter than normal human capacity. I acknowledge that this is conjecture and subject to innumerate physiological processes in the human body, many of which we do not yet fully understand. Nonetheless, current research on the use of graphene does support the possibility of the material playing an important role in future medicine. The recent advancements in genetic editing techniques, such as CRISPR/Cas9 (18, 30, 40), offer the possibility of modifying DNA to theoretically allow the body to fabricate new proteins, perhaps based on graphene, that could promote enhanced regenerative processes. While it is proposed that Colossus generates an impenetrable bio-graphene layer via a complicated protein-folding and self-assembly process, modifying human DNA using CRISPR/Cas9 to promote the assembly of such a layer is unquestionably a problematic task. For instance, recent research indicates that CRISPR/Cas9 can lead to frequent unintended genetic mutations far from the intended target site in DNA (47). Many of the scientific methods and technologies presented here, as well as in other similar superhero-themed articles (9, 24−26, 28, 63, 86), can be categorized as New and Emerging Science and Technology (NEST), a topic for which there has been notable ethical debate (55, 64, 65, 89). The impenetrable skin of Colossus and the hypothesized bio-graphene layer can also be used to differentiate between short- to mid-term ethics and speculative ethics. Short- to mid-term ethics refers to timely and relevant principles that can aid in the positive and practical integration of new technologies in society. For instance, GO is being considered for applications in wound healing to address health issues with pathogenic microbes (79a). Initial experiments with mice have shown that GO-mediated photothermal therapy may offer an alternative to antibiotic treatments. However, further laboratory research is naturally required, and relevant ethical guidelines are necessary to ensure its safe and responsible application in future nanomedicine. On the other hand, speculative ethics presupposes that a conjectured future world is inevitable where new technologies will facilitate unparalleled advancements in materials or human enhancement (64, 65). These speculations can be encapsulated with “if-and-then” statements, which starts with a possible technological advancement that then leads to a postulated “certain” future. For example, if humanity creates graphene-based bulletproof materials, then impenetrable wearable graphene armors will be developed and made widely available. This suggests that ethics to deal with their integration in the various facets of society are needed immediately. As pointed out by Nordmann and Rip (65) in relation to speculative ethics: “the imagined future overwhelms the present.” In other words, speculative ethics on technologies in their infancy diverts attention and resources away from more pressing technological developments that are in need of ethical consideration. Based on the arguments related to speculative ethics, I refrain from discussing the implications of impenetrable bio-graphene protein assemblies on society, given that their development is very much theoretical. However, other emergent technologies presented here, such as graphene-based wound-healing treatments, drug delivery technologies, and bulletproof materials, offer the real possibility for the synthetic enhancement of human physiology. Invariably, many of these technologies could be utilized in the military to augment the capabilities of soldiers. This motivates ethical and moral questions in relation to these technologies, such as: “Is the application of such technologies in military circles responsible?” “How should we decide who is morally capable of dealing with the consequences of using these technologies?” and “Can morality change with the introduction of these technologies?” It has been argued that morality can adapt to new technologies (90), but this will take time and is likely to be technology dependent. One possible way to choose those with suitable morality is a preselection test such as that used to identify Steve Rogers for a supersoldier program in Captain America: The First Avenger (10). However, such a test would have to be carefully constructed, since interaction with the new technologies could adversely change the moral viewpoint of a subject.

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Pedagogical Links While based on current scientific research, the self-assembly process for the construction of Colossus’ macroscale bio-graphene layers presented here does not account for a multitude of biological and physiological factors. For instance, issues related to pH, temperature, ionic strengths, and the composition of solutions for protein folding and self-assembly have not been addressed. Also, the biological pathways and neural activation control of the self-assembly process have not been described. Hence, Colossus and his proposed bio-graphene-impenetrable skin should be viewed as engaging content that can motivate classroom debate, student projects, and perhaps even research projects for the dedicated scientists. Adaptation of a popular culture character such as Colossus in the classroom could create a more engaging and fun learning environment, facilitate more open dialogue between educators and students, emphasize specific learning objectives, and promote critical thinking. The primary topics associated with Colossus are summarized in Fig. 6. In an introductory anatomy and physiology module, the integumentary system is typically a primary unit where learning objectives relate to topics such as the structure of skin and the function of skin, such as protecting against pathogens and aiding in the body’s thermoregulation processes. The following questions or investigations based on the integumentary system and Colossus could be posed to students in the classroom or as part of student projects: Fig. 6.Summary of the various topics in physiology and other disciplines that can be addressed in the classroom using Colossus. Download figureDownload PowerPoint 1. With reference to your own experiences of exercising and thermoregulation via sweating, speculate on the sensations a steel-covered Colossus might experience during prolonged physical exertion. 2. Dean Karnazes is a famed ultramarathon runner who has raced on numerous occasions in the Badwater ultramarathon, a 135-mile race that starts in Death Valley, CA, and has been heralded as “The World’s Toughest Footrace.” During the race, Dean and other runners often wear UV-proof suits to deflect heat from the summer sun. Explore if such a suit would be of benefit to Colossus when he exercises. 3. Rust, otherwise known as iron oxide, is formed when iron or an alloy of iron, such as steel, reacts with oxygen in the presence of water. When in his steel form, Colossus may accelerate the formation of iron oxide in his skin as sweat trapped in the dermal layers may promote rust formation. Given the hazardous nature of iron oxide, what health implications would this have for Colossus and the functionality of his integumentary system? 4. The dermis layer contains nerve endings that react to sensations such as heat, cold, touch, and pressure. How would the functionality of the nerves in Colossus’ dermis layer be affected when his skin is covered by steel? 5. Graphene as the perspective protective material for Colossus’ impenetrable skin offers a number of advantages over steel. How would graphene negatively affect Colossus’ physiology? In addition, for courses including protein folding and self-assembly, the following questions could be employed: 1. The potential for Colossus to generate a bio-graphene layer is dependent on protein folding and self-assembly, two processes that are integral for human life. Propose a detailed summary of the folding of a bio-graphene protein into a quarternary protein structure. 2. Outline in detail the process by which the bio-graphene proteins could self-assemble to form a 2D lattice dermal structure. As presented in this article, a study on Colossus’ physiology considers a number of disciplines, many of which intersect in a biomedical engineering course. First, the aforementioned physiology questions can be used to substantiate learning objectives in physiology modules linked to a biomedical engineering course. Second, the following questions in relation to biomedical electronic and medical devices can also be utilized: 1. A photoplethysmography (PPG) device quantifies heart rate by illuminating a tissue with light and then measuring how the intensity of the light changes due to scattering in the tissue. a. Would it be possible to use a PPG device to measure Colossus’ heart rate and other cardiovascular functions, even when his skin is covered by steel or graphene? b. What adaptations might be necessary to allow the PPG device to take measurements through steel or graphene? 2. In addition to imaging skin structure, OCT can be used to measure blood flow and heart rate as well as image fingerprints. How could OCT be integrated into a fingerprint reader to accurately image one of Colossus’ prints, even if he is covered in steel or graphene? Colossus can be also used to stimulate discussion on short- to mid-term ethics, speculative ethics, and morality with sample questions provided in the previous section. Additionally, Colossus can motivate the examination of both hard science and soft science topics in the classroom (26, 39). Hard science fiction, which relates to technological developments such as time machines or impenetrable materials, may appeal more to male students in physics, whereas soft science fiction, which relates to the application of scientific developments for the benefit of society, such as advanced medical treatments, may be of interest to mostly female students (39). The prospect of an impenetrable skin made of graphene is indicative of hard science, whereas the use of graphene in medical treatments for wound healing and bone regeneration are representative of soft science. Therefore, Colossus can function as a powerful exemplar that can help educators avoid neglecting specific groups in the classroom. Just like Hawkeye of the Avengers (26), I also employ Colossus in my public outreach activities. Although Colossus appeared in earlier X-Men films (99, 100, 102), there has been a recent surge of interest in the character due to his appearances in the Deadpool films (14, 15). In the presentations, I discuss many of the aspects presented in this paper, including the ethical and moral topics related to the character. Discussions on Colossus have been well received, with many people fascinated by the character’s impenetrable power and seeking further explanation of the scientific topics that I link to the character.

journals.physiology.org - The physiology of impenetrable skin: Colossus of the X-Men

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