Innocuous sterilant using hemocyanin and functionalized fullerenes with broad-spectrum intracellular and interstitial microbiocidal and radical scavenging effects for packaged matter, biologics and organics including liquids, gases, tissue, organs, cells, and limbs with copper mediated oxygenation for viability and preservation

Abstract

Tissue and organ transplantation success is chiefly dependent on the harvest and preservation techniques employed. Yield and quality enhancements are needed, because qualified patient demand far outpaces donors and ultimately, tissues and organs stored and transported so as to render them in acceptable condition for surgery. In the case of corneal transplants, tissue supplies help 1 out of 70 patients in need despite being among the most successful such procedures today. Transplant success has improved overall with enhanced storage and transport methods that have allowed for greater time and distances to reach patients in need, whether corneas, kidneys, hearts, lungs, livers or other tissues, limbs or cellular materials. While corneal tissues specifically benefit from oxygenation via direct air contact and normal tears in a healthy individual, once donated, the lack of corneal vascularization can uniquely accelerate the depletion of oxygen that occurs upon harvesting of all transplant materials. Directly infusing oxygen has had little impact, given partial pressure requirements, volatility and reactivity to many compounds, including self-affinity or agglomeration and bubbling. However, the hemocyanin, such as that of the horseshoe crab (Limulus polyphemus), can deliver oxygen directly to transplant tissues, while free radical scavenging by pristine fullerenes can help maintain cellular integrity. Horseshoe crabs have successfully evolved by developing two immunological mechanisms, amoebocytes and less characterized antiviral and gram-positive bactericidal properties ascribed to other hemolymph constituents. The amoebocytes have a specific, ultrasensitive affinity to gram-negative bacteria and fungi; they engulf and consume these microbes in an enzymatic coagulation process. However, hemocyanin is extracellular and non-toxic to mammalian tissue, notwithstanding its microbiocidal properties. As such, the viral-scale peptides appear to intracellularly invade the pathogens and disrupt replication without conferring a caustic or toxic effect on non-pathogenic tissue. Hence, hemocyanin appears to be non-immunogenic and thus applicable as a carrier molecule for some human therapeutics (e.g., hemocyanin from Keyhole Limpet snails). The present invention relates in the initial embodiment to the use of horseshoe crab hemocyanin and functionalized carbon nanostructures, halogenated fullerenes, pristine fullerenes and fullerene derivatives as an antimicrobial and antioxidant enrichment composition that is added to tissue storage and preservation media as a potent, broad-spectrum antimicrobial and tissue preservation composition for safe and effective storage and transport. Notably, methods for horseshoe crab aquaculture husbandry have been developed to achieve sustainable hemocyanin supplies for hemolymph harvest and biomedical applications. With regard to unique needs and benefits for the transplantation of corneal tissue, the aim of this patent is development of an antioxidant and broad-spectrum microbicidal that is benign to endothelial and endothelium cells to preserve and maximize the viability of a pathogen-free specimen.

Claims

1. An antimicrobial composition that has broad-spectrum biocidal activity to eradicate bacteria, viruses, and fungal pathogens, wherein the antimicrobial composition has oxygen transport activity, maintains the integrity of cells and does not comprise the integrity of packaged materials, the antimicrobial composition comprising: a. a biocidally effective amount of one or more halo fullerenes with the chemical formula of C.sub.60X.sub.6, C.sub.60X.sub.8, or C.sub.60X.sub.24, wherein X is a halogen selected from the group consisting of fluorine, chlorine, bromine, and iodine; b. a biocidally effective amount of one or more pristine fullerenes with the chemical formula of C.sub.60, C.sub.70, C.sub.76, C.sub.78, C.sub.80, C.sub.82, or C.sub.84; c. an effective amount of horseshoe crab or other invertebrate extracellular hemocyanin; d. optionally one or more organic or inorganic ions; and e. a buffer to maintain neutral pH of the composition.

2. The composition of claim 1, wherein the concentration of halo fullerenes is between 0.01% and 10.0%.

3. The composition of claim 1, wherein the concentration of pristine fullerenes is between 0.01% and 10.0%.

4. The composition of claim 1, wherein the concentration of extracellular hemocyanin is between 1.0 μg/mL and 1,000 μg/mL.

5. The composition of claim 1, which further comprises one or more of the following: raw hemolymph, proteins or peptides from the phylum Arthropoda or Mollusca.

6. The composition of claim 1, which further comprises one or more functionalized fullerene derivatives.

7. The composition of claim 1, wherein the organic or inorganic ions include one or more of the following: calcium, sodium, chloride, potassium, magnesium, lactate, urate, or another divalent ion, at a concentration that enhances hemocyanin and oxygen-binding affinities.

8. The composition of claim 1, wherein the halo fullerenes, pristine fullerenes, and hemocyanin are biocidal and neutralize bacterial, fungal and viral pathogens present in the packaged material.

9. The composition of claim 1, wherein the halo fullerenes, pristine fullerenes, and hemocyanin are antioxidants that scavenge free radicals present in the packaged material.

10. The composition of claim 1, wherein the buffer is sufficient to maintain a neutral or physiological pH during the period of contact.

11. The composition of claim 1, wherein the composition sustains the viability without damage to the packaged material.

12. A method of preserving and maintaining an organ, a tissue, a limb, a cell, a nutrient medium, a storage medium, a packaged material, or reusable instruments or devices, by contacting the organ, the tissue, the limb, the cell, or the other packaged material with a therapeutically effective amount of the composition of claim 1.

13. The method of claim 12, wherein the contacting comprises soaking, immersion, perfusion, flushing, washing, mixing, combining, or diluting.

14. The method of claim 12, wherein the contacting of the organ, tissue, limb, cell, nutrient medium, storage medium, or other packaged material with the composition of claim 1 sterilizes or neutralizes bacterial, viral, and fungal pathogens contained in or on the organ, the tissue, the limb, the cell, nutrient medium, storage medium, or other packaged material.

15. The method of claim 12, wherein the contacting of the organ, tissue, limb, cell, nutrient medium, storage medium, or packaged material with the composition of claim 1 preserves and maintains the integrity the organ, tissue, limb, cell, nutrient medium, storage medium, or packaged material with antioxidant, radical-scavenging properties.

16. The method of claim 12, wherein the organ, the tissue, the limb, cell or other material is intended for transplantation.

17. The method of claim 12, wherein the packaged material is a reusable instrument or device.

18. The method of claim 12, wherein contacting the reusable instruments and the devices with the composition of claim 1 cold sterilizes the reusable instruments and the devices.

19. A method for non-staining disinfection of a substrate with oxygen transport and free-radical scavenging properties, comprising contacting the substrate with an amount of the composition of claim 1 that is effective for disinfection.

20. The method of claim 19, wherein the substrate is skin, ocular regions, organs, tissues, or wounds.

Description

BRIEF DESCRIPTION OF THE DRAWING

(1) FIGS. 1A, 1B, and 1C are molecular representations of prototypical halo fullerenes of 60 carbon atoms functionalized with 4, 8, or 24 halogens (X).

DETAILED DESCRIPTION OF THE INVENTION

(2) The composition of the present invention includes a novel allotrope of carbon, the fullerenes, which are carbon nanospheres that typically measure less than 100 nm. Fullerenes possess a unique and powerful radical scavenging capability that takes up electrons from radicals (an ongoing organic decay process with microbes). The carbon cage of fullerenes can absorb electrons and disperse them through the 3D π-conjugated structure distributed over its surface. In the present invention, the fullerene cage is vital, because the mass is on the atomic scale (1-100 nm) but arranged in a geodesic configuration of hollow/empty space. The fullerenes draw pathogens toward the carbon nanomaterial orbitals during electron exchanges using atomic forces. These fullerene physical properties represent a stable and broad-spectrum molecule with high binding affinity to all pathogens (which typically possess negatively charged cell walls). Furthermore, the outer carbon cages of the fullerenes are capable of modification via chemical functionalization of side-chain moieties. The use of specific chemical reactions that result in the generation of a halo fullerene, or a functional fullerene cage possessing multiple side-chain halogens (e.g., iodine, bromine, chlorine and fluorine), renders a nano-scale antimicrobial agent. In the present invention, fullerenes are functionalized for aqueous relaxivity as well as bonding with potent halogen molecules to leverage their high pathogen affinity and create an inescapable antimicrobial atomic field. In turn, the halo fullerenes caustically eradicate microbes on contact, without lost energy or halogens, hence maintaining a continuous antimicrobial capability.

(3) A broad range of carbon shell fullerene modifications have been demonstrated via numerous, well-described chemical reactions [Yan, et al., 2015]. The halo fullerene of the present invention is comprised of a fullerene shell of C.sub.2n; whereby, n=10, 12, 13, 14, 15, . . . 360, such that multiple side-chain halogens can be attached to the core carbon cage. The halo fullerene of the present invention includes three carbon-60 nanomaterials of C.sub.60X.sub.6; C.sub.60X.sub.8, and C.sub.60X.sub.24; whereby, X=a halogen molecule (e.g., bromine, chlorine, iodine, or fluorine). In turn, the pristine (non-functionalized) fullerene or functionalized fullerene derivative component includes C.sub.60, C.sub.70, C.sub.76, C.sub.78, C.sub.80, C.sub.82, C.sub.84 and higher molecular weight molecules with or without the addition of functional groups on the outer shell of the core carbon cage. Functionalized fullerenes include common fullerenol (—OH) nanoparticles, as well as chemical functionalization with amino acids, proteins, peptides, carboxylic acids, polyhydroxyl groups, amphiphilic polymers, or any type of mono- or multi-addition to the external fullerene carbon topology via nucleophilic or pericyclic reactions, hydrogenation, oxidation, hydroxylation, electrophilic, carbene, or radical additions, etc.

(4) Halogen atoms are hydrophobic and lipophilic, which may be exploited for a myriad of applications. Lipophilicity enables halogens to cross and pass through cell membranes. In the parent patent, the halogen functional group applies these highly caustic atomic properties that induce cell rupture and destruction of vulnerable microbes, without a net loss of energy or efficacy, for ongoing caustic capabilities at the application or exposure site. Likewise, pristine fullerenes and functionalized fullerene derivatives have demonstrated iterative capture of free radicals, a dynamic radical scavenging mechanism for maintaining cellular viability, without a net loss of energy or efficacy, for continuous antioxidant capability at the application or exposure site.

(5) In addition to the antioxidative and caustic properties of the functionalized fullerenes in the proposed invention, the composition encompasses hemolymph derived from horseshoe crabs, which contains proteins and peptides associated with its primordial immunity. Among these is hemocyanin, a well characterized molecule found in the Arthropoda, Mollusca, and certain insect hemolymph.

(6) Hemocyanin is a multi-functional protein that provides oxygen transport and host immunity and is the predominant protein component (between 50% to 90%) of invertebrate hemolymph. In L. polyphemus, hemocyanins are the largest recorded and present in the highest concentrations, exceeding 100 mg/mL.sup.−1 (Coates et al., 2017; Coates et al., 2012 and Van Holde et al., 2001). The oxygen binding affinity of horseshoe crab hemocyanin is modulated by protons or various organic ions (e.g., calcium, sodium, chloride, potassium, magnesium, lactate, urate, etc.) with optimal binding at or around physiological pH and across a broad temperature range (4 to 37° C.). Unlike hemoglobin, which is contained in the red blood cells of vertebrates, hemocyanin is dissolved and distributed throughout the hemolymph. While oxygen-transport blood substitutes (i.e., perfluorocarbons, human red blood cells, artificial hemoglobin-based oxygen carriers) require limited, specific conditions, hemocyanin is stable and functional across a broad range of temperatures, including colder environments, which reduce cellular metabolism and are favored in transplant preservation protocols. In contrast to hemoglobin dependence on negative allosteric effectors for oxygen diffusion, L. polyphemus hemocyanin can passively release oxygen with a gradient that optimizes uniform delivery without requiring external allosteric interactions.

(7) Given their primary function to transport oxygen to respiring tissues within invertebrates, hemocyanins contain di-cupric groups that reversibly bind molecular oxygen (van Holde et al., 2001) and play a dual role in host immunity. However, L. polyphemus hemocyanin notably differs substantially from bulk or nanoscale copper used as a conventional healthcare antimicrobial agent. Likewise, most antibiotics require knowledge of the causative agent for effective application; whereas, this invention utilizes a composition of radical scavenging and antimicrobial halo fullerenes and hemocyanin for antioxidant and broad-spectrum biocidal capabilities across species, as well as mutated species.

(8) In addition to oxygen transport, hemocyanin is associated with numerous homeostatic and physiological processes, such as molting, hormone transport, osmoregulation, and protein storage (Adachi et al., 2005; Kuballa and Elizur, 2008; Glazer et al., 2013; Jaenicke et al., 1999; Paul and Pirow, 1998). Immunologically, hemocyanin is capable of converting into phenoloxidase-like (PO) enzymes upon physical disruption of the structural motifs surround the central coppers. These POs play vital roles in a broad-based response to pathogens (Coates et al., 2012); whereas, hemocyanin demonstrates greater activity than POs against bacteria and fungi. In addition to hemocyanin, horseshoe crab hemolymph contains a range of molecules with profound antimicrobial functionality (Iwanaga, 2002). See Table 1 (Iwanaga, 2002).

(9) TABLE-US-00001 TABLE 1 Defense Molecules Found in Amebocytes and Hemolymph Plasma of the Horseshoe Crab. Proteins and Peptides Mass (kDa) Function Specificity Localization Coagulation Factors Factor C 123 Serine protease L-granule Factor B 64 Serine protease L-granule Factor G 110 Serine protease L-granule Proclotting enzyme 54 Serine protease L-granule Coagulogen 20 Gelation L-granule Protease Inhibitors LICI-1 48 Serpin/Factor C L-granule LICI-2 42 Serpin/Clotting Enzyme L-granule LICI-3 53 Serpin/Factor G L-granule Trypsin Inhibitor 6.8 Kunitz Type Not Determined LTI 16 New Type Not Determined LEBP-PI 12 New Type L-granule Limulus Cystatin 12.6 Cystatin Family 2 L-granule a2-Macroglobulin Granule 180 Complement Plasma and L-granule Chymotrypsin Inhibitor 10 Not Determined Plasma Antimicrobial Substances Anti-LPS Factor 12 Gram Neg Bacteria L-granule Tachyplesins 2.3 Gram Neg/Gram Pos/Fungal S-granule Polyphemusins 2.3 Gram Neg/Gram Pos/Fungal S-granule Big Defensin 8.6 Gram Neg/Gram Pos/Fungal L-granule and S-granule Tachycitin 8.3 Gram Neg/Gram Pos/Fungal S-granule Tachystatins 6.5 Gram Neg/Gram Pos/Fungal S-granule Factor D 42 Gram Neg Bacteria L-granule Lectins TL-1 27 LPS (KDO), LTA L-granule TL-2 27 GlcNAc, LTA L-granule TL-3 15 LPS (O-antigen) L-granule TL-4 470 LPS (O-antigen), LTA Not Determined TL-5 380-440 N-acetyl Group Plasma Limunectin 54 PC L-granule 18K-LAF 18 Hemocyte Aggregation L-granule Limulin 300 HLA/PC, PE, SA, KDO Plasma LCRP 300 PC, PE Plasma tCRP-1 300 PE Plasma tCRP-2 330 HLA/PE, SA Plasma tCRP-3 340 HLA/SA, KDO Plasma Polyphemin Not Det. LTA, GlcNAc Plasma TTA Not Det. SA, GlcNAc, GalNAc Plasma Liphemin 400-500 SA Hemolymph Carcinoscorpin 420 SA, KDO Hemolymph GBP 40 Gal Hemolymph PAP 40 Protein A Hemolymph Others Transglutaminase 8.6 Crosslinking Cytosol 8.6 kDa Protein 8.6 TGase Substrate L-granule Pro-rich Protein 80 TGase Substrate L-granule Limulus Kexin 70 Precursor Processing Not Determined Hemocyanin 3,600 Oxygen Transporter (PO Activity) Plasma Toll-like Receptor (tToll) 110 Not Determined Hemocyte L1 11 Unknown L-granule L4 11 Unknown L-granule Gal: Galactose; GalNAc: N-Acetylgalactosamine; GBP: Galactose-binding Protein; GlcNAc: N-Acetylglucosamine; HLA: Hemolytic Activity; KDO: 3-deoxy-α-D-mannooctulosonic Acid; LAF: Limulus 18-kDa Agglutination-aggregation Factor; LCRP: Limulus C-reactive Protein; LEBP-PI: Limulus Endotoxin-binding Protein-Protease Inhibitor; L-granule: Large Granule; LICI: Limulus Intracellular Coagulation Inhibitor; LPS: Lipopolysaccharide; LTA: Lipoteichoic Acid; LTI: Limulus Trypsin Inhibitor; PAP: Protein A Binding Protein; PC: Phosphorylcholine; PE: Phosphorylethanolamine; PO: Phenoloxidase; SA: Sialic Acid; S-granule: Small Granule; TGase: Protein-glutamine γ-glutamyl-transferase; TL: Tachylectin; TTA: T. Tridentatus Agglutinin.

(10) The cellular material, or the amebocytes, in horseshoe crab hemolymph contains antimicrobial polypeptides and peptides stored in two granules, the small granule (S-granule) and the large granule (L-granule). The S-granule-derived antimicrobial peptides bind to chitin but not to other polysaccharides, such as cellulose, mannan, xylan, and laminarin (Iwanaga, 2002). Both the L- and S-granules contain a substance termed “Big Defensin” that is similar to mammalian neutrophil-derived defensins, but distinct in size (Lehrer, 1992; Ganz and Lehrer, 1995). Big Defensin strongly inhibits the growth of gram-negative and gram-positive bacteria, and fungi (including the cause of most corneal transplant fungal infections, C. albicans). Numerous lectins in both the plasma and granules, and several bacterial agglutinins, interact to protect horseshoe crabs from invading microbes and foreign substances (Iwanaga et al., 1998; Kawabata et al., 2003). Additionally, the hemolymph contains a class of C-reactive proteins (CRP) that exhibits cytolytic and opsonic activities against foreign cells and bears structural similarity to the complement system in mammals (Iwaki et al., 1999).

(11) The antimicrobial polypeptides and peptides of the horseshoe crab hemolymph also demonstrate peptide-membrane interactions and cellular uptake that provide an intracellular bulwark against bacteria, viruses or other pathogens that evade the extracellular environment and may act as latent reservoirs of infection harbored within the cell. Additionally, these peptides are capable of binding to cell receptors and prevent pathogen entry, as well as inhibiting cell-to-cell pathogen proliferation.

(12) The invention suspension is easily added to the packaged material alone or can be used in conjunction with nutrient dense media commonly used for the specific packaged biologics, whether organs or other organic material.

(13) The preferred embodiment of this invention includes caustic halo fullerenes (halogenated functionalized fullerenes), pristine fullerenes and/or a functionalized fullerene derivative, and horseshoe crab hemocyanin in a neutral, buffered, aqueous stabilizing suspension for use as an additive to media or as a standalone antimicrobial composition for package materials that require sterility. The extracellular hemocyanin is contained in the hemolymph collected from the horseshoe crab (or alternative Arthropoda or Mollusca hemocyanin source) and can be purified from the hemolymph. In addition to antimicrobial capabilities, the inclusion of horseshoe crab hemocyanin transports and delivers necessary oxygen. The composition is further understood to be nontoxic, neutral with respect to organic matter and otherwise innocuous to any non-pathogenic components. It is also understood that additional embodiments may include halo fullerenes, other functionalized fullerenes, pristine fullerenes, hemocyanin, and other components contained in hemolymph as a sterilant composition.

(14) The inclusion of halo fullerenes and hemocyanin provides broad protection from pathogens with multiple mechanisms to impede microbial mutation and resistance as alternatives to antibiotic additives. The invention would impart the intended antimicrobial benefits of antibiotics while averting risks of tissue cell wall or plasma membrane damage, disruption of DNA or RNA synthesis, inhibition of protein synthesis through binding of peptides with ribosomal subunits, or interfering with bacterial metabolic pathways. Apart from some broad-spectrum classes, antibiotic efficacy is also typically optimized with known species—but ineffective against viruses and fungi, all of which may be difficult to anticipate in tissue and organ transport or storage. Confounding possibilities also include those of co-colonization between two types of species (i.e., gram-negative and gram-positive bacteria) or across microorganisms (e.g., a fungus, gram-negative bacteria, and virus).

(15) In theory, including broad-spectrum antibiotics in combination with antifungal and antiviral material would represent the most comprehensive approach to ensuring sterile tissue and organ storage and transport. However, optimal concentrations of such additives have been shown to pose risks of cytotoxicity and lost specimen viability. Another consideration is temperature, as antibiotics are generally most effective at higher temperatures than those in typically cooler tissue and organ storage conditions.

(16) The combination of halo fullerenes and more broadly, radical scavenging pristine fullerenes and/or fullerene derivatives, and hemocyanin would establish a comprehensive antimicrobial solution to eradicate bacteria, viruses, or fungi, with a broader range of temperature and storage flexibility than antibiotics and antifungals. Furthermore, replacing traditional preservation additives such as surfactants, antiseptics, detergents, and disinfectants with halo and scavenging fullerenes and hemocyanin would provide sterility while further mitigating risks of cytotoxicity to improve specimen quality and viability, as well as potential longevity from antioxidant activity.

(17) Beyond antimicrobial attributes of the proposed composition, the use of hemocyanin is directed at stabilizing vulnerable organ, tissue, and cellular specimens after harvest. Oxygenation of donor material for a suitable duration that allows for transport and improves specimen storage and preservation are essential to maintaining the quality, prolonged viability, and ultimately, a successful transplantation. Hemocyanin offers functional oxygen transport advantages over use of human red blood cells, hemoglobin-based oxygen carriers, and perfluorocarbons, which rely on specific temperatures, pressurized environments, and allosteric effects to optimize oxygenation. Preferably the composition is delivered in a commercial aqueous stabilizing or preservation solution commonly used for the target transplant material including, but not limited to: BMPS Belzer®; Celsior®; Custodiol®; Euro-Collins®; IGL-1®; Optisol-GS™; Perfadex®; Plegisol®; SCOT 15 Multi Organes Abdominaux®; SCOT 30 Greffons Vasulaires®; Ringer Lactate®; Soltran®; Steen®; and Viaspan® solutions. The inclusion of stabilizing agents in the composition may also comprise organic or inorganic ions, including one or more of the following: calcium, chloride, lactate, magnesium, potassium, sodium, urate, or another divalent ion, at a concentration that enhances hemocyanin and oxygen binding affinities. It is further understood that the composition is suitable as a cold sterilization solution that is a non-corrosive and safe agent for medical instruments. Most commercial cold-sterilant solutions (i.e., Cidex®, Cidex Plus®, Cetylcide-G®, Banicide®) contain the active ingredient glutaraldehyde and have proven effectiveness against viruses, bacteria, fungi and tuberculosis; however, glutaraldehyde has been linked to a variety of maladies (e.g., asthma, breathing difficulties, respiratory irritation, skin rashes, etc.). The composition of the proposed invention would confer optimal sterilization capabilities with reduced hazards compared to such activated glutaraldehyde sterilants.

(18) In addition to use in storage, preservation and transport media, the proposed composition may be directly perfused into or over the donor organ or tissues before, awaiting and throughout the harvest procedure to prevent warm ischemia, and prior to cooling the harvested material. During this time, transplant material can rapidly deteriorate from oxygen and nutrient depletion and consequent necrosis. Mitigation of ischemia-reperfusion injury is key to prolonging viability and specimen integrity for transplant. As such, the solution may be administered with an arterial, venous, or a triple lumen catheter, or an extracorporeal membrane oxygenation (ECMO) system, or a similar technique to provide oxygenation to target organs or tissues.

(19) The invention composition may also be used as a non-staining antiseptic, biocidal, free radical antioxidant, and oxygen transporting topical or washing suspension. Iodine and derivative complexes (i.e., povidone iodine) are traditional medical biocidal disinfectants (Lepelletier et al., 2020). Notably, the use of iodine and the complexed forms with increased stability are known to cause epidermal irritation and staining. While the commercial use of iodine solutions as an antiseptic is well described, the active iodine ingredient concentration must remain low and requires solubilizing agents or carriers to prevent unwanted irritation, cytotoxicity, systemic distribution, tissue/cellular damage, and excessive staining (Flynn, 2003; Fumal, 2002; Balin and Pratt, 2002; Niedner, 1997; Van den Broek et al., 1982; Viljanto, 1980). Like many antiseptics, efficacy and potential risks are largely dependent on concentration; therefore, longer exposure (i.e., up to 36 hours) may be necessary for optimal results (Lawrence 1988a; Lawrence 1998b). The composition of the proposed invention would provide the microbiocidal benefits of iodine and complexed iodine-based antiseptics, while eliminating concentration dependent side effects and staining. It would also impart oxygen transport and free radical scavenging properties that could promote cellular integrity and healing when applied to a wound, such as a burn, ulcer, laceration, surgical site, etc.

(20) Additional embodiments would include obvious combinations of the above-mentioned approaches or complementary molecules to those that have been proposed and as described in the specification of the patent. Particularly, the invention would be relevant to blood banking storage of packed red blood cells and platelets. Red blood cells exist in a variety of shapes, contours, are increasingly “sticky,” and dense in both hemoglobin and iron. These cells represent the most abundant cell type in the blood (4.2-6.1×10.sup.9 cells mL.sup.−1) and play a role in innate immunity. In circulation, pathogens coexist and are recognized by the white cells as antigenic matter. However, the inventors have observed that gram-negative bacteria, and likely many other pathogens, have evolved to elude the host's immunological response through a red blood cell affinity, which conceals the pathogen from host defenses (likely similar in the case of platelets). Thus, in the case of blood banking, there is a significant risk that pathogenic material can be transferred when bound to red blood cells or platelets. The proposed invention establishes an innocuous sterilant with broad-spectrum antimicrobial properties for banked blood with enhanced cellular integrity that would promise significantly improved quality of donated and stored blood, one of the most common tissue transplants performed as transfusions, worldwide. As such, the various embodiments disclosed in this patent thus provide illustration, not limitations; the intended scope is therefore reflected in the following claims.