High dissolved oxygen fluid, aqueous solution, GEL, hydrogel, cream, micro-encapsulation, patch or aerosol spray for therapeutic applications for humans and animals

Abstract

This invention introduces the use of high dissolved oxygen levels exceeding 185%, delivered through fluids, aqueous solutions, gels, hydrogels, creams, aerosol sprays or micro- encapsulation, for treatment of wounds, burns, ischemia, neuropathy, skin-related conditions, skin grafts, and bovine mastitis. Unlike traditional Hyperbaric Oxygen Therapy (HBOT), this method provides a targeted, noninvasive approach, ensuring optimal oxygen availability at the cellular level. Unlike traditional HBOT the patient does not have to be referred for treatment and can self-treat at home. The composition enhances therapeutic outcomes by promoting cell proliferation, reepithelialization, collagen synthesis, and angiogenesis while reducing infection risks and inflammation. The vehicle stabilizes the high dissolved oxygen and facilitates its effective application. This invention addresses the limitations of existing oxygen delivery mechanisms, offering a versatile and efficient solution for tissue oxygenation and accelerated healing in both human and veterinary medicine.

Description

HIGH DISSOLVED OXYGEN FLUID, AQUEOUS SOLUTION, GEL, HYDROGEL, CREAM, MICRO-ENCAPSULATION, PATCH OR AEROSOL SPRAY FOR THERAPEUTIC APPLICATIONS FOR HUMANS AND ANIMALSFIELD OF THE INVENTION

[0001] The invention relates to the treatment of wounds, burns, ischemia, neuropathy, skin- related conditions, skin grafts, and bovine mastitis.BACKGROUND OF THE INVENTION

[0002] Oxygen is essential for cellular metabolism, particularly through oxygen-dependent enzymes that facilitate the production of ATP and NADPH. Elevated oxygen levels promote aerobic glycolysis, which induces a shift in the wound microbiome's species composition and population levels. This shift significantly enhances wound healing processes by increasing cell proliferation and reepithelialization, reducing endothelial gap closure, and by boosting fibroblast proliferation and the number of endothelial progenitor cells (EPCs).

[0003] Higher oxygen levels enhance collagen synthesis and tensile strength. Oxygen is necessary for the hydroxylation of proline and lysine in procollagen formation.

[0004] Elevated oxygen levels enhance antibacterial activities through the production of reactive oxygen species (ROS), which are crucial for oxidative burst and the removal of necrotic cellular debris. NADPH supports macrophage survival, and leukocyte activity is directly proportional to cellular oxygen levels.

[0005] Angiogenesis and revascularization are significantly promoted under elevated oxygen conditions, with increased blood vessel growth and collagen production. NOS contributes to promoting vascular tone and reducing vasoconstriction. Elevated oxygen levels also stimulate growth factor signaling transduction, leukocyte recruitment, and the production of vascular endothelial growth factor (VEGF) in macrophages and keratinocytes via H2O2.

[0006] Reactive oxygen species (ROS) are essential for regulating platelet-derived growth factor (PDGF) in cell division and growth, and they also influence keratinocyte growth factor, insulin-like growth factor, integrin function, cytokine production, and cell motility.

[0007] Hyperbaric Oxygen Therapy (HBOT) is a noninvasive technique that supplies 100% oxygen at pressures greater than 1 atmosphere absolute (ATA), in which oxygen penetrates the cutaneous tissue as proven by Trans Cutaneous Oxygen Measuring devices. It has become a well-established treatment modality for various conditions, including non-healing wounds, infections, and medical emergencies. Consequently, HBOT can induce a wide range of cellular, biochemical, and physiological changes throughout the body. Proven biological mechanisms through which HBOT exerts its beneficial effects include promoting angiogenesis, alleviating inflammation, enhancing antioxidant defenses, and stimulating stem cells. However, the uses of HBOT over extended periods can cause health risks and be economically and medically impractical.

[0008] There are limited effective delivery mechanisms capable of providing continuous high dissolved oxygen levels to adequately oxygenate the defined target area. This invention addresses this gap by enabling the achievement of dissolved oxygen levels exceeding 185%, matching or exceeding HBOT, delivered directly to treatment sites through fluids, aqueous solutions, gels, hydrogels, creams, aerosol sprays or micro-encapsulation.

[0009] The invention leverages the therapeutic benefits of oxygen by improving tissue hypoxia, enhancing perfusion, reducing edema, down regulating inflammatory cytokines, promoting fibroblast proliferation, collagen production, and angiogenesis.SUMMARY OF THE INVENTION

[0010] Forming one aspect of the invention is a novel approach to oxygen therapy, designed to accelerate the wound healing process by delivering continuous high levels of dissolved oxygen directly to the wound, burn, and skin graft.

[0011] Forming another aspect of the invention is a therapeutic and / or preventative approach for treatment of bovine mastitis, skin-related conditions, ischemia, and neuropathy.

[0012] According to another aspect of the invention, the level of dissolved oxygen is greater than 185% of saturation, and is delivered through fluids, aqueous solutions, gels, hydrogels, creams, aerosol sprays or micro-encapsulation.

[0013] Forming another aspect of the invention is a method for treating an injury in a body part, the method comprising: periodically immersing the body part in water containing dissolved oxygen at >185% saturation.

[0014] According to another aspect of the invention the body part is immersed in the water for about 25 minutes twice a day and the water contains dissolved oxygen at >300% saturation.

[0015] According to another aspect of the invention, the injury is selected from the group consisting of: wound, neuropathy and ischemia.

[0016] Forming another aspect of the invention is a method for treating a skin condition, the method comprising: periodically applying to the affected skin a composition comprising dissolved oxygen at >185% saturation, the composition selected from the group consisting of cream, lotion, gel, serum, balm and patch.

[0017] According to another aspect of the invention wherein the condition is selected from acne and sunburn.

[0018] Forming another aspect of the invention is a consumer package for treating an injury in a body part, the package comprising: a quantity of water containing dissolved oxygen at >185% saturation, the quantity being sufficient to immerse the body part twice a day for a week.

[0019] According to another aspect of the invention, the package can further comprise a container adapted for the immersion of the body part.

[0020] Forming another aspect of the invention is a composition comprising water and oxygen for application to the skin, the oxygen being present at >185% saturation, the composition being selected from the group consisting of cream, lotion, gel, serum and balm.

[0021] Advantages, features and characteristics of the invention will become evident to persons of ordinary skill with reference to the following detailed description with reference to the appended drawings, the latter being briefly described hereinafter.BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In the Figures:FIG. 1. tissue oxygenation measurements of a patientFIG 2. is a photograph of a wound at the commencement of treatmentFIG. 3 is a photograph of a wound after 2 days of treatmentFIG. 4 is a photograph of a wound after 4 days of treatmentFIG. 5 is a photograph of a wound after 6 days of treatmentFIG. 6 tissue oxygenation measurements of a patient with ischemia and neuropathyFIG. 7 tissue oxygenation measurements of a patient with ischemia and neuropathyFIG. 8 photographs of petri dishesFIG. 9 photographs of petri dishesFIG. 10 is a photograph of a container used during stability testingFIG. 11 is a photograph of measuring containers used during stability testingFIG. 12 shows average dissolved oxygen concentration during stability testingDETAILED DESCRIPTION

[0023] One embodiment of the invention involves the introduction of high dissolved oxygen greater than 185% into a suitable vehicle, such as a fluid, aqueous solutions, gels, hydrogels, creams, aerosol sprays or micro-encapsulation. Water can be oxygenated to this level using, for example, the device sold by BioTherm Hydronic Inc., under model no. DOS1, United States Patent No. 10,933,386. Water so oxygenated can be incorporated into gels, creams, sprays, etc., using conventional techniques.

[0024] Another embodiment of the invention is the use of the high dissolved oxygen solution for incorporation to commercially available treatments for wound care, burn injuries, ischemia, neuropathy, skin-related conditions, skin grafts, and bovine mastitis.

[0025] Another embodiment of the invention is the introduction of the high dissolved oxygen to remove debris and reducing microbial loads within the dermis, epidermis and sub-dermis.

[0026] Another embodiment of the invention is the application of the high dissolved oxygen solution delivered topically to the treatment area, ensuring direct contact with the affected area.

[0027] In another embodiment of the invention, the treatment area can be covered with a sterile dressing containing high dissolved oxygen and covered with a sealed dressing to maintain moisture and prolonged oxygen exposure.

[0028] Another embodiment of the invention is the application of the composition formulated for sprayable application, allowing for easy and even distribution over the desired surface.

[0029] Another embodiment of the invention are pre-formed hydrogel sheets, or films impregnated with the composition for consistent and controlled delivery of high dissolved oxygen.

[0030] Another embodiment of the invention is the use of devices designed for continuous oxygen delivery to the treatment area, to ensure consistent supply of high dissolved oxygen.

[0031] Another embodiment of the invention is the use of adhesive patches infused with high dissolved oxygen, to adhere directly to the skin surface, providing localized treatment.

[0032] Another embodiment of the invention, the composition is tailored with various additives and ingredients to address specific treatment conditions and patient needs, optimizing therapeutic outcomes.

[0033] Another embodiment of the invention, the composition can be integrated into routine treatment modalities, minimizing the time burden on patients and healthcare providers compared to traditional hyperbaric oxygen therapy.Tissue OxygenationBackground

[0034] To harness the benefits of oxygen therapy, ensuring precise delivery to targeted areas in humans or animals is crucial. Existing methods like hyperbaric chambers administer oxygen systemically rather than locally, potentially leading to systemic oxygen exposure and risks of hyperoxia across all bodily tissues. Moreover, hyperbaric oxygen therapy can be medically contraindicated and imposes significant resource costs.

[0035] To address these challenges, alternative delivery methods such as high-dissolved oxygen fluids, aqueous solutions, gels, hydrogels, creams, aerosol sprays, or micro-encapsulation have been proposed. All these vehicles are manufactured with a base of high-dissolved oxygen water (HDOW), which is the main component for the treatment of a specific target area.

[0036] Effective oxygenation in therapeutic applications requires penetration into the epidermis, dermis, and subdermal tissues, these layers encompass a width of up to 0.1cm in depth depending on the area. When no fat is present, the subcutaneous layer can be as thin as 1 millimeter, having a depth of less than 2cm for all tissues. Furthermore, some muscles are usually located at around 1.2cm deep from the skin.

[0037] Within animals, such as cows, there are three layers, the epidermis, dermis and hypodermis. The epidermis of a cow ranges on 1 to mm, and dermis can vary in thickness. The skin layers of other mammals may vary with species, but they all encompass an epidermis and a vascular dermis.

[0038] The case study evaluates the efficacy of high dissolved oxygen water in oxygenating tissues at 2.5cm depth, a depth typically encompassing all skin layers in both humans and animals. The study involved assessing tissue oxygenation on the right leg of a healthy male over a 50-minute period. High dissolved oxygen water was administered by immersing the patient’s right leg up to the knee, and tissue oxygenation levels were monitored using the Foresight 5 transcutaneous oxygen device.

[0039] The foresight 5 device utilizes near-infrared spectroscopy (NIRS) to analyze different optical spectra, enabling precise measurement of transcutaneous oxygen levels. According to the manufacturer’s specifications from Edwards, this device is capable of accurately measuring tissue oxygenation at a depth of 2.5cm. This allowed us to investigate the effectiveness of high dissolved oxygen water in enhancing tissue oxygenation at a clinically relevant depth.

[0040] The levels of high dissolved oxygen water that were delivered to the leg of the test subject were verified using a YSI Pro Solo probe.Experimental

[0041] A male patient presented. Patient right leg was submerged in HDOW at a concentration of 317.6% oxygen saturation. The leg was immersed for a total of 50 minutes, and continuous measurements for tissue oxygenation were made using the Foresight 5. The sensor of the Foresight device was placed above the water level to avoid direct contact of the sensors with the water.Table 1. NIRS measurements of healthy patient in response to high dissolved oxygen water treatments.

[0042] Figure l is a line graph that measures tissue oxygenation of the right leg of the patient. The results presented demonstrated an 8.59% (SE ± 0.48%) average increase in tissue oxygenation over the 50-minute treatment with high dissolved oxygen water. Tissue oxygenation increased by 7.8% at the 50-minute mark relative to the baseline StCE value.Discussion

[0043] The patient's leg demonstrated consistent and adequate oxygenation throughout the 50- minute trial. On average, tissue oxygenation increased by 8.59% (SE ± 0.48%) over the treatment period, achieved at less than three minutes of initial exposure to high dissolved oxygen water. These findings validate that HDOW as an effective method for delivering oxygen to the epidermis, dermis, and subcutaneous layers of human tissue. Moreover, similar efficacy can be expected in oxygenating tissues at in animals.

[0044] The results demonstrate effective tissue oxygenation leveraging HDOW for various therapeutic applications, including wound healing, oral wound treatment, infection reduction, burn care, skin graft integration, neuropathy, ischemia, nasal therapies, bovine mastitis, and management of skin-related conditions in specific areas. These benefits parallel those previously identified with the benefits of oxygen through hyperbaric oxygen therapy, highlight HDOW as a promising alternative for targeted and continuous oxygen delivery in medical treatments.Conclusion

[0045] The study conclusively demonstrates that high dissolved oxygen water serves as an effective vehicle and delivery mechanism for continuous oxygen supply to targeted areas. Moreover, it substantiates its efficacy in significantly enhancing tissue oxygenation at the intended site of application and provides a user-friendly treatment modality.Wound CareBackground

[0046] A wound is a break in the epithelial integrity that can be accompanied by the disruption of structure and function of underlying normal tissue. Wound healing studies are complex due to the nature of the wound environment and healing process. To study wounds, wounds can be monitored through non-invasive protocols such as photographic documentation. These record the wound area over time, and wound closure is calculated on wound size relative to their original dimensions, to calculate the wound healing rate. Furthermore, analysis of granulation bed characteristics including recruited call populations, vascularity and matrix alterations can further help to identify the benefits of wound healing treatments.

[0047] Oxygen is commonly used in wound care, especially through hyperbaric oxygen therapy (HBOT), which delivers 100% oxygen at pressures above 1 atmosphere absolute. This method is well-recognized for treating various conditions, including non-healing wounds and infections. However, prolonged use of HBOT can pose health risks and may become both economically and medically impractical.

[0048] High oxygen levels the biochemical pathways driven by cytokines, growth factors and immune cells. These changes significantly improve wound healing by promoting cell proliferation, re-epithelialization, shortening the time for endothelial gap closure, and increasing both fibroblast proliferation and the number of endothelial progenitor cells.

[0049] Additionally, higher oxygen levels boost collagen synthesis and tensile strength, which are crucial for effective wound healing. Oxygen is needed for the hydroxylation of proline and lysine during procollagen formation.

[0050] Moreover, reduced vasoconstriction, aided by nitric oxide synthase (NOS), leads to a ten-fold increase in collagen deposition. High oxygen levels also enhance antibacterial activities by producing reactive oxygen species (ROS), essential for oxidative burst and the removal of necrotic cellular debris.ExperimentalPatient 1

[0051] A 76-y ear-old male patient with no known significant medical history, no documentation of chronic diseases and a self-reported history of poor wound healing presented with a partial-thickness skin abrasion in the thigh following a fall. Various wound healing methods were attempted by the patient, including 2nd generation Cephalosporin in addition to Acetaminophen to reduce pain and inflammation, but the wound persisted. Without improvement and given the lack of progress with initial treatments and persistent signs of impaired healing, patient elected to undergo the recommended treatment using high dissolved oxygen water (HDOW) at 300% saturation twice a day for 25 minutes each application. Treatment was applied by submergence of the leg into a container containing HDOW. Treatment was applied for a length of six days, and photographic documentation was taken at day 0, 2, 4 and 6.

[0052] The results were as follows:Day 0 - FIG 2 wound exhibits signs of inflammation, infection and central dermal necrosisDay 2 - FIG 3 reduction in inflammation and erythema, and the resolution of central necrosis. The presence of granulation tissue indicated initial stages of skin regeneration and wound healing. The wound border no longer appears swollen, and erythema in the surrounding tissue has subsided.Day 4 - FIG 4 significant wound contraction and skin growth covering the previously necrotic area. There are no further signs of inflammatory fluid, and the wound surface shows the development of a new dermal layer.Day 6 - FIG 5 wound exhibits full recovery, with only residual post-inflammatory erythema remaining.Patient 2

[0053] A 28-year-old male patient with no significant medical history or chronic illnesses is presented with a contact abrasion sustained during an 8-hour work shift. Seeking immediate recovery to continue his duties, the patient consented to a novel treatment. Patient is administered a high dissolved oxygen hydrogel with a dissolved oxygen concentration of 317%, to be applied topically every hour for 12 hours. For comparative analysis, half of the wound was left untreated. During the manufacturing of the hydrogel, dissolved oxygen concentration was validated with the YSI Pro Solo DO Probe.

[0054] The results were as follows:Before HDOH treatment - wound measured approximately 5 inches in length and 2 inches in width. Clinical assessment revealed an area of poor clotting, moderate-to-severe partial thickness dermal abrasion, and large surrounding area of erythema. The injured area is significantly warm to palpation. The patient reported pain and discomfort in the affected area.After HDOW treatment - treated area demonstrated significant wound contraction, initiation of endothelial gap closure, and the presence of granulation tissue with mild erythema at the wound borders. The patient reported no pain in the treated area, increased comfort, and an absence of itchiness.Untreated area - wound remained open, non-contracted wound with edema, erythema, and inflammation around the borders. The patient reported discomfort and itchiness in the untreated area.Discussion

[0055] It is well documented that, typically, there are four stages in wound healing. After the initial stage of hemostasis, the inflammatory phase lasts 4-6 days and is characterized by the presence of inflammatory signs such as erythema, warmth, edema and pain. This inflammatory period can be prolonged in patients with underlying diseases or a history of poor wound healing. The third stage, proliferation, can last 6-21 days and is marked by the presence of granulation tissue and epithelialization. In this case study, granulation tissue appeared earlier, on Day 2, with epithelialization and wound contraction observed on Days 4 and 6.

[0056] In the second case study, after 12 hours of treatment, the wound appeared to have advances in the inflammatory phase as there were signs of mild erythema at the wound borders, and discomfort was minimized.

[0057] Both case studies demonstrated that high dissolved oxygen water and high dissolved oxygen hydrogel treatments can promote wound healing, similar to hyperbaric oxygen therapy. The evidence above demonstrates that HDOW and HDOH is effective in treating abrasions, non-healing and infected wounds, particularly in cases where traditional treatments, such as the use of oral antibiotics, have failed. In a patient with a history of poor wound healing, or in a healthy patient, HDOW treatment resulted in significant and rapid improvement, as evidenced by the resolution of necrosis, reduction of inflammation, and regeneration of dermal tissue.Conclusion

[0058] High dissolved oxygen is a safe method for treating wounds, as it is a non-invasive topical application that minimizes the risk of complications associated with more invasive procedures. This method reduces the likelihood of introducing additional infections, and can be easily administered, making it a practical option for a wide range of patients.Ischemia and Peripheral NeuropathyIntroduction

[0059] Hyperbaric oxygen therapy (HBOT) is acknowledged and validated by the Undersea and Hyperbaric Medical Society and the Centers for Medicare and Medicaid Services (CMS), as a treatment for acute traumatic peripheral ischemia and peripheral neuropathy. It acts as a complementary treatment to standard care, improving tissue oxygenation, aiming to prevent loss of function, limb damage, or death.

[0060] HBOT has been shown to protect neuronal and muscle structures, reduce inflammatory markers, and influence cell death and astrocyte activation. It works in conjunction with drug therapies, partly by increasing heme oxygenase- 1 and promoting autophagy, which helps to relieve neuropathic pain.

[0061] Furthermore, HBOT reduces neutrophil adhesion to injured blood vessels, thereby decreasing ischemia-reperfusion injury. Mechanistically, HBOT causes blood vessels to constrict, reducing fluid leakage while improving oxygen delivery through increased plasma oxygen solubility. This effect lessens swelling and increases oxygen diffusion. Despite its recognition for treating ischemia and peripheral neuropathy, HBOT can be costly and resource intensive. Additionally, it does not provide localized treatment to the affected area.

[0062] Ischemia and peripheral neuropathy were treated through the application of high- dissolved oxygen water (HDOW) via immersion. Transcutaneous oximetry was employed to measure oxygen partial pressure (StO2) at deeper tissue levels. The Foresight 5, a near-infrared spectroscopy (NIRS) was utilized as the method for TCOM. This device is a highly effective tool for quantifying changes in muscle tissue. NIRS operates by identifying the different optical spectra of oxygenated and de-oxygenated hemoglobin at a maximum of 2.5cm of depth.Consequently, NIRS was employed to assess tissue hypoxia and monitor muscle responsiveness to oxygenation.

[0063] There are numerous potential measurement sites for NIRS measurements; however, the preferred sites are those characterized by significant musculature, as this provides a homogenous tissue compartment for TCOM. The forearm is a predominant site of vasoconstriction in the presence of circulatory diseases, with vascular responses occurring earlier and more intensely than in other body regions. Consequently, the forearm was considered a suitable area for NIRS measurement. The gastrocnemius muscle was also chosen for monitoring due to its underlying musculature and the specific medical history of the patient.

[0064] Dissolved oxygen monitoring and water temperature were monitored using a YSI ProSolo device, with the aim of monitoring changes in dissolved oxygen in response to treatment time and temperature fluctuations.

[0065] A male patient with chronic spinal cord stenosis presented . The patient underwent treatment with HDOW at an initial concentration of 317 ± 1% oxygen saturation with a total immersion time of 50 minutes. Oxygen monitoring and HDOW re-oxygenation were conducted prior to immersion and at five-minute intervals throughout the treatment. Baseline StO? values were calculated by the average of four values. For the purposes of analysis, data is presented in tables of 5-minute intervals. The water temperature was maintained constant during each reoxygenation period for both the control and treatment groups.Experimental

[0066] A 64-year-old male with peripheral neuropathy, resulting from chronic spinal cord injury and stenosis, presented with chronic ischemia, reduced sensitivity in the lower limbs and Diabetes Type II. Due to his medical history, he has received lidocaine injections to manage pain and sensitivity, though without significant improvement.

[0067] For the study, the Foresight sensor was positioned in the forearm and secured with Tegaderm to protect it from water exposure. The forearm with the attached sensor was immersed in high-dissolved oxygen water, and StCE measurements were taken every 5 minutes, starting at a dissolved oxygen concentration of 317% oxygen saturation (Table 2). Dissolved oxygen concentration and water temperature were measured prior to the treatment start, at 20 minutes and at the end of the trial.Table 2. TCOM (StCE) measurements of 64-year-old patient in response to high dissolved oxygen water treatments in the forearm - Case study 1.

[0068] Fig 6. is a line graph demonstrating the forearm tissue oxygenation measurements over the course of the treatment and the ten-minute period following immersion. Tissue oxygenation increased by 9.37% relative to the ending StCE value at the 50-minute mark, having an 8.59% (SE ± 0.35%) average increase in tissue oxygenation over the treatment time, with levels remaining mostly constant less than 5 minutes since the treatment started. Notably, slight decreases in dissolved oxygen concentration did not affect tissue oxygenation over time.

[0069] A second treatment was performed in which the patient immersed the left leg, which suffers from mild to moderate peripheral neuropathy and loss of sensitivity due to spinal cord stenosis. The sensor was attached to the gastrocnemius muscle, and monitoring frequency was maintained at five-minute intervals, with an initial dissolved oxygen concentration of 317.6% oxygen saturation (Table 3).Table 3. TCOM (StCE) measurements of 65-year-old patient in response to high dissolved oxygen water treatments in the left gastrocnemius muscle - Case study 1.

[0070] Fig 7 is a line graph demonstrating tissue oxygenation of the gastrocnemius muscle of the 64-year-old patient measured over the immersion. During the second treatment, the patient exhibited an average of 19.14% (SE ± 0.64%) increased tissue oxygenation throughout the 50- minute immersion in high dissolved oxygen water. Tissue oxygenation increased by 15.02% at the 50-minute mark relative to the baseline StCE value. The patient’s StCE values experienced greater fluctuations, likely due to the history of peripheral neuropathy and ischemia in the treated leg. Changes in dissolved oxygen concentration did not affect tissue oxygenation over time.

[0071] The patient demonstrated increased tissue oxygenation in both the forearm and the gastrocnemius muscle. Tissue oxygenation was higher in the gastrocnemius muscle than in the forearm. These discrepancies can be due to a few factors including surface area exposure to HDOW, differences in muscle mass, and skin changes due to neuropathy. Trauma to the limb likely induced local hypoxic conditions caused by edema. Consequently, it can be theorized that the left leg exhibited greater tissue oxygenation as a response to the present ischemia.

[0072] Despite the history chronic spinal stenosis, neuropathy and aging, the 64-year-old patient showed successful tissue oxygenation at a depth of 2.5cm in both areas. Tissue oxygenation remained elevated from 5 minutes of treatment onwards, despite a reduction in the concentration of dissolved oxygen. A mean value of 67 StO2 can be considered normoxic in the lower limbs. This suggests that the baseline value of the lower limb of the 64-year-old male was likely below the mean value, and the treatment effectively enhanced oxygen levels in the affected area.

[0073] The case study demonstrated that, regardless of slight decreases in dissolved oxygen concentration in the water, tissue oxygenation continued to increase. These increases were dependent on the selected tissue and the patient’s condition in the treated area. This supports previous studies that demonstrate the effect of age, blood circulation, trauma, and any underlying diseases on tissue oxygenation.Conclusion

[0074] The study demonstrated that tissue oxygenation can be significantly increased, as in our case study up to 19.14% (SE ± 0.64%), by transcutaneous oxygen delivery using high dissolved oxygen water. This non-invasive transdermal method can be an effective tool in treating many diseases such as ischemic neuropathy and wound healing.AcneBackground

[0075] It is well-established that oxygen confers various dermatological benefits and has been employed in several delivery modalities, such as direct oxygen application to the skin and nebulized oxygen. However, these methods often yield suboptimal results and limit the penetration depth of oxygen. It is therefore recommended that dissolved oxygen can be administered through mediums such as fluids, gels, hydrogels, creams, and aerosol sprays to enhance its therapeutic efficacy.

[0076] Oxygen exhibits significant antibacterial properties through the generation of reactive oxygen species (ROS), which are highly toxic and capable of eliminating pathogenic microorganisms. This mechanism is particularly effective against acne-causing bacteria such as Cutibacterium acnes and inhibits the proliferation Staphylococcus aureus, and Streptococcus species. Additionally, oxygen helps maintain the skin’s acidic pH by releasing fatty acids that inhibit bacterial growth and disrupt bacterial cell membranes via lipid peroxidation.

[0077] For optimal results, oxygen must penetrate the epidermal surface, base of hair follicles, and reach at least the basal layer of the epidermis, this layer is of most importance due to its high oxygen demand and location of the sebaceous gland. Previous studies examining the oxygen delivery capabilities of our high-dissolved oxygen applications have demonstrated that oxygen can penetrate up to 2.5 cm, as measured by a Foresight 5 transcutaneous oxygen monitoring device.

[0078] In the present case study, a 28-year-old male patient was treated with a high-dissolved oxygen hydrogel (HDOH) formulated at a concentration of 317%, applied hourly over three days. The HDOH was applied to the affected areas with active acne, and the treatment continued until full absorption. Evidence was collected before and after the treatment period to document the outcomes.Experimental

[0079] The patient is a 28-year-old healthy male with a chronic history of acne. The patient reported a moderate case of acne outbreak, which occurred after using helmets during work. The patient typically experiences moderate acne outbreaks on the back of his neck near the hairline after weekly shifts. No other relevant medical history was reported relevant to the case study. The patient described that his acne usually presented with redness, itchiness, and multiple microcomedo and pustule formation. After experiencing no improvement with other interventions, the patient applied the high-dissolved oxygen hydrogel (HDOH). For comparative analysis, half of the affected area remained untreated (left side of the neck).

[0080] On the first day of treatment, there was a noticeable decrease in pustules. The pustules no longer had a white-colored tip and appeared moderately red and inflamed.

[0081] By the second day of treatment, the number of whiteheads had decreased, with only 10- 15 nodular lesions in the treated area compared to 15-20 in the untreated area. Erythema significantly decreased, and inflammation was minimized, with most remaining lesions located within the hair-bearing region.

[0082] On the third day of treatment, only one active pustule remained, with substantial improvement in the treated area. Inflammation was almost completely absent, with very few residual erythema. The patient reported no remaining itchiness.Discussion

[0083] It is well known that acne healing involves several stages, including reduced inflammation, clearing of pustules and skin debris, skin regeneration, and the restoration of the skin barrier. The high-dissolved oxygen hydrogel (HDOH) proved effective in treating moderate acne, as evidenced by a reduction in inflammation by day three, the absence of whiteheads, and fewer pustules in the treated area.

[0084] The HDOH demonstrated both antibacterial and anti-inflammatory properties, which minimized the presence of pustules within the treated area. The antibacterial effect as previously mentioned, can be attributed to the oxygen capacity of aiding in the generation of reactive oxygen species (ROS), which are highly toxic to pathogenic microorganisms.

[0085] The anti-inflammatory effect of HDOH was effective in reducing erythema and swelling. This reduction in inflammation helped to alleviate the discomfort and itchiness reported by the patient. Furthermore, HDOH aided in skin regeneration by promoting the repair and restoration of the skin barrier, which is essential for maintaining skin integrity and preventing future acne outbreaks in the area.

[0086] Moreover, the moisturizing properties of HDOD, effectively contributed to the effectiveness in reducing erythema. Hydration is vital for maintaining the skin’s barrier function and preventive dryness, which can exacerbate acne symptoms. The moisturizing effect of the HDOH likely helps to soothe the skin and support its natural healing processes.

[0087] Patient feedback indicated a significant reduction in itchiness and discomfort, highlighting the therapeutic benefits of HDOH. The combination of antibacterial, antiinflammatory, and moisturizing properties makes HDOH a promising treatment for moderate acne, providing comprehensive support for skin healing and improving patient outcomes.Conclusion

[0088] Overall, the application of HDOH resulted in a notable improvement in the patient's acne condition, demonstrating its potential as an effective treatment modality for managing moderate acne and enhancing skin health.Anti-Bacterial UsesIntroduction

[0089] Independent in-vitro tests were performed by BRI Frontage Laboratories to assess the effect of high dissolved oxygen water (HDOW) on bacterial viability and growth.Study 1

[0090] The first study was performed to assess bacterial contaminants present in the HDOW manufactured at different concentrations. The aim of the study was to identify present contaminants in the water, and if necessary, further develop new manufacturing procedures or storage mechanisms to reduce any bacterial contaminants present in HDOW. The HDOW initial oxygen concentrations tested included control tap water, 17.5ppm, 25ppm, 32.5ppm and 40ppm.Study 2

[0091] After refining manufacturing practices to reduce microbial contaminants present in water, a further study was performed. This assessed bacterial contamination testing of HDOW at control tap water, 30ppm and 40ppm initial oxygen concentration.Materials and MethodsStudy 1

[0092] lOOmL of HDOW at 17.5ppm, 25ppm, 32.5 ppm, and control tap water were filtered through a 45uM paper filter to capture any bacterial contaminants present. Filtered HDOW water was plated on both LB and Blood Agar plates to detect bacterial growth. Two volumes were tested, half plates with lOOuL of HDOW and full plates of 250uL.Study 2

[0093] lOOuL of Control tap water, 30ppm, and 40ppm HDOW were plated on blood agar plates, and sealed as quickly as possible with parafilm. Plates were placed agar side down until HDOW was completely absorbed (5 minutes), and then placed in the 30°C incubation chamber. Pictures were taken at 0, 6, 24 and 48 hours contact time.ResultsStudy 1

[0094] FIG 8 is photographs of petri dishes of bacterial contaminants pre-filtration and postfiltration, showing half plates and full plates. The HDOW oxygen concentration in ppm is indicated in parentheses.

[0095] The control water in FIG 8 exhibited two bacterial colonies before filtration, indicating signs of bacterial contamination in the control water used to produce the HDOW. After filtration, no colonies were found.

[0096] The 17ppm water demonstrated increased bacterial growth in the unfiltered sample, and no bacteria were noted after filtration. However, small yellow colonies were noted on the 17ppm filter.

[0097] The 25ppm water HDOW exhibited increased bacterial growth in the unfiltered sample as compared to the 17ppm unfiltered water. After filtration, no bacterial growth was noticed on the 25ppm samples; however, two bacterial colonies were found in the filter.

[0098] The 32.5ppm and 40ppm samples showed no bacterial growth prior to or after filtration. As indicated in Table 1, the 30ppm and 40ppm samples of HDOW demonstrated no bacterial growth regardless of filtration. In contrast, the lower concentrations, 17ppm and 25ppm benefited from using 0.45uM paper filters to reduce the bacterial contaminants present.Study 2

[0099] FIG 9 is photos of the petri dishes at 0, 6, 24 and 48 hours contact time for bacteria found in control water, 30ppm and 40ppm initial oxygen HDOW concentration water.

[0100] As delineated in FIG. 9 , bacterial contaminants were present in the control water and 30ppm samples from 24 hours onwards, with increased proliferation after 48 hours. Notably, in the 40ppm treatment cohort, no colonies were observed until 48 hours. Therefore, indicating a potential inhibitory effect of HDOW at 40ppm to bacterial growth.ConclusionsStudy 1

[0101] The findings indicate that filtration using a 45uM filter, has the potential to reduce contaminants found in water. However, it was observed that HDOW at 17ppm and 25ppm HDOW, led to increased bacterial growth prior to filtration and contamination in the filter. These results underscore the necessity for caution during manufacturing of 17-25ppm, as these concentrations may contribute to further bacterial growth and water contamination.

[0102] The results suggested that HDOW samples at concentration of 32.5ppm and 40ppm samples effectively impede bacterial growth, before and after filtration. However, it is important to note that the study did not record specific contact times for these concentrations. Thus, it is recommended to investigate varying contact times to ascertain the bactericidal potential of HDOW more comprehensively. It is noted that the selected study methodology did not include the incubation of the petri-dishes at any higher temperature. Samples were maintained at room temperature throughout the experiment, and no plates were regularly disturbed post-plating.Study 2

[0103] As indicated in the results bacterial contaminants became apparent in the control water from 24 hours onwards, showing more substantial growth by 48 hours. Similarly, bacterial growth was observed in the 30ppm treatment group from 24 hours onward, with prominent growth at 48 hours.

[0104] The 40ppm treatment group exhibited no colonies until 48 hours of contact time, suggesting a potential retardant effect on bacterial growth at this concentration. The study suggested that bacterial growth is contingent upon contact time, as evidenced by the bacterial proliferation in the 30ppm treatment group and control groups after 24 hours. Nevertheless, the 40ppm treatment group exhibited no colonies until 48 hours of contact time, suggesting a potential retardant effect on bacterial growth at this concentration.

[0105] The results suggest that bacterial growth retardation could take place at 40ppm HDOW over <24 hours of contact time. Similarly, at 30ppm, retardation of bacterial growth might occur until a maximum of 6 hours since contact time.General Conclusions

[0106] The study suggested that bacterial growth is contingent upon contact time, as evidenced by the bacterial proliferation in the 30ppm treatment group and control groups after 24 hours. 40ppm HDOW has a potential retardant effect on bacterial growth within 24 hours contact of time. For all concentrations, prolonged contact time with HDOW exceeding 24 hours may not reduce bacterial growth. Necessity for caution during manufacturing of 17-25ppm, as these concentrations may contribute to further bacterial growth of bacteria. For all applications utilizing HDOW careful consideration of contact time, concentration, and temperature is imperative to accurately determine the viability of the intended application. It is advisable to incorporate methods for re-oxygenating HDOW at intervals of less than 24 hours when used in various applications. Moreover, in environments where HDOW is exposed to elevated temperatures or stored in open spaces / containers, more frequent re-oxygenation is recommended to optimize the beneficial effects of oxygen, particularly concerning bactericidal activity.Oxygen administration in wound healing

[0107] Wound contaminants, to which bacteria are included, can originate from the environment, surrounding skin and endogenous sources such as mucous membranes. Bacterial infections are prone to develop in wounds of any type, mostly characterized by anaerobic bacteria that cause complications such as gangrene, post-operative infections, etc. As such, the administration of oxygen is known to both minimize bacterial colonization that results in infection and deliver oxygen to maximize blood perfusion. Current methods rely on the administration of oxygen gas to the patient and subsequent diffusion to the bloodstream and wound tissue, but no methods currently include the delivery of high-dissolved oxygen water or gels directly into the wound. Through HDOW application to the wound, opportunities could arise to decrease wound contaminants, and improve the bioavailability and benefits of oxygen to the wound.Oxygen administration in bovine mastitis

[0108] Bovine mastitis is a bacterial infection in cows which is characterized by an inflammation of the mammary gland. Pathogenic bacteria can be either environmental or contagious (host-to- host) and include both gram-positive and gram-negative bacteria. Existing treatments include disinfection of milking equipment, treatment of existing mastitis with antibiotics, and prevention through post-milking teat disinfection. However, the emergence of antibiotic-resistant pathogens, and rising costs associated with treatments urge for a novel solution for reduction of bovine mastitis incidences. HDOW delivery could aid to suppress bacterial intramammary infections, reduce opportunistic pathogens through disinfection of animal facilities, support dis-infection of milking equipment, and deliver oxygen to damaged milking tissues.Oxygen administration for acne treatments

[0109] Oxygen has demonstrated numerous beneficial effects on the skin, making it a valuable therapeutic agent for conditions such as acne vulgaris, rosacea, and aging skin. Oxygen enhances cellular metabolism, accelerates healing processes, reduces skin irritations, and exerts antiinflammatory effects through the production of cytokines such as IL- 12, IL-8, and TNF. Moreover, oxygen exhibits antibacterial properties by generating reactive oxygen species (ROS), which are highly toxic and capable of eliminating pathogenic microorganisms. This mechanism is particularly effective in reducing acne-causing bacteria like Cuti bacterium acnes and inhibiting the proliferation of Staphylococcus aureus and Streptococcus species. The application of wet oxygen through gels and creams can enhance oxygen penetration into the skin.Industrial ConsiderationsIntroduction

[0110] A four-week stability study was conducted to determine the changes in dissolved oxygen concentration in high dissolved oxygen water (HDOW) stored at an average refrigeration temperature of 5.01°C, following a circadian rhythm. The study included 15 samples (N=15) of HDOW, each initially produced at a concentration of 41.63 ppm. The dissolved oxygen concentration for each sample was monitored over a period of twenty-eight days.Materials and Methods

[0111] 5 liter transparent bags as shown in FIG. 10 were selected for the study. Container closure tests were conducted using closed containers.

[0112] Quantitative analyses included dissolved oxygen measurements in both parts per million (ppm) and percentage (%). Measurements for dissolved oxygen (ppm and %), temperature, and barometric pressure were recorded using a YSI Pro Solo Probe.

[0113] All samples were generated on January 1st, 2024. Samples were produced at an average of 41.63ppm, at a temperature of 10.35°C, 02 outflow of 4 and H2O flow of 55, as measured by the Biotherm Probe. After the manufacturing date on February 11th, a reduction in temperature was initiated to maintain sample storage at refrigeration temperature. This adjustment resulted in fluctuations, yielding an average temperature of 4.3°C.

[0114] Dissolved oxygen monitoring was conducted every three days. Sampling started at 1 :00pm daily.

[0115] The high dissolved oxygen water from each sample was poured into the same measuring container, namely, a 500ml glass jar covered with metallic tape into which the YSI probe is inserted. These are seen in FIG. 11. The water was poured from the container directly into the measuring container without taking the water bag lid off. No open space was left within the bag.Results

[0116] Standard errors (SE) were calculated for all sampling events for each sampling tool. To ensure data analysis accuracy, the YSI Pro Solo Probe measurements employed for analysis included ppm and temperature.

[0117] Figure 12 is a line graph of average dissolved oxygen concentration (%) for all 40ppm samples (N=15). Over the twenty-eight-day period, the change from the initial to the final dissolved oxygen concentration was 0.99ppm, corresponding to a 2% deviation from the starting concentration. The maximum concentration of 41.63ppm was recorded on the first day, while the minimum of 40.53 ppm was observed on the nineteenth day.Discussion

[0118] Dissolved oxygen concentrations, reflected in ppm, displayed analogous trends. Concentrations from all samples remained within 2% of their initial dissolved concentration value, experiencing minimum values around January 30th, possibly attributed to a decrease in temperature within the fridge.Conclusions

[0119] High dissolved oxygen samples demonstrated good stability over the three-week period, suggesting potential stability over longer durations. Maintaining a low initial temperature during the high dissolved oxygen manufacturing process is crucial to minimize the initial loss of dissolved oxygen concentration. Additionally, implementing external temperature controls is recommended for the storage of high dissolved oxygen samples.

[0120] Whereas specific embodiments are shown and described, it will be apparent that variations are possible. Accordingly, the invention should be understood to be limited only by the accompanying claims, purposively construed.

Claims

CLAIMS:

1. Use of high-dissolved oxygen, delivered through fluids, aqueous solutions, gels, hydrogels, creams, aerosol sprays or micro-encapsulation, wherein the composition is introduced to a human or animal treatment area.

2. Use according to claim 1, wherein the level of high dissolved oxygen is greater than 185% saturation.

3. Use according to claim 2, wherein the composition is used for treatment of wounds and ulcer healing in both humans and animals, including chronic and acute wounds, diabetic and vascular insufficiency wounds, surgical wounds, and traumatic injuries.

4. Use according to claim 2, wherein the composition is used for treating ischemia, where blood flow and oxygen supply are compromised, mitigating the effects of hypoxia, and supporting tissue repair.

5. Use according to claim 2, wherein the composition is used for treating chronic ulcers, including pressure sores and venous ulcers, providing sustained oxygenation to accelerate the healing process.

6. Use according to claim 2, wherein the composition is used in dental care to treat oral wounds, such as those resulting from extractions or oral surgeries, promoting faster healing and reducing the risk of infection.

7. Use according to claim 2, wherein the composition is used to treat burns ranging from first-degree to fourth-degree burns caused by heat, chemicals, or electrical sources, promoting tissue oxygenation, reducing edema, and enhancing healing.

8. Use according to claim 2, wherein the composition is used for enhancing outcomes of skin grafts used in cosmetic procedures, such as scar revision or reconstructive surgeries, by optimizing the graft environment, graft integration and minimizing complications.

9. Use according to claim 2, wherein the composition is used for targeting specific nerve clusters or pain points affected by peripheral neuropathy, providing localized relief, and promoting neuronal integrity.

10. Use according to claim 2, wherein the composition is used to promote accelerated healing postnasal turbinate reduction, septoplasty, or rhinoplasty.

11. Use according to claim 2, wherein the composition is used for intra-mammary and topical mammary applications to provide therapeutic antimicrobial activity and preventative measure against mastitis-causing environmental and contagious pathogens.

12. Use according to claim 2, wherein the composition is used to oxygenate the epidermis, dermis and sub-dermis for treatment of skin-related conditions, enhancing skin complexion and eliminating pathogenic microorganisms that lead to cutaneous diseases.

13. A method for treating an injury in a body part, the method comprising: periodically immersing the body part in water containing dissolved oxygen at >185% saturation.

14. The method of claim 13, wherein the body part is immersed in the water for about 25 minutes twice a day and the water contains dissolved oxygen at >300% saturation.

15. The method of claim 13, wherein the injury is selected from the group consisting of: wound, neuropathy and ischemia.

16. A method for treating a skin condition, the method comprising: periodically applying to the affected skin a composition comprising dissolved oxygen at >185% saturation, the composition selected from the group consisting of cream, lotion, gel, serum, balm and patch.

17. The method of claim 16, wherein the condition is acne.

18. A consumer package for treating an injury in a body part, the package comprising: a quantity of water containing dissolved oxygen at >185% saturation, the quantity being sufficient to immerse the body part twice a day for a week.

19. The package of claim 18, further comprising a container adapted for the immersion of the body part.

20. A composition comprising water and oxygen for application to the skin, the oxygen being present at >185% saturation, the composition being selected from the group consisting of cream, lotion, gel, serum and balm.

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