WO2013020136A2 - Treatment of traumatic brain injury - Google Patents

Abstract

The invention teaches a method and kit for treating traumatic brain injury, concussion and/or other conditions involving brain, head/neck and/or neurological injury. Specifically, the invention teaches the use of erythropoietin and stem cells to treat conditions including, but not limited to, quadriplegia due to traumatic subdural hematoma, pons hemorrhage, hypoxic brain injury, cerebral contusion, cerebral palsy, hemiplegia, subarachnoid hemorrhage, intracerebral hemorrhage, epidural hemorrhage, cerebral infarction, transverse myelitis, spinal cord injury, peripheral nerve injury, multisystem atrophy and combinations thereof.

Description

TREATMENT OF TRAUMATIC BRAIN INJURY

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from US Provisional Patent Application No. 61/515,272, filed on August 4, 2011, which is incorporated herein by reference in its entirety.

FIELD OF INVENTION

This invention generally relates to the treatment of traumatic brain injury, concussion and/or other conditions involving brain, head/neck and/or neurological injury.

SUMMARY OF THE INVENTION

In some embodiments, the invention teaches a method of treating a traumatic brain injury and/or a neurological injury in a subject, including administering erythropoietin and stem cells to the subject. In some embodiments, the stem cells are selected from the group consisting of: umbilical cord blood cells, embryonic stem cells, bone marrow cells, mesenchymal stem cells, adipose-derived stem cells, endothelial stem cells, dental pulp stem cells, placenta stem cells, olfactory ensheathing cells, neural precursor cells and combinations thereof. In some embodiments, the stem cells are autologous. In some embodiments, the stem cells are allogeneic. In some embodiments, the erythropoietin is selected from the group consisting of: recombinant erythropoietin, synthetic erythropoietin, chemically modified erythropoietin, darbepoieten, glycosylated erythropoietin, EPO-alpha, EPO-beta and combinations thereof. In some embodiments, rehabilitation therapy is also provided to the subject.

In various embodiments, the subject has a condition selected from the group consisting of: quadriplegia due to traumatic subdural hematoma, pons hemorrhage, hypoxic brain injury, cerebral contusion, cerebral palsy, hemiplegia, subarachnoid hemorrhage, intracerebral hemorrhage, epidural hemorrhage, cerebral infarction, transverse myelitis, spinal cord injury, peripheral nerve injury, multisystem atrophy and combinations thereof. In some embodiments, the erythropoietin and/or stem cells are administered to the subject within a period of from one minute to 100 years after traumatic brain injury and/or neurological injury. In some embodiments, the erythropoietin and stem cells are administered to the subject simultaneously. In some embodiments, the erythropoietin and stem cells are not administered simultaneously. In various embodiments, the invention teaches a kit for treating a traumatic brain injury and/or a neurological injury in a subject, including erythropoietin, stem cells and instructions for the use of the erythropoietin and stem cells for the treatment of the traumatic brain injury and/or the neurological injury. In some embodiments, the stem cells are selected from the group consisting of: umbilical cord blood cells, embryonic stem cells, bone marrow cells, mesenchymal stem cells, adipose-derived stem cells, endothelial stem cells, dental pulp stem cells, placenta stem cells, olfactory ensheathing cells, neural precursor cells and combinations thereof. In some embodiments, the stem cells are autologous. In some embodiments, the stem cells are allogeneic. In some embodiments, the erythropoietin is selected from the group consisting of: recombinant erythropoietin, synthetic erythropoietin, chemically modified erythropoietin, darbepoieten, glycosylated erythropoietin, EPO-alpha, EPO-beta and combinations thereof. In some embodiments, the subject has a condition selected from the group consisting of: quadriplegia due to traumatic subdural hematoma, pons hemorrhage, hypoxic brain injury, cerebral contusion, cerebral palsy, hemiplegia, subarachnoid hemorrhage, intracerebral hemorrhage, epidural hemorrhage, cerebral infarction, transverse myelitis, spinal cord injury, peripheral nerve injury, multisystem atrophy and combinations thereof. In some embodiments, the instructions indicate the erythropoietin and/or stem cells are to be administered to the subject within a period of from one minute to 100 years after traumatic brain injury and/or neurological injury. In some embodiments, the instructions indicate the erythropoietin and stem cells are to be administered to the subject simultaneously.

BACKGROUND

Traumatic brain injury (TBI), a form of acquired brain injury, occurs when a sudden trauma causes damage to the brain. TBI can result when the head suddenly and violently hits an object, or when an object pierces the skull and enters brain tissue. Symptoms of a TBI can be mild, moderate, or severe, depending on the extent of the damage to the brain. A person with a mild TBI may remain conscious or may experience a loss of consciousness for a few seconds or minutes. Other symptoms of mild TBI include headache, confusion, lightheadedness, dizziness, blurred vision or tired eyes, ringing in the ears, bad taste in the mouth, fatigue or lethargy, a change in sleep patterns, behavioral or mood changes, and trouble with memory, concentration, attention, or thinking. A person with a moderate or severe TBI may show these same symptoms, but may also have a headache that gets worse or does not go away, repeated vomiting or nausea, convulsions or seizures, an inability to awaken from sleep, dilation of one or both pupils of the eyes, slurred speech, weakness or numbness in the extremities, loss of coordination, and increased confusion, restlessness, or agitation.

Historically, little could be done to reverse the initial brain damage caused by trauma, so medical personnel typically try to stabilize an individual with TBI and focus on preventing further injury. Primary concerns include insuring proper oxygen supply to the brain and the rest of the body, maintaining adequate blood flow, and controlling blood pressure.

Imaging tests help in determining the diagnosis and prognosis of a TBI patient. Patients with mild to moderate injuries may receive skull and neck X-rays to check for bone fractures or spinal instability. For moderate to severe cases, the imaging test is often a computed tomography (CT) scan.

Approximately half of severely head-injured patients will need surgery to remove or repair hematomas (ruptured blood vessels) or contusions (bruised brain tissue). Disabilities resulting from a TBI depend upon the severity of the injury, the location of the injury, and the age and general health of the individual. Some common disabilities include problems with cognition (thinking, memory, and reasoning), sensory processing (sight, hearing, touch, taste, and smell), communication (expression and understanding), and behavior or mental health (depression, anxiety, personality changes, aggression, acting out, and social inappropriateness). More serious head injuries may result in stupor, an unresponsive state, but one in which an individual can be aroused briefly by a strong stimulus, such as sharp pain; coma, a state in which an individual is totally unconscious, unresponsive, unaware, and unarousable; vegetative state, in which an individual is unconscious and unaware of his or her surroundings, but continues to have a sleep-wake cycle and periods of alertness; and a persistent vegetative state (PVS), in which an individual stays in a vegetative state for more than a month.

Moderately to severely injured patients often receive rehabilitation that involves individually tailored treatment programs in the areas of physical therapy, occupational therapy, speech/language therapy, physiatry (physical medicine), psychology/psychiatry, and social support. Although important, traditional rehabilitative therapies typically yield relatively slow improvement.

Thus, there is a need in the art for a more effective medical treatment that can improve the rate and extent of rehabilitation in individuals with TBI. BRIEF DESCRIPTION OF THE DRAWINGS

Exemplary embodiments are illustrated in the referenced figures. It is intended that the embodiments and figures disclosed herein are to be considered illustrative rather than restrictive.

Figure 1 depicts, in accordance with one or more embodiments of the invention, some of the action mechanisms of erythropoietin (EPO) on the brain.

Figure 2 is an image of the brain of a subject (Part 1, Case 1) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. Encephalomalatic change is observed.

Figure 3 is an image of the brain of a subject (Part 1, Case 2) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. Bruises are observed in the two sides in the frontal lobe.

Figure 4 is an image of the brain of a subject (Part 1, Case 4) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. Pressure- sensitive status after resection is demonstrated. Existing bruises are detected at the two sides of the frontal lobe and left temporal lobe contusion of ephalomalacia (contusional encephalomalacia). The right basal ganglia, egg-shaped center, and an existing tear in the left temporal lobe with white matter infarction are observed.

Figure 5 is an image of the brain of a subject (Part 2, Case 1) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. In the left frontal and parietal regions of the upper part synergistic encephalomalacia (cavitary encephalomalacia) is observed. On the right side of the brain stem, Waller degeneration on the left is observed.

Figure 6 is an image of the brain of a subject (Part 2, Case 2) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. Overall, a small old cerebral hemorrhage with focal atrophy (diffuse brain atrophy) is observed.

Figure 7 is an image of the brain of a subject (Part 2, Case 3) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. Softening of the brain is observed from the cerebral atrophy.

Figure 8 is an image of the brain of a subject (Part 2, Case 4) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. Softening of the brain is observed from bilateral frontal and temporal lobes. State of the ventricle after the complex abdominal shunt - enlarged left ventricle is observed. Figure 9 is an image of the brain of a subject (Part 1, Case 5) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. Left frontal lobe, occipital lobe and left basal ganglia hemorrhage are observed with multiple bruises. Gray lines of right hemisphere with white matter substance regions of diffuse axonal injury are observed with small multiple hemorrhages. Intraventricular hemorrhage is small in this figure.

Figure 10 is an image of the brain of a subject (Part 2, Case 5) diagnosed with traumatic brain injury, treatable by one or more methods of the present invention. An existing bruise is observed on both sides of the frontal lobe.

Figure 11 is an image of the brain of a patient (Part 2, Case 3) before and after EPO and cord blood treatment.

Figure 12A and 12B are images of the brain of a patient (age 42, female) who has been diagnosed with traumatic brain injury and has not been treated with EPO or cord blood, but could be treated according to one or more methods of the invention. Figure 12A is an image of the patient's brain taken approximately four months after a traffic accident. The image shows somewhat reduced perfusion at the right side basal ganglia. There appears to be decreased perfusion, cerebellar diaschisis on the right side of the cerebellum. Figure 12B is an image of the patient's brain taken approximately 36 months after the traffic accident. No significant change can be identified at the right basal ganglia compared with the image of Figure 12 A. General, reduced perfusion can be observed in the left forehead, parietal, and temporal cortex. Reduced perfusion can also be observed in the right cerebellum r/o CCD.

Figure 13A and 13B are images of the brain of a patient (age 72, male) who has been diagnosed with traumatic brain injury and has not been treated with EPO or cord blood, but could be treated according to one or more methods of the invention. Figure 13A is an image of the patient's brain taken approximately three months after a traffic accident. The image shows perfusion defect at the left frontoparietal lobe and reduced perfusion around. Reduced perfusion can also be observed at the left temporal cortex, left basal ganglia and left thalamus. Reduced perfusion at the right cerebellum, and r/o crossed cerebellar diaschisis is suspected. Figure 13B is an image of the patient's brain taken approximately 10 months after the traffic accident. No significant change is identified at the left frontoparietal lobe and around. No significant change can be identified at the left temporal cortex. Interval improvements in perfusion can be observed at the left basal ganglia and left thalamus. Light perfusion reduction is observed at the right temporal cortex. Reduced perfusion at the right cerebellum, and crossed cerebellar diaschisis are suspected. Figure 14A and 14B are images of the brain of a patient (age 63, male) who has been diagnosed with traumatic brain injury and has not been treated with EPO or cord blood, but could be treated according to one or more methods of the invention. Figure 14A is an image of the patient's brain taken approximately three months after an accident in which the patient fell. Huge, heavy perfusion defect can be seen in the area from the right temporal to occipital lobe, and slightly reduced perfusion at the right basal ganglia. Figure 14B depicts an image of the patient's brain taken approximately 16 months after the accident. Slightly reduced perfusion can be identified in the left parietal lobe cortex. Right temporal lobe shows moderate or severe huge perfusion defect. Slightly reduced perfusion at the right basal ganglia and thalamus can be identified.

DESCRIPTION OF THE INVENTION

All references cited herein are incorporated by reference in their entirety as though fully set forth. Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Singleton et al., Dictionary of Microbiology and Molecular Biology 3^rd ed., J. Wiley & Sons (New York, NY 2001); March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 5^th ed., J. Wiley & Sons (New York, NY 2001); and Sambrook and Russel, Molecular Cloning: A Laboratory Manual 3rd ed., Cold Spring Harbor Laboratory Press (Cold Spring Harbor, NY 2001), provide one skilled in the art with a general guide to many of the terms used in the present application.

One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. Indeed, the present invention is in no way limited to the methods and materials described.

As used herein:

The acronym "TBI" means traumatic brain injury.

The acronym "SOD" means superoxide dismutase.

The acronym "XO" means xanthine oxidase.

The acronym "MDA" means malondialdehyde.

The acronym "NO" means nitric oxide.

The acronym "FGF" means fibroblast growth factor.

The acronym "VEGF" means vascular endothelial growth factor.

The acronym "EPO" means erythropoietin. The acronym "EPOR" means EPO receptor.

The acronym "rhEPO" means recombinant human erythropoietin.

The acronym "MCA" means middle cerebral artery.

The acronym "DTI" means diffusion tensor imaging.

The acronym "GMFCS" means gross motor functional classification system.

As used herein, "beneficial results" may include, but are in no way limited to reducing, inhibiting, or eliminating the pathologies associated with traumatic brain injury. Therefore, beneficial results may include but are in no way limited to improvements in speech, coordination, balance and reasoning. Beneficial results may also include, but are in no way limited to, improved general responsiveness, as well as improvements in hearing, taste, touch, smell, sight, and combinations thereof. Beneficial results may also include reduced depression and/or anxiety and an increase in a desire for socialization. Furthermore, "beneficial results" may include any improvement in any disability described herein, or resulting from any injury depicted in the brain images or patient profiles disclosed herein.

"Mammal" as used herein refers to a member of the class Mammalia, including, without limitation, humans as well as nonhuman primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, sheep, pigs, goats and horses; domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats and guinea pigs, and the like. The term does not denote a particular age or sex. Thus, newborn subjects and infant subjects, as well as fetuses, whether male or female, are intended to be included within the scope of this term.

In some embodiments, the numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth, used to describe and claim certain embodiments of the application are to be understood as being modified in some instances by the term "about." Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the application are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable.

By way of background, EPO is an endogenous 30.4-kDa hematopoietic glycoprotein, which can induce the homodimerization of two molecules of the EPO receptor (EPOR) on the cell surface. EPO is produced mainly by interstitial fibroblasts in the kidneys of adults and in the hepatocytes of fetuses (Fisher and Laron, 2003). EPO was initially identified for its role in erythropoiesis, regulating red blood cell production by preventing apoptosis of erythroid progenitor cells (Koury and Bondurant, 1992). To date, rhEPO has been clinically used to correct anemia arising from chronic renal failure and cancers in adults, and from prematurity in infants. Interestingly, rhEPO therapy for chronic renal failure revealed its role in enhancing neuromuscular functions (Sobh et al, 1992), where rhEPO also showed neuroprotective effects in both in vitro and in vivo studies (Spandou et al, 2004; Yu et al, 2005; Wen et al, 2006; Kellert et al, 2007). Administration of rhEPO after ischemia, injected either peripherally or centrally, suggested its beneficial effects on brain edema by delaying neuronal death (Calapai et al. , 2000), resulting in functional improvement of neurons in both adult and neonatal animal models (Wen et al., 2006; Li et al., 2007). Post-ischemic treatment with erythropoietin or carbamylated erythropoietin has been reported to reduce infarction and improve neurological outcome in a rat model of focal cerebral ischemia (Wang, Y. et al., 2007).

Some studies indicate that optimal neuroprotection from EPO administration is observed when EPO is given immediately following brain insult. On the other hand, reduced neuroprotection was observed when EPO administration was delayed (van der Kooij et al, 2008). In fact, the therapeutic window for EPO treatment after an ischemic stroke has been reported to be 3 hours (Lapchak et al, 2008).

As demonstrated herein, through clinical experiences and experimentation, the inventors made several important discoveries, including, but not limited to: 1) a longer therapeutic window exists for EPO treatment of TBI than what was previously accepted; 2) EPO is an effective therapy for TBI patients of a wide-range of ages, from infants to the elderly; 3) there is significant therapeutic synergy when EPO administration is combined with stem cell therapy to treat patients with TBI.

In certain embodiments, the invention teaches a method of treating traumatic brain injury (TBI) in a subject, including: administering EPO to a subject having traumatic brain injury. In certain embodiments, the method also includes administering stem cells to the subject. In some embodiments, the stem cells administered are selected from the group consisting of: umbilical cord blood cells, embryonic stem cells, bone marrow cells, mesenchymal stem cells, adipose-derived stem cells, endothelial stem cells, dental pulp stem cells, placenta stem cells, olfactory ensheathing cells, neural precursor cells and combinations thereof. In certain embodiments, the stem cells transplanted are autologous. In some embodiments, the stem cells transplanted are allogeneic.

In certain embodiments, the erythropoietin used in the inventive method is recombinant erythropoietin. In certain embodiments, the recombinant erythropoietin is recombinant human erythropoietin. In certain embodiments, the erythropoietin is derived from an animal. In certain embodiments, the erythropoietin is synthetic. In some embodiments, an analog of erythropoietin with substantially the same therapeutic properties with respect to treating TBI is used. In certain embodiments chemically modified erythropoietin is used. In certain embodiments, glycosylated erythropoietin is used. In some embodiments the glycosylated erythropoietin is selected from the group consisting of: EPO- alpha, EPO-beta, or combinations thereof. In certain embodiments, darbepoieten is used. In certain embodiments, the dosage of erythropoietin administered is 50 IU/kg, 100 IU/kg, 150 IU/kg, 200 IU/kg, 250 IU/kg, 300 IU/kg, 350 IU/kg, 400 IU/kg, 500 IU/kg or more. In an embodiment, the dosage is 250 IU/kg. In an embodiment, the dosage is 500 IU/kg. In some embodiments, erythropoietin is administered one or more times per year. In some embodiments, erythropoietin is administered one or more times per month. In some embodiments, erythropoietin is administered one or more times per week. In certain embodiments, erythropoietin is administered twice per week, subcutaneously. In an embodiment, 250 IU/kg is administered subcutaneously two to three times per week. In another embodiment, 500 IU/kg is administered subcutaneously two to three times per week. In certain embodiments, erythropoietin is administered by alternate routes of administration, as disclosed herein. In certain embodiments, the subject's blood is drawn and a complete blood cell count is performed one or more times per week during the course of therapy. In certain embodiments, the subject's hemoglobin levels are tested one or more times per week. In certain embodiments erythropoietin administration is stopped if hemoglobin levels detected in the subject reach 15 g/dl. In certain embodiments, erythropoietin administration is stopped if hemoglobin levels detected in the subject reach 16 g/dl. In certain embodiments, erythropoietin administration is stopped if the subject's platelet count reaches 400,000/μ1. In certain embodiments, erythropoietin administration is resumed when hemoglobin and/or platelet levels return to normal.

In certain embodiments, the subject treated according to one or more of the inventive methods is an animal. In certain embodiments, the subject is a mammal. In certain embodiments, the subject is a human. In some embodiments, the subject is an infant. In other embodiments, the subject is a child. In some embodiments, the subject is an adult. In certain embodiments, the subject treated according to one or more of the inventive methods, has sustained a type of TBI consistent with one or more of the images demonstrated in figures 2-14. In certain embodiments, the subject has been diagnosed with quadriplegia due to traumatic subdural hematoma. In certain embodiments, the subject has been diagnosed with pons hemorrhage. In certain embodiments, the subject has been diagnosed with hypoxic brain injury. In certain embodiments, the subject has been diagnosed with a cerebral contusion. In certain embodiments, the subject has been diagnosed with cerebral palsy. In certain embodiments, the subject has been diagnosed with hemiplegia. In certain embodiments, the subject has been diagnosed with subarachnoid hemorrhage. In come embodiments, the subject has been diagnosed with intracerebral hemorrhage. In certain embodiments, the subject has been diagnosed with epidural hemorrhage. In certain embodiments, the subject has been diagnosed with a condition or injury selected from the group consisting of: cerebral infarction, transverse myelitis, spinal cord injury, peripheral nerve injury, Parkinson's disease, Alzheimer's disease, Parkinson Syndrome, dementia by various causes, multiple sclerosis, multisystem atrophy and combinations thereof. In certain embodiments, the subject has been diagnosed with one or more of the injuries and/or conditions demonstrated in figure 9.

In various embodiments, erythropoietin is administered immediately after injury. In some embodiments, erythropoietin is administered within a month after injury. In some embodiments erythropoietin is administered more than one month after injury. In certain embodiments, erythropoietin is administered to a subject a year or more after surgery. In certain embodiments, erythropoietin is administered to a subject with a chronic injury or condition described herein.

In various embodiments, the stem cell therapy is administered immediately after injury. In an embodiment, stem cell therapy is administered after the subject's medical and surgical condition is stabilized. In some embodiments, erythropoietin is administered between 1 hour and 2.5 or more days before stem cell therapy. In an embodiment, erythropoietin is administered between 3 hours and 2.5 days or more before stem cell treatment. In certain embodiments, erythropoietin is administered two or more times per week prior to stem cell transplantation, according to the dosage recommendations disclosed herein. In an embodiment, stem cell therapy is repeated as tolerated by the subject. In certain embodiments, stem cell therapy is repeated when immunosupression and/or related effects from the prior stem cell therapy subsides. In various embodiments, stem cell therapy is repeated after 6 months. In various embodiments, stem cell therapy can be administered before or after treatment with erythropoietin.

In various embodiments, erythropoietin may be provided as a pharmaceutical composition including a pharmaceutically acceptable excipient along with a therapeutically effective amount of the erythropoietin. "Pharmaceutically acceptable excipient" means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, nontoxic, and desirable, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients may be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous.

In various embodiments, the pharmaceutical compositions according to the invention may be formulated for delivery via any route of administration. "Route of administration" may refer to any administration pathway known in the art, including but not limited to aerosol, nasal, oral, transmucosal, transdermal or parenteral. "Transdermal" administration may be accomplished using a topical cream or ointment or by means of a transdermal patch. "Parenteral" refers to a route of administration that is generally associated with injection, including intraorbital, infusion, intraarterial, intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intrathecal, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal. Via the parenteral route, the compositions may be in the form of solutions or suspensions for infusion or for injection, or as lyophilized powders. Via the enteral route, the pharmaceutical compositions can be in the form of tablets, gel capsules, sugar-coated tablets, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres or lipid vesicles or polymer vesicles allowing controlled release. Via the parenteral route, the compositions may be in the form of solutions or suspensions for infusion or for injection. Via the topical route, the pharmaceutical compositions based on compounds according to the invention may be formulated for treating the skin and mucous membranes and are in the form of ointments, creams, milks, salves, powders, impregnated pads, solutions, gels, sprays, lotions or suspensions. They can also be in the form of microspheres or nanospheres or lipid vesicles or polymer vesicles or polymer patches and hydrogels allowing controlled release. These topical-route compositions can be either in anhydrous form or in aqueous form depending on the clinical indication.

The pharmaceutical compositions according to the invention can also contain any pharmaceutically acceptable carrier. "Pharmaceutically acceptable carrier" as used herein refers to a pharmaceutically acceptable material, composition, or vehicle that is involved in carrying or transporting a compound of interest from one tissue, organ, or portion of the body to another tissue, organ, or portion of the body. For example, the carrier may be a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, or a combination thereof. Each component of the carrier must be "pharmaceutically acceptable" in that it must be compatible with the other ingredients of the formulation. It must also be suitable for use in contact with any tissues or organs with which it may come in contact, meaning that it must not carry a risk of toxicity, irritation, allergic response, immunogenicity, or any other complication that excessively outweighs its therapeutic benefits.

The pharmaceutical preparations are made following the conventional techniques of pharmacy involving milling, mixing, granulation, and compressing, when necessary, for tablet forms; or milling, mixing and filling for hard gelatin capsule forms. When a liquid carrier is used, the preparation will be in the form of syrup, elixir, emulsion or an aqueous or non-aqueous suspension. Such a liquid formulation may be administered directly p.o. or filled into a soft gelatin capsule.

The pharmaceutical compositions according to the invention may be delivered in a therapeutically effective amount. The precise therapeutically effective amount is that amount of the composition that will yield the most effective results in terms of efficacy of treatment in a given subject. This amount will vary depending upon a variety of factors, including but not limited to the characteristics of the therapeutic compound (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (including age, sex, disease type and stage, general physical condition, responsiveness to a given dosage, and type of medication), the nature of the pharmaceutically acceptable carrier or carriers in the formulation, and the route of administration. One skilled in the clinical and pharmacological arts will be able to determine a therapeutically effective amount through routine experimentation, for instance, by monitoring a subject's response to administration of a compound and adjusting the dosage accordingly. For additional guidance, see Remington: The Science and Practice of Pharmacy (Gennaro ed. 20th edition, Williams & Wilkins PA, USA) (2000).

Typical dosages of an effective amount of the erythropoietin and/or stem cells can be as indicated to the skilled artisan by the in vitro responses or responses in animal models. The actual dosage will depend upon the judgment of the physician, the condition of the patient, and the effectiveness of the therapeutic method.

The present invention is also directed to a kit to treat TBI in a mammal in need thereof. The kit is useful for practicing the inventive method of treating TBI. The kit is an assemblage of materials or components, including at least one of the inventive compositions. Thus, in some embodiments the kit contains a composition including erythropoietin and/or stem cells as described above.

The exact nature of the components configured in the inventive kit depends on its intended purpose. For example, some embodiments are configured for the purpose of treating TBI. In one embodiment, the kit is configured particularly for the purpose of treating mammalian subjects. In another embodiment, the kit is configured particularly for the purpose of treating human subjects. In another embodiment, the kit is configured for treating adolescent, child, or infant human subjects. In further embodiments, the kit is configured for veterinary applications, treating subjects such as, but not limited to, farm animals, domestic animals, and laboratory animals.

Instructions for use may be included in the kit. "Instructions for use" typically include a tangible expression describing the technique to be employed in using the components of the kit to effect a desired outcome, such as to treat TBI. Instructions for use may include instructions to administer a dose of erythropoietin from 1-10 times per week. Particularly, instructions for use may include instructions to administer 250 IU/kg rhEPO, subcutaneously, twice per week. Alternatively, instructions for use may include instructions to administer 500 IU/kg rhEPO. For example, total dosage should be administered as almost the same amount to patients with chronic kidney. In some cases, the instructions may indicate a dosage that varies according to an established treatment protocol in which not all doses are the same. Optionally, the kit also contains other useful components, such as, diluents, buffers, pharmaceutically acceptable carriers, syringes, catheters, applicators, pipetting or measuring tools, bandaging materials or other useful paraphernalia as will be readily recognized by those of skill in the art.

The materials or components assembled in the kit can be provided to the practitioner stored in any convenient and suitable ways that preserve their operability and utility. For example the components can be in dissolved, dehydrated, or lyophilized form; they can be provided at room, refrigerated or frozen temperatures. The components are typically contained in suitable packaging material(s). As employed herein, the phrase "packaging material" refers to one or more physical structures used to house the contents of the kit, such as inventive compositions and the like. The packaging material is constructed by well-known methods, preferably to provide a sterile, contaminant-free environment. As used herein, the term "package" refers to a suitable solid matrix or material such as glass, plastic, paper, foil, and the like, capable of holding the individual kit components. Thus, for example, a package can be one or more glass vials or plastic containers used to contain suitable quantities of erythropoietin. The packaging material generally has an external label which indicates the contents and/or purpose of the kit and/or its components.

One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. Indeed, the present invention is in no way limited to the methods and materials described.

The following examples are for illustrative purposes only and are not intended to limit the scope of the disclosure or its various embodiments in any way.

EXAMPLES

Example 1

Case Reports of Erythropoietin Use Only (Part I)

Case number 1 (Figure 2)

1. Sex/age: female/4 (body weight 14 kg)

2. Diagnosis: quadriplegia due to traumatic subdural hematoma (onset date: June 5 ^th, 2010)

3. Perinatal history: intrauterine period 37 weeks; no specific problem

4. Present illness

The previously healthy child had a car accident on June 5 ^th, 2010 when she was about 3 years 8 months old. In that accident, the car turned over and the child was thrown out of the car. She was transferred to a hospital. Brain CT showed acute bilateral frontal subdural hematoma and cerebral contusion. She was cared for at the intensive care unit for 2 weeks. After that, quadriplegia developed and rehabilitation treatment was started.

5. MRI findings on December 11^th, 2010

A. Multifocal small old brain contusion with old hemorrhages at bilateral frontal, right temporal and parietal lobes.

B. Small volume of fiber tracts with defects at bilateral frontal regions.

6. Visiting record

A. First visit on October 11^th, 2010.

B. Erythropoietin therapy began on November 11^th, 2010. At each treatment, she was given 3500 IU (250 IU/kg, 14kg) and 8 injections were completed by administering injections twice per week.

C. Follow up on January 14^th, 2011. 7. Improvements after erythropoietin administration

A. Increased range of eye movement.

B. Increased right hand activity, which was near none at initial therapy session.

C. Improved pathologic gait pattern such as circumduction and toe gait.

D. Increased language expression.

8. Evaluation

A. Initial GMFM (Gross Motor Function Measurement) on November 4^th, 2010.

1. A: 100%, B: 98%, C: 100%, D: 79%, E: 56%

B. Follow up GMFM in December 17^th, 2010.

2. A: 100%, B: 98%, C: 100%, D: 95%, E: 64%

C. Initial GMPM (Gross Motor Performance Measurement) on November 4¹ 2010.

1. Dissociated movement 20 (57%)

2. Coordination 27 (54%)

3. Alignment 47 (49%)

4. Weight shift 24 (48%)

5. Stability 30 (55%)

Total score: 52.6 %

D. Follow up GMPM on December 17^th, 2010

1. Dissociated movement 21 (67%)

2. Coordination 30 (60%)

3. Alignment 56 (59%)

4. Weight shift 30 (60%)

5. Stability 33 (60%)

Total score: 59.8 %

E. Bayley Scales of Infant Development-II on November 4^th, 2010

1. Mental raw score: 140, Motor raw score: 91

F. Follow up Bayley Scales of Infant Development-II on December 17^th, 2010

1. Mental raw score: 147, Motor raw score: 91

Case number 2 (Figure 3)

1. Sex/age: male/11

2. Diagnosis: quadriplegia due to traumatic brain injury (onset date: August 25^th, 2007).

3. Perinatal history: intrauterine period 40 weeks, no developmental problem. Present illness

The previously healthy child had a pedestrian traffic accident on August 25^th, 2007 when he was about 7 years 2 months old. He was transferred to a hospital and the brain CT showed basal skull fracture and intracranial hemorrhage. He was cared for at the intensive care unit for 4 weeks. He was suspected to have suffered from hypoxic brain injury due to acute pharyngitis during his stay in the ICU. After that, quadriplegia developed and he started to receive rehabilitation treatment.

MRI finding on November 22^th, 2010

A. Old contusions at the base of bilateral frontal lobes

Visiting records

A. First visit on March 25^th, 2008

B. Erythropoietin therapy began on December 20^th, 2010. At each treatment, he was given 9,000 IU (250 IU/kg, 39kg) each time and 6 injections were completed by administering injections twice per week.

C. Follow up on January 27^th, 2011

D. The second erythropoietin treatment began on May 19^th, 2011. At each treatment, he was given 10,000 IU (250 IU/kg, 40 kg) each time, and 6 injections were completed by administering injections twice per week.

E. Follow up on June 13^th, 2011

Improvements after erythropoietin administration

A. Much improved hearing capacity

B. His knees were straightened, which had been flexed a little when sitting on the floor.

C. Increased motor strength: he became able to flex his hips about 10 degrees for 5 seconds when sitting on a chair, which had been impossible for him before.

Evaluation

A. Muscle strength by manual muscle testing (from December 20^th, 2010→ July 8^th, 2011)

1. Right lower extremity: hip flexor/knee extensor/ankle dorsiflexor: fair/fair/fair— > fair+/fair+/fair

2. Left lower extremity: hip flexor/knee extensor/ankle dorsiflexor: fair/fair/fair— > fair+/fair+/fair Case number 3

1 Sex/age: male/4 (body weight 13 kg)

2 Diagnosis: cerebral palsy, hemiplegia, right

3 Perinatal history

Intrauterine period 37 weeks full term, birth body weight 3.1 kg, cesarean section

4 Present illness

He had been in the course of normal development from birth without any specific perinatal problem. However, when he was hit by a big toy car on one hundred days after birth, seizure-like activity was shown. At two days after the insult, right side muscle weakness was observed. The parents took him to the general hospital and, brain CT revealed left hemisphere injury and brain edema. The infant had an emergency operation. Afterwards, he showed developmental delay.

5. Visiting record

A. First visit on November 18^th, 2010

B. Erythropoietin therapy began on December 2^nd, 2010. At each treatment, he was given 3,000 IU (250 IU/kg, 13 kg) each time and 6 injections were completed by administering injections twice per week.

C. Follow up in January 27^th, 2011

6. Improvements after erythropoietin administration

A. Appearance of imitating words; for example, he made a sound "A" for the Korean word "A-juh-ssi", which literally means "uncle" in English. Other examples were making honking sound instead of saying a "car" and "Ha" for "Hal-mu-ni", that is an equivalent Korean word for grandmother.

B. When a caregiver told him to sit down, he followed the order.

C. He became able to move sitting 50 days after the first EPO injection. In addition, sitting crossed-legged and kneeling became possible.

Case number 4 (Figure 4)

1. Sex/age: female/68 (body weight 46 kg)

2. Diagnosis: quadriplegia due to traumatic subdural hematoma, subarachnoid hemorrhage, intracerebral hemorrhage (onset date: July 1^st, 2009)

3. Past history: hypertension, DM, operation history: coronary artery bypass on May, 2009 Present illness

The patient fell down from the second floor of her house to the first floor on July 1^st, 2009. Brain CT showed traumatic subdural hematoma, subarachnoid hemorrhage, intracerebral hemorrhage and she had hematoma evacuation and craniectomy. Her mentality was drowsy to stupor and incommunicable even without meaningful eye contact.

MRI findings on December 10^th, 2009

A. S/P decompressive craniectomy, left

B. Old contusional encephalomalacia at bilateral frontal and left temporal lobes

C. Old lacunar infarct at right basal ganglia, centrum semiovale, and left temporal white matter

D. Mastoiditis, bilateral

E. Decreased fiber volume at left frontal and parietal regions on 3D DTI (diffusion tensor imaging)

Visiting record

A. First visit on September 17^th, 2009

B. Erythropoietin treatment was started on April 23^th, 2010. For the first four sessions, she was given 3000 IU (65 IU/kg, 46 kg) each time, and the next 2 sessions, she received 9,000 IU (200 IU/kg) each time. In May 13^th, 2010, erythropoietin therapy was started again at 9,000 IU (200 IU/kg) each time. 12 injections were completed by administering injections twice per week.

C. Follow up on July 12^th, 2011

Improvements

A. Improved eye opening

B. Became more alert

C. Increased interdigital muscle thickness

Case number 5 (Figure 9)

1. Sex/age: male/7 years 4 months

2. Diagnosis: quadriplegia due to traumatic brain injury, traumatic subarachnoid hemorrhag (onset date: Aug. 9^th, 2011)

3. Past history: non-significant before traffic accident

4. Present illness He was previously healthy. He had a pedestrian traffic accident on Aug. 9 , 2011. Brain CT showed traumatic subarachnoid hemorrhage, and intracerebral hemorrhage at left basal gangalia. The patient had hematoma evacuation and hospitalized with intubation.

Visiting record

A. First visit on Aug. 9^th, 2011

B. First combined Erythropoietin treatment on Aug. 16^th.

1. 6,500IU (250IU/kg) each time, 20 injections were completed by administering injections twice per week.

C. Follow up on Nov. 24^th, 2011

D. Follow up on March 22^nd, 2011

MRI findings on Aug. 15^th, 2011

A. Multicentric, contusiform cerebral hemorrhage at left frontal lobe and left basal ganglia.

B. Small multicentric hemorrhage at right hemisphere- gray substance and white substance - diffuse axonal injury suspected.

C. Small intraventricular hemorrhage.

Improvements

A. Reaction to first erythropoietin treatment

1. From bed rest state when transferred to rehabilitation on Sep.

8^th, 2011, to fast walking without assistance, at discharge from the hospital on Nov. 10^th 2011.

2. At the early phase after transfer, conversation was not possible due to inability in pronunciation, however, there was no difficulty in verbal communication at the time of leaving the hospital.

3. Eyes only at the initial phase of transfer

B. Evaluation

1. GMFCS (Gross Motor Function Classification System) level evaluation (Sep. 8^th, 2011→ Nov. 9^th, 2011) 5→ 1

2. GMFM evaluation (Sep. 8^th, 2011→ Nov. 9^th, 2011) 13%→ 96% 3. MMT evaluation (Sep. 8^ffi, 201 1→ Nov. 9^m, 201 1) - Improved from "left leg and arm good, right leg and arm trace"→ "left leg and arm good, right leg and arm good"

4. MMSE (Oct. 5^th, 201 1→ Nov. 9^th, 201 1) 16→ 24

5. Korean Bayley Scale of Infant Development-II (Sep. 9^th, 201 1 → Nov. 9^th, 201 1) - recognition 56 (5 months), motion 26 (4 months) → recognision 173 (42 months), motion 96 (30 months)

6. WeeFIM evaluation (Sep. 9^th, 201 1 → Nov. 9^th, 201 1) 19→ 1 14

7. MAS evaluation (Sep. 9^th, 201 1→ Nov. 9^th, 201 1)

8. Hand evaluation (Sep. 9^th, 201 1→ Nov. 10^th, 201 1)- evaluation impossible→ grasp power 7 kg / 8 kg, lateral pinch 1.5 kg / 3 kg, tripod pinch 2 kg / 3.5 kg.

Case number 6 (Figure 10) - Same subject as Part 2, Case number 5. Changes after EPO treatment and before stem cell treatment

1. Sex/age: male/ 1 1 years

2. Diagnosis: quadriplegia due to traumatic brain injury and traumatic brain damage and hypoxic brain injury (onset date: Aug. 25^th, 2007).

3. Past history: non-significant before the traffic accident

4. Present illness

He was previously healthy. He was involved in a pedestrian traffic accident on Aug. 25^th, 2007 (7 years 2 months). Brain CT showed skull fracture and intracerebral hemorrhage. The hypoxic brain injury was thought to be caused by acute pharyngitis while hospitalized in intensive care room. Rehabilitation therapy was applied, due to reduced muscle power in the limbs.

5. Visiting record

A. First visit on March 25^th, 2008

B. First combined Erythropoietin treatment on Dec. 20^th, 2010

1. 9,000 IU (250 IU/kg) each time; 6 injections were completed by administering injections twice per week.

D. Follow up on Jan. 17^th, 201 1

E. Second combined erythropoietin treatment on May 19^th, 201 1 1. 10,000 IU (250 IU/kg) each time, 6 injections were completed by administering injections twice per week.

F. Follow up on June 13^th, 2011

G. Cord blood treatment via femoral artery, on June 30^th, 2011

MRI findings on Nov. 22^nd, 2010

A. Old bruise on the bottom of both frontal lobes

B. DTI showed reduced fibrin in both frontal lobes and parietal lobe

Improvements

A. Reaction to the first and second erythropoietin treatments:

1. Hearing ability has greatly improved.

2. When seated on floor, both knees were bent by 10-20 degrees straightened perfectly

3. While seated on a chair, the patient can remain for 5 seconds with both hip joints bent by about 10 degrees. Previously, hip joints could not be bent in that posture at all.

4. The muscular force ratings of the hip joints, knees and ankle joints have been improved from fair to fair plus.

Example 2

Case Reports of Combined Erythropoietin and Stem Cell Therapy (Part II)

Case number 1 (Figure 5)

1. Sex/age: male/62 (body weight 75 kg)

2. Diagnosis: right hemiplegia due to traumatic subdural hematoma, and intracerebral hemorrhage at left basal ganglia (onset date: August 19^th, 2008).

3. Past history: hypertension

4. Present illness

The patient was healthy except for taking antihypertensive medication for 10 years. After he fell down while carrying stuffs, he was transferred to the hospital. Brain CT revealed traumatic subdural hematoma and hemorrhage at left basal ganglia. He had surgery, and afterwards, right side hemiplgeia was developed.

5. MRI findings on September 24^th, 2009 A. Cavitary encephalomalacia at superomedial portion of left frontal and parietal lobes.

B. Wallerian degeneration at left-side brainstem.

C. Fiber defect at left cerebral hemisphere, preserving corpus callosum genu and anterior frontal white matter on 3D DTI. Fiber defects were seen in left hemisphere including corticospinal tract and thalamo-cortical radiations.

6. Visiting record

A. First visit in September 8^th, 2009

B. Autologous bone marrow stem cell transplantation on September 25^th, 2009.

Combined erythropoietin treatment: 18,750 IU each time (250 IU/kg body weight 75 kg), 2 injections were completed by administering injections twice per week.

C. Alcohol motor point block and botulinum toxin injection at right biceps, finger flexors on September 28^th, 2009

D. Follow up on October 16^th, 2009

7. Improvements (September 8^th, 2009→October 16^th, 2009)

A. Improved muscle power

1. Right upper extremity: Shoulder abductor/elbow flexor/wrist extensor: fair/poor/trace→ fair/poor+/poor+

2. Right lower extremity: Hip flexor/knee extensor/ankle dorsiflexor: fair+/fair/trace→ fair+/fair+/trace

B. Grasp power: uncheckable→ 3.5 kg

C. Increased gait speed Case number 2 (Figure 6)

1. Sex/age: male/32 (body weight 61 kg)

2. Diagnosis: quadriplegia due to traumatic brain injury, traumatic subdural hematoma, epidural hemorrhage, subarachnoid hemorrhage (onset date: Aug. 20^th, 2006). Diffuse axonal injury, dysphagia, aphasia and neurogenic bladder bowel.

3. Past history: non-significant before traffic accident

4. Present illness

He got into a traffic accident as a pedestrian on August 20^th, 2006 and was transferred to a hospital. Brain CT showed traumatic subdural hematoma, epidural hematoma and subarachnoid hemorrhage and the patient had an emergency operation. After that accident, the patient became quadriplegic and incommunicable.

MRI findings on May 25^th, 2010

A. Progressed diffuse brain atrophy

B. Multifocal small old hemorrhages with contusion at bilateral frontal, right temporal lobes, corpus callosum, right mesial temporal lobe, left thalamus, and midbrain.

C. Defect at bilateral cerebral hemisphere, preserving thin fiber tracts at bilateral posterior frontal regions.

Visiting record

A. First visit on September 30^th, 2006

B. Allogeneic umbilical cord blood stem cell transplantation on June 14^th, 2010.

Combined erythropoietin treatment: 30,000 IU each time (500 IU/kg body weight 61 kg); 3 injections were completed by administering injections twice per week.

C. On March 18^th, 2011, the same cord blood stem cells were implanted + EPO injection with the same dose.

D. Follow up examination: showed almost no improvement July 2^nd, 2010, Feb.

6^th, 2011, and April 14^th, 2011. General body weakness and immunosuppression had made the condition of the patient worse. Only the arm stiffness had been improved by one grade from MAS 2 to MAS 1 , from the second cord blood treatment to discharge from the hospital. On June 12^th, 2012, functional improvement was identified in the evaluation at the Myeongji Hospital.

Improvements

A. Whether he likes or not became more apparent.

B. He became able to express exactly whether it is feces or urine after urination or defecation.

C. He could not stand initially, but became able to stand holding side rails at a parallel bar with back support for roughly 10 minutes.

D. Improved muscle power (June 14^th, 2010→July 2^nd, 2010) -> June 12^th, 2012:

He was evaluated at another hospital (Myeongji Hospital)

1. Right upper extremity: Shoulder abductor/elbow flexor/wrist extensor: poor/poor/trace→ fair/fair/poor fair+/fair+/fair+ 2. Right lower extremity: Hip flexor/knee extensor/ankle dorsiflexor: poor/poor/trace → poor+/poor+/trace good/good/good

3. Left lower extremity: Hip flexor/knee extensor/ankle dorsiflexor: trace/trace/trace→ poor/poor/trace.

E. Evaluation of daily living motion (modified Barthel Index Score): Zero (June & July, 2010, Feb. & April, 2011) -> evaluated to be 7 on June 12^th, 2012.

Case number 3 (Figure 7)

1. Sex/age: male/20 (body weight 62 kg)

2. Diagnosis: quadriplegia due to traumatic brain injury, traumatic subdural hematoma at left fronto-temporo-parietal lobe (onset date: March 6^th, 2010)

3. Past history : non-significant before traffic accident

4. Present illness

He was hit by a car as a pedestrian on March 6^th, 2010. After being transferred to a hospital, brain CT revealed acute subdural hematoma at left fronto-temporo-parietal lobe. He underwent neurosurgical operation including a craniectomy and removal of subdural hematoma. However he was unable to be conscious and remained in a vegetative status.

5. MRI findings on September 22^th, 2010

A. Encephalomalacia with atrophy of entire cerebrum.

B. No fiber tract visible by DTI.

6. Visiting record

A. First visit on August 12^th, 2010

B. First allogeneic umbilical stem cell transplantation on September 17^th, 2010

C. First combined erythropoietin treatment: 27,000 IU (500 IU/kg 54 kg) each time; 2 injections were completed by administering injections twice per week.

D. 13,500 IU (250 IU/kg 54 kg) each time; 4 injections were completed by administering injections twice per week.

E. Follow up on December 23^th, 2010

F. Second allogeneic umbilical stem cell transplantation on February 25^th, 2011

G. Second combined erythropoietin treatment

H. 31,000 IU (500 IU/kg 62 kg) each time; 2 injections were completed by administering injections twice per week. I. 15,500 IU (250 IU/kg 62 kg) each time, and 1 injection was administered. J. Follow up on March 7^th, 2011

Improvements

A. Response to first stem cell and erythropoietin treatment

1. The patient showed a new response to a certain sound.

Especially he responded to the bamboo flute sound made by his father.

2. Decreased spasticity (modified Ashworth scale) within one month

i. Right elbow flexor : Gl→ GO

ii. Left elbow flexor : Gl→ GO

iii. Right knee flexor: G2→ G1+

iv. Left knee flexor : G2→ G1+

3. No interval change in muscle power

i. zero grade in terms of MMT (manual muscle testing)

4. Significant disappearance of abnormally high body temperature without known origin and normalized pulse rate from tachycardia.

B. Response to second stem cell and erythropoietin treatment

1. He showed some responses to certain kinds of familiar voices or sounds. For example, when he listened to the voice of his brother through a cell phone, he made a certain sound. Case number 4 (Figure 8)

1. Sex/age: female/23

A. Diagnosis: quadriplegia due to traumatic brain injury, traumatic subdural hematoma (onset date : August 3^rd, 2006)

2. Past history: non-significant before traffic accident

3. Present illness

She was previously healthy, but was involved in a pedestrian traffic accident on August 3^rd, 2006. Brain CT showed traumatic subdural hematoma, and left brain swelling. She had an emergency operation - craniectomy and hematoma removal. Brain CT taken on October 23 , 2006 revealed hydrocephalus, and she had an operation- ventriculo-peritoneal shunting.

Visiting records

A. First visit on November 28^th, 2007

B. First allogeneic umbilical stem cell transplantation on May 17^th, 2010

C. First combined erythropoietin treatment

1. 27,500 IU (500 IU/kg) each time; 2 injections were completed by administering injections twice per week.

2. 13,750 IU (250 IU/kg) each time, 8 injections were completed by administering injections twice per week.

D. Follow up on December 23^th, 2010

E. Second allogeneic umbilical stem cell transplantation on March 5^th, 2011

F. Second combined erythropoietin treatment

1. 27,500 IU (500 IU/kg) each time; 2 injections were completed by administering injections twice per week.

2. 13,750 IU (250 IU/kg) each time; 8 injections were completed by administering injections twice per week.

G. Follow up on May 19^th, 2011

MRI findings on May 15^th, 2010

A. Encephalomalacia at bilateral frontal and temporal lobes

B. S/P VP shunt

C. Enlarged left ventricles

D. Fiber defects at bilateral frontal and parietal regions

E. Artifact at right cerebral vertex.

Improvements

A. Response to first stem cell and erythropoietin treatment

1. She had shown severe aggressiveness before the treatment, but she began to try to control aggression. Her attention span was increased.

2. Visual perception function was improved, for example, drawing along lines and color matching.

3. Increased right hand use which was not involved during any activities. 4. Improved standing balance in one month. Static balance:

poor→fair , dynamic balance: zero→poor

5. Evaluation

i. Score of modified Barthel index (March 5^th, 2009→ June 21^th, 2010) 0→ 26

ii. Fugl-Meyer assessment (April 27^th, 2010 → June 21^st, 2010). Right upper extremity 15→30 iii. Motor assessment scale score 14 (March 4^th, 2009)→22 (January 29^th, 2011)→24 (April 8^th, 2011)

B. Response to second stem cell and erythropoietin treatment

1. She became able to stand up straight from the wheel chair. She did not bite her clothes any more.

2. She had not been able to walk, despite the aid of the therapist.

However, after the treatment, she could walk with moderate assistance.

Case number 5 (Figure 10)- Same patient as Part 1, Case number 6, but including changed clinical aspects after stem cell and third EPO treatments.

1. Sex/age: male/ 11 years

2. Diagnosis: quadriplegia due to traumatic brain injury and traumatic brain damage and hypoxic brain injury (onset date: Aug. 25^th, 2007).

3. Past history: non-significant before the traffic accident

4. Present illness

He was previously healthy, but was involved in a pedestrian traffic accident on Aug. 25^th, 2007 (7 years 2 months). Brain CT showed a skull fracture and intracerebral hemorrhage. The hypoxic brain injury was thought to be caused by acute pharyngitis while he was hospitalized in the intensive care room. Rehabilitation therapy was applied due to reduced muscle power in the limbs.

5. Visiting record

A. First visit on March 25^th, 2008

B. First combined erythropoietin treatment on Dec. 20^th, 2010

1. 9,000 IU (250 IU/kg) each time; 6 injections were completed by administering injections twice per week. C. Follow up on Jan. 17^ffi, 2011

D. Second combined erythropoietin treatment on May 19^th, 2011

1. 10,000 IU (250 IU/kg) each time, 6 injections were completed by administering injections twice per week.

E. Follow up on June 13^th, 2011

F. Cord blood treatment via femoral artery, on June 30^th, 2011

6. MRI findings on Nov. 22^nd, 2010

A. Old bruise on the bottom of both frontal lobes

B. DTI showed reduced fibrin in both frontal lobes and parietal lobe

7. Improvements

A. Reaction to the first and second erythropoietin treatments

1. Hearing ability has greatly improved.

2. When seated on the floor, both knees could be straightened perfectly, from bent by 10 to 20 degrees previously.

3. While seated on a chair, the patient can remain for 5 seconds with both hip joints bent by about 10 degrees. Previously, hip joints could not be bent in that posture at all.

4. The muscular force ratings of the hip joints, knees and ankle joints have been improved from fair to fair plus.

B. Reaction to the first cord blood and third Erythropoietin treatments:

1. Upon leaving the hospital on July 9^th, 2011, the patient could lift up his legs by bending his hip joints, and could hold his leg off of the floor for about 5 seconds, due to strengthened muscle power of the legs.

2. By Aug 25^th, 2011, his legs could move more easily. When getting on a car, he stepped on one foot first and then followed with another, in time. While lying down, he could lift his head above his shoulders.

Example 3

Tables of Patient Data

Table 1A

F ua rpega Hypoxc 5-5- None conserve

Table IB

Table 2A

Table 2B

Table 3A

Table 3B

The various methods and techniques described above provide a number of ways to carry out the invention. Of course, it is to be understood that not necessarily all objectives or advantages described can be achieved in accordance with any particular embodiment described herein. Thus, for example, those skilled in the art will recognize that the methods can be performed in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other objectives or advantages as taught or suggested herein. A variety of alternatives are mentioned herein. It is to be understood that some preferred embodiments specifically include one, another, or several features, while others specifically exclude one, another, or several features, while still others mitigate a particular feature by inclusion of one, another, or several advantageous features.

Furthermore, the skilled artisan will recognize the applicability of various features from different embodiments. Similarly, the various elements, features and steps discussed above, as well as other known equivalents for each such element, feature or step, can be employed in various combinations by one of ordinary skill in this art to perform methods in accordance with the principles described herein. Among the various elements, features, and steps some will be specifically included and others specifically excluded in diverse embodiments.

Although the application has been disclosed in the context of certain embodiments and examples, it will be understood by those skilled in the art that the embodiments of the application extend beyond the specifically disclosed embodiments to other alternative embodiments and/or uses and modifications and equivalents thereof.

In some embodiments, the terms "a" and "an" and "the" and similar references used in the context of describing a particular embodiment of the application (especially in the context of certain of the following claims) can be construed to cover both the singular and the plural. The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (for example, "such as") provided with respect to certain embodiments herein is intended merely to better illuminate the application and does not pose a limitation on the scope of the application otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the application.

Preferred embodiments of this application are described herein, including the best mode known to the inventors for carrying out the application. Variations on those preferred embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. It is contemplated that skilled artisans can employ such variations as appropriate, and the application can be practiced otherwise than specifically described herein. Accordingly, many embodiments of this application include all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the application unless otherwise indicated herein or otherwise clearly contradicted by context.

All patents, patent applications, publications of patent applications, and other material, such as articles, books, specifications, publications, documents, things, and/or the like, referenced herein are hereby incorporated herein by this reference in their entirety for all purposes, excepting any prosecution file history associated with same, any of same that is inconsistent with or in conflict with the present document, or any of same that may have a limiting affect as to the broadest scope of the claims now or later associated with the present document. By way of example, should there be any inconsistency or conflict between the description, definition, and/or the use of a term associated with any of the incorporated material and that associated with the present document, the description, definition, and/or the use of the term in the present document shall prevail.

In closing, it is to be understood that the embodiments of the application disclosed herein are illustrative of the principles of the embodiments of the application. Other modifications that can be employed can be within the scope of the application. Thus, by way of example, but not of limitation, alternative configurations of the embodiments of the application can be utilized in accordance with the teachings herein. Accordingly, embodiments of the present application are not limited to that precisely as shown and described.

Claims

CLAIMS What is claimed is:

1. A method of treating a traumatic brain injury and/or a neurological injury in a subject, comprising administering erythropoietin and stem cells to the subject.

2. The method of claim 1, wherein the stem cells are selected from the group consisting of: umbilical cord blood cells, embryonic stem cells, bone marrow cells, mesenchymal stem cells, adipose-derived stem cells, endothelial stem cells, dental pulp stem cells, placenta stem cells, olfactory ensheathing cells, neural precursor cells and combinations thereof.

3. The method of claim 1, wherein the stem cells are autologous.

4. The method of claim 1, wherein the stem cells are allogeneic.

5. The method of claim 1, wherein the erythropoietin is selected from the group consisting of: recombinant erythropoietin, synthetic erythropoietin, chemically modified erythropoietin, darbepoieten, glycosylated erythropoietin, EPO- alpha, EPO-beta and combinations thereof.

6. The method of claim 1, further comprising providing rehabilitation therapy to the

subject.

7. The method of claim 1, wherein the subject has a condition selected from the group consisting of: quadriplegia due to traumatic subdural hematoma, pons hemorrhage, hypoxic brain injury, cerebral contusion, cerebral palsy, hemiplegia, subarachnoid hemorrhage, intracerebral hemorrhage, epidural hemorrhage, cerebral infarction, transverse myelitis, spinal cord injury, peripheral nerve injury, multisystem atrophy and combinations thereof.

8. The method of claim 1, wherein the erythropoietin and/or stem cells are administered to the subject within a period of from one minute to 100 years after traumatic brain injury and/or neurological injury.

9. The method of claim 1, wherein the erythropoietin and stem cells are administered to the subject simultaneously.

10. The method of claim 1, wherein the erythropoietin and stem cells are not administered simultaneously.

11. A kit for treating a traumatic brain injury and/or a neurological injury

in a subject, comprising erythropoietin, stem cells and instructions for the use of the erythropoietin and stem cells for the treatment of the traumatic brain injury and/or the neurological injury.

12. The kit of claim 11, wherein the stem cells are selected from the group consisting of: umbilical cord blood cells, embryonic stem cells, bone marrow cells, mesenchymal stem cells, adipose-derived stem cells, endothelial stem cells, dental pulp stem cells, placenta stem cells, olfactory ensheathing cells, neural precursor cells and combinations thereof.

13. The kit of claim 12, wherein the stem cells are autologous.

14. The kit of claim 12, wherein the stem cells are allogeneic.

15. The kit of claim 11, wherein the erythropoietin is selected from the group consisting of: recombinant erythropoietin, synthetic erythropoietin, chemically modified erythropoietin, darbepoieten, glycosylated erythropoietin, EPO- alpha, EPO-beta and combinations thereof.

16. The kit of claim 11, wherein the subject has a condition selected from the group consisting of: quadriplegia due to traumatic subdural hematoma, pons hemorrhage, hypoxic brain injury, cerebral contusion, cerebral palsy, hemiplegia, subarachnoid hemorrhage, intracerebral hemorrhage, epidural hemorrhage, cerebral infarction, transverse myelitis, spinal cord injury, peripheral nerve injury, multisystem atrophy and combinations thereof.

17. The kit of claim 11, wherein the instructions indicate the erythropoietin and/or stem cells are to be administered to the subject within a period of from one minute to 100 years after traumatic brain injury and/or neurological injury.

18. The kit of claim 11, wherein the instructions indicate the erythropoietin and stem cells are to be administered to the subject simultaneously.