Molybdenum diselenide nanoparticles for neuroprotection

Abstract

The present disclosure provides compositions and methods for treating a subject afflicted with or at risk of developing a neurological condition, including a synucleinopathy such as Parkinson's disease, increasing mitochondrial function, reducing alpha-synuclein aggregation, and decreasing cellular oxidative stress. Aspects of the disclosure further relate to molybdenum diselenide nanomaterial compositions and combinatorial therapies for neuroprotection.

Description

TITLE OF THE INVENTIONMOLYBDENUM DISELENIDE NANOPARTICLES FOR NEUROPROTECTION CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority of U.S. Provisional Appl. Ser. No. 63 / 773,832, filed March 18, 2025, the entire disclosure of which is incorporated herein by reference.STATEMENT OF GOVERNMENT RIGHTS

[0002] This invention was made with government support under GM142869 awarded by the National Institute of Health. The government has certain rights in the invention.INCORPORATION OF SEQUENCE LISTING

[0003] A sequence listing containing the file named “TAMC092WO_ST26.xml” which is 37,862 bytes (measured in MS-Windows®) and created on March 11, 2026, and comprises 42 sequences, is incorporated herein by reference in its entirety.FIELD OF THE INVENTION

[0004] This present disclosure relates to the field of biotechnology and more specifically to methods and compositions for the treatment or prevention of neurological conditions associated with decreased mitochondrial function or increased cellular oxidative stress.BACKGROUND OF THE INVENTION

[0005] Neurological conditions are pathologies that affect central and peripheral nervous systems. There are over 600 known neurological diseases that can be broadly categorized into three subtypes: neurotraumatic, neurodegenerative, and neuropsychiatric. Neurological conditions exhibit excitotoxicity, oxidative stress, and neuroinflammation linked to irreversible changes in mitochondrial homeostasis. Consequently, mitochondria have long been direct targets for therapeutic treatments that aim to balance mitochondrial metabolism and mitophagy. maintain calcium ion concentration, and regulate mitochondrial dynamics. Mitochondrial dysfunction is an expected cause of etiology and progression in numeroushuman neurological pathologies, including stroke, Alzheimer's disease, and Parkinson’s disease. Therefore, neuroprotective treatments are an urgent and unmet need.

[0006] Synucleinopathies are a group of neurodegenerative conditions characterized by the abnormal accumulation of alpha-synuclein (a-syn) protein aggregates in neurons and glial cells. The most prevalent synucleinopathy is Parkinson's disease (PD), which is the fastest increasing neurological disorder, with an estimated 25.2 million diagnoses worldwide projected by 2050. Other synucleinopathies include dementia with Lewy bodies, multiple system atrophy, and pure autonomic failure. The onset and progression of PD is driven by the pathological aggregation of a-syn and propagation of cytotoxic aggregates from cell to cell. Upon endocytosis, a-syn aggregates damage cell mitochondria and the endoplasmic reticulum, triggering an unfolded protein response (UPR), increased reactive oxygen species (ROS), stress granule formation, reduced autophagy, suppressed exocytosis, and increased mitophagy, ultimately leading to cell death. Decades of research have investigated therapeutic approaches to treat PD, including inhibiting amyloid fibril formation, reducing amyloid burden in the brain using antibodies, and alleviating mitochondrial dysfunction using agents such as Coenzyme Q10 and idebenone. However, these strategies have not achieved expected neuroprotection, catalyzing the search for novel neuroprotective therapeutic approaches capable of decelerating the progression of PD.

[0007] Transition metal dichalcogenide nanoflowers (TMD NFs), including molybdenum disulfide (M0S2) and molybdenum diselenide (MoSe2), are novel nanomaterials that exhibit high surface-to-volume ratio via self-organization of individual sheets into higher coordinated structures. Although optical, electronic, and photocatalytic properties of TMD NFs are well-characterized, the biological activity of these nanostructures remains poorly understood. The present disclosure demonstrates that molybdenum disulfide (M0S2) and molybdenum diselenide (MoSe2) TMD NFs lower reactive oxygen species (ROS) levels, reduce mitochondrial impairment, and increase mitochondrial biogenesis in neurons and astrocytes. The neuroprotective effects of both TMD NFs result from upregulation of the PGC-la pathway, the biological system responsible for mitochondrial biogenesis. Furthermore, the present disclosure demonstrates that administration of TMD NFs to C. elegans extends the lifespan of the nematodes. These results indicate that TMD NFs can be used as novel neuroprotective therapeutic agents against acute and chronic neurological conditions associated with mitochondrial dysfunction. Furthermore, the present disclosure demonstrates that MoSe2 NFs rescue neurons and astrocytes from the cytotoxic effects of alpha-synucleinfibrils, suppress alpha-synuclein-induced unfolded protein response, upregulate autophagy and exocytosis of alpha-synuclein aggregates, reverse stress granule formation, and significantly reduce alpha-synuclein aggregate accumulation and extend lifespan in an in vivo Parkinson's disease model. Notably, MoSe? NFs exhibit superior neuroprotective efficacy compared to M0S2 NFs across all tested endpoints.

[0008] In yet another aspect, the present disclosure provides a method of reducing alpha-synuclein aggregation in a subject in need thereof, the method comprising administering an effective amount of a nanomaterial structure comprising a transition metal dichalcogenide to the subject. In one embodiment, the transition metal di chalcogenide is selected from the group consisting of molybdenum diselenide (MoSe2), molybdenum disulfide (MoS2), tungsten disulfide, tungsten diselenide, and combinations thereof. In another embodiment, the nanomaterial structure is defined as a nanoflower structure. In yet another embodiment, the nanomaterial structure comprises a high surface area-to-volume ratio. In still yet another embodiment, the subject is afflicted with or at risk of developing a synucleinopathy. In one embodiment, the synucleinopathy is selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, and pure autonomic failure.SUMMARY OF THE INVENTION

[0009] In one aspect, the present disclosure provides a method of treating a subject afflicted with or at risk of developing a neurological condition comprising administering an effective amount of a nanomaterial structure comprising molybdenum diselenide (MoSe2) to the subject.

[0010] In another aspect, the present disclosure provides a method of increasing longevity in a subject comprising administering an effective amount of a nanomaterial structure comprising molybdenum diselenide (MoSe2) to the subject. In one embodiment, the subject has at least about a 1%, about a 2%, about a 3%, about a 4%, about a 5%, about a 6%. about a 7%, about an 8%, about a 9%. about a 10%, about an 11%, about a 12%, about a 13%, about a 14%, about a 15%, or about a 20% increase in life expectancy compared to a control subject, including all ranges and values derivable therebetween.

[0011] In yet another aspect, the present disclosure provides a method of decreasing cellular oxidative stress in a subject in need thereof, the method comprising administering an effective amount of a nanomaterial structure comprising a selenium moiety active site to the subject, wherein said selenium moiety active site acts as an active site of reaction to reduce cellular oxidative stress. In one embodiment, the nanomaterial structure comprises a transition metalselected from the group consisting of: titanium, vanadium, zirconium, niobium, molybdenum, hafnium, tantalum, and tungsten. In another embodiment, the nanomaterial structure comprises molybdenum diselenide (MoSe2). In yet another embodiment, the nanomaterial structure comprises a plurality of selenium moiety active sites. The subject, in still yet another embodiment, is afflicted with or at risk of developing a disease or condition associated with increased cellular oxidative stress. The subject, in one embodiment, is afflicted with or at risk of developing a neurological condition.

[0012] In still yet another aspect, the present disclosure provides a pharmaceutical composition comprising a nanomaterial structure comprising a selenium moiety active site and a mitochondrial therapeutic agent or a neurotherapeutic agent. In one embodiment, the nanomaterial structure comprises a transition metal selected from the group consisting of: titanium, vanadium, zirconium, niobium, molybdenum, hafnium, tantalum, and tungsten. In another embodiment, the nanomaterial structure comprises molybdenum diselenide (MoSe2). The nanomaterial structure, in yet another embodiment, comprises a plurality of selenium moiety7active sites.

[0013] In one embodiment, the neurological condition may be a neurotraumatic condition, a neurodegenerative condition, or a neuropsychiatric condition. Non-limiting examples of neurotraumatic conditions include a brain injury, a traumatic brain injury, a stroke, a brain aneurysm, a brain hemorrhage, a brain tumor, a concussion, a skull fracture, a hematoma, a spinal trauma, a spinal cord injury, carbon monoxide poisoning, and encephalitis. A neurodegenerative condition, may include but is not limited to, Alzheimer’s disease, a memory7disorder, dementia, ataxia, Huntington’s disease, Parkinson’s disease, a motor neuron disease, amyotrophic lateral sclerosis, progressive bulbar palsy, primary lateral sclerosis, progressive muscular atrophy, spinal muscular atrophy, Kennedy’s disease, post-polio syndrome, multiple system atrophy, progressive supranuclear palsy, a tauopathy, corticobasal degeneration, frontotemporal dementia, globular glial tauopathy, Pick disease, argyrophilic grain disease, primary age-related tauopathy, a prion disease, Creutzfeldt-Jakob disease (CJD). variably protease-sensitive phonopathy (VPSPr), Gerstmann-Straussler-Scheinker disease (GSS), Kuru, and fatal insomnia. In certain embodiments, the neurodegenerative condition is a synucleinopathy. Non-limiting examples of synucleinopathies include Parkinson's disease, dementia with Lewy7bodies, multiple system atrophy, and pure autonomic failure. Nonlimiting examples of a neuropsychiatric condition include a seizure disorder, an attention deficit disorder, a cognitive deficit disorder, migraine headaches, a substance abuse disorder,an eating disorder, a depressive disorder, an anxiety disorder, schizophrenia, bipolar disorder, post-traumatic stress disorder, obsessive-compulsive disorder, panic disorder, insomnia, and epilepsy.

[0014] In another embodiment, the nanomaterial structure comprises a high surface area-to-volume ratio. The nanomaterial structure, in yet another embodiment, is defined as a nanoflower structure. In still yet another embodiment, the treating or administering increases a mitochondrial function. The mitochondrial function, in one embodiment, is selected from the group consisting of mitochondrial biogenesis, mitochondrial encoded gene expression, nuclear encoded mitochondrial gene expression, mitochondrial protein expression, mitochondrial respiratory capacity, and adenosine triphosphate production. In another embodiment, the treating or administering decreases accumulation of a reactive oxygen species. The reactive oxygen species, in yet another embodiment, is selected from the group consisting of hydrogen peroxide (H2O2), superoxide anions (O2 ), and hydroxyl radicals ('OH). In still yet another embodiment, the treating or administering increases neuron or astrocyte proliferation. The treating or administering, in one embodiment, increases neuron or astrocyte survival.

[0015] In certain embodiments, the treating or administering reduces alpha-synuclein-induced cytotoxicity in neurons, astrocytes, or both. In one embodiment, the treating or administering reverses alpha-synuclein-induced mitochondrial membrane potential depolarization. In another embodiment, the treating or administering increases mitochondrial biogenesis in neurons, astrocytes, or both. In certain embodiments, mitochondrial biogenesis may be measured by upregulation of succinate dehydrogenase subunit A (SDH-A), cytochrome c oxidase subunit I (COX-I), or both. In yet another embodiment, the treating or administering reduces alpha-synuclein-induced stress granule formation. In still yet another embodiment, the treating or administering downregulates at least one unfolded protein response (UPR) gene. Non-limiting examples of UPR genes that may be downregulated include protein kinase R-like endoplasmic reticulum kinase (PERK), activating transcription factor 6 (ATF6), C / EBP homologous protein (CHOP), and X-box binding protein 1 (XBP1). In one embodiment, the treating or administering upregulates autophagy of alpha-synuclein aggregates. The upregulation of autophagy' of alpha-synuclein aggregates, in some embodiments, may be measured by upregulation of microtubule-associated protein 1 light chain 3 beta (LC3B), sequestosome 1 (p62), or both. In another embodiment, the treating or administering upregulates exocytosis. In some embodiments, the upregulation of exocytosis may bemeasured by upregulation of at least one exosomal marker selected from the group consisting of cluster of differentiation 81 (CD81) and cluster of differentiation 63 (CD63). In yet another embodiment, the treating or administering downregulates mitophagy. In certain embodiments, downregulation of mitophagy may be measured by downregulation of PTEN-induced putative kinase 1 (PINK1), parkin (PRKN), or both. In still yet another embodiment, the treating or administering reduces accumulation of alpha-synuclein aggregates in vivo.

[0016] The nanomaterial structure, in another embodiment, further comprises a targeting molecule. In yet another embodiment, the targeting molecule is cell-specific or tissue-specific. The targeting molecule, in still yet another embodiment, is brain-specific, astrocyte-specific, or neuron-specific. In one embodiment, the nanomaterial structure further comprises a therapeutic agent or a detectable label. In another embodiment, the therapeutic agent is a mitochondrial therapeutic agent or a neurotherapeutic agent. The detectable label, in yet another embodiment, is a paramagnetic ion, a radioactive isotope, a fluorochrome, an NMR-detectable agent, or an X-ray imaging agent. Non-limiting examples of mitochondrial therapeutic agents include CoQlO (ubiquinone), idebenone, riboflavin, dichloroacetate, thiamine, creatine, lipoic acid, glutathione, N- acetylcysteine, cysteamine, EPI-743 (para-benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide. bezafibrate, resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule. A neurotherapeutic agent, may include but is not limited to, a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, clomethiazole, D-JNKI-1, dexanabinol. ebselen, edaravone, epigallocatechin gallate, huperzine A, N-(3-propylcarbamoyloxirane-2-carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742, tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

[0017] In one embodiment, administration may comprise injection, microneedle administration, inhalation, transnasal application, oral administration, buccal administration, vaginal administration, intraosseous administration, topical administration, transdermalapplication, or rectal administration. In another embodiment, the subject is an animal subject, a mammalian subj ect, a rodent subj ect. or a human subj ect. A method of the present disclosure, in yet another embodiment, may comprise administering a pharmaceutical composition comprising the effective amount of the nanomaterial structure to the subject. In still yet another embodiment, the pharmaceutical composition comprises the nanomaterial structure comprising molybdenum diselenide (MoSe2) and a mitochondrial therapeutic agent or a neurotherapeutic agent. The pharmaceutical composition, in one embodiment, comprises the nanomaterial structure comprising the selenium moiety active site and a mitochondrial therapeutic agent or a neurotherapeutic agent. In one embodiment, a method of the present disclosure may further comprise administering a second therapeutic agent to the subject. The second therapeutic agent, in another embodiment, is a mitochondrial therapeutic agent or a neurotherapeutic agent.BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0019] FIG. 1 shows SEM images of both M0S2 (Panels A-D) and MoSe2 (Panels E-H) nanoflowers (NFs). The magnification of each image increases from 3,000 X (Panel A) to 50.000 X (Panel D) for M0S2. and from 2,700 X (Panel E) to 20.000 X (Panel H) for MoSe2. Each NF type displays the characteristic ridged surface that lends to a very high surface area-to-volume ratio.BRIEF DESCRIPTION OF THE SEQUENCES

[0020] SEQ ID NO: 1 - A representative sense primer sequence for amplification of SIRT 3.

[0021] SEQ ID NO:2 - A representative anti-sense primer sequence for amplification of SIRT 3.

[0022] SEQ ID NO:3 - A representative sense primer sequence for amplification of ERRa.

[0023] SEQ ID NO:4 - A representative anti-sense primer sequence for amplification of ERRa.

[0024] SEQ ID NO:5 - A representative sense primer sequence for amplification of ERRft.

[0025] SEQ ID NO:6 - A representative anti-sense primer sequence for amplification of ERRJ3.

[0026] SEQ ID NO: 7 - A representative sense primer sequence for amplification o NRF2.

[0027] SEQ ID NO:8 - A representative anti-sense primer sequence for amplification <A'NRF2.

[0028] SEQ ID NO:9 - A representative sense primer sequence for amplification of PPARa.

[0029] SEQ ID NO: 10 - A representative anti-sense primer sequence for amplification of PPARa.

[0030] SEQ ID NO: 11 - A representative sense primer sequence for amplification of PPARy.

[0031] SEQ ID NO: 12 - A representative anti-sense primer sequence for amplification of PPARy.

[0032] SEQ ID NO: 13 - A representative sense primer sequence for amplification of PPARA

[0033] SEQ ID NO: 14 - A representative anti-sense primer sequence for amplification of PPARa

[0034] SEQ ID NO: 15 - A representative sense primer sequence for amplification of GCN5.

[0035] SEQ ID NO: 16 - A representative anti-sense primer sequence for amplification of GCN5.

[0036] SEQ ID NO: 17 - A representative sense primer sequence for amplification of SRC3.

[0037] SEQ ID NO: 18 - A representative anti-sense primer sequence for amplification of SRC3.

[0038] SEQ ID NO: 19 - A representative sense primer sequence for amplification of GAPDH.

[0039] SEQ ID NO:20 - A representative anti-sense primer sequence for amplification of GAPDH.

[0040] SEQ ID NO:21 - A representative sense primer sequence for amplification of ATF6.

[0041] SEQ ID NO:22 - A representative anti-sense primer sequence for amplification of ATF6.

[0042] SEQ ID NO:23 - A representative sense primer sequence for amplification of LC3B.

[0043] SEQ ID NO:24 - A representative anti-sense primer sequence for amplification of LC3B.

[0044] SEQ ID NO:25 - A representative sense primer sequence for amplification of P62.

[0045] SEQ ID NO:26 - A representative anti-sense primer sequence for amplification of P62.

[0046] SEQ ID NO:27 - A representative sense primer sequence for amplification of CD81.

[0047] SEQ ID NO:28 - A representative anti-sense primer sequence for amplification of CD81.

[0048] SEQ ID NO:29 - A representative sense primer sequence for amplification oiPERK.

[0049] SEQ ID NO:30 - A representative anti-sense primer sequence for amplification of PERK.

[0050] SEQ ID NO:31 - A representative sense primer sequence for amplification of XBPL

[0051] SEQ ID NO:32 - A representative anti-sense primer sequence for amplification of XBPE

[0052] SEQ ID NO:33 - A representative sense primer sequence for amplification of CD63.

[0053] SEQ ID NO:34 - A representative anti-sense primer sequence for amplification of CD63.

[0054] SEQ ID NO:35 - A representative sense primer sequence for amplification of CHOP.

[0055] SEQ ID NO:36 - A representative anti-sense primer sequence for amplification of CHOP.

[0056] SEQ ID NO:37 - A representative sense primer sequence for amplification of PINKP

[0057] SEQ ID NO:38 - A representative anti-sense primer sequence for amplification of PINKP

[0058] SEQ ID NO:39 - A representative sense primer sequence for amplification of PRKN.

[0059] SEQ ID NO:40 - A representative anti-sense primer sequence for amplification of PRKN.

[0060] SEQ ID NO:41 - A representative sense primer sequence for amplification of GAPDH.

[0061] SEQ ID NO:42 - A representative anti-sense primer sequence for amplification of GAPDH.DETAILED DESCRIPTION OF THE INVENTION

[0062] The present disclosure provides compositions and method for treating a subject afflicted with or at risk of developing a neurological condition, including a synucleinopathy such as Parkinson's disease, increasing mitochondrial function, reducing alpha-synuclein-induced cytotoxicity, reducing alpha-synuclein aggregation, and decreasing cellular oxidative stress. The present disclosure provides a significant advance in the art by providing high efficacy, mitochondria-targeted therapeutics demonstrating neuroprotective properties.

[0063] Neurological conditions are the second leading cause of death and are the leading cause of disability worldwide. Proper mitochondrial function is vital for cellular homeostasis. In a dysfunctional state, mitochondria cause deleterious effects on the cell, including oxidative stress, secondary excitotoxicity. and insufficient production of ATP, which in turn, can trigger apoptosis. The present disclosure provides a long unmet need by providing high efficacy, mitochondria-targeted therapeutics demonstrating neuroprotective properties.

[0064] To meet this need, two structurally different NFs were synthesized. Cell biogenesis assays demonstrate that MoSe? increases biogenesis in neurons more so than M0S2. A strong decrease in the magnitude of mitochondrial impairment and ROS levels was observed in all cells exposed to M0S2 and MoSe2 NFs. The increase in mitochondrial biogenesis and C. elegans lifespan demonstrate the TMD NFs' neuroprotective potential. The magnitude of the beneficial effects varies between NFs, and this observation further demonstrates that the neuroprotective effects of TMD NFs are determined by their structure. In some embodiments, neuroprotective properties may be obtained using any TMD NF oxide, including but not limited to, nanomaterial structures comprising M0S2. MoSe2, tungsten disulfide, tungsten diselenide, or combinations of any thereof.

[0065] The present disclosure further demonstrates that MoSe2 NFs rescue neurons and astrocytes from alpha-synuclein (a-syn)-induced cytotoxicity, including reversal of a-syn-induced mitochondrial membrane potential depolarization, stimulation of mitochondrial biogenesis, reduction of a-syn-induced reactive oxygen species, and reversal of a-syn-induced stress granule formation. Mechanistic investigation revealed that MoSe2 NFs downregulate the unfolded protein response (UPR) in the endoplasmic reticulum, upregulate autophagy and exocytosis of a-syn aggregates, and downregulate milophagy in neurons exposed to a-syn fibrils. In an in vivo Parkinson's disease C. elegans model, MoSe2 NFs significantly reduced a-syn aggregate accumulation and extended lifespan by up to 37.84%. These results demonstrate the therapeutic potential of MoSe2 NFs as neuroprotective agents capable of decelerating the progression of synucleinopathies, including Parkinson's disease. In some embodiments, rescue from alpha-synuclein (a-syn)-induced cytotoxicity may be obtained using any TMD NF oxide, including but not limited to, nanomaterial structures comprising M0S2, MoSe2, tungsten disulfide, tungsten diselenide, or combinations of any thereof.A. Nanomaterial Structures

[0066] The present disclosure provides nanomaterial structures comprising a selenium moiety or a sulfur moiety active site. In certain embodiments of the present disclosure, a nanomaterial structure of the present disclosure may be referred to as a nanoflower. As used herein the term “nanoflowef ’ refers to a nanomaterial structure with a high surface area-to-volume ratio. In certain embodiments, a nanoflower may resemble a flower, or in some cases a tree, when viewed using electron microscopy. In some embodiments, the surface area-to-volume ratio for a nanoflower of the present disclosure may be about 5 to about 1000.

[0067] In some embodiments of the present disclosure a nanomaterial structure, as described herein, may comprise a transition metal and a chalcogen. In certain embodiments, the transition metal may be a transition metal dichalcogenide. Non-limiting example of transition metals that may be included in a nanomaterial structure of the present disclosure include titanium, vanadium, zirconium, niobium, molybdenum, hafnium, tantalum, and tungsten. Nonlimiting examples of chalcogens that may be used according to the present disclosure include sulfur, selenium, and tellurium. In one embodiment, the nanomaterial of the present disclosure may include a selenium, a sulfur, or a tellurium moiety active site. In another embodiment, a nanomaterial of the present disclosure may comprise M0S2, MoSe2, tungsten disulfide, tungsten diselenide, or a combination of any thereof.

[0068] In particular embodiments, a nanomaterial structure of the present disclosure is about 5 nm to about 1500 nm, about 100 nm to about 1500 nm, about 200 nm to about 1500 nm, about 300 nm to about 1500 nm, about 400 nm to about 1500 nm, about 500 nm to about 1500 nm, about 600 nm to about 1500 nm, about 700 nm to about 1500 nm, about 100 nm to about 1200 nm, about 200 nm to about 1200 nm, about 300 nm to about 1200 nm about 400 nm to about 1200 nm, about 500 nm to about 1200 nm, about 600 nm to about 1200 nm, about 700 nm to about 1200 nm, about 800 nm to about 1200 nm, about 900 nm to about 1100 nm, about 100 nm to about 1000 nm, about 200 nm to about 900 nm, about 200 nm to about 800 nm about 200 nm to about 700 nm. or about 300 nm to about 600 nm in length, width, depth, or diameter, including all ranges and values derivable therebetween.

[0069] The nanomaterial structures of the present disclosure may be produced using any method known in the art. In one embodiment, the nanomaterial structures of the present disclosure may be produced using a hydrothermal synthesis reaction.B. Nanomaterial Compositions and Methods

[0070] In certain aspects, the present disclosure provides pharmaceutical and therapeutic compositions comprising the nanomaterial structures of the present disclosure. In some embodiments, the nanomaterial structures of the present disclosure may be combined with a pharmaceutically acceptable carrier. As used herein, a ‘‘pharmaceutically acceptable carrier,” “pharmaceutically acceptable adjuvant,” or “adjuvant” refers to reagents, cells, compounds, materials, compositions, and / or dosage forms that are not only compatible with the nanomaterial structures or other agents to be administered therapeutically, but also are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other complication commensurate with a reasonable benefit / risk ratio. Also included may be an agent that modifies the effect of other agents and is useful in preparing a therapeutic compound or composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable. Such an agent may be added to a therapeutic composition or pharmaceutical composition to modify for example the cellular target, cellular localization, or cellular uptake of a nanomaterial structure as described herein. Such an agent may include any excipient, diluent, carrier, or adjuvant that is acceptable for pharmaceutical use. Such an agent may be non-naturally occurring, or may be naturally occurring, but not naturally found in combination with other agents in the therapeutic or pharmaceutical composition.

[0071] As used herein, a “therapeutic compound” or “therapeutic composition” refers to a composition comprising a nanomaterial structure of the present disclosure. In some embodiments, a therapeutic composition has the activity of increasing mitochondrial function in a subject or decreasing cellular oxidative stress in a subject as described herein. In one embodiment, the composition is capable of increasing mitochondrial biogenesis, mitochondrial encoded gene expression, nuclear encoded mitochondrial gene expression, mitochondrial protein expression, mitochondrial respiratory capacity, or adenosine triphosphate production. In another embodiment, the therapeutic composition has the activity of decreasing ROS in a subject. Non-limiting examples of ROS that may be reduced by a therapeutic composition of the present disclosure include hydrogen peroxide (H2O2), superoxide anions (O2 ), and hydroxyl radicals ('OH). Such a compound or composition is meant to encompass a composition suitable for administration to a subject, such as a mammal, particularly a human subj ect. In yet another embodiment, the therapeutic composition has the activity of reducing alpha-synuclein-induced cytotoxicity' in neurons, astrocytes, or both. Instill yet another embodiment, the therapeutic composition has the activity of reducing accumulation of alpha-synuclein aggregates in a subject. In one embodiment, the therapeutic composition has the activity of downregulating the unfolded protein response, upregulating autophagy, upregulating exocytosis, or downregulating mitophagy in cells exposed to alpha-synuclein aggregates.

[0072] In general, a therapeutic composition is sterile, and preferably free of contaminants that are capable of eliciting an undesirable response within the subject (e.g., the compound(s) in the composition are pharmaceutical grade). Therapeutic compositions may be designed for administration to subjects in need thereof via a number of different routes of administration including oral, intravenous, intraarticular, intraarterial, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, subcutaneous, inhalation, vaginal, intraosseous, transnasal, injection, microneedle, topical, and transdermal. The appropriate dosage of a composition, as described herein, may be determined based on the type of disease to be treated, the severity and course of the disease, the clinical condition of the individual, clinical history', response to the treatment, and the discretion of the attending physician. In some embodiments, therapeutic compositions provided by the present disclosure may include various "‘unit doses.” A unit dose is defined as containing a predetermined quantity of the therapeutic composition. The quantity to be administered, and the particular route and formulation, is w ithin the skill of determination of those in the clinical arts. A unit dose need not be administered as a single injection but may comprise continuous infusion over a set period of time. In some aspects, a unit dose comprises a single administrable dose.

[0073] Precise amounts of the therapeutic composition also depend on the judgment of the practitioner and are peculiar to each individual. Factors affecting dose include physical and clinical state of the patient, the route of administration, the intended goal of treatment (alleviation of symptoms versus cure) and the potency, stability7and toxicity' of the particular therapeutic substance or other therapies a subject may be undergoing.

[0074] As used herein, “subject” or “patient” refers to animals, including humans, who are treated with the therapeutic compounds or compositions or in accordance with the methods described herein. For diagnostic or research applications, a wide variety of animals may be suitable subjects, including nematodes, rodents (e g., mice, rats, hamsters), rabbits, primates, and swine, such as inbred pigs and the like. In particular embodiments, a subject in need of therapy may be any subject who comprises a cell that exhibits decreased mitochondrial function or increased cellular oxidative stress as described herein. In another embodiment,the subject may be afflicted with or at risk of developing a disease or condition associated with decreased mitochondrial function or increased cellular oxidative stress. Non-limiting examples of such diseases or conditions include a neurological condition, a neurotraumatic condition, a neurodegenerative condition, a neuropsychiatric condition, a brain injury, a traumatic brain injury, a stroke, a brain aneurysm, a brain hemorrhage, a brain tumor, a concussion, a skull fracture, a hematoma, a spinal trauma, a spinal cord injury’, carbon monoxide poisoning, encephalitis, Alzheimer’s disease, a memory disorder, dementia, ataxia. Huntington’s disease, Parkinson’s disease, a synucleinopathy, dementia with Lewy bodies, pure autonomic failure, a motor neuron disease, amyotrophic lateral sclerosis, progressive bulbar palsy, primary' lateral sclerosis, progressive muscular atrophy, spinal muscular atrophy, Kennedy's disease, post-polio syndrome, multiple system atrophy, progressive supranuclear palsy, a tauopathy, corticobasal degeneration, frontotemporal dementia, globular glial tauopathy, Pick disease, argyrophilic grain disease, primary' age-related tauopathy, a prion disease, Creutzfeldt- Jakob disease (CJD), variably protease-sensitive prionopathy (VPSPr), Gerstmann-Straussler-Scheinker disease (GSS), Kuru, fatal insomnia, a seizure disorder, an attention deficit disorder, a cognitive deficit disorder, migraine headaches, a substance abuse disorder, an eating disorder, a depressive disorder, an anxiety' disorder, schizophrenia, bipolar disorder, post-traumatic stress disorder, obsessive-compulsive disorder, panic disorder, insomnia, and epilepsy.

[0075] As used herein, the term ‘‘synucleinopathy” refers to a group of neurodegenerative diseases characterized by the abnormal accumulation and aggregation of alpha-synuclein (a-syn) protein in neurons, nerve fibers, or glial cells. Non-limiting examples of synucleinopathies include Parkinson's disease, dementia with Lewy' bodies, multiple system atrophy, and pure autonomic failure. Alpha-synuclein is a 140-amino acid protein encoded by the SNCA gene that facilitates trafficking of presynaptic vesicles releasing neurotransmitters into the synaptic cleft. Under pathological conditions, a-syn misfolds and aggregates into beta-sheet-rich oligomers and fibrils that are cytotoxic to neurons and astrocytes.

[0076] As used herein, ‘‘al pha-synucl ein-induced cytotoxicity ” refers to the deleterious cellular effects caused by the uptake and accumulation of a-syn aggregates. Non-limiting examples of such deleterious cellular effects include, but are not limited to, mitochondrial membrane potential depolarization, increased reactive oxygen species, suppression of mitochondrial biogenesis, activation of the unfolded protein response (UPR) in theendoplasmic reticulum, formation of stress granules, suppression of autophagy, reduction of exocytosis. and increased mitophagy.

[0077] In certain embodiments of the present disclosure, the unfolded protein response (UPR) refers to a cellular stress response activated in the endoplasmic reticulum upon accumulation of misfolded proteins. Non-limiting examples of UPR genes that may be modulated according to the methods of the present disclosure include protein kinase R-like endoplasmic reticulum kinase (PERK), activating transcription factor 6 (ATF6), C / EBP homologous protein (CHOP), and X-box binding protein 1 (XBP 1 ).

[0078] In certain embodiments, autophagy of alpha-synuclein aggregates may be measured by changes in the expression of microtubule-associated protein 1 light chain 3 beta (LC3B) and sequestosome 1 (p62). In other embodiments, exocytosis may be measured by changes in the expression of exosomal markers, including but not limited to cluster of differentiation 81 (CD81) and cluster of differentiation 63 (CD63). Mitophagy , in certain embodiments, may be measured by changes in the expression of PTEN-induced putative kinase 1 (PINK1) and parkin (PRKN).

[0079] In certain embodiments, stress granules are dynamic ribonucleoprotein condensates that form in the cytosol upon cellular stress. In one embodiment, alpha-synuclein-induced stress granule formation may be detected by tracking the subcellular localization of T-cell intracellular antigen 1 -related (TIA-R) protein, which translocates from the nucleus to the cytosol upon activation by cellular stress. In another embodiment, the administering or treating reverses the translocation of TIA-R from the cytosol back to the nucleus, indicating reduction of cellular stress.

[0080] In certain aspects, the present disclosure provides a method of reducing alpha-synuclein aggregation in a subject in need thereof, the method comprising administering an effective amount of a nanomaterial structure comprising a transition metal dichalcogenide to the subject. As used herein, "reducing alpha-synuclein aggregation7’ refers to a measurable decrease in the quantity or accumulation rate of alpha-synuclein aggregates in cells, tissues, or a subject in vivo. In certain embodiments, the method of reducing alpha-synuclein aggregation may be practiced in a subject afflicted with or at risk of developing a synucleinopathy. including but not limited to Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, and pure autonomic failure.

[0081] A composition, as described herein, may include, in particular embodiments, a combination of therapeutic agents. In some embodiments, a composition as described here may be administered as a single composition or as more than one composition. Different compositions as provided herein, in certain embodiments, may be administered by the same route of administration or by different routes of administration.

[0082] A pharmaceutical composition of the present disclosure may comprise, in certain embodiments, a nanomaterial structure comprising a selenium moiety active site and a mitochondrial therapeutic agent or a neurotherapeutic agent. The nanomaterial structure, in particular embodiments, may comprise a plurality of selenium moiety active sites. In one embodiment, the nanomaterial structure comprises a transition metal selected from the group consisting of: titanium, vanadium, zirconium, niobium, moly bdenum, hafnium, tantalum, and tungsten. In one embodiment, the nanomaterial structure comprises molybdenum diselenide (MoSe2). Any mitochondrial therapeutic agent known in the art may be used according to the composition and methods of the present disclosure. Non-limiting examples of such mitochondrial therapeutic agents include CoQlO (ubiquinone), idebenone, riboflavin, dichloroacetate, thiamine, creatine, lipoic acid, glutathione, A-acetylcysteine, cysteamine, EPI-743 ( / wra-benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide, bezafibrate, resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule. A therapeutic peptide or nucleic acid used according to the present disclosure may be any therapeutic peptide or nucleic acid known in the art to increase mitochondrial function, decrease cellular oxidative stress, improve neurological function, slow the decline of neurological function, alleviate neurological symptoms, slow the progression of a neurological condition, improve cognition, improve or preserve memory function, or delay the onset of neurological condition or neurological decline. In particular embodiments, the therapeutic peptide or therapeutic nucleic acid may function in or encode a protein associated with cellular metabolism and respiration. Any neurotherapeutic agent known in the art may be used according to the compositions and methods of the present disclosure. Non-limiting examples of such neurotherapeutic agents include a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutj ric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, clomethiazole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A. N-(3-propylcarbamoyloxirane-2-carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742, tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

[0083] A pharmaceutical composition of the present disclosure may comprise, in some embodiments, a targeting molecule. In one embodiment, the targeting molecule may be cellspecific or tissue-specific. In another embodiment, the targeting molecule may be brainspecific, neuron-specific, or astrocyte-specific. Numerous such targeting molecules are known in the art and any such targeting molecule may be used according to the present disclosure. In certain embodiments, a nanomaterial structure of the present disclosure may be modified with or conjugated to a peptide, a protein, a colloidal molecule, or a polymer to facilitate delivery or adsorption. The pharmaceutical composition of the present disclosure, in some embodiments, may be serum-free, endotoxin-free, or sterile.

[0084] A peptide or polynucleotide molecule for use according to the compositions of the present disclosure may, in some embodiments, be a recombinant peptide or nucleic acid. As used herein, the term “recombinant"’ refers to a polynucleotide molecule, protein, or cell that is not naturally present, or is not naturally present in the same form or structure and was created by human intervention. In one embodiment, a recombinant polynucleotide may be a DNA molecule or may be an RNA molecule. A recombinant polynucleotide molecule or a recombinant polypeptide molecule or protein may comprise, in certain embodiments, a combination of two or more polynucleotide or polypeptide sequences that do not naturally occur together in the same manner, such as a polynucleotide molecule or protein that comprises at least two polynucleotide or protein sequences that are operably linked but heterologous with respect to each other. As used herein the term “heterologous” refers to a polynucleotide molecule or protein that is not naturally present or is not naturally present in the same form or structure and was created by human intervention. For example, a heterologous polynucleotide molecule or protein may not naturally occur in the cell being transformed or may be expressed in a manner or genomic context that differs from the natural expression pattern or genomic context found in the cell being transformed. The heterologous polynucleotide molecule or protein, in some embodiments, may be overexpressed in the cell being transformed. In certain embodiments, a recombinant polynucleotide molecule, protein, construct, or vector maycomprise any combination of two or more polynucleotide or protein sequences in the same molecule which are heterologous to one another, such that the combination is man-made and not normally found in nature. As used herein, the phrase “not normally found in nature’’ means not found in nature without human intervention. A recombinant polynucleotide or protein molecule, may comprise, for example, polynucleotide or protein sequences that are separated from other polynucleotide or protein sequences that exist in proximity to each other in nature. A recombinant polynucleotide or protein molecule may also comprise, for example, polynucleotide or protein sequences that are adjacent to or contiguous with other polynucleotide or protein sequences that are not naturally in proximity with each other. Such a recombinant polynucleotide molecule, protein, or expression construct may also refer to a polynucleotide or protein molecule or sequence that has been genetically engineered or constructed outside of a cell. For example, a recombinant polynucleotide molecule may comprise any engineered or man-made plasmid, vector, or expression construct, and may include a linear or circular DNA molecule. Such plasmids, vectors, and expression constructs may comprise, for example, various maintenance elements including, but not limited to, a heterologous promoter sequence, a prokaryotic origin of replication, or a selectable marker.

[0085] In certain aspects, a therapeutic composition of the present disclosure may comprise a nanomaterial structure of the present disclosure and a detectable label. Non-limiting examples of detectable labels that may used according to embodiments of the present disclosure include a paramagnetic ion, a radioactive isotope, a fluorochrome, an NMR-detectable agent, and an X-ray imaging agent.

[0086] In certain embodiments, the compositions and methods for treating an individual described herein may be combined with any other composition or method of treatment known in the art. The compositions and methods may be administered in any suitable manner known in the art. For example, a first and a second mitochondrial or neurotherapeutic treatment may be administered sequentially (at different times) or concurrently (at the same time). In some aspects, a first and a second mitochondrial or neurotherapeutic treatment may be administered in separate compositions. In certain embodiments, a first and a second treatment may be administered in the same composition.

[0087] Non-limiting examples of additional treatment modalities that may be included in combination with the compositions and methods provided herein include a therapeutic agent or surgery. In specific embodiments, the methods and compositions of the present disclosure may be combined with other therapies directed towards increasing mitochondrial function,decreasing cellular oxidative stress, improving neurological function, slowing the decline of neurological function, alleviating neurological symptoms, slowing the progression of a neurological condition, improving cognition, improving or preserving memory function, or delaying the onset of neurological condition or neurological decline as described herein. C. Detection and Therapeutic Agents

[0088] In some aspects, the present disclosure provided methods and compositions for detection and therapeutic labeling of the nanomaterial structures of the present disclosure. Methods for labeling and detection of such structures are well-known in the art, and any such method known in the art may be used to label or detect the nanomaterial structures described herein. As anon-limiting example, nanomaterial structures may be labeled with a detectable moiety, such as a radioactive atom, a chromophore, a fluorophore, or the like, and then detected using methods known in the art. Such labeled nanomaterial structures may be used, in some embodiments, for in vivo or in vitro diagnostic techniques.

[0089] As used herein, the term “label” refers to a directly or indirectly detectable compound or composition that is conjugated directly or indirectly to the composition to be detected. In certain embodiments, a nanomaterial structure, a polynucleotide molecule, protein, or cell may be labeled to generate a labeled composition. In particular embodiments, labeled compositions also include sequences which are conjugated a polynucleotide molecule that will provide a signal upon expression of the inserted sequences, such as green fluorescent protein (GFP) and the like. The label may be detectable by itself (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition that is detectable. Labels may be suitable for small scale detection or for high-throughput screening. As such, suitable labels include, but are not limited to radioisotopes, fluorochromes, chemiluminescent compounds, dyes, and proteins, including enzymes. Labels may be simply detected or may be quantified. In certain embodiments, labels that may be quantified provide numerically reportable value. In luminescence or fluorescence assays, the detectable response may be generated directly using a luminophore or fluorophore associated with an assay component involved in binding, or indirectly using a luminophore or fluorophore associated with another (e.g., reporter or indicator) component.

[0090] The term “about” is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and / or” unless explicitly indicated to refer to alternativesonly or the alternatives are mutually exclusive. When used in conjunction with the word “comprising’" or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have,” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes,” and “including,” are also open-ended. For example, any method that “comprises,” “has.” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any system or method that “comprises,” “has,” or “includes” one or more components is not limited to possessing only those components and covers other unlisted components.

[0091] Other objects, features, and advantages of the present disclosure are apparent from detailed description provided herein. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. Any embodiment of the present disclosure may be used in combination with any other embodiment described herein.

[0092] All references herein are incorporated herein by reference in their entirety.EXAMPLES

[0093] The following examples are included to illustrate embodiments of the present disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventor to function well in the practice of the invention. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.Example 1: Synthesis and Characterization of M0S2 and MoSei Nanoflowers

[0094] M0S2 and MoSe2 transition metal dichalcogenide nanoflowers (TMD NFs) were synthesized using hydrothermal synthesis in Teflon-lined autoclaves. A scanning electron microscope (SEM) revealed that both TMD NFs were assembled into dense nanoflower-like structures with characteristically high surface area-to-volume ratio, with M0S2 measured between 300-600 nm and MoSe2 measured at less than ~1 pm (FIG. 1. Panels A-H).

[0095] Cellular uptake of NFs was confirmed by both Raman spectroscopy and fluorescent imaging. Cells were treated with 10% solutions of respective concentrations of M0S2 and MoSe2 and allowed to incubate for 24 h. Raman spectra were acquired with a 532 nm laser from individual cells exposed to NFs and the dry NFs material. Raman spectra of dry M0S2 exhibited two peaks centered at 385 and 405 cm'1that correspond to E2gand Aigvibrations of M0S2. The same vibrational bands were observed in Raman spectra acquired from cells exposed to M0S2. These results indicate that the analyzed cells possessed M0S2 NFs. Fluorescent imaging confirmed the presence of NFs in N27 neurons, DI astrocytes, and CTX astrocytes.Example 2: M0S2 and MoSe2 Nanoflowers Increase Neuron and Astrocyte Proliferation

[0096] Cellular growth curve assays determined the extent to which TMD NFs altered cell proliferation. The number of N27, DI, and CTX cells was determined at 24 h, 48 h, and 72 h post-treatment with TMDs. N27, DI, and CTX cells treated with M0S2 experienced a significant boost in cell count within the first 24 h. Specifically, rat N27 neurons exhibited a 93.2% increase in cell number, while both the rat DI TNC 1 and CTX TNA2 astrocytes showed a smaller increase in the rate of cell proliferation, 29.2 % and 27.4 %, respectively. Although all cell types initially underwent a significant increase in the cell count within the first 24 h, no significant changes in the rate of cell proliferation were observed following 48 h and 72 h. These results indicate that M0S2 NFs strongly enhanced the cell proliferation only within the first 24h after introduction to N27, DI, and CTX cells.

[0097] Similarly, N27, DI, and CTX cells treated with MoSe2 experienced a significant initial increase in cell count. N27 neurons exhibited a 34.7% increase, while DI TNC1 astrocytes demonstrated a 14.2% increase in total cell number within the first 24 h. The cellular grow th curve assay revealed the highest boost (36.9% increase) in the proliferation of CTX TNA2 astrocytes after the first 24 h of exposure. Similarly to M0S2, none of the tested cell types experienced a significant difference in total cell number after 48 h of exposure to MoSe2TMDs. After 72 h, a significant increase in total number of cells was observed only for CTX astrocytes, whereas no significant changes in the cell counts were evident for N27 and DI cells. These results indicate that both neurons and astrocytes exhibited similar increases in cell proliferation within the first 24 h after addition of M0S2 and MoSe2 NFs to the cell media. However, observed differences in the cell proliferation of CTX astrocytes exposed to M0S2 andMoSe2 at 24 h and 72 h demonstrates that the magnitude of cell proliferation was linked to the chemical nature of TMDs.

[0098] An alamarBlue™ assay was used to assess changes in the proliferation of N27, DI, and CTX cells after 6 h of after administration of both M0S2 and MoSe2. The assay revealed no significant changes in the reducing potential of N27, DI, or CTX cells exposed to M0S2 compared to control cells. This result indicates that M0S2 NFs did not exhibit any detrimental effects on the cells. On average, N27, DI, and CTX cells duplicate every 12 hours. Consequently, the results also indicate that M0S2NFS enhance viability and, consequently, the potential to replicate between 6 h and 12 h after cell exposure to TMDs.

[0099] Similarly to M0S2, neuronal cells exposed to low concentrations of MoSe2 showed no changes in the reducing potential within the first 6 h. However, at a 1.0 mg / rnL concentration of MoSe2, a decrease of 11.2% of the cell reducing potential was observed in N27 cells. These results indicate that high concentration of MoSe2 can cause slight detrimental effects on cell proliferation potential. These effects were not observed for DI astrocytes. At the same time, CTX astrocytes positively responded on MoSe2 treatment exhibiting an increase in the reducing potential at the 0.1 mg / ml and 1.0 mg / ml concentrations. These results demonstrate that different cell types exhibit slightly different responses on treatment with TMDs.Example 3: M0S2 and oSei Nanoflowers Decrease ROS and Improve Mitochondrial Function in Neurons and Astrocytes

[0100] Cells that received M0S2 treatments exhibited substantially lower ROS levels compared to the control. Specifically. N27 neurons exposed to 0.1 mg / mL of M0S2 demonstrated a 25.8% decrease in ROS levels. As the concentration of M0S2 in the cell media increased to 0.5 and 1.0 mg / mL, there was a reciprocal decrease of ROS in N27 neurons by 43.8% and 74.2%, respectively. In DI astrocytes, a 9.7% decrease in ROS was observed for the lowest concentration of M0S2. The cells treated with 0.5 and 1.0 mg / mL of M0S2 exhibited a greater magnitude of decrease in the ROS levels (35.5% and 38.8%, respectively). CTX astrocytes treated with 0.1 mg / mL M0S2 experienced a 11.4% decrease in the ROS,whereas the cells treated with 0.5 mg / mL and 1.0 mg / mL of M0S2 had 36.2% and 51.3% lower ROS levels compared to the control.

[0101] Cells treated with MoSe2 also experienced suppressed production of ROS. Compared to other cell types, N27 cells experienced the strongest suppression of ROS levels. When treated with 0.1 mg / mL of MoSe2, N27 neurons exhibited 37.5% decrease in ROS compared to the control. Furthermore, 70.3% and 80.4% suppression in ROS levels was observed in neurons treated with 0.5 and 1.0 mg / mL MoSe2, respectively. In DI astrocytes, a trend of decreasing ROS was observed. When treated with 0.1 mg / mL MoSe2, there was a 12.9% decrease in measured ROS. As the concentration of administered NFs increased to 0.5 and 1.0 mg / mL, the cells demonstrated 43.8% and 59.8% decreases in ROS levels, respectively. After receiving the lowest dose of MoSe2, the CTX cells experienced a reported 32.6% decrease in overall ROS. As the dosage concentration increased to 0.5 and 1.0 mg / mL MoSe2, the cells showed significant 46.6% and 67.9% decreases in ROS, respectively.

[0102] The extent of mitochondrial impairment in neurons and astrocytes was measured using JC-1. Administration of M0S2 treatment lowered mitochondrial impairment in N27 neurons. Cells exposed to 0.1 mg / mL of M0S2 exhibited a 21.4% decrease in damage of cell mitochondria compared to the control. JC-1 assays revealed an 83.7% and 98.9% suppression of mitochondrial impairment in N27 neurons exposed to 0.5 mg / mL and 1.0 mg / mL of M0S2, respectively. Thus, M0S2 reduced the progression of naturally occurring mitochondrial damage. In DI astrocytes, the low est concentration (0.1 mg / mL) resulted in a 11.7% decrease in mitochondrial impairment. DI astrocytes treated with 0.5 mg / mL of M0S2, experienced a 49.8% decrease in the mitochondrial damage, whereas a 90.8% decrease in the organelle impairment was observed for DI astrocytes treated with 1.0 mg / mL of M0S2. CTX astrocytes treated with 0.1 and 0.5 mg / mL exhibited much lower magnitude of mitochondrial survival equal to 4.1% and 10.9%, respectively. At the highest concentration of M0S2 (1.0 mg / mL), however, there was a significant 50.7% decrease in mitochondrial impairment in CTX astrocytes.

[0103] Similarly, N27 neurons that received MoSe2 treatments exhibited 20.6% increase in mitochondrial survival compared to the control, whereas cells exposed to 0.5 and 1.0 mg / mL of MoSe2 demonstrated 87.3% and 99.1% suppression of mitochondrial damage. DI astrocytes did not exhibit a significant change in the mitochondrial impairment when treated with the lowest concentration of MoSe2, but a significant decrease in the mitochondrial damage (72.8% and 98.5%, respectively) was observed in DI astrocytes treated with 0.5 and1.0 mg / mL of MoSez. CTX astrocytes did not exhibit any significant changes when treated with 0.1 and 0.5 mg / mL of MoSe2. However, an 85.2% increase in the suppression of mitochondrial impairment was observed in CTX astrocytes treated with 1.0 mg / mL of MoSe2.

[0104] The experiments described above demonstrate the phenotypic effects that TMD NF treatments induce. To better elucidate the mechanistic effects resulting from TMD NF exposure, enzy me-linked immunosorbent assays (ELIS As) and quantitative polymerase chain reactions (qPCR) were performed.

[0105] The utilized in-cell ELISA detects treatment-induced effects on mitochondrial biogenesis via measurement of two mitochondrial proteins: nuclear encoded succinate dehydrogenase subunit A (SDH- A) and mitochondrial encoded cytochrome c oxidase subunit 1 (COX-I). N27 neurons experienced an increase in both SDH-A and COX-I levels for all tested concentrations of M0S2 NFs. Treatments with 0.1 mg / mL M0S2 yielded the smallest increase in both SDH-A and COX-I, with only a 2.9% and 9.8% increase, respectively. The higher concentration treatments induced much higher increases, with the 0.5 mg / mL resulting in 13.4% and 37.0% increases in SDH-A and COX-I, respectively, and the 1.0 mg / mL treatment resulting in 7.2% and 38.2% increases, respectively. The DI astrocytes that received M0S2 treatments did not exhibit any significant changes in SDH-A or COX-I expression levels. The CTX astrocytes that received M0S2 experienced increases in both SDH-A and COX-I levels at every test concentration, similarly to the N27 neurons. At the 0.1 mg / mL concentration, there were minor increases of 0.5% and 6.6% in SDH-A and COX-I, respectively. As the treatment concentrations increased, there were significant increases of 4.3% and 12.4%, respectively, for the 0.5 mg / mL group and 7.2% and 19.1%, respectively, for the 1.0 mg / mL group.

[0106] When treated with MoSez NFs, N27 experienced a dosage dependent increase in both SDH-A and COX-I levels. At the lowest concentration of 0.1 mg / mL, there was a measured increase of 1.9% and 1.4% in SDH-A and COX-I, respectively. As the concentration increased to 0.5 mg / mL, there were significant increases of 6.5% and 10.1%, respectively. At the highest concentration of 1.0 mg / mL, the highest significant increases of 18.2% and 35.1%, respectively, were observed. The DI astrocytes treated with MoSe2 also experienced a dosage dependent increase of SDH-A and COX-I levels. At the lowest concentration of 0.1 mg / mL, there was only slight 0.3% and 1.5% respective increase in SDH-A and COX-I. When treated with the 0.5 mg / mL concentration, there was a 5.8% and 9.1% increase, respectively. The only significant increases were measured when treated with 1.0 mg / mL MoSe2. with a positivechange of 14.0% and 22.6%, respectively. The measured effects of MoSez were greater in CTX astrocytes compared to the M0S2 treatments. At the 0.1 mg / mL concentration, there was a slight increase of 2.5% and 7.7% in SDH-A and COX-I, respectively. At the 0.5 mg / mL concentration, there were significant increases of 10.5% and 18.2%, respectively. At the highest concentration of 1.0 mg / mL, the significant increases in protein levels were measured at 28.5% and 44.6%. respectively.Example 4: M0S2 and MoSei Nanoflowers Regulate the Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1 Alpha (PGC-la) Pathway

[0107] The PGC-la pathway is directly responsible for mitochondrial biogenesis in the cell and thus, plays key roles in other processes such as ROS regulation, glucose utilization, fatty acid oxidation, and antioxidant detoxification. Therefore, quantitative polymerase chain reactions (qPCR) were utilized to quantify changes in the expression of proteins in this pathway when exposed to both M0S2 and MoSe2 NFs.

[0108] The central component of the pathway. PGC-la. is regulated by the sirtuin 1 (SIRT1) protein. SIRT1 and PGC-la expression is upregulated in response to M0S2 administration. In another upstream arm of the pathway, steroid receptor coactivator-3 (SRC-3) monitors energy excess and acts as a coactivator of nuclear receptors and transcription factors. SRC3 is positive feedback regulator of non-depressible 5 (GCN5) that inhibits PGC-la and downregulates its expression through acetylation. Both SRC -3 and GCN5 were downregulated when administered M0S2 and MoSe2 NFs, with M0S2 resulting in a greater downregulation in both genes. SRC-3 only had 36.0% and 81.3% expression after exposure to M0S2 and MoSe2. GCN5 expression was reduced to 76.5% with M0S2 and did not change in response to MoSe2.

[0109] PGC-la suppresses ROS activity by activation of sirtuin 3 (SIRT3) and estrogen related receptor alpha (ERRa). MoSe2 suppressed ROS more efficiently than M0S2 NFs in neurons. Indeed, qPCR revealed that SIRT3 was 5% and 31% upregulated in cells exposed to M0S2 and MoSe2 NFs, respectively. Consequently, ERRa experienced an 89% and 7.8-fold upregulation when administered M0S2 and MoSe2 NFs.

[0110] PGC-la regulates multiple cellular processes, such as uncoupling proteins, fatty acid oxidation, glucose utilization, antioxidants detoxification, and mitochondrial biogenesis via peroxisome proliferator-activated receptor alpha, gamma, and delta (PPAR a / y / 8), estrogen-related receptors alpha, beta, and gamma (ERR a / p / y), and nuclear respiratory factor 1 and 2 (NRF 1 / 2). qPCR revealed that in neurons exposed to TMD NFs, the expression of these protein groups was strongly upregulated to varying magnitudes. Analysis of qPCR for PPARa and PPAR8 showed that MoSe2 triggered a greater upregulation of these genes versus M0S2. Specifically, PPARa experienced a 3.9-fold and 3.4-fold increase, respectively, while PPAR8 experienced 2.2-fold and 27% increase, respectively. PPARy had the inverse effect, with M0S2 causing a higher upregulation than MoSe2. Analysis revealed 6.2-fold and 3.9-fold increases in expression, respectively. As previously stated, ERRa experienced a higher upregulation in response to MoSe2 over M0S2. Contrarily, ERR experienced a higher upregulation when administered M0S2 compared to MoSe2, with 74.6% and 40.1% measured increases. Lastly, investigation into changes in the expression of NRF2 was conducted. When administered M0S2, NRF2 had a modest upregulation of only 7.5%, but had a steeper upregulation of 51.9% when treated with MoSe2.Example 5: MoSe2 Nanoflowers Increase Survival Expectancy in Caenorhabditis elegans

[0111] Caenorhabditis elegans (C. elegans) are a well-established, multicellular model organism broadly utilized to investigate neurological effects and toxicity of molecular analytes, to track developmental stages of neuronal networks in embryos, and to investigate molecular mechanisms of neurodegenerative diseases. The usage of C. elegans nematodes as a simplistic in vivo model has been well established. The relatively quick lifespan allows for robust analysis of the effects of any macronutrient or therapeutic. A wild-type N2 C. elegans strain was used to investigate the extent to which TMD NFs on could alter the lifespan of the worms.

[0112] Administration of both M0S2 and MoSe2 NFs to the model organism was accomplished via supplemented diets. The wild-type N2 C. elegans were housed on nematode growth media (NGM) agar plates that contained UV-killed OP50 E. coli, the main food source of nematodes. As the C. elegans consumed the plated E. coli, the inoculated NFs were also consumed. The C. elegans are also allowed to navigate through the food source, allowing for uptake through the hypodermis. The hypodermis of C. elegans is translationally similar to human skin and is a metabolically active organ that allows for adsorption of various materials.

[0113] The C. elegans that received M0S2 supplementation did not experience a great shift in survival expectancy p=50%. The M0S2 supplemented worms had a calculated survival expectancy of 18 ± 2 days. The 0.1 mg / mL M0S2 group had a slightly lower p=50% at 16days. As the concentration of M0S2 increased, the 0.5 mg / mL and 1.0 mg / mL groups experienced higher p=50% values of 19 days and 20 days, respectively. These two groups exceeded the survival expectancy of the healthy control worms (18 days).

[0114] The C. elegans that received MoSe2 supplementation experienced a much greater shift in survival expectancy. The MoSe2 supplemented worms had a calculated survival expectancy of 22 ± 1.7 days. All concentrations of MoSe2 supplementation resulted in an increase in survival expectancy over the healthy control group. Both the 0.1 mg / mL and 0.5 mg / mL had a calculated p=50% value of 21 days, 3 days higher than the control. The group that experienced the greatest change in p=50% was the 1.0 mg / mL MoSe2 worms, with a survival expectancy of 24 days, a significant 33% increase over the control p=50% value of 18 days. This represents an increase of 6 days over the healthy wild-ty pe C. elegans. The p=50% value represents the time at which fifty percent of the worms are expected to still be alive. Therefore, MoSe2 treatment extended the expected benchmark by one-third. The above reported life-expectancy experiments were conducted in healthy, wild-type N2 C. elegans. This is important to note because MoSe2 had the innate ability to significantly increase life expectancy in perfectly healthy nematodes. Furthermore, a very low mortality was observed during the first 15 days for C. elegans exposed to 0.5 and 1.0 mg / mL MoSe2. These results demonstrate that MoSe2 drastically extends the lifespan of C. elegans and strongly reduces nematode death during the first half of their life.Example 6: Materials and Methods

[0115] Materials: All chemicals received were utilized without further purification. MilliQ water was utilized in the synthesis of nanomaterials. Ammonium molybdate tetrahydrate ((NH4)6MO?O24) 4H2O), thiourea 99% (NH2CSNH2), selenium powder (Se), and sodium borohydride (NaBH4) were purchased from Sigma-Aldrich (SigmaAldrich. St. Louis, MO).

[0116] Synthesis of Nanoflowers: Molybdenum disulfide (M0S2) nanoflowers (NFs) were synthesized using a hydrothermal synthesis reaction. For this, 50 mL solution of 0.01 mol (NH4)eMo7O24 and 0.06 mol NH2CSNH2 in MilliQ water was kept for 30 min at room temperature prior to its transfer into 50-mL Teflon lined autoclave reactors. The autoclaves were exposed to 200 °C for 24 h. After autoclave reactors were allowed to cool to room temperature overnight, the resulting black precipitate was collected and centrifuged at 7,000 rpm for 5 mins. The samples were washed with MilliQ water and centrifuged twice at 7,000 rpm for 5 mins. Next, samples were washed with ethanol; placed on a glass petri dish andkept overnight in a dry hot air oven at 60 °C. The desiccated nanoflowers were collected from the petri dish and stored. Molybdenum diselenide (MoSe2) NFs were synthesized using a hydrothermal synthesis reaction. (NH- SMOTCLJ (0.003 mol) and NaBFL (0.1g) were dissolved in 50 mL solution of MilliQ water and pure ethanol at a 1 : 1 ratio. Separately, 0.006 mol of Se was dissolved in 10 mL of NaBFL; added to the solution of (NH- gMoyC r and stirred for 30 minutes at room temperature. The obtained solution was transferred into two 50-mL Teflon lined autoclave reactors and placed at 200 °C for 48 h. After the heating was turned off, the reactor was allowed to cool to room temperature. The resulting black precipitate was collected and centrifuged at 7, 000 rpm for 5 min, washed with MilliQ water and centrifuged three times. The washed nanoparticles were placed on a glass petri dish and allowed to dry in a hot air oven at 60 °C overnight. The desiccated nanoflowers were collected and stored.

[0117] Scanning Electron Microscopy: M0S2 and MoSe2 nanoflowers were imaged with a scanning electron microscope (SEM) at the Material Characterization Facility (MCF) at Texas A&M University.

[0118] Cell Culture: Dopaminergic midbrain N27 rat neurons were purchased from ATCC and grown in RPMI 1640 Medium (Thermo Fisher Scientific, Waltham, Massachusetts) supplemented with 10% heat inactivated fetal bovine serum (FBS) (Invitrogen, Waltham, Massachusetts). CTX TNA2 and DI TNC1 rat astrocyte cells were purchased from ATCC and grown in DMEM supplemented with 10% heat inactivated fetal bovine serum (FBS) (Invitrogen, Waltham, Massachusetts). All cells were cultured in T-75 flasks purchased from (Thermo Fisher Scientific, Waltham. Massachusetts) and incubated at 37 °C in 5 % CO2 environment until utilized in experiments. Cells were passaged at approximately 90 % confluency. All cell types were used prior to the tenth passage to ensure reliable results.

[0119] Nanoflower Treatments’. Cells were seeded based on the quantity necessary for the assay being performed (10,000-100,000 cells per well). The cells were allowed to fully adhere overnight. After adhering, ten percent of the media was removed. A two milligram per milliliter (2 mg / mL) stock of the nanoflower was made in phosphate buffer solution (PBS). The 2 mg / mL stock was probe sonicated for 30 s to suspend the nanoflowers completely. The 2 mg / mL stock was then serially diluted into the three tested concentrations: 1.0 mg / mL, 0.5 mg / mL, and 0.1 mg / mL. The same volume of removed media was then replaced by an equivalent volume of the tested concentration of nanoflowers. The control cells that did not receive any nanoflowers got an equivalent volume of PBS. The final volume of nanoflowersfor each sample is 10 % solution consistent across all cell types and prepared concentrations of nanoflowers.

[0120] Raman Spectroscopy: Confirmation of cellular uptake of nanoflowers was conducted using Raman spectroscopy. The Raman spectra were collected using a TE-2000U Nikon inverted confocal microscope equipped with a solid-state laser generated continuous wavelength (CW) 532 nm light. Electromagnetic radiation was directed towards the sample directed using a 50 / 50 beam splitter and focused by 20X Nikon objective. Scattered light was collected using the same objective and directed into an IsoPlane-320 spectrometer (Princeton Instruments) equipped with a 1200 groove / nm grating. Prior to entering the spectrograph, elastically scattered photons were removed using a long-pass filter (Semrock). The inelastically scattered photons were collected using PIX-400B CCD (Princeton Instruments). For the measurements. 10.000 cells were seeded in glass-bottom 96-well plate and allowed to adhere overnight. Nanoflower treatments were administered as previously described, and cells were incubated for 24 h. After incubation, the 96-well plate was mounted on the confocal Raman spectroscope. Individual cells were selected and placed in the focal plane of the laser. The Raman spectra were collected using 10 s acquisition time and 6.85 mW of light. In parallel, reference Raman spectra were collected from dry nanoflowers.

[0121] Proliferation Assays: Growth assays for each cell type with each NF were conducted. Cells were placed in a 24 well plate and allowed to adhere for 24 h. Cells then either received a 10 % concentration of PBS as a negative control, or a 10% concentration treatment of 0.5 mg / mL of M0S2 or MoSe2. Cells were counted after 24, 48, and 72 h with an Invitrogen Countess 3 Automated Cell Counter (Thermo Fisher Scientific, Waltham, Massachusetts) after NF treatment and compared. A proliferation assay was also conducted using alamarBlue™ HS Cell Viability Reagent (Thermo Fisher Scientific, Waltham, Massachusetts). After allowing 20,000 cells to adhere to the plate overnight, each well was treated with either PBS or the specified NF at each concentration. After two hours, the alamarBlue reagent was added to the wells. The color change was observed, and the fluorescence was measured after 6 h.

[0122] Mitochondrial Health Assays: Cells were plated in 96 well plates and treated with NFs according to parameters previously detailed. To assess the levels of reactive oxygen species present in the cells, the wells were treated with CellROX™ Deep Red Reagent (Thermo Fisher Scientific, Waltham, Massachusetts). The treated cells were scanned and measured using an LSRII BD flow cytometer (BD, San Jose, California). The data wasanalyzed, and cell toxicity was calculated using LSRII software. To assess the health of mitochondria in the NF treated cells, the wells were treated with MitoProbe™ JC-1 (Thermo Fisher Scientific, Waltham, Massachusetts). The treated cells were scanned and measured using an LSRII BD flow cytometer (BD, San Jose, California). The data was analyzed, and cell toxicity was calculated using LSRII software.

[0123] Enzyme-linked Immunoassay (ELISA): Mitochondrial biogenesis was quantified using the MitoBiogenesis™ In-Cell ELISA Colorimetric Kit (Abeam, Cambridge, United Kingdom). Cells were treated according to the NF parameters previously stated. After 24 h of treatment, the cells were fixed to the plate using BD Cytofix™ fixation buffer (BD Biosciences, Franklin Lakes, New Jersey). The cells were then blocked, permeabilized, and treated with primary and secondary antibodies. The cells were then treated with AP development solution and absorbance was measured using a plate reader to quantify expression of SDH-A. The media was removed, and the cells were treated with HRP development solution and absorbance was measured using a plate reader to quantify expression of COX-1. Data was saved, collected, and analyzed for reporting.

[0124] Polymerization Chain Reaction (PCR): PCR for genes with roles in the mitochondrial biogenesis pathway was conducted, as well as mitochondria copy number, using GAPDH as a housekeeping gene. All primers were designed using the known rat gene sequences in the NCBI database and created using the IDT custom oligonucleotide PCR primer generation software. All primers were ordered and received from IDT (Integrated DNA Technologies, Morrisville, North Carolina). Cells were treated according to treatments described above. The cells were harvested and centrifuged to form a pellet. The pellet was resuspended in TRIzol Reagent (Thermo Fisher Scientific, Waltham, Massachusetts) and chloroform (Avantor, Radnor, Pennsylvania). The solution was centrifuged, and RNA was extracted from the aqueous phase. The RNA was then converted to complementary DNA (cDNA) to be used in the PCR reaction. The utilized primers are listed in Table 1.Table 1: qPCR Primers for Genes with Roles in the Mitochondrial Biogenesis Pathway

[0125] C. elegans in vivo modeling: Wild type N2 C. elegans strain was acquired as a kind gift from Dr. Michael Polymenis, Texas A&M University. The C. elegans were raised on NGM plates seeded with OP50 E. coll and maintained at a constant 20°C until reaching an egg-producing age. Age synchronization was conducted by collecting all worms and eggs and bleaching the solution to remove all adult worms, according to Sutphin & Kaeberl ein’s protocol. Synchronized worms were allowed to reach "‘Day 1 Adult” age before being transferred onto experimental plates. Ten individual worms were moved onto each conditioned agar plate. The conditioned plates were produced by plating E. coli supplemented with TMD NFs. NF supplementation was performed by mixing concentrated 2% stocks with 1 Ox concentrated OP50 E. coli at a 1 : 1 ratio before plating, quickly drying, and UV irradiating. Survival of the C. elegans was determined by counting the number of alive and dead organisms on each plate until no surviving worms were counted, then calculated using the Kaplan-Meier equation.

[0126] Fluorescent Imaging: Fluorescent imaging was conducted in an EVOS M5000 microscope (Thermo Fisher Scientific, Waltham, Massachusetts). To image the cellular skeleton, CellMask™ Green Actin Tracking Stain and CellMask™ Deep Red Actin Trackingstains were utilized. To locate and image mitochondria, MitoTracker™ Orange CM-H2TMRos were used. The nuclei were stained using NucBlue™ Live Cell Stain ReadyProbe™ reagent. To enable visualization of ROS levels and the extent of mitochondrial impairment, CellROX™ Deep Red reagent and MitoProbe™ JC-1 reagent were used, respectively. All fluorescent stains and reagents were acquired from Thermo Fisher Scientific, Waltham. Massachusetts. To image NFs in the cells, N27 neurons, DI astrocytes, and CTX astrocytes were captured when incubated without NFs, as well as with 0.5 mg / mL M0S2, and with 0.5 mg / mL MoSe? using green channel.Example 7: TMD NFs Rescue Neurons and Astrocytes from u-Synuclein-Induced Mitochondrial Impairment and Boost Mitochondrial Biogenesis

[0127] To investigate the neuroprotective properties of TMD NFs in a Parkinson's disease (PD) model, the cytotoxic effects of alpha-synuclein (a-syn) fibrils were modeled in neurons and astrocytes. a-Syn aggregates were added to N27 neurons and DI TNC1 and CTX TNA2 astrocytes that had been cultured to approximately 90% confluency. Fibrils were sonicated for 10 min prior to administration and added to the cell media at a final concentration of 40 pM. Cells were allowed to incubate for 12 h with the aggregates before the addition of TMD NFs. After 12 h induction with a-syn, a 2 mg / mL stock of M0S2 or MoSe2 NFs was resuspended in PBS, probe sonicated for 30 s and serially diluted into three tested concentrations: 1.0 mg / mL, 0.5 mg / mL, and 0.1 mg / mL. Equivalent volumes of each tested concentration of NFs were administered to the cells, while control cells received equivalent volumes of PBS.

[0128] A JC-1 assay was used to determine the extent to which TMD NFs could reverse a-syn-induced mitochondrial membrane potential depolarization. An uptake of a-syn fibrils induced significant mitochondrial damage in both neurons and astrocytes. Compared to the control, N27 neurons experienced a 257% increase, while DI astrocytes experienced a 327% increase in mitochondrial damage upon exposure to a-syn aggregates. Similarly, a-syn fibrils triggered a 251% increase in mitochondrial damage in CTX astrocytes. The subsequent (12 h) treatment of N27 neurons with different concentrations of M0S2 NFs resulted in 15.8% to 55.4% decreases in the magnitude of a-syn-induced mitochondrial damage. DI and CTX astrocytes treated with M0S2 NFs also experienced strong dose-dependent therapeutic relief, which resulted in 31.5% to 62.7% (DI) and 31.9% to 49.7% (CTX) decrease in mitochondrial impairment. Similar effects were observed for MoSe2 NFs in both neurons and astrocytes. N27 neurons exposed to a-syn fibrils and subsequently treated with MoSe2 NFs exhibited33.5% to 64.0% reduction in mitochondrial impairment. In DI and CTX astrocytes exposed to a-syn fibrils, MoSe2 NFs caused a strong concentration-dependent reduction in mitochondrial damage ranging from 24.9% to 63.7%. These results indicate that both MoSe2 and M0S2 NFs efficiently rescue cell mitochondria from cytotoxic effects caused by a-syn aggregates.

[0129] To determine whether TMD NFs also facilitate the synthesis of novel mitochondria in the a-syn-exposed cells, changes in the expression of two mitochondrial proteins, succinate dehydrogenase subunit A (SDH-A) and cytochrome c oxidase subunit I (COX-I), were analyzed by ELISA. Exposure of neurons and astrocytes to a-syn fibrils caused a strong downregulation in the expression of both SDH-A and COX-I. N27 neurons experienced only a small, 1.34% and 1.39% decrease in the expression of SDH-A and COX-I protein levels, respectively. However, a substantially higher magnitude of downregulation of mitochondrial biogenesis was observed in DI astrocytes (4.32% for SDH-A and 5.60% for COX-I) as well as in CTX astrocytes (7.48% for SDH-A and 7.82% for COX-I). These results indicate that endocytosis of a-syn aggregates strongly suppresses mitochondrial biogenesis in both neurons and astrocytes.

[0130] A dose-dependent upregulation in mitochondrial biogenesis was observed in N27 neurons after administration of M0S2 and MoSe2 NFs. As the concentration of M0S2 NFs increased from 0.1 to 1.0 mg / mL, SDH-A was upregulated from 0.51% to 5.49%, and COX-I was upregulated from 1.57% to 12.35%. When treated with MoSe2 NFs, the expression of SDH-A was upregulated from 7.63% to 13.11%, while COX-I expression was upregulated from 5.38% to 8.39%. Similarly to neurons. DI astrocytes underwent a dose-dependent upregulation of mitochondrial biogenesis upon receiving TMD NFs subsequently to a-syn aggregates. As the concentration of M0S2 NFs increased from 0.1 to 1.0 mg / mL, an upregulation of SDH-A from 1.40% to 3.74% was observed, and COX-I upregulation was found to increase from 2.76% to 8.60%. When treated with MoSe2 NFs, SDH-A experienced an upregulation from 5.68% to 12.43%, while COX-I expression was alleviated from 4.16% to 7.22%. CTX astrocytes treated with M0S2 NFs had little to no change in the levels of SDH-A expression, with only a 1.37% increase observed at the highest concentration of M0S2NFS. At the same time, a significant upregulation (1.81% to 7.43%) of COX-I expression was observed in CTX astrocytes exposed to M0S2 NFs. CTX astrocytes treated with MoSe2 NFs showed a more prominent dose-dependent response for both mitochondrial proteins. The expression of SDH-A was elevated by 6.44%, 10.80%, and 13.02%, and the COX-I proteinexperienced an increase in expression from 5.09% to 8.03% as the concentration of MoSez NFs increased. These results indicate that M0S2 and MoSe2 NFs not only protect mitochondria from a-syn aggregates but also boost mitochondrial biogenesis in both neurons and astrocytes.Example 8: TMD NFs Reduce a-Synuclein-Induced ROS and Reverse Stress Granule Formation in Neurons and Astrocytes

[0131] A flow cytometry-based ROS assay was used to determine the extent to which TMD NFs altered ROS levels in neurons and astrocytes exposed to a-syn aggregates. An exposition of N27 neurons, DI, and CTX astrocytes to a-syn aggregates caused a significant increase in the ROS levels. Specifically, N27 neurons experienced a 213% increase, while DI and CTX astrocytes had 132% and 59.3% increase in ROS, respectively.

[0132] A subsequent (12 h) treatment of neurons and astrocytes exposed to a-syn aggregates with M0S2 NFs resulted in a dose-dependent decrease in the cellular ROS levels. N27 neurons exposed to the lowest concentration of M0S2 NFs (0.1 mg / mL) experienced a moderate (13.4%) ROS reduction. Higher concentrations of M0S2 NFs (0.5 mg / mL and 1.0 mg / mL) caused a substantially larger (24.8% and 43.8%, respectively) decrease in the ROS levels. Similar results were observed for DI astrocytes, where the lowest concentration of M0S2 NFs resulted in 7.6% reduction, while administration of 0.5 mg / mL and 1.0 mg / mL M0S2 NFs led to significantly larger decrease (31.5% and 41.8%, respectively) in ROS levels. Similar magnitudes of M0S2 NFs-triggered decrease in ROS levels in CTX astrocytes were also observed, with a decrease to 4.9%, 23.6%, and 36.1% in the cells exposed to 0.1, 0.5, and 1.0 mg / mL M0S2 NFs, respectively.

[0133] A much larger magnitude in the suppression of ROS levels was observed in neurons and astrocytes exposed to MoSe2 NFs. Specifically, N27 neurons exposed to 0.1 mg / mL MoSe2 NFs experienced a 22.5% reduction in ROS levels, while at 0.5 and 1.0 mg / mL, reductions of 63.0% and 71.6% were recorded. A similar magnitude in the reduction of ROS was observed in DI astrocytes exposed to MoSe2 NFs, where the lowest concentration of NFs resulted in 8.9% reduction of ROS, and medium and high (0.5 and 1.0 mg / mL) concentrations of MoSe2 NFs caused a suppression of ROS levels of 57.6% and 77.8%, respectively. A gradual decrease in ROS levels (11.1%. 60.3%, and 74.3%) in CTX astrocytes was also observed with a gradual increase in the concentration of MoSe2 NFs (0.1, 0.5 and 1.0 mg / mL). It should be noted that for all cell types, the highest concentration of NFs reduced ROS levelsto the baseline observed in the cells that experienced no a-syn exposure. These results show that both MoSe2 and M0S2 NFs drastically reduce a-syn-induced ROS levels in neurons and astrocytes.

[0134] In order to probe the effects TMD NF therapeutics have on cellular stress mechanisms, immunofluorescent tracking of T-cell intracellular antigen 1 -related (TIA-R) protein was conducted. TIA-R is a nucleic acid binding protein localized inside the nucleus under normal functioning conditions but vacates to the cytosol upon activation via cellular stress in pathological environments. To ensure that M0S2 and MoSe2 NFs caused no cellular stress, neurons and astrocytes were treated with TMD NFs alone. Immunofluorescent staining of these cells revealed confined localization of TIA-R in the nuclei, indicating that TMD NFs alone did not induce any stress in neurons and astrocytes.

[0135] Immunofluorescent staining revealed the appearance of stress granules in the cytosol of neurons and astrocytes exposed to a-syn aggregates. From all analyzed cells, neurons had the most drastic response to a-syn aggregates with nearly complete evacuation of TIA-R from the nuclei. CTX astrocytes were also observed to form high levels of stress granules in the cytosol, while DI astrocytes did not respond with the same degree of magnitude, forming fewer stress granules in response to a-syn aggregates. All cell types treated with M0S2 and MoSe2 NFs experienced a significant reduction in stress granule formation. It was found that TMD NFs administration resulted in the re-localization of TIA-R back to the nuclei, with very few fluorescent signals still present in the cytosol. Cells receiving MoSe2 NFs treatments experienced even greater stress relief compared to their M0S2 NFs counterparts. Thus, TIA-R imaging confirms that TMD NFs provide neuroprotection via regulating the stress response in neurons and astrocytes exposed to a-syn aggregates.Example 9: TMD NFs Modulate Unfolded Protein Response, Autophagy, Exocytosis, and Mitophagy Pathways in Neurons Exposed to u-Synuclein Aggregates

[0136] Using quantitative polymerase chain reaction (qPCR), changes in the expression of genes involved in the unfolded protein response (UPR), autophagy, exocytosis, and mitophagy were determined in N27 neurons exposed to a-syn fibrils followed by TMD NFs treatment, as well as a-syn fibrils alone (control). Accumulation of aggregated a-syn in neuronal cells triggers UPR in the endoplasmic reticulum (ER), reduction of exocytosis, and increased mitophagy.

[0137] Administration of M0S2 NFs downregulated all four UPR genes (PERK, ATF6, CHOP, and XBP1) after 18 hours of TMD NFs administration. ATF6 experienced a 62% reduction, while XBP1 underwent a 95% downregulation in the cells exposed to a-syn fibrils followed by M0S2 NFs treatment. PERK and CHOP each experienced significant downregulation, with 90% and 78% reductions, respectively. Administration of MoSe2 NFs similarly resulted in downregulation of all four chosen UPR genes at the 18 h timepoint. ATF6 experienced a 25% reduction, while the other UPR genes exhibited very strong downregulations with 94%, 94%, and 80% reductions for XBP1, PERK, and CHOP, respectively.

[0138] In neurons, accumulation of aggregated proteins also first triggers downregulation of autophagy genes, microtubule-associated protein 1 light chain 3 beta (LC3B) and sequestosome 1 (p62). This initial suppression is followed by upregulation of these genes as neurons degrade and autophagy amyloid aggregates. M0S2 NFs upregulated both LC3B and p62 after 3 h and 6 h of treatment, indicating their innate ability to advance cell autophagy and quickly mitigate amyloid-induced stress. LC3B w as found to be subsequently downregulated at 18 h, while p62 maintained upregulated. MoSe2 NFs triggered upregulation of p62 2.22-fold at the 3 h timepoint and then tapered off for the remaining timepoints as the presence of aggregates decreased.

[0139] Cluster of differentiation 81 and 63 (CD81 and CD63) are exosomal markers that can be used to quantify changes in cell exocytosis activity. Both CD81 and CD63 were strongly downregulated in the cells exposed to a-syn fibrils alone. In the neurons exposed to a-syn fibrils followed by M0S2 NFs treatment, CD81 was unchanged at the two early time points and then experienced a downregulation at the 9 h timepoint. From that point forward, there was a great upregulation, indicating an increase in the cell's ability to exocytose. CD63 was initially upregulated at the 3 h timepoint and remained upregulated. In the cells exposed to MoSe2 NFs, CD81 was downregulated at the early timepoints, then experienced an upregulation up to 2.57-fold at 18 h. CD63 was upregulated much faster, with upregulation occurring at the 9 h, 12 h, and 18 h timepoints, with a maximum upregulation of 1.74-fold at the 18 h timepoint.

[0140] Two major players involved in mitophagy are the PTEN-induced putative kinase 1 (PINK1) and parkin (PRKN) proteins. qPCR revealed that neurons exposed to a-syn fibrils alone experienced strong upregulation of both PINK1 and PRKN. M0S2 NFs strongly affected the regulatory' elements of the mitophagy' pathway. At the 9 h, 12 h, and 18 h timepoints, both PINK1 and PRKN were significantly downregulated, indicating decreased need for mitophagydue to the presence of healthier mitochondria. Administration of MoSe2 NFs also led to downregulation of mitophagy. At the 18 h timepoint, PINK1 experienced a 74% reduction. The PRKN gene experienced a reduction at the 6 h, 9 h, 12 h, and 18 h timepoints, culminating at an 89% reduction for the 18 h timepoint. Both M0S2 and MoSe2 NFs were thus able to down regulate the UPR and mitophagy responses, while upregulating autophagy and exocytosis pathways in a neuroprotective manner.Example 10: TMD NFs Reduce a-Synuclein Aggregation and Extend Lifespan in a Parkinson's Disease C. elegans Model

[0141] To probe the neuroprotective effects of TMD NFs in vivo, transgenic C. elegans that overexpress the a-syn protein (strain NL5901, genotype pkls2386 [unc-54p::alphasynuclein::YFP + unc-119(+)]) were utilized. The overexpressed a-syn was tagged with green fluorescent protein allowing amyloid aggregates to form puncta and be tracked using fluorescent microscopy. M0S2 or MoSe2 NFs were supplemented in the nematodes' diet. The quantification of fluorescent puncta was used to track the rate of a-syn accumulation in nematodes, and C. elegans lifespan was analyzed for further determination of TMD NF neuroprotective properties.

[0142] C. elegans receiving M0S2 NFs in their diet exhibited a slight increase in life expectancy compared to the control group. The control nematodes had a calculated survival expectancy (p=50%) of 18.5 days. C. elegans supplied with 0.1 mg / mL MoS2 NFs in their diet did not experience a shift in the p=50% (18.5 days). While nematodes receiving 0.5 mg / mL supplementation of the same nanostructures had an increase in p=50% of one day (19.5 days), the nematodes receiving the highest concentration of MoS2 NFs (1.0 mg / mL) demonstrated an extension of p=50% of two days (20.5 days).

[0143] Upon investigation of the quantity of puncta present in each group of worms on Day 5, Day 10, and Day 15, a clear dose-dependent reduction in the accumulation rate of a-syn was observed. On Day 5, control nematodes had an average of 20.8 counted puncta (20.8 ± 6.34). There was a reduction of 9.62%, 17.31%, and 36.54% when M0S2 was administered at 0.1 mg / mL (18.8 ± 3.96), 0.5 mg / mL (17.2 ± 4.21), and 1.0 mg / mL (13.2 ± 2.95), respectively. On Day 10, the control group had an average of 22.8 puncta present (22.8 ± 4.21), and a dosedependent reduction of 21.05%, 28.95%, and 31.58% was observed for the 0.1 mg / mL (18.0 ± 2.74), 0.5 mg / mL (16.2 ± 2.28), and 1.0 mg / mL (15.6 ± 3.21) groups, respectively. The greatest reductions in puncta quantity were observed on Day 15. The control group had 22.4± 4.16 puncta count. When administered 0.1 mg / mL M0S2, there was a 43.75% reduction in the puncta count (12.6 ± 3.85). Nematodes supplemented with 0.5 mg / mL had a reduction of 52.68% (10.6 ± 2.41) in puncta present. The greatest reduction was observed in nematodes receiving the highest concentration (1.0 mg / mL) of M0S2 NFs, with puncta count reduced by 58.93% (9.2 ± 3.11).

[0144] MoSe2 NFs exhibited a significant increase in life expectancy when administered to the PD C. elegans. Nematodes administered 0.1 mg / mL and 0.5 mg / mL both experienced an increase in p=50% of three days (21.5 days), representing a 16.22% increase in life expectancy. The nematodes receiving the highest dosage of MoSe2 (1.0 mg / mL) experienced the largest extension in life expectancy, demonstrating a seven-day extension with a p=50% value of 25.5 days.

[0145] Analysis of fluorescent puncta in the nematodes receiving MoSe2 NF treatments revealed a significant reduction in amyloid burden. On Day 5, a clear dose-dependent reduction in puncta was observed, with reductions of 21.15%, 38.46%, and 46.15% as the MoSe2 NF concentrations increased from 0.1 to 1.0 mg / mL (16.4 ± 2.07, 12.8 ± 3.56, 11.2 ± 2.77), respectively. At the next timepoint, Day 10, C. elegans that received MoSe2 NFs continued to exhibit a drastic decrease in the number of puncta, with decreases of 32.46%, 49.12%, and 45.61% as the concentration of NFs increased from 0.1 mg / mL (15.4 ± 4.34) to 0.5 mg / mL (11.6 ± 2.07) and 1.0 mg / mL (12.4 ± 5.03). Day 15 revealed the most drastic reduction of puncta in nematodes administered MoSe2 NF treatment. Upon receiving 0.1 mg / mL MoSe2 NFs, there was a 50.0% reduction in the counted puncta (11.2 ± 3.11). The nematodes treated with 0.5 mg / mL MoSe2 experienced a 50.89% reduction in aggregates (11.0 ± 2.45). The highest magnitude of reduction was found in C. elegans exposed to 1.0 mg / mL MoSe2 NFs, with a 67.86% decrease in the puncta present at Day 15 (7.2± 1.92). These results demonstrate that MoSe2 and M0S2 NFs exert similar therapeutic effects; however, MoSe2 exhibits increased neuroprotective properties in the PD C. elegans model.Example 11 : Additional Materials and Methods for a-Synuelein Studies

[0146] Expression and Purification of a-Synuclein. To express a-syn, the pET21a-a-synuclein plasmid was transformed into BL21 Escherichia coli (E. coll) strain Rosetta. The transformed bacteria were incubated at 37 °C in Luria broth (LB) media until reaching OD600 of 0.8-1.2. Next, cells were exposed to 1 mM of IPTG to induce expression of a-syn. Induced bacteria were incubated for additional 4-5 h under the same experimental conditions. Bacteriawere collected by centrifugation at 8,000 rpm for 10 min. Bacterial pellets were resuspended in Tris lysis buffer (10 mM EDTA, 150 mM NaCl, 50 mM Tris HC1, pH 7.4) and placed in a water bath pre-heated to 78 °C for 30 min. Next, bacterial suspensions were centrifuged at 16,000 rpm for 40 min to pellet cellular debris that were discarded. Supernatant was collected and treated with streptomycin sulfate (10% solution, 136 pL / mL) in glacial acetic acid (228 pL / mL) followed by centrifugation at 16,000 rpm for 20 min to precipitate bacterial proteins, lipids, and nucleic acids. a-Syn present in the supernatant was precipitated by saturated ammonium sulfate ((NH4)2SO4). The pellet was then resuspended and washed with ammonium acetate (100 mM NH4(CH3COO)) and absolute ethanol. Harvested a-syn was resuspended in phosphate buffer solution (PBS). Protein samples were concentrated using centrifugation in Amicon Ultra 10 kDa protein filters (Merck Millipore). Prior to size exclusion chromatography (SEC), protein samples were centrifuged at 14,000 g for 30 min to remove aggregates and impurities. For each SEC cycle, 500 pL of concentrated protein was syringe injected into an AKTA pure FPLC system (GE Healthcare) equipped with a gelfiltration column (Superdex 200 10 / 300) that was maintained at 4 °C. Proteins were eluted isocratically using PBS, pH 7.4 at a flow rate of 0.5 mL / min. Collected a-syn was reconcentrated in Amicon Ultra 10 kDa protein filters (Merck Millipore) with a final concentration of 400 pM.

[0147] a-Synuclein Aggregation. A 500 pL solution of a-syn was aggregated in conical microcentrifuge tubes at 37 °C at 510 rpm for 5 days. Fibril formation was confirmed using infrared (IR) spectroscopy and atomic force microscopy (AFM). Mature fibrils were stored at -20 °C until further utilization.

[0148] qPCR for a-Synuclein Studies. All primers were designed based on rat gene sequences documented in the NCBT database and developed using the SnapGene custom oligonucleotide PCR primer generation software. All primers were fabricated by Integrated DNA Technologies (IDT). RNA was extracted from the cell pellet using a GeneJET RNA Purification Kit (Thermo Scientific). RNA concentration was measured using a NanoDrop One instrument. Next, RNA was converted to complementary DNA (cDNA) using SuperScript II Reverse Transcriptase (Invitrogen) with random primers (Invitrogen). Synthesized cDNA, constructed primers, and Luna Universal qPCR Master Mix (New England Biolabs) were mixed and placed in a QuantStudio 7 Flex Real-Time PCR System (Thermo Scientific). Changes in gene expression levels were calculated using the comparative CTmethod (2A-AACT). GAPDH was used as a housekeeping gene to calculate 2A-AACT. qPCR primer sequences are shown in Table 2.Table 2: qPCR Primers for Genes with Roles in the Unfolded Protein Response (UPR), Autophagy, Exocytosis, and Mitophagy

[0149] Stress Granule Imaging. After washing with PBS, cells were fixed with 100 yiL BD Cytofix Fixation Buffer (BD Biosciences). Next, cell membranes were permeabilized with Triton-X and blocked with bovine serum albumin (BSA) to limit unspecific protein binding. Administration of T-cell intracellular antigen 1 -related protein (TIA-R) fluorescent primary antibodies was conducted at 4 °C overnight to ensure complete binding. Treated cells werewashed to remove unbound fluorescent antibodies. Cells were imaged using an EVOS M5000 microscope (Thermo Fisher Scientific) to track the presence of induced stress granules.

[0150] Pl) C. elegans Assays. Transgenic fluorescently tagged alpha-synuclein strain NL5901. with genotype pkls2386 [unc-54p::alphasynuclein::YFP + unc-119(+)], was purchased from University of Minnesota Caenorhabditis Genetic Center (CGC). C. elegans were kept on nematode grow th medium (NGM) plates seeded with OP50 E. coli and at 20 °C until reaching an egg-producing age. To ensure accurate lifespan tracking, age synchronization was conducted by collecting all worms and eggs and bleaching the solution to remove all adult worms. Three days after synchronization, nematodes were transferred onto experimental NGM plates doped with fluorodeoxyuridine (FUDR) to prevent eggs from hatching. A total of ten nematodes w ere transferred to each experimental plate. The plates w ere seeded with E. coli supplemented with different concentrations of TMD NFs. NF supplementation was conducted by mixing concentrated 2% stocks with lOx concentrated OP50 E. coli at a 1 : 1 ratio before plating, quickly drying, and UV irradiating for a final concentration of 1% TMD NFs. Daily counts of the number of alive and dead nematodes were conducted to generate a Kaplan-Meier survival curve and calculate statistical analyses.

[0151] Fluorescent Imaging of C. elegans Puncta. Fluorescent imaging of endogenous a-syn puncta was conducted via an EVOS M5000 microscope (Thermo Fisher Scientific). Individual nematodes were placed in 5 pL of 50 mM sodium azide (NaN3) for immobilization. The nematodes were imaged in the green fluorescent protein (GFP) light channel. Five nematodes were imaged for each experimental condition at each given time point. Images were analyzed using ImageJ software to quantify the number of fluorescent puncta present in each worm. The average number of puncta across the five nematodes was recorded.

[0152] Data Analysis and Statistics. ROS and JC-1 analysis via flow cytometry was calculated relative to negative and positive controls using LSRII software. COX-I and SDH-A quantification via fluorescent ELISA was completed with a Tecan plate reader. Calculated values were subsequently analyzed using two-w ay ANOVA followed by Kruskal -Wallis and Dunn's post hoc tests to report significant differences across experimental conditions. Quantification of the relative expression of key genes was calculated using the established comparative CT method (2A-ACT). Real-time PCR data was processed in Microsoft Excel for Microsoft 365 (Microsoft Corporation, USA). Average CT values were reported as a heat map across experimental conditions and across time points. In vivo data was analyzed by Kaplan-Meier survival curve, reporting 95% confidence interval (CI), followed by two-way ANOVA,log-rank (Mantel-Haenszel), Kruskal-Wallis, and Dunn's post hoc tests to report significant differences in life expectancy across the experimental conditions. Aggregated a-syn quantification via fluorescent imaging was analyzed with ImageJ. Calculated values were analyzed using two-way ANOVA followed by Kruskal -Wallis and Dunn's post hoc tests. Statistical analyses were conducted in the JASP software (JASP Team, 2026).* * *

[0153] All of the methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments or aspects, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit, and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Claims

CLAIMSWhat is claimed is:

1. A method of treating a subject afflicted with or at risk of developing a neurological condition comprising administering an effective amount of a nanomaterial structure comprising molybdenum diselenide (MoSe2) to the subject.

2. The method of claim 1, wherein the neurological condition is selected from the group consisting of a neurotraumatic condition, a neurodegenerative condition, and a neuropsychiatric condition.

3. The method of claim 2, wherein:a) the neurotraumatic condition is selected from the group consisting of a brain injury, a traumatic brain injury7, a stroke, a brain aneurysm, a brain hemorrhage, a brain tumor, a concussion, a skull fracture, a hematoma, a spinal trauma, a spinal cord injury, carbon monoxide poisoning, and encephalitis; b) the neurodegenerative condition is selected from the group consisting of a synucleinopathy, Parkinson’s disease, dementia with Lewy bodies, pure autonomic failure, multiple system atrophy, Alzheimer’s disease, a memory disorder, dementia, ataxia, Huntington’s disease, a motor neuron disease, amyotrophic lateral sclerosis, progressive bulbar palsy, primary lateral sclerosis, progressive muscular atrophy, spinal muscular atrophy, Kennedy's disease, post-polio syndrome, progressive supranuclear palsy, a tauopathy, corticobasal degeneration, frontotemporal dementia, globular glial tauopathy, Pick disease, argy rophilic grain disease, primary age-related tauopathy, a prion disease. Creutzfeldt-Jakob disease (CJD), variably protease-sensitive phonopathy (VPSPr), Gerstmann-Straussler-Scheinker disease (GSS), Kuru, and fatal insomnia; orc) the neuropsychiatric condition is selected from the group consisting of a seizure disorder, an attention deficit disorder, a cognitive deficit disorder, migraine headaches, a substance abuse disorder, an eating disorder, a depressive disorder, an anxiety disorder, schizophrenia, bipolar disorder, post-traumatic stress disorder, obsessive-compulsive disorder, panic disorder, insomnia, and epilepsy.

4. The method of claim 1, wherein the nanomaterial structure comprises a high surface area-to-volume ratio.

5. The method of claim 1, wherein the nanomaterial structure is defined as a nanoflower structure.

6. The method of claim 1, wherein said treating:a) reduces alpha-synuclein-induced cytotoxicity in neurons, astrocytes, or both neurons and astrocytes:b) reverses alpha-synuclein-induced mitochondrial membrane potential depolarization;c) increases mitochondrial biogenesis in neurons, astrocytes, or both neurons and astrocytes;d) reduces alpha-synuclein-induced stress granule formation;e) downregulates at least one unfolded protein response (UPR) gene;f) upregulates autophagy of alpha-synuclein aggregates;g) upregulates exocytosis; orh) downregulates mitophagy.

7. The method of claim 1, wherein said treating increases a mitochondrial function.

8. The method of claim 7, wherein said mitochondrial function is selected from the group consisting of mitochondrial biogenesis, mitochondrial encoded gene expression, nuclear encoded mitochondrial gene expression, mitochondrial protein expression, mitochondrial respirators’ capacity, and adenosine triphosphate production.

9. The method of claim 1, wherein said treating decreases accumulation of a reactive oxygen species.

10. The method of claim 9, wherein the reactive oxygen species is selected from the group consisting of hydrogen peroxide (H2O2), superoxide anions (Ch-), and hydroxyl radicals (’OH).

11. The method of claim 1, wherein said treating increases neuron or astrocyte proliferation.

12. The method of claim 1, wherein said treating increases neuron or astrocyte survival.

13. The method of claim 1, wherein the nanomaterial structure further comprises a targeting molecule.

14. The method of claim 13, wherein the targeting molecule is cell-specific or tissuespecific.

15. The method of claim 13, wherein the targeting molecule is brain-specific, astrocytespecific, or neuron-specific.

16. The method of claim 1, wherein the nanomaterial structure further comprises a therapeutic agent or a detectable label.

17. The method of claim 16, wherein:a) the therapeutic agent is a mitochondrial therapeutic agent or a neurotherapeutic agent; orb) the detectable label is a paramagnetic ion, a radioactive isotope, a fluorochrome, an NMR-detectable agent, or an X-ray imaging agent.

18. The method of claim 17, wherein:a) the mitochondrial therapeutic agent is selected from the group consisting of CoQlO (ubiquinone), idebenone, riboflavin, di chloroacetate, thiamine, creatine, lipoic acid, glutathione, A-acetylcysteine, cysteamine, EPI-743 (para- benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide, bezafibrate. resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule; or b) the neurotherapeutic agent is selected from the group consisting of a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, clomethiazole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A. N-(3-propylcarbamoyloxirane-2- carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742, tempol, tenocyclidine,topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

19. The method of claim 1, wherein said administering comprises injection, microneedle administration, inhalation, transnasal application, oral administration, buccal administration, vaginal administration, intraosseous administration, topical administration, transdermal application, or rectal administration.

20. The method of claim 1, wherein the subject is an animal subject, a mammalian subject, a rodent subject, or a human subject.

21. The method of claim 1, the method comprising administering a pharmaceutical composition comprising the effective amount of the nanomaterial structure to the subject.

22. The method of claim 21. wherein the pharmaceutical composition comprises the nanomaterial structure comprising molybdenum diselenide (MoSe2) and a mitochondrial therapeutic agent or a neurotherapeutic agent.

23. The method of claim 1, further comprising administering a second therapeutic agent to said subject.

24. The method of claim 23, wherein the second therapeutic agent is a mitochondrial therapeutic agent or a neurotherapeutic agent.

25. The method of claim 24, wherein:a) the mitochondrial therapeutic agent is selected from the group consisting of CoQlO (ubiquinone), idebenone, riboflavin, dichloroacetate, thiamine, creatine, lipoic acid, glutathione, A-acetylcysteine, cysteamine. EPI-743 (para- benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide, bezafibrate, resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule; or b) the neurotherapeutic agent is selected from the group consisting of a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone,methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, cl omethi azole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A, N-(3-propylcarbamoyloxirane-2- carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742. tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

26. A method of increasing longevity in a subject comprising administering an effective amount of a nanomaterial structure comprising molybdenum diselenide (MoSe2) to the subject.

27. The method of claim 26, wherein the nanomaterial structure comprises a high surface area-to-volume ratio.

28. The method of claim 26, wherein the nanomaterial structure is defined as a nanoflower structure.

29. The method of claim 26, wherein said administering:a) reduces alpha-synuclein-induced cytotoxicity in neurons, astrocytes, or both neurons and astrocytes:b) reverses alpha-synuclein-induced mitochondrial membrane potential depolarization;c) increases mitochondrial biogenesis in neurons, astrocytes, or both neurons and astrocytes;d) reduces alpha-synuclein-induced stress granule formation;e) downregulales at least one unfolded protein response (UPR) gene;f) upregulates autophagy of alpha-synuclein aggregates;g) upregulates exocytosis: orh) downregulates mitophagy.

30. The method of claim 26, wherein said administering increases a mitochondrial function.

31. The method of claim 30, wherein said mitochondrial function is selected from the group consisting of mitochondrial biogenesis, mitochondrial encoded gene expression, nuclear encoded mitochondrial gene expression, mitochondrial protein expression, mitochondrial respirator}' capacity, and adenosine triphosphate production.

32. The method of claim 26, wherein said administering decreases accumulation of a reactive oxygen species.

33. The method of claim 32, wherein the reactive oxygen species is selected from the group consisting of hydrogen peroxide (H2O2), superoxide anions (Ch-), and hydroxyl radicals (’OH).

34. The method of claim 26, wherein the subject has at least about a 5%, about a 10%, or about a 15% increase in life expectancy compared to a control subject.

35. The method of claim 26. wherein the nanomaterial structure further comprises a targeting molecule.

36. The method of claim 35, wherein the targeting molecule is cell-specific or tissuespecific.

37. The method of claim 26. wherein the nanomaterial structure further comprises a therapeutic agent or a detectable label.

38. The method of claim 37, wherein:a) the therapeutic agent is a mitochondrial therapeutic agent or a neurotherapeutic agent; orb) the detectable label is a paramagnetic ion, a radioactive isotope, a fluorochrome, an NMR-detectable agent, or an X-ray imaging agent.

39. The method of claim 38, wherein:a) the mitochondrial therapeutic agent is selected from the group consisting of CoQlO (ubiquinone), idebenone, riboflavin, di chloroacetate, thiamine, creatine, lipoic acid, glutathione, JV-acetylcysteine, cysteamine, EPI-743 (para- benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide, bezafibrate. resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule; orb) the neurotherapeutic agent is selected from the group consisting of a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, cl omethi azole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A, N-(3-propylcarbamoyloxirane-2- carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742, tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

40. The method of claim 26, wherein said administering comprises injection, microneedle administration, inhalation, transnasal application, oral administration, buccal administration, vaginal administration, intraosseous administration, topical administration, transdermal application, or rectal administration.

41. The method of claim 26, wherein the subject is an animal subject, a mammalian subject, a rodent subject, or a human subject.

42. The method of claim 26. the method comprising administering a pharmaceutical composition comprising the effective amount of the nanomaterial structure to the subject.

43. The method of claim 42, wherein the pharmaceutical composition comprises the nanomaterial structure comprising molybdenum diselenide (MoSe2) and a mitochondrial therapeutic agent or a neurotherapeutic agent.

44. The method of claim 26, further comprising administering a second therapeutic agent to said subject.

45. The method of claim 44, wherein the second therapeutic agent is a mitochondrial therapeutic agent or a neurotherapeutic agent.

46. The method of claim 45, wherein:a) the mitochondrial therapeutic agent is selected from the group consisting of CoQlO (ubiquinone), idebenone, riboflavin, dichloroacetate, thiamine, creatine, lipoic acid, glutathione, A-acetylcysteine, cysteamine. EPI-743 (para- benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide, bezafibrate, resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule; or b) the neurotherapeutic agent is selected from the group consisting of a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a \-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, cl omethi azole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A, N-(3-propylcarbamoyloxirane-2- carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742. tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

47. A method of decreasing cellular oxidative stress in a subj ect in need thereof, the method comprising administering an effective amount of a nanomaterial structure comprising a selenium moiety active site to the subject, wherein said selenium moiety active site acts as an active site of reaction to reduce cellular oxidative stress.

48. The method of claim 47, wherein the nanomaterial structure comprises a transition metal selected from the group consisting of: titanium, vanadium, zirconium, niobium, molybdenum, hafnium, tantalum, and tungsten.

49. The method of claim 47, wherein the nanomaterial structure comprises molybdenum diselenide (MoSe2).

50. The method of claim 47, wherein the nanomaterial structure comprises a plurality of selenium moiety active sites.

51. The method of claim 47, wherein the nanomaterial structure further comprises a targeting molecule.

52. The method of claim 51, wherein the targeting molecule is cell-specific or tissuespecific.

53. The method of claim 47. wherein the nanomaterial structure further comprises a therapeutic agent or a detectable label.

54. The method of claim 53. wherein:a) the therapeutic agent is a mitochondrial therapeutic agent or a neurotherapeutic agent; orb) the detectable label is a paramagnetic ion, a radioactive isotope, a fluorochrome, an NMR-detectable agent, or an X-ray imaging agent.

55. The method of claim 54, wherein:a) the mitochondrial therapeutic agent is selected from the group consisting of CoQlO (ubiquinone), idebenone, riboflavin, di chloroacetate, thiamine, creatine, lipoic acid, glutathione, JV-acetylcysteine, cysteamine, EPI-743 (para- benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide, bezafibrate. resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule; or b) the neurotherapeutic agent is selected from the group consisting of a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, clomethiazole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A. N-(3-propylcarbamoyloxirane-2- carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742, tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Aspfluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

56. The method of claim 47, wherein the subject is afflicted with or at risk of developing a disease or condition associated with increased cellular oxidative stress.

57. The method of claim 47, wherein the subject is afflicted with or at risk of developing a neurological condition.

58. The method of claim 57, wherein the neurological condition is selected from the group consisting of a neurotraumatic condition, a neurodegenerative condition, and a neuropsychiatric condition.

59. The method of claim 47, wherein said administering comprises injection, microneedle administration, inhalation, transnasal application, oral administration, buccal administration, vaginal administration, intraosseous administration, topical administration, transdermal application, or rectal administration.

60. The method of claim 47, wherein the subject is an animal subject, a mammalian subject, a rodent subject, or a human subject.

61. The method of claim 47. the method comprising administering a pharmaceutical composition comprising the effective amount of the nanomaterial structure to the subject.

62. The method of claim 61, wherein the pharmaceutical composition comprises the nanomaterial structure comprising the selenium moiety active site and a mitochondrial therapeutic agent or a neurotherapeutic agent.

63. The method of claim 47, further comprising administering a second therapeutic agent to said subject.

64. The method of claim 63, wherein the second therapeutic agent is a mitochondrial therapeutic agent or a neurotherapeutic agent.

65. The method of claim 64, wherein:a) the mitochondrial therapeutic agent is selected from the group consisting of CoQlO (ubiquinone), idebenone. riboflavin, di chloroacetate, thiamine, creatine, lipoic acid, glutathione, iV-acetylcysteine, cysteamine, EPI-743 (para- benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide,bezafibrate, resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule; or b) the neurotherapeutic agent is selected from the group consisting of a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, cl omethi azole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A, N-(3-propylcarbamoyloxirane-2- carbonyl)-isoleucyl-proline, nerinetide, propentofylline. rasagiline, remacemide, nluzole, rivastigmine, selegiline, SGS-742, tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

66. A pharmaceutical composition comprising a nanomaterial structure comprising a selenium moiety active site and a mitochondrial therapeutic agent or a neurotherapeutic agent.

67. The pharmaceutical composition of claim 66, wherein the nanomaterial structure comprises a transition metal selected from the group consisting of: titanium, vanadium, zirconium, niobium, molybdenum, hafnium, tantalum, and tungsten.

68. The pharmaceutical composition of claim 66, wherein the nanomaterial structure comprises molybdenum diselenide (MoSez).

69. The pharmaceutical composition of claim 66, wherein the nanomaterial structure comprises a plurality of selenium moiety active sites.

70. The pharmaceutical composition of claim 66, wherein the nanomaterial structure further comprises a targeting molecule.

71. The pharmaceutical composition of claim 70, wherein the targeting molecule is cellspecific or tissue-specific.

72. The pharmaceutical composition of claim 66, wherein the targeting molecule is brainspecific, astrocyte-specific, or neuron-specific.

73. The pharmaceutical composition of claim 66, wherein the nanomaterial structure further comprises a detectable label.

74. The pharmaceutical composition of claim 73, wherein the detectable label is a paramagnetic ion, a radioactive isotope, a fluorochrome, an NMR-detectable agent, or an X-ray imaging agent.

75. The pharmaceutical composition of claim 66, wherein:a) the mitochondrial therapeutic agent is selected from the group consisting of CoQlO (ubiquinone), idebenone, riboflavin, di chloroacetate, thiamine, creatine, lipoic acid, glutathione, JV-acetylcysteine, cysteamine, EPI-743 (para- benzoquinone analog), arginine, citrulline, cardiolipin, elamipretide, bezafibrate. resveratrol, AICAR (aminoimidazole carboxamide ribonucleoside), epicatechin, RTA 408 (synthetic isoprenoid), decanoic acid, a therapeutic peptide, and a therapeutic polynucleotide molecule; or b) the neurotherapeutic agent is selected from the group consisting of a calcium channel antagonist, a sodium channel antagonist, an immunosuppressant, a growth factor, a protease inhibitor, a y-aminobutyric acid agonist, a thiopental glucocorticoid, diazepam, magnesium sulfate, gabapentin, methylprednisolone, methylprednisolone hemisuccinate, melatonin, minocycline, a statin, trazodone, lithium, valproate, lamotrigine, eliprodil, enadoline, 7-nitroindazole, almitrine, cilostazol, clomethiazole, D-JNKI-1, dexanabinol, ebselen, edaravone, epigallocatechin gallate, huperzine A. N-(3-propylcarbamoyloxirane-2- carbonyl)-isoleucyl-proline, nerinetide, propentofylline, rasagiline, remacemide, riluzole, rivastigmine, selegiline, SGS-742, tempol, tenocyclidine, topiramate, vasoactive intestinal peptide, vinpocetine, Z-Val-Ala-Asp fluoromethyl ketone, ziconotide, a therapeutic peptide, and a therapeutic polynucleotide molecule.

76. A method of reducing alpha-synuclein aggregation in a subject in need thereof, the method comprising administering an effective amount of a nanomaterial structure comprising a transition metal di chalcogenide to the subject.

77. The method of claim 76, wherein the transition metal dichalcogenide is selected from the group consisting of molybdenum diselenide (MoSe2). molybdenum disulfide (M0S2), tungsten disulfide, tungsten diselenide, and combinations thereof.

78. The method of claim 76, wherein the nanomaterial structure is defined as a nanoflower structure.

79. The method of claim 76, wherein the nanomaterial structure comprises a high surface area-to-volume ratio.

80. The method of claim 76, wherein the subject is afflicted with or at risk of developing a synucleinopathy.

81. The method of claim 80, wherein the synucleinopathy is selected from the group consisting of Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, and pure autonomic failure.

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