A MAGNETIC FIELD EXPOSURE SYSTEM AND USES THEREOF
20230372726 · 2023-11-23
Assignee
Inventors
Cpc classification
International classification
Abstract
The present invention relates to a magnetic field exposure system for use in exposing organic cells, cellular tissue or at least a part of a subject to a low frequency magnetic field. The system includes a magnetic field generator and amplifier feeding one or more electromagnetic coils configured to produce a magnetic field that varies according to an amplitude modulated signal with a carrier frequency of from 360 to 450. Hz, wherein the modulation frequency is from 0.5 to 100. Hz; and wherein the MF has a field strength of from 0.5 to 250 μT. The invention also relates to a method for improving wellbeing, reducing stress, enhancing concentration, reducing anxiety, reducing skin aging, enhancing tissue regeneration, enhancing wound healing, enhancing immune function and/or inhibiting senescent related conditions in a subject by exposing cellular tissue of at least a part of the subject to said low frequency magnetic field and to a method for enhancing in vitro proliferation of stem, progenitor and other primary cells by exposing the cells to said low frequency magnetic field.
Claims
1. A magnetic field exposure system for use in exposing organic cells, cellular tissue or at least a part of a subject to a low frequency magnetic field, said system comprising a magnetic field generator comprising one or more coils configured to produce an magnetic field that varies according to an amplitude modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT.
2. The magnetic field exposure system according to claim 1, wherein said one or more coils are configured to at least partially surround the subject to be exposed to said low frequency magnetic field.
3. The magnetic field exposure system of claim 1, wherein the system is a chair comprising a hollow egg-shaped body forming a seat and a backrest, and wherein said one or more coils are integrated in the hollow egg-shaped body of the chair.
4. The magnetic field exposure system according to claim 3, wherein the hollow egg-shaped body of the chair is electromagnetically shielded, preferably the hollow egg-shaped body of the chair comprises a lining formed of a mu-metal.
5. The magnetic field exposure system according to claim 3, wherein at least two orthogonal copper coils are integrated in the hollow egg-shaped body of the chair.
6. The magnetic field exposure system according to claim 1, wherein the system is in the form of a cell culture system, said system comprising cell culture vessel defining a cell culture cavity within the vessel and a magnetic field generator with one or more coils configured to produce a magnetic field within the cell culture cavity that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT.
7. A cosmetic method for treating a condition associated with skin stem cells or skin progenitor cells in a subject, the method comprising exposing cellular tissue of at least a part of the subject to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT.
8. The cosmetic method according to claim 7, wherein the condition associated with skin stem cells or skin progenitor cells is selected from maintaining skin homeostasis, reducing skin aging, enhancing skin rejuvenation, preventing or reducing skin wrinkles, increasing skin elasticity, preventing or reducing skin stretch marks, preventing or reducing cellulitis, increasing skin hydration, reducing skin roughness, reducing skin pore size, inducing hair growth, preventing or reducing hair loss, inhibiting hair greying, inducing hair re-pigmentation, maintaining skin pigmentation, and inducing nail growth.
9. A therapeutic method for treating a condition associated with stem cells or progenitor cells in a subject, the method comprising exposing cellular tissue of at least a part of the subject to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT.
10. The therapeutic method according to claim 9, wherein the condition is selected from tissue regeneration, wound healing, bone regeneration, an inflammatory disease, cardiovascular disease, a neurodegenerative condition, a cognitive disorder, an autoimmune disease, osteoarthritis, tissue fibrosis, periodontal disease, or a dermatological disorder.
11. The therapeutic method according to claim 9, wherein the condition is wound healing, for example healing of surgical wounds, trauma wounds, abrasions, burns, blisters, ulcers or preventing or minimising scarring.
12. A method for enhancing proliferation of organic cells, the method comprising exposing the cells to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT.
13. The method according to claim 12, wherein the method is an in-vivo method, for example an in-vivo method for enhancing tissue regeneration in a subject.
14. The method according to claim 12, wherein the method is an in-vitro method for enhancing proliferation of cells.
15. A method for tissue regeneration in a subject comprising: (i) culturing one or more stem cells or progenitor cells from the subject in vitro, wherein the cells are exposed to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT during at least part of the culturing, thereby expanding the population of stem or progenitor cells; (ii) harvesting the cells; and (iii) administering the cells to the subject.
16. An in vitro method for inhibiting cellular senescence comprising exposing organic cells to a LF-MF that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the MF has a field strength of from 0.5 to 250 μT.
17. The method according to claim 16, wherein the organic cells are stem cells or progenitor cells.
18. The method according to claim 12, wherein the stem cells are adult human stem cells selected from muscle stem cells, hematopoietic stem cells, epithelial stem cells, neural stem cells, mesenchymal stem cells, mammary stem cells, intestinal stem cells, mesodermal stem cells, endothelial stem cells, skin stem cells, melanocyte stem cells, hair follicle stem cells, olfactory stem cells, neural crest stem cells and dental pulp stem cells.
19. The method according to claim 12, wherein the stem cells or progenitor cells are human keratinocyte stem cells or human keratinocyte progenitor cells.
20. A method for treating a senescence associated disease in a subject, comprising exposing cellular tissue of at least a part of the subject to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT.
21. The method of claim 20, wherein the senescence associated disease is selected from a cardiovascular disorder; a pulmonary disease; osteoarthritis; a senescence-associated dermatological disease or disorder, a neurodegenerative disease, an ocular disease, diabetes, a liver disease, sarcopenia and prostatic hyperplasia.
22. A method for improving wellbeing, reducing stress, enhancing concentration and/or reducing anxiety in a subject by exposing at least a part of a subject to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the magnetic field has a field strength of from 0.5 to 250 μT.
23. The method of claim 22, wherein the method is a non-therapeutic method for reducing psychological stress, inducing relaxation, enhancing concentration, increasing focus, increasing alertness, enhancing cognition and/or reducing anxiety.
24. The system of claim 1, wherein the magnetic field varies according to an amplitude modulated signal with a carrier frequency within of from 400 to 450 Hz and a modulation frequency of 3 to 30 Hz.
25. The system of claim 1, wherein the carrier frequency is 432 Hz.
26. The system of claim 1, wherein the resonance frequency is from 3 to 28 Hz.
27. The system of claim 1, wherein the modulation frequency corresponding to a Schumann resonance mode.
28. The system of claim 1, wherein the modulation frequency is within the range of 7.50 to 8.00 Hz.
29. (canceled)
30. The system of claim 1, wherein the carrier frequency is 432 Hz and the modulation frequency is 7.83 Hz.
31. The system of claim 1, wherein the maximum magnetic field strength is 200 μT.
32. The system of claim 1, wherein the modulation frequency is sinusoidal.
33. The system of claim 1, wherein the low frequency magnetic field is not pulsed.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0096] Embodiments of the invention are further described hereinafter with reference to the accompanying drawings, in which:
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DETAILED DESCRIPTION
[0113] As used herein, the term “subject” refers to a human or an animal (e.g. non-human primates; domestic and farm animals, such as dogs, horses, cats, cattle sheep, mice, rats, or rabbits). Suitably the subject is a human.
[0114] As used herein, the term “a healthy subject” means a subject (e.g. a human) absent of any clinical signs of physical diseases and infections.
The term “stem cell” is used herein to refer to a cell that has the ability both to self-renew, and to generate differentiated progeny (see Morrison et al. (1997) Cell 88:287-298). Generally, stem cells also have one or more of the following properties: an ability to undergo asynchronous, or asymmetric replication; i.e., where the two daughter cells after division can have different phenotypes; extensive self-renewal capacity; capacity for existence in a mitotically quiescent form; and clonal regeneration of all the tissue in which they exist, for example the ability of hematopoietic stem cells to reconstitute all hematopoietic lineages. “Progenitor cells” differ from stem cells in that they typically do not have the extensive self-renewal capacity, and often can only regenerate a subset of the lineages in the tissue from which they derive. In certain embodiments the stem cells are multipotent stem cells, which include adult stem cells that are capable of differentiating into specific cell types, examples include hematopoietic stem cells, mesenchymal stem cells and neural stem cells. In certain embodiments the stem cells are pluripotent stem cells, cells which can differentiate into any type of cell including embryonic stem cells, perinatal stem cells (umbilical cord stem cells) and induced pluripotent stem cells (iPSCs). In certain embodiments the stem cell is a multipotent stem cell or an induced pluripotent stem cell. In certain embodiments the stem cell is a not an embryonic stem cell.
[0115] The stem cells or progenitor cells may be mammalian stem or progenitor cells, where the term “mammalian” refers to any animal classified as a mammal, including humans; non-human primates; domestic and farm animals (e.g. dogs, cats, horses, cattle, sheep, mice, rats or rabbits). Suitably the stem cell is a human stem cell.
[0116] Stem cells include adult stem cells. Adult stem cells are also referred to as somatic or tissue stem cells. Adult stem cells are resident in differentiated tissue, but retain the properties of self-renewal and ability to give rise to multiple cell types, usually cell types typical of the tissue in which the stem cells are found. Numerous examples of somatic stem cells are known to those of skill in the art, including muscle stem cells, hematopoietic stem cells, epithelial stem cells, neural stem cells, mesenchymal stem cells, mammary stem cells, intestinal stem cells, mesodermal stem cells, endothelial stem cells, olfactory stem cells, dental pulp stem cells or neural crest stem cells.
[0117] In some embodiments, the stem cell is a hematopoietic stem cell (HSC). HSCs are mesoderm-derived cells that can be isolated from bone marrow, blood, cord blood, foetal liver and yolk sac. HSCs can repopulate the erythroid, neutrophil-macrophage, megakaryocyte and lymphoid hematopoietic cell lineages in vivo. In vitro, HSCs can be induced to undergo at least some self-renewing cell divisions and can be induced to differentiate to the same lineages as is seen in vivo. As such, HSCs can be induced to differentiate into one or more of erythroid cells, megakaryocytes, neutrophils, macrophages, and lymphoid cells.
[0118] In other embodiments, the stem cell is a neural stem cell (NSC). Neural stem cells (NSCs) are capable of differentiating into neurons, and glia (including oligodendrocytes, and astrocytes). A neural stem cell is a multipotent stem cell which is capable of multiple divisions, and under specific conditions can produce daughter cells which are neural stem cells, or neural progenitor cells that can be neuroblasts or glioblasts, e.g., cells committed to become one or more types of neurons and glial cells respectively.
[0119] In other embodiments, the stem cell is a mesenchymal stem cell (MSC). MSCs originally derived from the embryonal mesoderm and isolated from adult bone marrow, can differentiate to form muscle, bone, cartilage, fat, marrow stroma, and tendon.
[0120] In a preferred embodiment the stem cell is a keratinocyte stem cell.
[0121] In certain embodiment the stem cells of are aged adult stem cells. For example, an aged adult stem cell is an adult stem cell obtained from, or present in, a human individual greater than 30 years, greater than 40 years, greater than 50 years, greater than 60 years, or greater than 70 years. For example, an aged adult stem cell obtained from, or present in, a human individual who is aged from 30 years to 70 years.
[0122] In certain embodiments the stem cells are young adult stem cells, for example adult stem cells obtained from, or present in, a human individual less than 10 years old, or less than 6 years old for example aged from 0 month to 10 years, or from 1 month to 6 years.
[0123] Reference to “primary cells” refers to cells that have been obtained from a subject. Examples of primary cells include epithelial cells, endothelial cells, fibroblasts, melanocytes, keratinocytes, neurons, astrocytes, hepatocytes, skeletal muscle cells, smooth muscle cells, osteoblasts, myocytes, chondrocytes, adipocytes, synoviocytes, hair cells, blood cells stem cells or progenitor cells. Primary cells may be obtained using well known methods, for example via a tissue biopsy followed separation/isolation of the primary cells of interest. Cell separation and isolation methods are well known and include immunomagnetic cell separation, fluorescent-activated cell sorting, centrifugation methods (e.g. density gradient centrifugation, immunodensity cell separation, cell sedimentation, cell adhesion methods or microfluidic cell separation).
[0124] As used herein, the terms “treatment,” “treating,” and the like, refer to obtaining a desired pharmacologic and/or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and/or may be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease. “Treatment,” as used herein, covers any treatment of a disease in a mammal, e.g., in a human, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., causing regression of the disease.
[0125] Reference to a condition being “associated with” stem or progenitor cells refers to the condition being caused directly or indirectly by stem or progenitor cells, for example wherein the disease or condition results from, or is mitigated by, reduced stem cell proliferation, stem cell or progenitor cell senescence, stem cell or progenitor cell stress, stem cell or progenitor cell age, and/or reduced stem cell or progenitor cell differentiation.
[0126] Unless stated otherwise, references herein to “magnetic field strength”, “MF strength”, “magnetic field intensity” or “field strength intensity” refer to the root mean square (rms) of the magnetic flux density (B) produced by the system described herein and is a vector quantity that determines the force on a moving charge or charges (electric current). Magnetic flux density is expressed in tesla (T).
[0127] The term “about” herein refers to +/−10% of the figure quoted.
[0128] Throughout the description and claims of this specification, the words “comprise” and “contain” and variations of them mean “including but not limited to”, and they are not intended to (and do not) exclude other components, integers or steps.
[0129] Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.
[0130] Features, integers, characteristics, or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and/or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and/or steps are mutually exclusive. The invention is not restricted to the details of any foregoing embodiments. The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.
[0131] The reader's attention is directed to all papers and documents which are filed concurrently with or previous to this specification in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference.
Magnetic Field Exposure System
[0132] Disclosed herein is a MF exposure system for use in exposing organic cells, cellular tissue or at least a part of a subject to a LF-MF. For example, the system may be a ring or cylinder configured to surround, for example, an arm or leg of the subject to be exposed, or a chair, a room or a chamber.
[0133] The system comprises a magnetic field generator comprising one or more coils configured to produce a magnetic field that varies according to an amplitude modulated signal with a carrier frequency within the range of 360 to 450 Hz (e.g. 400 to 450 Hz), and wherein the magnetic field has a maximum magnetic field strength within the range of 0.5 to 250 μT (e.g. 10 to 200 μT). The magnetic field generator may comprise an electric generator and an amplifier feeding one or more coils configured to produce a MF that varies according to an amplitude-modulated signal with a carrier frequency within the range of 360 to 450 Hz (e.g. 400 to 450 Hz) and a MF strength within the range of 0.5 to 250 μT (e.g. 0.5 to 200 μT, or 10 to 200 μT). The amplitude modulated magnetic field has a modulation frequency of from 0.5 to 100 Hz, for example from 3 to less than 30 Hz, or from 3 to 28 Hz.
[0134] In a preferred embodiment, the system comprises a magnetic field generator comprising an electric generator and an amplifier feeding one or more coils configured to produce a MF that varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz, a modulation frequency corresponding to a Schumann resonance mode (preferably 7.83 Hz), and a MF strength within the range of 0.5 to 250 μT (e.g. 0.5 to 200 μT, or 10 to 200 μT).
[0135] The one or more coils of the MF exposure system may be configured to at least partially surround the subject to be exposed to said LF-MF.
[0136] The one or more coils of the MF exposure system may be one or more copper coils.
[0137] The size, shape and type of coil system can be decided according to the biological system to be exposed. Circular coil systems, such as Helmholtz coil, may be used for generating MFs over a small volume whereas square coil systems such as Merritt and Ruben coils, may generate MF over a large volume, for example for whole body exposure or within a room or chamber. For whole body exposure, it is preferred that the generated MF is substantially uniform.
[0138] Numerous coil configurations and geometries may be used to generate the modulated LF-MF. The specific coil geometry used will be determined by the desired application and volume of LF-MF required. The design and optimisation of the coil geometries and orientations may be carried out using known coil configurations and methods (e.g. as described in Kirschvink J L Uniform magnetic fields and double-wrapped coil systems: improved techniques for the design of bioelectromagnetic experiments. Bioelectromagnetics 13:401-411). In certain embodiments the coil is a Helmholtz coil, wherein two or more circular coils of substantially the same diameter are coaxially spaced apart. In other embodiments Merritt coils may be used wherein square coils are arranged to provide a uniform magnetic field within the coils (Merritt et. al., (1983) Uniform magnetic-field produced by three, four, and five square coils. Rev Sci Instrum 54:879-882). For example Merritt coils may be arranged orthogonally with e.g. 2 to 5 (or more) coils per axis spaced apart from one another. This arrangement provides an inner volume defined by the coils in which the modulated LF magnetic field produced by a current passed through the coils is substantially uniform.
[0139] In certain embodiments the one or more coils is a Helmholtz coil. The Helmholtz coil comprises two substantially identical circular coils that are co-axially spaced apart, wherein the spacing between the coils is the substantially the same as the radius of the coils. The coils are supplied with the same input electrical current, which results in the generation of a substantially uniform magnetic field within the space between the two coils. The use of a Helmholtz coil in the system enables the subject (or part of the subject), or the sample (e.g. cells) to be placed in the space between the coils such that the subject or sample is exposed to a substantially uniform MF.
[0140] The maximum MF strength of the MF generated by the MF generator of the MF exposure system may be set within the acceptable limits according to national recommended exposure levels. For example, as set out in the International Commission on Non-Ionizing Radiation Protection's (ICNIRP's) guidelines 1998 and 2010. Suitably the MF strength of the LF-MF is within the range of 0.5 to 200 μT when the system is used to expose a human subject to the modulated LF-MF such as within the range of 1 to 200 μT, 5 to 200 μT, 10 to 200 μT, 25 to 200 μT, 50 to 200 μT, 100 to 200 μT, 150 to 200 μT, 1 to 40 μT, 1 to 20 μT, 1 to 15 μT, or 1 to 11 μT. In certain embodiments, the maximum magnetic field strength of the LF-MF is 200 μT. In other embodiments the maximum magnetic field strength is lower, for example a maximum magnetic field of about 100 μT, about 30 μT, about 30 μT, about 20 μT, about 15 μT, about 12 μT, or about 11 μT. Where the system is used to, for example expose cells to the modulated LF-MF ex-vivo (e.g. to enhance cell proliferation in vitro), a higher maximum magnetic field strength may be used, for example any of the magnetic field strengths disclosed herein up to 250 μT. The maximum magnetic field refers to the root mean square (rms) value of the magnetic field strength. The magnetic field strength may be determined using known methods, for example using a suitable Gauss meter.
[0141] Reference herein to the modulated LF-MF, refers to the modulated magnetic field produced by the coil(s) when a temporally-varying current is passed through the coil(s). Thus references to the magnetic field-strength herein refer to the magnetic field produced by electrical current passing through the coil(s) of the systems described herein. As will be appreciated by the skilled person other electrical and magnetic fields may also be present as a result of for example the earth's magnetic field and background fields field from electric equipment. In certain embodiments the effects of external electrical and/or magnetic background may be reduced or eliminated by, for example, locating the system in a Faraday cage and/or use of p-metal layers surrounding the system to screen out unwanted external electrical and/or magnetic background.
[0142] The MF exposure system as disclosed herein may be configured to expose at least 50%, such as 50-75%, of the subject's body surface to said MF. For example, the MF exposure system may be configured to expose the upper body, including head and torso, of the subject to said MF. In some embodiments, the MF exposure system may be configured for whole body exposure of the subject. In such embodiments, the MF exposure system provides cranial as well as peripheral MF exposure of the subject.
[0143] Alternatively, the MF exposure system as disclosed herein may be configured to provide only cranial MF exposure of the subject. In such embodiments, only the brain region of the subject is exposed to said MF.
[0144] Still alternatively, the MF exposure system as disclosed herein may be configured to provide only peripheral (transcutaneous) MF exposure of the subject. In such embodiments, the brain of the subject is not exposed to said MF.
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[0146] As will be well understood by the skilled person, when a current passes through the coil 104, a MF having an intensity proportional to the magnitude of the current is generated within and around the coil. Passage of a temporally-varying current through the coil therefore generates a temporally-varying MF.
[0147] In particular, the coil 104 is arranged such that, when a current is passes through the coil 104, a MF is generated in and around region 106A, in which at least a portion of a subject's head is likely to be located when the subject is sat on the chair 100.
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[0150] Particularly, at least parts (such as head and/or torso) of a subject sitting in any of chairs 100A, 100B or 1000 are exposed to the MF that varies according to an amplitude-modulated signal with a carrier frequency within the range as defined herein, for example, 400 to 450 Hz (e.g. 432 Hz), a modulation frequency corresponding to a Schumann resonance mode (e.g. 7.83 Hz), and a MF strength within the range of 10 to 200 μT. More particularly, at least parts (such as head and/or torso) of a subject sitting in any of chairs 100A, 100B or 1000 are exposed to a homogenous MF that varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz (e.g. 432 Hz), a modulation frequency corresponding to a Schumann resonance mode (e.g. 7.83 Hz), and a MF strength within the range of 0.5 to 250 μT (e.g. 10 to 200 μT).
[0151] Although the chair 1000 shown in
[0152] Furthermore, in some embodiments a cavity in which at least part of a subject can be located could be defined by a single curved wall having one or more coils located therein. A chair including such a cavity may be referred to as an egg-pod chair.
[0153] In an embodiment, the system is a chair comprising a hollow egg-shaped body forming a seat and a backrest, and one or more coils are integrated in the hollow egg-shaped body of the chair to generate the LF-MF.
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[0155] In some embodiments, the parts of chair that define a cavity in which a user may sit (e.g. an egg-pod chair as shown in
[0156] Thus, in a further embodiment, the system is a chair comprising a hollow egg-shaped body forming a seat and a backrest, one or more coils are integrated in the hollow egg-shaped body of the chair to generate the LF-MF, and the hollow egg-shaped body of the chair is electromagnetically shielded.
[0157] In a still further embodiment, the system is a chair comprising a hollow egg-shaped body forming a seat and a backrest, one or more coils are integrated in the hollow egg-shaped body of the chair to generate the LF-MF, and the hollow egg-shaped body comprises a lining formed of a mu-metal.
[0158] In another embodiment, the system is a chair comprising a hollow egg-shaped body forming a seat and a backrest, wherein at least two orthogonal copper coils are integrated in the hollow egg-shaped body of the chair to generate the LF-MF. The hollow egg-shaped body of the chair may be electromagnetically shielded, particularly the egg-shaped body may comprise a lining formed of a mu-metal.
[0159] In a still further embodiment, the system is a chair comprising a hollow egg-shaped body forming a seat and a backrest, wherein a first plurality of copper coils (e.g. 2-5 copper coils) and a second plurality of copper coils (e.g. 2-5 copper coils) are integrated in the hollow egg-shaped body of the chair, said first plurality of copper coils extending orthogonal to said second plurality of copper coils. The hollow egg-shaped body of the chair may be electromagnetically shielded, particularly the egg-shaped body may comprise a lining formed of a mu-metal.
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[0161] The coils may be arranged such that the direction of the MF passing through the coils at a given point in time is in the same direction (i.e. vertically up or down) for all of the coils, thereby reducing spatial variations in the MF. A subject present in the room or chamber 200 is exposed to a MF that varies according to an amplitude modulated signal with a carrier frequency within the range described herein (e.g. 400 to 450 Hz (e.g. 432 Hz)) and a MF strength within the range of 0.5 to 250 μT (e.g. 10 to 200 μT). Particularly, a subject present in the room or chamber 200 is exposed to a MF that varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz (e.g. 432 Hz), a modulation frequency corresponding to a Schumann resonance mode (e.g. 7.83 Hz), and a MF strength within the range of 10 to 200 μT. More particularly, a subject present in the room or chamber 200 may be exposed to a homogenous MF that varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz (e.g. 432 Hz), a modulation frequency corresponding to a Schumann resonance mode (e.g. 7.83 Hz), and MF strength within the range of 0.5 to 250 μT (e.g. 10 to 200 μT).
[0162] The walls, floor and ceiling of the room 200 may all be provided with an outer shielding layer, to prevent interference from ambient EMFs originating outside the room. Again, the shielding layer may be formed of a mu-metal (p-metal).
[0163] Although the embodiments shown in
[0164] In further embodiments the system comprises a horizontal platform (e.g. a bed) and one of more coils that surround the platform such that when a temporally-varying current is passed through the coils a corresponding temporally-varying MF is generated around the horizontal platform. In use when a subject lies on the bed, at least part of the subject is exposed to the modulated LF-MF. The system may be configured such that a plurality of coils surrounds the horizontal platform. For example a plurality of coils, (e.g. circular coils) surrounding the horizontal platform may be coaxially located along the horizontal axis of the platform, thereby exposing at least part, or preferably the whole body of a subject lying on the platform to the LF-MF. In further embodiments the system comprises a horizontal platform (e.g. a bed) and one of more coils located above and/or below the platform.
[0165] For exposing smaller areas of the body the system may comprise a housing with one of more coils located within the housing such that when a temporally-varying current is passed through the coils a corresponding temporally-varying MF is generated. The housing can then be positioned to expose a specific body part or tissue of a subject to the LF-MF. It may be that the housing is hand held such that the LF-MF can be positioned manually to direct the LF-MF to a target area on the subject. Alternatively, the housing my be, or comprise a frame that can be moved to direct the LF-MF to a particular part of the subject.
[0166] In a further embodiment the system is in the form of a cell culture system, said system comprising cell culture vessel defining a cell culture cavity within the vessel and a MF generator with one or more coils configured to produce a MF within the cell culture cavity that varies according to an amplitude-modulated signal with a carrier frequency of from 360 to 450 Hz, wherein the modulation frequency is from 0.5 to 100 Hz; and wherein the MF has a field strength of from 0.5 to 250 μT.
[0167] The cell culture cavity is a volume within the cell culture vessel that is suitable for containing cells to be proliferated (cultured). Thereby in use the cells contained within the cell culture cavity are exposed to the modulated LF-MF described herein.
[0168] The cell culture vessel may be any suitable vessel for culturing cells such as stem and progenitor cells, for example the vessel may be a petri-dish, a flask, multi-well plate (e.g. a 96-well plate), a bottle, a conical tube, a stacked culture vessel, a 2D tissue culture vessel or a 3D suspension culture vessel, or a larger scale bioreactor vessel. The one or more coils are located in a position such that when a current is applied to the coils, a LF-MF is generated within the cell culture cavity. In use, cells are contained within the cell culture cavity and are exposed to the LF-MF. In certain embodiments a pair of coils may be coaxially spaced apart (e.g. as a Helmholtz coil) and the reaction vessel is located between the coils such that in use the cell culture cavity in the cell culture vessel is exposed to a substantially uniform LF-MF when a current is passed through the coils. In a further embodiment the one or more coils are attached to the walls of the cell culture vessel and are configured to produce a LF-MF in the cell culture cavity. It may be that the one or more coils are embedded in the walls of the cell culture vessel. In a further embodiment the cell culture system further comprises one or more organic cells (e.g. one or more stem or progenitor cells or primary cells) in the cell culture cavity and optionally further comprises a cell culture medium in the cell culture cavity.
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[0170] Electric generator 306 is fed via power source 310, and the output frequency is controlled by the amplifier 304. The power source 310 may be a source of DC power, such as a battery, or a source of AC power, such as a mains connection.
[0171] Electric generator 306 is operable, under the control of the amplifier 304, to output a predefined, constant, temporally-varying voltage or current signal. In particular, the electric generator 306 and amplifier 304 may be configured to output an amplitude-modulated AC signal, having a carrier frequency as described herein (e.g. 400 to 450 Hz) and a sinusoidal modulation frequency, as illustrated in
[0172] The first Schumann resonance mode occurs at approximately 7.83 Hz, although any frequency within a range of approximately 7.0-10.0 Hz, more particularly 7.5-8.0 Hz, may be considered to correspond to the first Schumann resonance within the scope of the present disclosure. Further Schumann resonance frequencies occur at least within the following frequency ranges: 14.0 to 14.5 Hz, more particularly 14.1 Hz; 20.0 to 21.0 Hz, more particularly 20.3 Hz; and 26.0 to 27.5 Hz, more particularly 26.4 Hz.
[0173] In some embodiments the MF may vary in a substantially continuous manner. Accordingly, the voltage or current signal will generally also be non-pulsed and continuously varying.
[0174] In addition to determining the current or voltage waveform, the amplifier 304 may also be configured to determine the length of time for which the system exposes the subject to the temporally-varying MF. For example, the MF may be produced only during a user-defined time period each day, for example a time period of 15 minutes to 1 hour per day. Alternatively, the system may be configured to produce the MF substantially continuously for a pre-determined period of time.
[0175] The respective amplitudes of the carrier signal and the amplitude modulation (also called amplitude variation) signal may be selected to produce a field having maximum MF intensity or flux density of 200 μT, according to ICNIRP recommendations 2010, in the target region in which the subject is expected to be located (e.g. regions 106A-C as shown in
[0176] In embodiments, the respective amplitudes of the amplitude modulation signal and the carrier signal may be selected such that the peak amplitude of the amplitude-modulated signal, as illustrated in
[0177] The MF exposure system as disclosed herein, for example the chair as illustrated in any one
[0178] The Examples herein demonstrate that the modulated low frequency MF described herein enhances stem cell and progenitor cell (e.g. human epidermal stem and progenitor cell) proliferation and survival. The herein disclosed MF exposure system is therefore believed to be useful for reducing skin aging, for enhancing tissue regeneration, for enhancing wound healing, treating cancer and/or for enhancing immune function in a subject by exposing at least a part of the subject to said low frequency MF.
[0179] The modulated low frequency magnetic field (LF-MF) exposure system may be used for any of the methods or uses described herein. The system may be for use in enhancing in-vitro proliferation of stem and progenitor cells or other primary cells. The system may be for use in enhancing in-vivo proliferation of stem and progenitor cells. The system may be for use in the cosmetic treatment of a condition associated with skin stem cells or skin progenitor cells in a subject. The system may be for use in reducing skin aging. The system may be for use in the method for treating a medical condition associated with stem cells or progenitor cells in a subject. The system may be for use in enhancing tissue regeneration. The system may be for use in enhancing wound healing. The system may be for use in enhancing tissue regeneration. The system may be for use in the prevention or treatment of a cancer. The system may be for use in the method for inhibiting cell senescence. The system may be for use in the method for treating a senescence associated disease in a subject. The system may be for use in the treatment of a condition selected from an autoimmune disease, a neurodegenerative disease, an inflammatory disease, a cognitive disorder, and a psychiatric disorder.
[0180] The system, methods and uses described herein provides non-invasive methods and uses for the treatment or prevention of therapeutic and non-therapeutic conditions, including those described herein, by exposing at least part of the subject to the modulated LF-MF.
Cell Proliferation
[0181] The MF exposure system, methods and uses disclosed herein are useful for enhancing in vitro proliferation of organic cells, particularly stem cells, progenitor cells or other primary cells, more particularly human epidermal stem and progenitor cells by exposing the cells to said LF-MF.
[0182] Stem cell and progenitor cell proliferation and differentiation are essential for tissue homeostasis and tissue regeneration, for example in injury or wound healing. Stem/progenitor cell activation, proliferation, maintenance and differentiation can become dysregulated in certain diseases and conditions, particularly age-related conditions. The modulated LF-MF described herein provide beneficial effects on stem and progenitor cells, for example by enhancing proliferation, reducing stem/progenitor cell aging and/or reducing stem/progenitor cell stress. Accordingly, exposing stem cells and progenitor cells to the modulated LF-MF described herein is expected to be beneficial in many therapeutic and non-therapeutic/cosmetic applications, including those described above in the brief description of the invention. In particular the modulated LF-MF is expected to be beneficial in the treatment or prevention of conditions associated with keratinocyte stem and progenitor cells.
Wound Healing
[0183] The system, methods and uses of the modulated LF-MF described herein are expected to be beneficial in wound healing.
[0184] Wound healing is a complex and dynamic process involving, amongst other parameters interactions between dermal and epidermal dermal cells. Shortly after a wound is incurred, typically 2 to 10 days after wounding keratinocyte proliferation is increased and the formation of a continuous keratinocyte monolayer is rate limiting for successful wound healing (Harding, B M J. 2002 Jan. 19; 324(7330): 160-163). The increased proliferation of keratinocyte stem and progenitor cells exposed to the modulated LF-MF described herein is therefore expected to be beneficial in this process. For example, the modulated LF-MF may increase wound closure rate and hence prevent or reduce wound contracture, prevent or reduce scarring and/or enhance tissue re-modelling and finally reduce the time required for wound healing. It might also stimulate the rare event of hair follicle neogenesis (Ito, M., Nature. 2007 May 17; 447(7142): 316-320).
[0185] The modulated LF-MF may be applied to substantially all of the subject to promote wound healing. However, preferably the modulated LF-MF is applied locally to the wounded region, thereby exposing cells and cellular tissue at and close to the site of a wound to the modulated LF-MF described herein.
[0186] In certain embodiments wound healing may be treated by locally applying to a wound stem or progenitor cells that have been cultured ex vivo according to a method described herein, wherein stem cell and progenitor cells are exposed ex vivo to the modulated LF-MF to enhance proliferation and optionally differentiation of the cells. The cells may be applied directly to the wound site, for example by topical application or by injection. The cells may for example be applied to the wound using a suitable wound dressing or patch coated or impregnated with the cells. The cells may also be introduced into a wound together with a scaffold or matrix (e.g. a biodegradable polymer matrix) to further promote tissue regeneration in the wound. The scaffold of matrix may be impregnated or coated with the stem/progenitor cells. It is also possible to culture the stem/progenitor cells in the presence of the scaffold or matrix and the modulated LF-MF, thereby promoting growth of the cells directly on or within the scaffold or matrix.
[0187] The systems, methods and uses that utilise the modulated LF-MF described herein are expected be beneficial in promoting wound healing in a broad range of tissue and is not limited to epidermal wounds. For example the systems, methods and uses described herein that utilise the modulated LF-MF described herein may be useful for promoting wound healing (e.g. surgical wounds, injuries or other trauma) in tissues selected from muscle tissue, connective tissue, joint tissue, epithelial tissue, endothelial tissue, nervous tissue, fat tissue, skin tissue, lung tissue, liver tissue, bladder tissue, kidney tissue, cardiac tissue, pancreatic tissue, pancreatic tissue, stomach tissue, intestinal tissue, spinal tissue, brain tissue, eye tissue, fibrous tissue, dentin, bone or bone marrow. Thus in certain embodiments the systems, methods and uses described herein may be for promoting wound healing in a subject that has undergone neurosurgery, wherein the subject is exposed to the modulated LF-MF described herein to promote healing of brain tissue or nerve tissue. Suitably in this embodiment when the subject has undergone brain surgery, the head of the subject is exposed to the modulated LF-MF described herein. Also contemplated is localised application of the modulated LF-MF, wherein a particular part of the brain or nerve tissue is exposed to the modulated LF-MF.
In Vitro Cell Culturing
[0188] As discussed in the brief description of the invention, cells, particularly stem cells, progenitor cells or other primary cells may be cultured in vitro wherein the cells (e.g. step or progenitor cells are exposed to the modulated LF-MF during culturing and expansion of the cell population.
[0189] In vitro cell culture methods and suitable culture media are well known to the skilled person and are illustrated in the Examples herein. Cell culture media suitable for the proliferation and/or differentiation of stem or progenitor cells are well known (Dakhore, S. et al., Human Pluripotent Stem Cell Culture: Current Status, Challenges, and Advancement. Stem cells international, 2018, 7396905). For example, the expansion medium may be a modified basal culture medium based on minimum essential medium (MEM), Dulbecco's modified Eagle's medium (DMEM) or minimum essential medium Eagle alpha modification (α-MEM). The stem cell expansion medium is suitable a serum-free stem cell expansion medium. Stem cell expansion media are commercially. For differentiation of the stem or progenitor cells a differentiation medium may be used, alternatively the expansion medium is supplemented with growth factors to promote differentiation of the stem or progenitor cells.
[0190] The cells are suitably cultured and grown in a suitable culture vessel. The cell culture cavity is a volume within the cell culture vessel that contains the cells to be proliferated, for example a cell culture medium comprising the cells. In use the cells contained within the cell culture cavity are exposed to the modulated LF-MF described herein.
[0191] The culture vessel may be any vessel suitable for the culture of stem or progenitor cells, for example a 2D tissue culture vessel or a 3D suspension culture vessel. In some embodiments the culture vessel is selected from a petri-dish, a multi-well plate, stacked cell culture vessel, a conical tube, or a 3D-suspension culture vessel.
[0192] In 2D culture vessels the vessels are typically coated with a suitable protein such as cadherin, laminin or vitronectin. The stem cells attach to the coated surface and grow in colonies on the surface. The cells are passaged before the colonies become too dense, for example when reaching greater than 50%, 60%, 70%, 80% or 90% confluence, and/or when the cells exhibit increased differentiation. Upon reaching the desired confluence the cells are disassociated from the surface of the vessel using a suitable method (e.g. shaking, mechanical scraping or enzymatic disassociation (e.g. trypsin-EDTA)). The collected cells are then re-seeded at a lower surface density in a vessel together with a culture medium to continue growth. The passaging may be performed numerous times to provide the required population of cells.
[0193] In 3D culture vessels the cells are suspended in a suitable culture medium and may form aggregates which are typically spherical as a suspension in the culture medium. 3D culturing can also be performed using a suitable gel or scaffold in which the cells are suspended. In 3D culture vessels the cells are passaged when the average size of the cell aggregates reaches 100 μm to 500 μm, and/or when the cells reach 50%, 60%, 70%, 80% or 90% confluence, and/or when the cells exhibit increased differentiation. The cell aggregates are collected and dissociated (e.g. using a suitable enzyme) and are diluted to the required density/concentration for continued growth.
[0194] In certain embodiments the cells are passaged multiple times whilst being exposed the modulated LF-MF as described herein. Thus it may be that the cells (e.g. stem or progenitor cells are passaged at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times or at least 8 times. For example the cells may be passaged 3 to 8 times or 3 to 6 times, wherein the cells are exposed to the modulated LF-MF during each passage.
[0195] Proliferation of the cells according to the methods described herein may be monitored using conventional methods, for example optical microscopy to determine the degree of confluence.
[0196] Stem and progenitor cells and differentiated cells produced according to the method described herein may also be used to develop in-vitro models of tissue for in vitro modelling of diseases and the testing and screening drug candidates for efficacy and/or toxicity. Such cellular models or “diseases in a dish” have been used to study cardiomyocytes for cardiac disease models, and the modelling of neurological disorders.
Treatment of Autoimmune Disease, Neurodegenerative Disease, Inflammatory Disease, Cardiovascular Disease, Cognitive Disorders and Psychiatric Disorders
[0197] As set out in the brief description of the invention the review by Guerriero et al., suggests that low frequency magnetic fields may be beneficial in the treatment of a number of conditions, including autoimmune disease, neurodegenerative disease, inflammatory disease, cognitive disorders, psychiatric disorders, spinal injuries, ocular diseases.
[0198] The modulated LF-MF described herein may be for the treatment of a neurodegenerative disease, for example selected from Alzheimer's disease; amyotrophic lateral sclerosis; motor neurone disease; motor disorders; Parkinson's disease; Huntington's disease, or multiple sclerosis.
[0199] The modulated LF-MF described herein may be for the treatment of cognitive impairment, for example cognitive impairment associated with any of the neurodegenerative diseases above.
[0200] The modulated LF-MF described herein may be for the treatment of a disorder selected from a mood disorder, post-traumatic stress syndrome, attention deficit disorder attention deficit hyperactivity disorder, depression, or a depressive disorder.
[0201] The modulated LF-MF may be suitable for the treatment of bullous diseases of the skin such as autoimmune diseases of the pemphigus complex or inherited diseases like Epidermolysis simplex.
[0202] The modulated LF-MF may be suitable for the treatment of spinal cord injuries. For example to regenerate new nerve cells and promote growth of new nerve fibres and/or to improve nerve function.
[0203] The modulated LF-MF may be suitable for the treatment of ocular diseases, for example to regenerate damage to cornea, retinal optic nerve, treatment of age-related macular degeneration (AMD), glaucoma or retinitis pigmentosa.
[0204] The modulated LF-MF may be suitable for the treatment of autoimmune disorders, for example the treatment of type 1 diabetes to replenish pancreatic p-cells; or the treatment of multiple sclerosis (e.g. to regenerate neurons with a myelin sheath).
[0205] The modulated LF-MF may be suitable for the treatment of a cardiovascular condition, for example to regenerate cardiac muscle, for example resulting from cardiac ischemia; or to restoring cardiac blood vessels.
Modulation of Brain Waves
[0206] Electroencephalogram (EEG) studies indicate that brain waves are divided into five different bandwidths. Alpha-band waves have a frequency of 8.5 to 12.5 Hz and are known to reduce psychological stress and induce a relaxed mood state, increased attention and improve memory (Klimesch, W. alpha-band oscillations, attention, and controlled access to stored information. Trends Cogn. Sci 16, 606-617, doi:10.1016/j.tics.2012.10.007 (2012); Hammerschlag, Biofield Physiology: A Framework for an Emerging Discipline., Glob Adv Health Med, 4, 35-41, doi:10.7453/gahmj.2015.015.suppl (2015)).
Alpha-band waves also increase serotonin production and reduce cortisol levels boosting the immune system (Yu, Activation of the anterior prefrontal cortex and serotonergic system is associated with improvements in mood and EEG changes induced by Zen meditation practice in novices. Int J Psychophysiol 80, 103-111, doi:10.1016/j.ijpsycho.2011.02.004 (2011); Puig, M. V. & Gulledge, A. T. Serotonin and prefrontal cortex function: neurons, networks, and circuits. Mol Neurobiol 44, 449-464, doi:10.1007/s12035-011-8214-0 (2011); and Lozano-Soldevilla, D. On the Physiological Modulation and Potential Mechanisms Underlying Parieto-Occipital Alpha Oscillations. Front Comput Neurosci 12, 23, doi:10.3389/fncom.2018.00023 (2018)).
[0207] Beta-band waves have a frequency of 12.5 to 30 Hz and are associated with alertness, focus and cognition. The Examples herein show that both alpha and beta-band waves are increased when subjects were exposed to the modulated LF-MF described herein. Subjects undergoing the tests described an increased state of relaxation, which is consistent with increased alpha-band activity. Normally alpha-band waves are not detected when the eyes are open. Surprisingly it was found that in subjects exposed to the modulated LF-MF, alpha-band activity was also increased and persisted in particular in the Parietal Occipital cortex, even with the eyes open.
[0208] The observed increase in both alpha-band and beta-band activity resulting from exposure to the modulated LF-MF suggest that the modulated LF-MF may be useful in reducing anxiety and stress and/or increasing alertness, and/or increasing focus and/or enhancing cognition. In a particular embodiment the LF-MF is used for the reduction of physiological stress.
[0209] The modulated LF-MF described herein may therefore be useful in the treatment or prevention of an attention deficit disorder (e.g. attention deficit hyperactivity disorder (ADHD)), post-traumatic stress syndrome, cognitive impairment, or and anxiety disorder.
[0210] In some embodiments the modulated LF-MF is for use in the treatment or prevention of an anxiety disorder. Anxiety is a feeling of apprehension or fear that lingers due to an individual's perception of persistent and unrelenting stress. Anxiety is typically accompanied by various physical symptoms including twitching, trembling, muscle tension, headaches, sweating (e.g., night sweats), dry mouth, or difficulty swallowing. Examples of anxiety disorders include separation anxiety disorder, selective mutism, specific phobia, social anxiety disorder (social phobia), panic disorder, panic attack, agoraphobia, post-traumatic stress disorders (PTSD), generalized anxiety disorder, substance/medication-induced anxiety disorder, anxiety disorder due to another medical disorder or an obsessive compulsive disorder.
[0211] Additionally, the examples herein show that exposure of cells to the modulated LF-MF reduced markers for cell stress and cell aging, suggesting that the LF-MF will provide beneficial effects in subjects exposed to the modulated LF-MF.
[0212] Furthermore, the herein disclosed MF exposure system is believed to be useful for improving wellbeing, for reducing stress, for enhancing concentration and/or for reducing anxiety in a subject by exposing at least a part of the subject to said LF-MF. The system may be for use in non-therapeutic improvement of well being, reducing stress, including psychological stress, inducing relaxation, enhancing concentration, increasing focus, increasing alertness, enhancing cognition add/or reducing anxiety.
[0213] Thus also provided is a non-therapeutic method for improvement of well being, reducing stress, including psychological stress, inducing relaxation, enhancing concentration, increasing focus, increasing alertness, enhancing cognition add/or reducing anxiety in a subject, the method comprising exposing at least a part of the subject to the LF-MF.
[0214] In the embodiments herein relating to the modulation of brain-waves, it is preferred that at least the brain of the subject is exposed to the LF-MF.
FURTHER EMBODIMENTS
[0215] The invention is further illustrated by the following numbered clauses. [0216] 1. A magnetic field exposure system for use in exposing organic cells, cellular tissue or at least a part of a subject to a low frequency magnetic field, said system comprising a magnetic field generator comprising one or more coils configured to produce an magnetic field that varies according to an amplitude modulated signal with a carrier frequency within the range of 400 to 450 Hz, and wherein the magnetic field has a maximum magnetic field strength within the range of 10 to 200 μT. [0217] 2. A magnetic field exposure system according to clause 1, wherein the magnetic field varies according to an amplitude modulated signal with a carrier frequency within the range of 400 to 450 Hz and a modulation frequency within the range of 3 to 30 Hz. [0218] 3. A magnetic field exposure system according to clause 1 or clause 2, wherein the carrier frequency is 432 Hz. [0219] 4. A magnetic field exposure system according to any of the preceding clauses, wherein the maximum magnetic field strength is 200 μT. [0220] 5. A magnetic field exposure system according to any of the preceding clauses, wherein the magnetic field varies according to an amplitude modulated signal with a carrier frequency within the range of 400 to 450 Hz and a modulation frequency corresponding to a Schumann resonance mode. [0221] 6. A magnetic field exposure system according to any of clauses 2-5, wherein the modulation frequency is within the range of 7.50 to 8.00 Hz, such as 7.83 Hz. [0222] 7. A magnetic field exposure system according to any of clauses 2-5, wherein the modulation frequency is within the range of 14.0 to 14.5 Hz, such as 14.1 Hz; or within the range of 20.0 to 21.0 Hz, such as 20.3 Hz; or within the range of 26.0 to 27.5 Hz, such as 26.4 Hz. [0223] 8. A magnetic field exposure system according to any of clauses 2-7, wherein the modulation frequency is sinusoidal. [0224] 9. A magnetic field exposure system according to any of the preceding clauses, wherein said one or more coils are configured to at least partially surround the subject to be exposed to said low frequency magnetic field. [0225] 10. A magnetic field exposure system according to any of the preceding clauses, wherein the system is a chair comprising a hollow egg-shaped body forming a seat and a backrest, and wherein said one or more coils are integrated in the hollow egg-shaped body of the chair. [0226] 11. A magnetic field exposure system according to clause 10, wherein the hollow egg-shaped body of the chair is electromagnetically shielded, preferably the hollow egg-shaped body of the chair comprises a lining formed of a mu-metal. [0227] 12. A magnetic field exposure system according to any one of clause 10-11, wherein at least two orthogonal copper coils are integrated in the hollow egg-shaped body of the chair. [0228] 13. A method for improving wellbeing, reducing stress, enhancing concentration and/or reducing anxiety in a subject by exposing at least a part of a subject to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz and magnetic field strength within the range of 10 to 200 μT, preferably the magnetic field varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz and a modulation frequency within the range of 3 to 30 Hz. [0229] 14. A method for reducing skin aging in a subject by exposing cellular tissue of at least a part of the subject to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz and magnetic field strength within the range of 10 to 200 μT, preferably the magnetic field varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz and a modulation frequency within the range of 3 to 30 Hz. [0230] 15. A method for enhancing in vitro proliferation of human epidermal stem and progenitor cells by exposing the cells to a low frequency magnetic field that varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz and magnetic field strength within the range of 10 to 200 μT, preferably the magnetic field varies according to an amplitude-modulated signal with a carrier frequency within the range of 400 to 450 Hz and a modulation frequency within the range of 3 to 30 Hz.
EXAMPLES
[0231] Unless explicitly described otherwise, the following applies: [0232] Pooled young (below the age of 8; 3 donors) or adult primary human stem and progenitor keratinocytes (major cell type in the epidermis) were seeded in 6-well plates at an initial cell density of 4,000 cells/cm.sup.2 containing 2 ml CnT-07 (CnT-BM.1 (basal cell media with low calcium concentration (0.07 mM)) supplemented with CnT-07.S). [0233] Pooled young primary human fibroblasts (major cell type found in the dermis) were seeded in 6-well plates at an initial cell density of 2,000 cells/cm.sup.2 containing 2 ml CnT-Prime F. [0234] All cells, media and complementary products were purchased from CELLnTEC Advanced Cell Systems AG (Switzerland). [0235] The culture media were free from antibiotics or antimycotics and were changed every 2 to 3 days. [0236] The cells were cultured at 37° C. 0.1° C., 90%±5% humidity in 5%±0.1% CO.sub.2. [0237] All groups contain at least 3 or more sample values (at least triplicates) from 3 or more independent experiments. [0238] Upon any group reaching 80% confluency (Phase 1, described in
[0252]
[0253] Keratinocytes in the stratum basale intermittently proliferate, exit the cell cycle and go on to differentiate while migrating to the suprabasal layer during skin homeostasis.
[0254] Key events at cell cycle exit involve the suppression of c-Myc, which in turn sets the starting point for onset of differentiation. Consequently, expression of differentiation markers like keratin 10, Dsg1, involucrin and loricrin increase in cultured cells and in the epidermis during the terminal differentiation (Y. Poumay el al, J Invest Dermatol 104:271-276 (1995); Kolly, C. et al, J Invest Dermatol 124, 1014-1025 (2005)). The increase of extracellular calcium (switch) accelerates the process.
[0255] Under standard conditions, keratinocytes stop proliferating at 100% confluency following contact inhibition and cell cycle exit. In contrast, uncontrolled growth overriding contact inhibition is a hallmark of cancer and other hyper proliferative diseases.
Example 1: Effect of 432/7.83 Hz MF on Stem Cell and Progenitor Cell Proliferation
[0256] Primary young and adult donor keratinocytes were seeded and then exposed in a coil without generated MF (coil Off) or 432/7.83 Hz MF.
[0257] This experiment was continued for a total of 6 passages to compare short-term and long-term effects of LF-MF on cell proliferation. Extrapolated cell numbers (
The significant increase in cell count for primary young (
Example 2: Effect of 432/7.83 Hz Modulated MF Compared to 7.83 Hz Alone
[0258] Primary young and adult donor keratinocytes were seeded and then exposed to coil Off or 7.83 Hz or 432/7.83 Hz MF.
[0259] After one passage of continuous MF exposure, a significant increase in the number of young cells (
[0260] The effects of continuous MF exposure were investigated in parallel on primary adult donor keratinocytes after one passage (
[0261] As observed with the primary young cells (
[0262] Furthermore, similar to the primary young donor cells, the 432/7.83 Hz exposed cells had the additional beneficial effect of increasing the cell survival rate on average by 45% (see
[0263] Regarding morphology, the adult donor keratinocytes also exhibited improvements when exposed to 7.83 Hz and 432/7.83 Hz, with the healthiest morphology seen in the 432/7.83 Hz exposed group.
[0264] The results in primary adult donor cells corroborated the observations in primary young donor cells, that the 432/7.83 Hz MF may increase cell proliferation when compared to the 7.83 Hz and Off groups.
[0265]
Example 3: 432/7.83 Hz Modulated MF Compared to Non-Modulated Driver Frequency of 432 Hz Alone on Stem Cell and Progenitor Cell Proliferation
[0266] Primary young donor keratinocytes were seeded and then exposed to either coil Off, 432 Hz or 432/7.83 Hz MF.
[0267] After 1 passage, the 432 Hz exposed keratinocytes exhibited a significantly higher cell count than the coil Off control group. However, the 432/7.83 Hz MF group exhibited higher proliferation (
[0268] Over the first two passages, there was a small increase in cell count for the 432 Hz driver frequency exposed group, however, this turned into a negative effect after the third passage and the number of live cells started to decline in this group. After 6 passages of exposure cells in the 432 Hz showed a significant decrease in cell count compared to the coil Off control group. In contrast cells in the 432/7.83 Hz MF group showed a significant increase in proliferation compared to the coil Off control (
[0269] These results show that the modulated 432/7.83 Hz MF results in higher proliferation of primary young donor keratinocytes compared to cells exposed to the 432 Hz driver frequency alone.
Example 4: Effects of Modulation Frequency, Driver Frequency and Magnetic Field Intensity on Stem and Progenitor Cell Proliferation
[0270] Primary young and adult donor keratinocytes were seeded and then exposed to the following MF conditions to assess the effects of modulation frequency, driver frequency and magnetic field intensity on proliferation.
Example 4.1: Effects of Modulator Frequency on Proliferation
[0271] Cells were exposed to the following conditions during cell culturing:
Coil Off (control)
432 Hz amplitude modulated at 3 Hz
432 Hz amplitude modulated at 7.83 Hz
432 Hz amplitude modulated at 10 Hz
432 Hz amplitude modulated at 30 Hz
[0272] For primary young donor keratinocytes, modulation frequencies of 432/30 Hz, 432/50 Hz and 432/100 Hz resulted in a significant increase in proliferation compared to the control (coil Off) (
Example 4.2: Effects of Driver Frequency on Proliferation
[0273] Cells were exposed to the following conditions during cell culturing:
Coil Off (control)
368 Hz amplitude modulated at 7.83 Hz
400 Hz amplitude modulated at 7.83 Hz
432 Hz amplitude modulated at 7.83 Hz
464 Hz amplitude modulated at 7.83 Hz
[0274] Significant increases in proliferation were observed in the primary young donor keratinocytes exposed to 368/7.83 Hz and 400/7.83 Hz. A highly significant increase in proliferation was observed in the primary young donor cells exposed to 432/7.83 Hz. A non-significant effect was observed in the 464/7.83 Hz group (
[0275] In primary adult donor keratinocytes significant increases in proliferation were observed in the 400/7.83 Hz and 432/7.83 Hz groups. (
Example 4.3: Effects of Magnetic Field Intensity on Proliferation
[0276] Cells were exposed to the following conditions during cell culturing:
Coil Off (control)
10 μT, 432 Hz amplitude modulated at 7.83 Hz at 55 mV pp
30 μT, 432 Hz amplitude modulated at 7.83 Hz at 550 mV pp
200 μT, 432 Hz amplitude modulated at 7.83 Hz at 1050 mV pp
250 μT, 432 Hz amplitude modulated at 7.83 Hz at 1250 mV pp
[0277] Significant increases in proliferation were observed in all groups in the primary young donor keratinocytes compared to control cells (
Example 5: Effects of MF Treatment on Cell Survival
[0278] The same primary young and adult donor keratinocytes described in Example 8 below were seeded and then exposed to coil Off or 7.83 Hz or 432/7.83 Hz MF.
[0279] After one passage of continuous MF exposure to 432/7.83 Hz the survival rate the of young cells was significantly increased (
[0280] The keratinocytes also displayed better morphology in both 7.83 Hz and 432/7.83 Hz groups, with the healthiest looking cells observed in the 432/7.83 Hz group.
[0281] Similar to the young donor cells, the 432/7.83 Hz exposure significantly increased the survival rate of adult cells (
[0282] The primary adult donor keratinocytes also exhibited improvements in cell morphology when exposed to 7.83 Hz and 432/7.83 Hz frequencies, with the healthiest morphology seen in the 432/7.83 Hz group.
[0283] The results in adult donor cells corroborated the observations in primary young donor cells, that the 432/7.83 Hz MF significantly improves cell survival rate when compared to the 7.83 Hz and coil Off groups.
[0284]
Example 6: Effects of MF Treatment on Contact Inhibition
[0285] A high throughput assay was set up to measure cell numbers based on metabolic activity (MTS) throughout Phase 1 (80% confluency), Phase 2 (100% confluency) and Phase 3 (5 days after 100% confluency was reached and the calcium concentration was increased to 1.2 mM) (
[0286] Growth correlates with metabolic activity and was assessed by MTS assay.
[0287] Primary young and adult donor keratinocytes were seeded and then exposed to coil Off or 7.83 Hz or 432/7.83 Hz MF.
[0288]
[0289] Significant increases in metabolic activity in Phase 1 of primary young donor keratinocytes exposed to 432/7.83 Hz or 7.83 Hz MF and in adult keratinocytes exposed to 432/7.83 Hz MF were observed after one passage (
[0290] When the time course was continued into Phase 2 and 3 by letting the cells become confluent (Phase 2) and enter differentiation (Phase 3), a reduction in the metabolic turnover in Phase 2 and no difference between the treatment groups (
[0291] Taken together the results shown in
Example 7: Efficacy of MF Treatment on Senescence
[0292] Telomere shortening is a readout of senescent cells (Zhao et al., Telomere length maintenance, shortening, and lengthening. J. Cell Physiol. 229, 1323-1329 (2014). Cell senescence (cessation of cell division) is believed to be implicated in a number of diseases and medical conditions, for example carcinogenesis, aging and tissue repair.
[0293] The number of telomere repeats in human stem and progenitor keratinocytes was measured using quantitative PCR using a method analogous to that described in Cawthon, R. M. Telomere measurement by quantitative PCR. Nucleic Acids Res 30, e47 (2002). This approach represents the most frequently used method to assess the number of repeats reflecting telomere length.
[0294] In this experiment, it was also observed that different stocks of keratinocytes from the same donor and passage may exhibit different reactivity to 432/7.83 Hz MF in Phase 1. Without being bound by theory it is thought that there may be some initial variability in responsiveness to the MF associated with the initial thawing of a sample.
[0295] To further assess the effect of 432/7.83 Hz MF on telomere length, early (Passage 1) and late time-points (Passage 6) were compared. Using an analogous method to that described in Example 1 young donor pooled keratinocytes were lysed after passage 1 and after passage 6 in Phase 1 and DNA isolated. After 6 passages, the control (Off) featured a decrease in telomere length of more than 50% compared to the first passage. In contrast, the decrease in telomere length for the 432/7.83 Hz MF treated keratinocytes was not significant (
[0296] These results suggest that the 432/7.83 Hz MF treatment provides protective effect on telomere length.
Example 8: Effect of MF on Cellular Stress
[0297] Primary young donor keratinocytes were seeded, then cultured in either normal medium (CnT-07) or starved medium (25% CnT-07, 75% PBS), and exposed to coil Off or 432/7.83 Hz MF.
[0298] All samples were harvested for In-Cell Western analysis, staining for heat shock protein 47 (Hsp47) and 90 (Hsp90), both being stress markers (Scieglinska et al., Heat shock proteins in the physiology and pathophysiology of epidermal keratinocytes; Cell Stress Chaperones 24, 1027-1044 (2019)).
[0299] The group cultured in 25% media without exposure to MF (Off) displayed an approximately three-fold increase in relative Hsp47 expression level compared to cells cultured in the 100% medium. Hsp90 expression was also increased compared to the 100% medium. In contrast Hsp47 (
Summary of Examples 1 to 8
[0300] These studies demonstrated that the 432/7.83 Hz amplitude-modulated MF has a significant effect on enhancing human epidermal stem and progenitor cell proliferation and survival. The effect on proliferation of human epidermal stem and progenitor cells was shown to be superior to the use of 7.83 Hz alone, the use of 432 Hz alone, varied amplitude modulation frequencies, varied driver frequencies and varied average field intensities.
[0301] Further, the 432/7.83 Hz amplitude-modulated MF showed additional advantages such as increasing cell survival across all epidermal maturation phases.
The use of 432/7.83 Hz amplitude-modulated MF, but not the use of 7.83 Hz alone, was further demonstrated to prevent telomere shortening. Telomere maintenance is a characteristic feature of stem cell-mediated tissue regeneration.
Example 9: EEG Study
[0302] To assess the effect of modulated 432/7.83 Hz MF on the entrainment of brain waves, a one-blinded crossover pilot study was performed. In the study human brain states were investigated by measuring electroencephalogram (EEG) activity of subjects before and after 15 minutes of exposure to sham (no MF) or verum (432/7.83 Hz MF) in a chair wherein the subject was unaware of the exposure conditions and was seated in a cavity between Helmholtz coils located in the sidewalls of the chair such the head and substantially all the torso of the subject was exposed to the MF (
[0303] A total of 4 healthy subjects (3 females, 1 male) were measured in the study in order to assess the potential of the modulated 432/7.83 Hz MF to systematically alter brain functional states (namely anxiety, stress and arousal) and of using multichannel resting state EEG to reliably detect these changes (Cantonal Ethics Committee Bern, approval Req-2020-00895).
[0304] The subjects were comfortably sat in an isolated room with adequate lighting and background noise. Blinded cross-over sessions were repeated multiple times (n=18 sessions) with sham (no MF) or verum (432/7.83 Hz MF) conditions to compare 5 minutes before and 5 minutes after exposure, with both eyes closed and eyes opened at an interval of 30 seconds, to investigate the effect on brain wave activity. In detail, each EEG was recorded with 32 electrodes, positioned according to the international 10-10 system. All recordings were conducted using commercially available and safety certified systems (BrainVision Analyzer, Vers. 2.2.0, Brain Products GmbH, Gilching, Germany; recording reference FCz, analogue bandpass filter from 0.1 to 200 Hz, sampling rate 500 Hz).
[0305] The EEG data was then processed in a Brain Vision Analyzer. First, channels with bad signal, in terms of high noise to signal ratio, were excluded. Pre-processing of EEG data involved band pass filtering using a low cut-off of 0.1 Hz and a high cut-off of 30 Hz. An Independent Component Analysis (ICA) was then used to identify and eliminate eye movement components. The EEG was then visually inspected for residual artefacts and re-referenced to the average reference. Separately for the pre- and post-period and for eyes open and eyes closed, artefact free 2-second epochs were then extracted and submitted to an FFT. These single epoch FFT results were averaged within pre-defined frequency bands (Delta: 0.5-3.5 Hz, Theta 1: 3.5-5.5 Hz, Theta 2: 5.5-8.5 Hz, Alpha 1: 8.5-10.5 Hz, Alpha 2: 10.5-12.5 Hz, Beta 1: 12.5-18 Hz, Beta 2: 18-21 Hz and Beta 3: 21-30 Hz). The results were then averaged within conditions, i.e. separately for eyes open and eyes closed, both in the pre- and the post-stimulation periods and post and pre sham or verum values subtracted. The standard deviation of these means was also retained.
[0306] After one session of 15 minutes 432/7.83 Hz MF exposure, but not after sham, the subjects described an increased state of relaxation. This effect persisted after each following sessions.
[0307] Shown in
[0308] To summarize the EEG data, t-maps averaged across all subjects for alpha 1, alpha 2 and beta 1 waves are shown in
[0309] The alpha 1 waves increased during verum compared to sham in EC, this was also the case in EO. The alpha 2 waves also increased during verum compared to sham in both EC and EO. Both alpha 1 and 2 increased activities in verum indicates reduced anxiety and stress. The beta 1 waves increased in verum EO and not EC, indicating increased alertness and focus.
[0310] Normally alpha waves are mostly present during EC as EO suppresses their activity. Interestingly the verum increases alpha waves activity in both EC and even in EO, when alpha waves are normally not detected. This indicates that the alpha waves persist after 432/7.83 Hz MF to such a strong extent that even opening the eyes doesn't suppress their activity.
[0311] Taken together the results of the EEG study, when comparing multiple sessions across multiple days, the modulated MF exposure significantly increased both alpha and beta waves activities (
[0312] Studies were conducted with a field strength of 30 μT (in compliance with the ICNIRP 2010 recommendations).