WO2022066845A1

WO2022066845A1 - Novel genetic vector approaches for aesthetics, aesthetic plastic surgery, cosmetic dermatology, and other uses

Info

Publication number: WO2022066845A1
Application number: PCT/US2021/051661
Authority: WO
Inventors: Aaron Thomas Tabor
Original assignee: Aaron Thomas Tabor
Priority date: 2020-09-23
Filing date: 2021-09-23
Publication date: 2022-03-31

Abstract

The present invention provides methods for co-administration of individual or combinations of monocistronic and / or bicistronic / multicistronic non-viral (including, but not limited to living cells and phages) and / or viral vectors and / or nucleic acids and / or any nucleic acid (modified or unmodified) effector or non-effector of cells, extracellular matrix (ECM) or biological tissue (collectively "vectors"), with any drug and / or non-drug combination (including, but not limited to aesthetic injectables), in or on the mammal for any aesthetic change or scar aesthetic improvement purpose.

Description

Novel Genetic Vector Approaches for Aesthetics, Aesthetic Plastic Surgery, Cosmetic Dermatology, and Other Uses

Related Application

This application claims priority to U.S. Provisional Application No. 63/082,304 (filed on September 23, 2020), the contents of which are hereby incorporated by reference.

Background of the Invention

A variety of cosmetic procedures exist for improving the cells involved in skin tissue maintenance and repair, as well as for reducing fat accumulation. However, there is a need in the art for new and improved methods. Accordingly, it is an object of the present invention to provide such methods.

Summary of the Invention

This invention broadly contemplates the novel and unique use for a mammal of coadministration, by injection, topically or other administration method, in or on the mammal, individual or cocktail combinations of monocistronic and / or bicistronic / multicistronic non- viral (including, but not limited to living cells and phages) and / or viral vectors and / or nucleic acids and / or any nucleic acid (modified or unmodified) effector or non-effector of cells, extracellular matrix (ECM) or biological tissue (collectively “vectors”), with any drug and / or non-drug combination (including, but not limited to aesthetic injectables), in or on the mammal for any aesthetic change or scar aesthetic improvement purpose, including but not limited to: 1) injectable fillers of all types, human and non-human derived, including but not limited to hyaluronic acid aesthetic filler fragments, collagen, fat transfers, ECM components, proteoglycans, and biocompatible synthetic polymers or biopolymers (e.g. Acellular Dermal Matrix, Poly-L-lactic acid / Polylactic acid, Polymethyl-methacrylate microspheres (PMMA), Collagen, Fat, Polyalkylimide, and Calcium hydroxyapatite), of any and all molecular weights; 2) neuromodulation / neurotoxin agents like muscle paralyzing toxins; 3) enzymes (e.g. lipolytic enzymes that break down cellulite and submental fat); 4) platelet-rich plasma (PRP) with or without “microneedling” or other injection methods that can be applied to the body, skin or the scalp, and other similar blood- or biologically derived products including but not limited to platelet-rich fibrin matrix (PRFM) that can be applied to scars, aesthetic surgical sites and “third generation” PRP, also called CGF or concentrated growth factors; 5) extracellular / intracellular vesicles and exosomes; 6) stem cell or other cellular therapies; and the like. The present invention also contemplates use of any injectable, biological or non-biological, effector or noneffector of cells, autologous or allogeneic. The present invention also contemplates administration once or repeatedly, and sequentially in any order, with or without a delivery vehicle. The present invention also contemplates use of the vectors used in any combination, method of delivery, frequency and sequence (including optimized time frames), with medical devices for aesthetic and scar aesthetic purposes. The medical devices may employ any type of force or energy, including but not limited to negative or positive pressure, suction, hot or cold temperatures (e.g., heating or freezing cells), dermabrasion, electromagnetic (e.g. laser, light, radio waves, or microwaves), magnetic, radiation, radio frequency, sound / ultrasound, and combinations thereof. For example, such medical devices include monopolar, bipolar, and ultrasonic energy, dynamic heat engines, electrostatic direct collectors, solar cells, and/or magnetic converters.

The present invention also contemplates use of the vectors used in any combination, method of delivery, frequency and sequence, with surgical procedures for aesthetic and scar aesthetic purposes. The present invention also contemplates injection, or topical application, by any method, of the vectors not in combination with the aforementioned procedures, materials and devices. In some circumstances, the mammal can self-administer in the clinic or at home.

In certain embodiments, the vectors can encode for numerous cell, extracellular matrix (ECM), or biological tissue effectors including but not limited to growth factors, structural and ECM molecules like collagen, elastin and hyaluronic acid aesthetic filler fragments, enzymes (like hyaluronidases and collagenases/MMPs) that breakdown hyaluronic acid and collagen respectively), cytokines (including anti-inflammatory and pro-inflammatory interleukins and chemokines), antioxidants (like catalase and superoxide dismutases), and antimicrobial peptides (AMPs).

In other embodiments, the vectors can encode both sense and antisense nucleic acids so that beneficial effects can be achieved. For example, antioxidants can be encoded and expressed to prevent inflammatory and immunogenic responses of both muscle paralyzers and fillers giving the mammal a more pleasant clinical experience with less pain, reduced recovery time, extended time of effect, and smoother, more acceptable aesthetic results. Similarly, sense and / or antisense nucleic acids for hyaluronidases (digestion of HA) and hyaluronan synthases (synthesis of HA) can be titrated in a favorable ratio to prevent an initial breakdown of the injected hyaluronic acid aesthetic filler fragments and natural hyaluronic acid synthesis could be temporarily boosted, providing a better outcome with less chance of fibrotic nodule formation.

In one embodiment, the vectors can encode proteins to prevent the binding of low molecular weight hyaluronic acid aesthetic filler fragments to CD44 receptors, a known risk factor for skin cancer.

In another embodiment, the vectors can encode for an artificial cocktail mimetic blend of PRP comprised of a mixture of growth factors, cytokines, interleukins, chemokines, plasma and blood proteins, and the like, for scar aesthetic and / or aesthetic purposes. This would obviate the need to draw blood from the mammal as done in a traditional PRP process. This combination cocktail can, for example, be applied topically or injected.

In another embodiment, the vector comprises a nucleic acid that encodes a single or combination cocktail treatment of growth factors, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids and other biological molecules to restore growth activity turned off in the skin, hair, and /or nails by nucleic acid methylation patterns, e.g., DNA methylation and / or histone acetylation.

In another embodiment, the vectors can be combined with multiple injected materials for example hyaluronic acid aesthetic filler fragments and stem cells in combination for better aesthetic or scar aesthetic results.

In yet another embodiment, lyophilized vector(s) powder(s) can be administered, with or without mixing into a delivery vehicle and / or combined with a dermabrasion material, to a scar or intact skin for aesthetic purposes. In certain embodiments, the lyophilized vector(s) can be coadministered, in any sequence or frequency, with any vehicle and / or penetration enhancer. In certain embodiments, a bandage with or without additives (e.g. petrolatum, gel, hydrogel or liposomal gel) can be placed over the top of the area of application for occlusion and protection.

In another embodiment, a topical neurotoxin can also be used with the injected or noninjected vectors.

In another embodiment, stem cells, cells, or cellular vesicles are transfected and / or loaded with the vector prior to administration (not co-administered separately). In certain embodiments, autologous and / or “off-the-shelf’ allogeneic stem cells, cells and cellular vesicles are used.

In certain embodiments, DNA sequencing and epigenetic methylation measurement can be used to determine what growth factors and molecules and antioxidants a mammal is deficient in, or abnormal in, to determine what gene therapies are needed going forward so this is personalized gene therapy for aesthetics including scars. Accordingly, in another embodiment, the vector can encode a molecule that restores growth activity turned off in the skin, hair, and nails by nucleic acid methylation patterns including, but not limited to, DNA methylation and/or histone acetylation. For example, one or more vectors can be combined which encode a single or combination of growth factors, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids and other biological molecules to restore growth activity turned off in the skin, hair and nails by nucleic acid methylation patterns, such as DNA methylation and / or histone acetylation.

In certain embodiments, injection of the vector(s) is used.

In another embodiment, the vectors can be combined with plastic or cosmetic surgery, ablative and / or non-ablative laser resurfacing, laser vein treatment, sclerotherapy, injection lipolysis, cryotherapy, non-surgical fat reduction, chemical peel, hair removal, hair restoration, cellulite reduction procedure, liposuction procedure, intense pulsed light (IPL), skin tightening procedure, fat reduction procedure, microdermabrasion and / or non-invasive cosmetic dermatology procedure, in combination for better aesthetic or scar aesthetic enhancement.

In other embodiments, topical administration of vector(s) is used.

In other embodiments, topical administration of the vector(s) is used in combination with injection of the vector(s).

In other embodiments, medical devices are used in combination with injection of the vector(s) and / or topical administration of the vector(s).

In preferred embodiments, PRP is three-step (3-step) procedure that involves (1) having a person’s blood drawn from their arm, (2) the person’s blood is placed into a machine that separates the platelets from the rest of their blood, and (3) then re-injected into the person (only the part of their blood that contains a high concentration of platelets). Examples of potential applications include, but are not limited to, use of the technology of the present invention in scars or dermal fillers that remodel facial aesthetics after trauma. In one embodiment, the vector or plasmid is administered via a topical route of administration, in a suitable formulation, co-formulation and/or vehicle, at individual dosages within a range of about 1 to about 2000 micrograms/milliliter (pg/ml), e.g., 5 to 1500 pg/ml; preferably at individual dosages within a range of about 50 to about 1000 pg/ml; and more preferably at individual dosages within a range of about 100 to about 500 pg/ml. According to one example, sub-therapeutic microdermabrasion is performed for enhancement of KGF-1 pDNA delivery to porcine skin; and KGF-1 pDNA is delivered to the porcine skin at individual dosages within a range of about 100 to about 500 pg/ml of the KGF-1 pDNA. According to one embodiment, plasmid is delivered at concentrations of 100, 250, 500, 750, 1000, 1500, and 2000 micrograms/milliliter (pg/ml).

For example, vector constructs may be applied as naked DNA doses that range from about 1 ng/cm² to 1 mg/cm² of tissue area, or from about 10 ng/cm² to 10 mg/cm² of tissue area, or from about 100 ng/cm² to 100 pg/cm² of tissue area, or from about 500 ng/cm² to 10 pg/cm² of tissue area, or from about 1 pg/cm² to 5 pg/cm² of tissue area, or from about 10 pg/cm² to 2 pg/cm² of tissue area or ranges within these ranges may be used.

For topical application of the polynucleotide, in alternate embodiments, one or more doses at from about 1 ng/cm³ to 1 mg/cm² of tissue area is administered (e.g., 2, 3, 4, or more doses at about 1 ng/cm³ to 1 mg/cm² of tissue area), or from about 10 ng/cm² to 100 pg/cm² of tissue area, or from about 100 ng/cm² to 10 pg/cm² of tissue area, or from about 500 ng/cm² to 10 pg/cm² of tissue area, or from about 1 pg/cm² to 5 pg/cm² of tissue area, or from about 1 pg/cm² to 2 pg/cm² of tissue area may be used.

In another embodiment, for systemic administration, e.g., intraperitoneal or intravenous the dose may range from about 1 ng/kg/ day to 100 mg/kg/ day, or from about 10 ng/kg/day to 10 mg/kg/day, or from about 100 ng/kg/day to 5 mg/kg/day, or from about 1 pg/kg/day to 1 mg/kg/ day, or from about 100 pg/kg/ day to 500 pg/kg/ day, or from about 10 pg/kg/day to 100 pg/kg/day. Alternatively, ranges within these ranges may be used.

In alternate embodiments, the polynucleotide may provide the equivalent to a dose administration of polypeptide that ranges from about 1 ng/cm² to 1 mg/cm² of tissue area, or from about 10 ng/cm² to 100 pg/cm² of tissue area, or from about 100 ng/cm² to 10 pg/cm² of tissue area, or from about 500 ng/cm² to 10 pg/cm² of tissue area, or from about 1 pg/cm² to 5 pg/cm² of tissue area, or from about 10 pg/cm² to 2 pg/cm² of tissue area, or ranges within these ranges may be used.

In studies for evaluating the genetic modification of cells having cosmetic function, a plasmid may be administered at any dosage range that produces a desired effect on cosmetic function in the cells. For instance, a plasmid may be administered via a topical route of administration, in a suitable formulation, co-formulation and/or vehicle, at individual dosages within a range of about 5 to about 2000 micrograms/milliliter (pg/ml); preferably at individual dosages within a range of about 50 to about 1000 pg/ml; and more preferably at individual dosages within a range of about 100 to about 500 pg/ml. According to one example, sub- therapeutic microdermabrasion is performed for enhancement of KGF-1 pDNA delivery to porcine skin; and KGF-1 pDNA is delivered to the porcine skin at individual dosages within a range of about 100 to about 500 pg/ml of the KGF-1 pDNA. According to one embodiment, plasmid is delivered at concentrations of 100, 250, 500, 750, 1000, 1500, and 2000 micrograms/milliliter (pg/ml).

Detailed Description of the Invention

In order that the present description may be more readily understood, certain terms are first defined. Additional definitions are set forth throughout the application.

As used herein, a “subject” is a mammal who may require treatment to maintain and/or to improve the condition of any cells having cosmetic function. Such subjects include humans and animals (e.g., animals who may have a skin condition). The subject may be a human. The subject may comprise a post-menopausal female. Alternatively, the subject may be a non-human mammal as for example, pets that may have a need for treatment of the skin, nails, hair, and/or fur.

As used herein, “cells that have cosmetic function” include skin, fat, muscle, connective tissue, and nerve cells found in the epidermis, dermis and subcutaneous layers, including the nail root or nail bed, nail matrix and nail plate, and scalp, hair follicles and hair strands, as well as muscles found under the subcutaneous fat layer and the tongue; cells in the teeth and gums; cells in the bones, including facial bones; and cells found in the eye including the iris and stroma covering the iris. As used herein, "skin" is composed of the epidermis and the dermis. The outermost epidermis in skin consists of stratified squamous epithelium with an underlying basement membrane. The epidermis in skin does not contain any blood vessels, but receives nutrients by diffusion from the dermis. The main types of cells that make up the epidermis are keratinocytes. Also, melanocytes and Langerhans cells are present. The epidermis can be further subdivided into the following layers from outermost to innermost: comeum, lucidum, granulosum, spinosum, and basal. The dermis lies below the epidermis and includes blood vessels, nerves, hair follicles, smooth muscle, glands and lymphatic tissue. Also, fibroblasts are commonly found in the dermis and secrete an extracellular matrix rich in collagen, elastin, hyaluronic acid and other macromolecules. Below the dermis lies the subcutaneous layer containing fat-filled cells called adipose cells, larger blood vessels and larger nerves. Muscles are found below the subcutaneous fat. The skin structure is attached via connective tissues to the muscles. Connective tissues also anchor the skin, fat and muscles to underlying bone tissues. Excessive fat in the subcutaneous layer in the attachment areas causes a dimpled "cellulite" appearance.

As used herein, "epidermal tissue" comprises tissue derived from the ectoderm. The ectoderm is the outermost germ layer of metazoan embryos, developing into epidermal and nervous tissue. Epidermal tissue includes skin, nails, and hair.

As used herein, “cosmetic procedure” includes all procedures devoted to the improvement or alteration of the appearance of a human or mammal, such as a neuromodulator toxin procedure, a filler or fat transfer procedure, a platelet-rich plasma (PRP) procedure, a scar aesthetic procedure, an aesthetic surgery procedure, a fat-reducing procedure, a skin-tightening procedure, a laser resurfacing procedure, microdermabrasion, cryotherapy, chemical peel, a laser vein procedure, a cellular procedure, a hair removal procedure, and / or a hair restoration procedure.

As used herein, a “hair restoration procedure” refers to the restoration of hair due androgenetic alopecia (AGA) (e.g., wherein the AGA is male pattern baldness or female pattern baldness), skin injury (e.g., wherein the injury is a result of trauma or chemotherapy), and include, but is not limited to, hair loss in the scalp or eyebrow of said subject and / or in scarred skin tissue of said subject.

The term “vector,” as used herein, is intended to refer collectively to monocistronic and / or bicistronic / multicistronic non-viral (including, but not limited to living cells and phages) and / or viral vectors and / or nucleic acids and / or any nucleic acid (modified or unmodified) effector or non-effector of cells, extracellular matrix (ECM) or biological tissue. In one embodiment, “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid,” which refers to a circular double stranded DNA loop into which additional DNA segments may be ligated. Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “recombinant expression vectors” (or simply, “expression vectors”). Expression vectors useful in recombinant DNA techniques include plasmids. As used herein, “plasmid” and “vector” may be used interchangeably, as the plasmid is the most commonly used form of vector. However, also included are other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.

The term “nucleic acid molecule,” as used herein, is intended to include DNA molecules and RNA molecules. A nucleic acid molecule may be single- stranded or double- stranded, and may be cDNA. A nucleic acid molecule may be sense or antisense.

The term “cDNA” refers to a non-naturally occurring nucleic acid molecule that has been created or derived from mRNA, i.e., the non-coding regions have been removed.

The term “mRNA” or “messenger RNA” is a nucleic acid intermediate that specifies the amino acid sequence of a polypeptide during translation.

As used herein, a “growth factor” is a diffusible signaling protein that stimulates cell growth, differentiation, survival, inflammation, and tissue repair.

A “polypeptide” refers to a chain comprising at least two consecutively linked amino acid residues, with no upper limit on the length of the chain. One or more amino acid residues in the protein may contain a modification such as, but not limited to, glycosylation, phosphorylation or a disulfide bond. A “protein” may comprise one or more polypeptides. As used herein, “administering” refers to the physical introduction of a therapeutic agent to a subject, using any of the various methods and delivery systems known to those skilled in the art. “Administering” includes, for example, administration to a human patient by another, such as, for example, one or more healthcare providers, and self-administration by the human patient. Various routes of administration described herein include intravenous, intraperitoneal, intramuscular, subcutaneous, spinal or other parenteral routes of administration, for example by injection or infusion. The phrase “parenteral administration” as used herein means modes of administration other than enteral and topical administration, such as by injection, and includes, without limitation, intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrastemal injection and infusion, as well as in vivo electroporation. Alternatively, administration can be via a non-parenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically. Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.

As used herein, “adjunctive” or “combined” administration (coadministration) includes simultaneous administration of treatment methods and / or compounds in the same or different dosage form, or separate administration of the compounds (e.g., sequential administration). “Combination” therapy, as used herein, means administration of two or more therapeutic agents or therapeutic modalities in a coordinated fashion, and includes, but is not limited to, concurrent dosing, with or without a medical device used for aesthetic and scar aesthetic purposes. Specifically, combination therapy encompasses both co-administration (e.g. administration of a co-formulation or simultaneous administration of separate therapeutic compositions) and serial or sequential administration, provided that administration of one therapeutic agent is conditioned in some way on administration of another therapeutic agent, with or without a medical device used for aesthetic and scar aesthetic purposes. For example, one therapeutic agent may be administered only after a different therapeutic agent has been administered and allowed to act for a prescribed period of time. (See, e.g., Kohrt et al. (2011) Blood 117:2423).

The term “effective dose” or “effective dosage” is defined as an amount sufficient to achieve or at least partially achieve a desired effect. A “therapeutically effective amount” or “therapeutically effective dosage” of a drug or therapeutic agent is any amount of the drug that, when used alone or in combination with another therapeutic agent, or with an aesthetic medical device, promotes disease regression evidenced by a decrease in severity of disease symptoms, an increase in frequency and duration of disease symptom- free periods, or a prevention of impairment or disability due to the disease affliction. A “prophylactically effective amount” or a “prophylactic ally effective dosage” of a drug is an amount of the drug that, when administered alone or in combination with another therapeutic agent, or with an aesthetic medical device, to a subject at risk of developing a disease or of suffering a recurrence of disease, prevents the development or recurrence of the disease. The ability of a therapeutic agent to promote disease regression or of a prophylactic agent to prevent the development or recurrence of the disease can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays.

As used herein, “and/or” is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and/or” as used in a phrase such as “A and/or B” includes “A and B,” “A or B,” “A” alone, and “B” alone. Likewise, the term “and/or” as used in a phrase such as “A, B, and/or C” encompasses each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A alone; B alone; and C alone.

It is understood that wherever aspects are described herein with the language “comprising,” otherwise analogous aspects described in terms of “consisting of’ and/or “consisting essentially of’ are also provided.

Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Moreover, all ranges disclosed herein are to be understood to encompass any and all subranges subsumed therein. For example, a stated range of "1 to 10" should be considered to include any and all subranges between (and inclusive of) the minimum value of 1 and the maximum value of 10; that is, all subranges beginning with a minimum value of 1 or more, e.g., 1 to 6.1, and ending with a maximum value of 10 or less, e.g., 5.5 to 10. Additionally, any reference referred to as being "incorporated herein" is to be understood as being incorporated in its entirety.

As used herein, the singular forms "a," "an," and "the" include plural referents unless expressly and unequivocally limited to one referent. The term "or" is used interchangeably with the term "and/or" unless the context clearly indicates otherwise.

As used herein, the term “about” means approximately, roughly, around, or in the region of. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term “about” can modify a numerical value above and below the stated value by a variance of, e.g., 10 percent, up or down (higher or lower).

Units, prefixes, and symbols are denoted in their Systeme International de Unites (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, nucleotide sequences are written left to right in 5’ to 3’ orientation. Amino acid sequences are written left to right in amino to carboxy orientation.

The present invention contemplates a method of improving cells having a cosmetic function (e.g., skin cells, hair cells, adipose cells, muscle cells, and / or nail cells) of a subject undergoing a cosmetic procedure, comprising administering a vector in an effective amount to result in improvement of the cells compared to the cells without administration of the vector. In preferred embodiments, the procedure is a neuromodulator toxin procedure, a filler or fat transfer procedure, a platelet-rich plasma (PRP) procedure, a scar aesthetic procedure, an aesthetic surgery procedure, a fat-reducing procedure, a skin-tightening procedure, a laser resurfacing procedure, microdermabrasion, cryotherapy, chemical peel, a laser vein procedure, a cellular procedure, a hair removal procedure, or a hair restoration procedure. In one embodiment, the hair restoration procedure is to restore the loss of hair in the scalp, eyebrow, and / or in scarred skin tissue of the subject which is due to androgenetic alopecia (AGA) (e.g., wherein the AGA is male pattern baldness or female pattern baldness), skin injury (e.g., wherein the injury is a result of trauma or chemotherapy).

The present invention further contemplates the method of improving the cells (i.e., cells having a cosmetic function) of a subject undergoing a cosmetic procedure, wherein the neuromodulator toxin procedure comprises administering botulinum toxin. The present invention further contemplates wherein the fat transfer procedure comprises administering hyaluronic acid aesthetic filler fragments, collagen, an ECM component, a proteoglycan, and / or a biocompatible synthetic polymer (e.g., Acellular Dermal Matrix, Poly-L- lactic acid / Polylactic acid, Polymethyl-methacrylate microspheres (PMMA), polyalkylimide, calcium hydroxyapatite).

The present invention further contemplates wherein the fat-reducing procedure targets cellulite, submental fat, and / or fat around the waist.

The present invention further contemplates wherein the skin-tightening procedure comprises a medical device that emits radio frequency energy, electromagnetic waves, or ultrasound, or produces negative pressure or coldness.

The present invention further contemplates wherein the skin-tightening procedure comprises microneedling and / or a facelift.

The present invention further contemplates wherein the cosmetic procedure comprises a device that employs a force or energy, e.g., negative or positive pressure, suction, hot or cold temperatures (e.g., heating or freezing cells), dermabrasion, electromagnetic (e.g. laser, light, radio waves, or microwaves), magnetic, radiation, radio frequency, sound / ultrasound, and combinations thereof.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, wherein the vector comprises one or more combinations of monocistronic and / or bicistronic / multicistronic non-viral (e.g., a living cell or a phage) and / or viral vector and / or one or more nucleic acid and / or one or more effector or non-effector of cells (e.g., extracellular matrix (ECM).

The present invention further contemplates wherein the vector encodes an extracellular matrix (ECM) molecule, a biological tissue effector (e.g., a growth factor), collagen, elastin, hyaluronic acid aesthetic filler fragments, an enzyme (e.g., a hyaluronidase, a collagenase, metallomatrix proteinase), a cytokine (e.g., an anti-inflammatory interleukin, a pro-inflammatory interleukin, a chemokine), an antioxidant (e.g., a catalase, a superoxide dismutase), superoxide dismutase 3 (SOD3), and / or an anti-microbial peptide (AMP).

The present invention further contemplates wherein the vector comprises a nucleic acid that includes both sense and antisense nucleic acids.

The present invention further contemplates wherein the vector comprises a nucleic acid that encodes a protein that prevents the binding of low molecular weight hyaluronic acid aesthetic filler fragments to CD44 receptors.

The present invention further contemplates wherein the vector comprises a nucleic acid that encodes an artificial cocktail mimetic blend of platelet-rich plasma (PRP) comprising a mixture of growth factors, cytokines, interleukins, chemokines, plasma, and / or blood proteins.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the method comprises further administering hyaluronic acid aesthetic filler fragments and stem cells.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the vector is lyophilized and applied topically to a scar, or intact skin.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the vector is administered according to any sequence or frequency and with a vehicle and / or penetration enhancer.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the vector is transfected into a cell or cellular vesicle (e.g., an autologous and / or allogeneic stem cell) prior to administration.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further comprising a diagnostic step to determine a deficiency in the subject. The present invention further contemplates wherein the diagnostic step determines the amount or level of growth factors, molecules, and / or antioxidants in the subject. The present invention further contemplates wherein the diagnostic step comprises DNA sequencing and / or measuring the epigenetic methylation.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the vector is injected, administered topically, and / or administered using a medical device.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure is a neuromodulator toxin procedure and the vector is a non-viral bicistronic plasmid encoding superoxide dismutase 3 (SOD3) and catalase. The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises administering a hyaluronic acid aesthetic filler fragments and the vector is a non-viral plasmid encoding superoxide dismutase 3 (SOD3).

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises administering a collagen filler and the vector comprises antisense nucleic acid capable of inhibiting a collagen-degrading MMP and, optionally, a second vector which is a non-viral plasmid encoding for TIMP-1, TIMP-2, TIMP-3, and/or TIMP-4.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further comprising the step of administering a vector which encodes a growth factor (e.g., Keratinocyte Growth Factor 1 (KGF1)).

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises a medical device that reduces cellulite, submental fat, and / or fat accumulation around the waist and the vector comprises a non-viral plasmid encoding lipase, or other fat degrading enzymes.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises a medical device that tightens and firms skin through the use of radio frequency, electromagnetic waves (for example lasers), ultrasound, coldness, or negative pressure and the vector comprises a non-viral plasmid encoding encoding KGF1 (Keratinocyte Growth Factor 1).

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises microneedling and the vector comprises one or more non-viral plasmid encoding KGF1 (Keratinocyte Growth Factor 1), EGF (Epidermal Growth Factor), and VEGF (Vascular Endothelial Growth Factor) applied to the face and scalp.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises a microdermabrasion substance which is mixed with lyophilized non-viral plasma DNA encoding one or more growth factors, structural molecules, and / or antioxidants (e.g., KGF1, SOD3, elastin, and / or collagen) and physically scrubbed into the subject’s skin and bald scalp.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises cosmetic or aesthetic plastic surgery, ablative or non-ablative laser resurfacing, chemical peel, and / or microdermabrasion and the vector comprises a non-viral plasmid encoding OD-1, SOD- 2, and / or SOD-3 which is applied for up to 14 days after the skin procedure.

The present invention further contemplates a method of improving the cells of a subject undergoing a cosmetic procedure, as described herein, further wherein the procedure comprises a fat-transfer injection and the vector comprises an antisense vector or antisense encoding vector targeting lipase.

The present invention further contemplates a method of improving the effects of a platelet-rich plasma (PRP) procedure in a subject, comprising administering a vector in an effective amount to result in improvement of the PRP procedure compared to the procedure without administration of the vector, wherein the vector comprises of one or more non-viral plasmid encoding KGF1 (Keratinocyte Growth Factor 1), EGF (Epidermal Growth Factor), and VEGF (Vascular Endothelial Growth Factor) added to the PRP before application to a subject’s microneedled skin and scalp.

The present invention further contemplates wherein the vector is administered according to any sequence or frequency and with a vehicle and / or penetration enhancer. The present invention further contemplates wherein the vector is transfected into a cell or cellular vesicle (e.g., an autologous and / or allogeneic stem cell) prior to administration. The present invention also contemplates wherein the vector is injected, administered topically and / or administered using a medical device.

The present invention further contemplates the methods as described herein, further comprising a diagnostic step to determine a deficiency in the subject. The present invention also contemplates wherein the diagnostic step determines the amount or level of growth factors, molecules, and / or antioxidants in the subject. The present invention also contemplates wherein the diagnostic step comprises DNA sequencing and / or measuring the epigenetic methylation. The present disclosure is further illustrated by the following examples, which should not be construed as further limiting. The contents of all references cited throughout this application are expressly incorporated herein by reference.

EXAMPLES

Example 1: Neuromodulator toxin treatment in combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, and / or a nucleic acid that acts as a gene silencer or interferes with the expression of a gene that encodes an enzyme that breaks the toxin down (e.g., an antisense nucleic acid, siRNA, shRNA, etc.)

In a study, subjects are administered by injection anywhere in or on the body a neuromodulator toxin treatment in combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids to enzymes that break the toxin down. Subjects experience less side effects and adverse events, less inflammation, less pain, faster recovery time, longer duration of toxin effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered the toxin alone or placebo, results are superior. In similar studies, a) the toxin is administered by injection and the vector(s) are administered by non-injection; b) the toxin is administered by noninjection and the vector(s) are administered by non-injection; and, c) the toxin is administered topically and the vector(s) are administered topically. In other similar studies, the vector(s) are administered with optimized frequency, sequence and time frames. Results are superior to toxin alone.

Example 2: Filler or fat transfer treatment in combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, enzymes that synthesize the filler, and / or antisense nucleic acids to enzymes that degrade the filler and / or fat

In a study, subjects are administered by injection anywhere in or on the body a filler or fat transfer treatment in combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, enzymes that synthesize the filler, and / or antisense nucleic acids to enzymes that degrade the filler and / or fat. Subjects experience less side effects and adverse events, less inflammation, less fibrotic nodules, less pain, faster recovery time, longer duration of filler or fat transfer effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered the filler or fat transfer alone or placebo, results are superior. In similar studies, a) the filler or fat transfer is administered by injection and the vector(s) are administered by non-injection; b) the filler or fat transfer is administered by non-injection and the vector(s) are administered by non-injection; and, c) the filler or fat transfer is administered topically and the vector(s) are administered topically. In other similar studies, the vector(s) are administered with optimized frequency, sequence and time frames. Results are superior to filler or fat transfer alone.

Example 3: PRP type treatment in combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, growth factors and other biological molecules found in blood, plasma, or biological tissues, and / or antisense nucleic acids

In a study, subjects are administered by injection anywhere in or on the body any PRP type treatment in combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, growth factors and other biological molecules found in blood, plasma, or biological tissues, and / or antisense nucleic acids. Subjects experience less side effects and adverse events, less inflammation, less pain, faster recovery time, longer duration of effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered any PRP type or placebo alone, results are superior. In similar studies, a) the PRP is administered by injection and the vector(s) are administered by non-injection; b) the PRP is administered by non-injection and the vector(s) are administered by non-injection; and, c) the PRP is administered topically and the vector(s) are administered topically. In other similar studies, the vector(s) are administered with optimized frequency, sequence and time frames. Results are superior to any PRP type alone. Example 4: Administration of vectors encoding a synthetic PRP mimetic combination cocktail treatment of important growth factors and other biological molecules found in blood, plasma, or other tissues, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids

In a study, subjects are administered by injection anywhere in or on the body vectors encoding a synthetic PRP mimetic combination cocktail treatment of important growth factors and other biological molecules found in blood, plasma, or other tissues, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids. Subjects experience less side effects and adverse events, less inflammation, less pain, faster recovery time, longer duration of effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered placebo alone, results are superior. In similar studies, a) the synthetic PRP is administered by injection and the vector(s) are administered by non-injection; b) the synthetic PRP is administered by non-injection and the vector(s) are administered by non-injection; and, c) the synthetic PRP is administered by injection and the vector(s) are administered by injection. In other similar studies, the vector(s) are administered with optimized frequency, sequence and time frames. Results are superior to placebo alone.

Example 5: Administration (from any source) stem cells, cells, extracellular / intracellular vesicles, including exosomes and the like, treatment(s) in any combination, combined with one or more vectors injected encoding growth factors, structural molecules, antioxidants, antiinflammatory cytokines, growth factors and other biological molecules, and / or antisense nucleic acids

In a study, subjects are administered by injection anywhere in or on the body, from any source, stem cells, cells, extracellular / intracellular vesicles, including exosomes and the like, treatment(s) in any combination, combined with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, growth factors and other biological molecules, and / or antisense nucleic acids. Subjects experience less side effects and adverse events, less inflammation, less pain, faster recovery time, longer duration of effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered the cells, cellular vesicles or placebo alone, results are superior. In similar studies, a) the cellular and / or cellular vesicle treatment(s) is administered by injection and the vector(s) are administered by noninjection; b) the cellular and / or cellular vesicle treatment(s) is administered by non-injection and the vector(s) are administered by non-injection; and, c) the cellular and / or cellular vesicle treatment(s) is administered topically and the vector(s) are administered topically. In other similar studies, the vector(s) are administered with optimized frequency, sequence and time frames. Results are superior to cellular and / or cellular vesicle treatment(s) alone.

Example 6: Administration of vectors encoding a single or combination cocktail treatment of growth factors, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids and other biological molecules to restore growth activity turned off in the skin, hair and nails by nucleic acid methylation patterns including but not limited to DNA methylation and histone acetylation.

In a study, subjects’ skin is analyzed for methylation patterns in promoter regions shutting down important cosmetically beneficial gene expression pathways. Subjects are then treated by topical application or injection, alone in combination with a medical device, vectors encoding a single or combination cocktail treatment of epigenetic-modifying enzymes. Subjects experience skin regeneration unexpected subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered placebo alone, results are superior.

In a related study, ten-eleven translocation (TET) family enzymes including TET1 to demethylate targeted gene promoter regions plus histone acetyltransferases (HATs) to relax chromatin for more transcription are applied topically to subjects having aging skin, balding areas, and / or brittle nail beds to produce rejuvenation. Skin is analyzed as described immediately above and compared to subjects administered a placebo alone.

Example 7 ; An aesthetic medical device treatment or protocol treatment, and administered by injection anywhere in or on the body vectors encoding a synthetic combination cocktail treatment of important aesthetic growth factors, structural molecules, antioxidants, antiinflammatory cytokines, and / or antisense nucleic acids

In a study, subjects are treated anywhere in or on the body with an aesthetic medical device treatment or protocol treatment, and administered by injection anywhere in or on the body vectors encoding a synthetic combination cocktail treatment of important aesthetic growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids. Subjects experience less side effects and adverse events, less inflammation, less pain, faster recovery time, longer duration of effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered placebo alone, results are superior. In similar studies, the vector(s) are administered by non-inj ection. In other similar studies, the vector(s) are administered in combinations by injection and by non-injection with optimized frequency, sequence and time frames. The vector(s) are administered in any sequence, frequency and time frame relative to treatment with the aesthetic medical device or protocol treatment. Results are superior to placebo alone.

Example 8: Aesthetic plastic or cosmetic surgery, ablative and / or non-ablative laser resurfacing, laser vein treatment, sclerotherapy, injection lipolysis, cryotherapy, chemical peel, hair removal, cellulite reduction procedure, liposuction procedure, intense pulsed light (IPL), skin tightening procedure, fat reduction procedure, microdermabrasion and / or non-invasive cosmetic dermatology procedure, and administered by injection anywhere in or on the body vectors encoding a synthetic combination cocktail treatment of important aesthetic growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids

In a study, subjects are treated anywhere in or on the body with aesthetic plastic or cosmetic surgery, ablative and / or non-ablative laser resurfacing, laser vein treatment, sclerotherapy, injection lipolysis, cryotherapy, chemical peel, hair removal, cellulite reduction procedure, liposuction procedure, intense pulsed light (IPL), skin tightening procedure, fat reduction procedure, microdermabrasion and / or non-invasive cosmetic dermatology procedure, and administered by injection anywhere in or on the body vectors encoding a synthetic combination cocktail treatment of important aesthetic growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids. Subjects experience less side effects and adverse events, less inflammation, less pain, faster recovery time, longer duration of effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered placebo alone, results are superior. In similar studies, the vector(s) are administered by noninjection. In other similar studies, the vector(s) are administered in combinations by injection and by non-injection with optimized frequency, sequence and time frames. The vector(s) are administered in any sequence, frequency and time frame relative to the aforementioned aesthetic surgical and non-surgical treatments. Results are superior to placebo alone.

Example 9: Administration of a combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids

In a study, subjects are administered by injection anywhere in or on the body a combination with one or more vectors injected encoding growth factors, structural molecules, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids. Subjects experience less side effects and adverse events, less inflammation, less pain, faster recovery time, longer duration of effect, less immunogenic response and better subjective or objective outcomes as determined by the administrator and subject self-assessment for scars and / or aesthetic outcomes. Compared to subjects administered placebo, results are superior. In similar studies, the vector(s) are administered by non-injection. In other similar studies, the vector(s) are administered in combinations by injection and by non-injection with optimized frequency, sequence and time frames. Results are superior to placebo alone.

Example 10: Administration of vectors encoding a single or combination cocktail treatment of growth factors, antioxidants, anti-inflammatory cytokines, and / or antisense nucleic acids and other biological molecules found in hair follicles and the scalp

In a study, subjects are administered by topical application or injection, alone in combination with a medical device, anywhere on the body where hair growth is desired, vectors encoding a single or combination cocktail treatment of growth factors, antioxidants, antiinflammatory cytokines, and / or antisense nucleic acids. Subjects experience hair restoration, fullness, thickness, sheen and manageability subjective or objective outcomes as determined by the administrator and subject self-assessment. Compared to subjects administered placebo alone, results are superior. In a related study, keratinocyte Growth Factor 1 plus superoxide dismutase 3 (SOD3) are applied topically to subjects having balding scalp areas producing hair growth and reduction in follicular inflammation. Outcomes and comparisons with subjects receiving a placebo are analyzed as described above.

Equivalents

Those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, many equivalents of the specific embodiments described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

I claim:

1. A method of improving cells that have a cosmetic function in a subject undergoing a cosmetic procedure, comprising administering one or more vectors in an effective amount which result in improvement of the cells compared to the cells without administration of the vector.

2. The method of claim 1, wherein the cells are skin cells, hair cells, adipose cells, muscle cells, and / or nail cells.

3. The method of claim 1 or 2, wherein the procedure is a neuromodulator toxin procedure, a filler or fat transfer procedure, a platelet-rich plasma (PRP) procedure, a scar aesthetic procedure, a cosmetic or aesthetic plastic surgery procedure, a fat-reducing procedure, a skin-tightening procedure, a laser resurfacing procedure, microdermabrasion, cryotherapy, chemical peel, a laser vein procedure, a cellular procedure, a hair removal procedure, or a hair restoration procedure.

4. The method of claim 3, wherein the hair restoration procedure is to restore the loss of hair in the scalp, eyebrow, and / or in scarred skin tissue of the subject which is due to androgenetic alopecia (AGA) (e.g., wherein the AGA is male pattern baldness or female pattern baldness), skin injury (e.g., wherein the injury is a result of trauma or chemotherapy), or other.

5. The method of claim 3, wherein the neuromodulator toxin procedure comprises administering botulinum toxin.

6. The method of claim 3, wherein the fat transfer procedure comprises administering hyaluronic acid aesthetic filler fragments, collagen, an ECM component, a proteoglycan, and / or a biocompatible synthetic polymer (e.g., Acellular Dermal Matrix, Poly-L- lactic acid / Polylactic acid, Polymethyl-methacrylate microspheres (PMMA), polyalkylimide, calcium hydroxyapatite).

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7. The method of claim 3, wherein the fat-reducing procedure targets cellulite, submental fat, and / or fat around the waist.

8. The method of claim 3, wherein the skin-tightening procedure comprises a medical device that emits radio frequency energy, electromagnetic waves, or ultrasound, or produces negative pressure or coldness.

9. The method of claim 3, wherein the skin-tightening procedure comprises microneedling and / or a facelift.

10. The method of any one of claims 1-9, wherein the cosmetic procedure comprises a device that employs a force or energy, e.g., negative or positive pressure, suction, hot or cold temperatures (e.g., heating or freezing cells), dermabrasion, electromagnetic (e.g. laser, light, radio waves, or microwaves), magnetic, radiation, radio frequency, sound / ultrasound, and combinations thereof.

11. The method of any one of claims 1-10, wherein the vector comprises one or more combinations of monocistronic and / or bicistronic / multicistronic non-viral (e.g., a living cell or a phage) and / or viral vector and / or one or more nucleic acid and / or one or more effector or non-effector of cells (e.g., extracellular matrix (ECM).

12. The method of any one of claims 1-11, wherein the vector encodes an extracellular matrix (ECM) molecule, a biological tissue effector (e.g., a growth factor), collagen, elastin, hyaluronic acid aesthetic filler fragment, an enzyme (e.g., a hyaluronidase, a collagenase, metallomatrix proteinase), a cytokine (e.g., an anti-inflammatory interleukin, a pro- inflammatory interleukin, a chemokine), an antioxidant (e.g., a catalase, a superoxide dismutase), superoxide dismutase 3 (SOD3), and / or an anti-microbial peptide (AMP).

13. The method of claim 12, wherein the vector comprises a nucleic acid that includes both sense and antisense nucleic acids.

14. The method of any one of claims 1-13, wherein the vector comprises a nucleic acid that encodes a protein that prevents the binding of low molecular weight hyaluronic acid aesthetic filler fragments to CD44 receptors.

15. The method of any one of claims 1-13, wherein the vector comprises a nucleic acid that encodes an artificial cocktail mimetic blend of platelet-rich plasma (PRP) comprising a mixture of growth factors, cytokines, interleukins, chemokines, plasma, and / or blood proteins.

16. The method of any one of claims 1-13, wherein the vector comprises a nucleic acid that encodes a single or combination cocktail treatment of growth factors, antioxidants, antiinflammatory cytokines, and / or antisense nucleic acids and other biological molecules to restore growth activity turned off in the skin, hair, and /or nails by nucleic acid methylation patterns, e.g., DNA methylation and / or histone acetylation.

17. The method of any one of claims 1-16, comprising further administering hyaluronic acid aesthetic filler fragments and stem cells.

18. The method of any one of claims 1-17, wherein the vector is lyophilized and applied topically to a scar, or intact skin.

19. The method of any one of claims 1-18, wherein the vector is administered according to any sequence or frequency and with a vehicle and / or penetration enhancer.

20. The method of any one of claims 1-19, wherein the vector is transfected into a cell or cellular vesicle (e.g., an autologous and / or allogeneic stem cell) prior to administration.

21. The method of any one of claims 1-20, further comprising a diagnostic step to determine a deficiency in the subject.

22. The method of claim 21, wherein the diagnostic step determines the amount or level of growth factors, molecules, and / or antioxidants in the subject.

23. The method of claims 21 or 22, wherein the diagnostic step comprises DNA sequencing and / or measuring the epigenetic methylation (e.g., DNA methylation or histone acetylation).

24. The method of any one of claims 1-23, wherein the vector is injected, administered topically, and / or administered using a medical device.

25. The method of claim 1, wherein the procedure is a neuromodulator toxin procedure and the vector is a non- viral bicistronic plasmid encoding superoxide dismutase 3 (SOD3) and catalase.

26. The method of claim 1, wherein the procedure comprises administering a hyaluronic acid aesthetic filler fragments and the vector is a non- viral plasmid encoding superoxide dismutase 3 (SOD3).

27. The method of claim 1, wherein the procedure comprises administering a collagen filler and the vector comprises antisense nucleic acid capable of inhibiting a collagen-degrading MMP and, optionally, a second vector which is a non-viral plasmid encoding for TIMP-1, TIMP- 2, TIMP-3, and/or TIMP-4.

28. The method of any one of claims 25-27, further comprising the step of administering a vector which encodes a growth factor (e.g., Keratinocyte Growth Factor 1 (KGF1)).

29. The method of claim 1, wherein the procedure comprises a medical device that reduces cellulite, submental fat, and / or fat accumulation around the waist and the vector comprises a non-viral plasmid encoding lipase, or other fat degrading enzyme.

30. The method of claim 1, wherein the procedure comprises a medical device that tightens and firms skin through the use of radio frequency, electromagnetic waves (e.g., lasers),

26 ultrasound, negative pressure, or cold temperature (e.g., freezing the cells) and the vector comprises a non- viral plasmid encoding encoding KGF1 (Keratinocyte Growth Factor 1).

31. The method of claim 1, wherein the procedure comprises microneedling and the vector comprises one or more non-viral plasmid encoding KGF1 (Keratinocyte Growth Factor 1), EGF (Epidermal Growth Factor), and VEGF (Vascular Endothelial Growth Factor) applied to the face and scalp.

32. The method of claim 1, wherein the procedure comprises a microdermabrasion substance which is mixed with lyophilized non-viral plasma DNA encoding one or more growth factors, structural molecules, and / or antioxidants (e.g., KGF1, SOD3, elastin, and / or collagen) and physically scrubbed into the subject’s skin and bald scalp.

33. The method of claim 1, wherein the procedure comprises cosmetic or aesthetic plastic surgery, ablative or non-ablative laser resurfacing, chemical peel, and / or microdermabrasion and the vector comprises a non-viral plasmid encoding SOD-1, SOD-2, and / or SOD-3 which is applied for up to 14 days after the skin procedure.

34. The method of claim 1, wherein the procedure comprises a fat-transfer injection and the vector comprises an antisense vector or antisense encoding vector targeting lipase.

35. A method of improving the effects of a platelet-rich plasma (PRP) procedure in a subject, comprising administering a vector in an effective amount to result in improvement of the PRP procedure compared to the procedure without administration of the vector, wherein the vector comprises of one or more non-viral plasmid encoding KGF1 (Keratinocyte Growth Factor 1), EGF (Epidermal Growth Factor), and VEGF (Vascular Endothelial Growth Factor) added to the PRP before application to a subject’s microneedled skin and scalp.

36. The method of any one of claims 25-35, wherein the vector is administered according to any sequence or frequency and with a vehicle and / or penetration enhancer.

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37. The method of any one of claims 25-36, wherein the vector is transfected into a cell or cellular vesicle (e.g., an autologous and / or allogeneic stem cell) prior to administration.

38. The method of any one of claims 25-37, further comprising a diagnostic step to determine a deficiency in the subject.

39. The method of claim 38, wherein the diagnostic step determines the amount or level of growth factors, molecules, and / or antioxidants in the subject.

40. The method of claims 38 or 39, wherein the diagnostic step comprises DNA sequencing and / or measuring the epigenetic methylation.

41. The method of any one of claims 25-40, wherein the vector is injected, administered topically and / or administered using a medical device.

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