JP2013527756A - GENE SIGNATURE AND GENE CHIP, AND THEIR METHODS AND TREATMENTS RELATED TO ADMINISTRATION OF ELECTRIC CONDUCTIVE HIGH FREQUENCY CURRENT - Google Patents

Abstract

  A gene panel comprising genes that are regulated in mammalian skin in response to the generation of a high frequency current sufficient to heat the tissue volume to a treatment temperature within the tissue volume of mammalian skin, A gene panel wherein at least one gene is selected from Table 1 or Table 2 and at least one gene is selected from Table 3. Furthermore, there are methods for providing benefits to mammalian skin, which benefit in the absence of an inflammatory cytokine response that damages the skin, within the dermal layer of mammalian skin, and / or collagen formation. Or inducing remodeling of the dermis, wherein the method generates high frequency currents within a tissue volume of mammalian skin for a treatment cycle sufficient to heat the tissue volume to a treatment temperature, Avoiding upregulation of the expression of the genes listed in Table 3.

Description

  The present invention relates to a cosmetic skin treatment regimen that involves the administration of a high frequency current that is electrically conducted through the skin to provide benefits to the skin, and to the monitoring, screening, and optimization of the skin treatment regimen. Further to the genomics-based assessment of benefits and treatments.

  Skin treatment to avoid or reduce the effects of intrinsic aging of the skin and extrinsic environmental aging is a multi-billion dollar commercial industry, which represents a new technology development and validation It is also supported by large investments. Administration of electromagnetic (EM) energy to the skin via application to the surface of the skin has been known for decades and has been implemented in a wide variety of forms and through a variety of delivery devices. In general, EM-based skin treatment methods can be categorized as excision procedures and non-ablation procedures, both of which utilize the thermal melting effect of the application of EM energy.

  A variety of products are available to consumers to improve skin condition and delay and / or prevent typical signs of aging. Such signs include, for example, fine wrinkles, wrinkles, hyperpigmentation, ruddyness, sagging, dark circles under the eyes, swollen eyes, uneven skin tone, enlarged pores, epidermal cell turnover. Reduced speed and abnormal delamination or desquamation. However, for some consumers, the wide variety of available products and advances in skin care technology still cannot produce the desired results and consumers feel the need to shift to more invasive medical procedures Some people.

  Ablation procedures, such as ablation lasers, have proven to be an effective method for surface replacement or removal of the skin's macroscopic morphology, such as during scar and tattoo removal procedures, as well as aging and light. It has also been found effective to treat and improve the appearance of damaged skin. The ablation procedure is effective to improve the appearance of fine lines and wrinkles in the cosmetic skin around the cosmetically vulnerable perioral and periorbital area, but the major disadvantage is an unnecessarily long period of time. Over time, there is long-term healing and recovery that forces consumers to have a significantly impaired cosmetic appearance. Furthermore, the potential side effects of infection, scarring, and pigmentation irregularities that may result may be cosmetically unacceptable to consumers, especially when involving the facial skin. Are often considered to be

  Recent research and development efforts have therefore focused on providing consumers with cheaper, easier and safer, non-resective, skin anti-aging and rejuvenation treatments. Cosmetic regimens involving the administration of heat energy to the skin aimed at promoting improved skin appearance are well known in the art. Electromagnetic energy supply technology in the form of handheld devices intended for consumer home use has been commercially available for almost 10 years. Improvements and advances in this technology maximize the thermally induced benefit to deeper target skin tissue while minimizing unwanted damage to the target tissue and both surrounding and surface tissue. Centered on. Selective thermal treatment induces new collagen formation, and selective thermal damage is thought to induce, among other things, dermal matrix remodeling. However, currently available techniques are known to result in undesirable and sustained negative side effects of problematic damage, including overheating, burnout, erythema, and pigment irregularities. .

  In the past, the effectiveness of these devices and compositions has been determined only by external methods. That is, if there is a visible improvement in the outer skin layer, the energy delivery device should be functional. However, there are few methods for determining the optimal time, temperature, and composition for these energy delivery devices. Furthermore, it is generally accepted that many of the visible changes that occur on the surface of the skin are the result of changes that occur below the surface in the dermal and epidermal layers of the skin. It is difficult to know exactly how to control and maximize device performance without knowing how the device affects the lower layers of the skin.

  Selective photothermal melting of skin tissue is a form of cosmetic treatment that has been widely practiced, particularly in treatments involving the administration of monochromatic laser energy and broad spectrum intense pulsed light (IPL) energy. In these techniques, light energy is applied directly to the surface of the skin, and penetration depends on penetration through the epidermis and absorption within the dermis. Dark skin, and hyperpigmentation on the epidermis can impede permeation by absorbing energy, impairing the effectiveness of the treatment, and blistering, burning, and other cosmetically undesirable It can also result in overheating of the pigmented area, which has an effect.

  As an alternative to EM-based thermal melting, electrically conducted radio frequency (RF) current is also being considered as a modality for cosmetic skin treatment. Although the use of RF current and pulsed radio frequency (pRF) in pharmaceutical technology is known, the use of RF current as a non-resectant skin rejuvenation technique for self-supply by consumers is relatively still innovative. It is. In application of RF current to the skin, transmission of bipolar RF current occurs through two electrodes applied directly to the skin. This current therefore flows directly through the dermis and is conducted from electrode to electrode, so it is focused through the epidermis and is limited by factors that affect the wavelength penetration depth. Distinguished from application.

  The administration of RF current theoretically appears to provide significant skin treatment advantages over the application of EM energy. Unlike electromagnetic energy, for example, electrically conducted RF energy is chromophore-independent, which leads to the existence of an absorption difference between pigmented and non-pigmented skin. The associated complexity and effectiveness issues and the resulting problems in treating darker skin with more energy absorbing melamine and lighter skin that can reflect light wavelengths Is avoided. In both cases, the consistency of the results is compromised and there remains a problem with thermal control in the skin containing chromophores.

  However, administration of RF currents to the skin at an energy level that can provide thermal treatment effectiveness allows the investigator to optimize the parameters to achieve the desired benefit in the absence of undesirable skin damage. Have a problem that can not be converted. RF current is supplied through the dermal tissue below the skin surface, but effect monitoring by temperature or moisture sensors is limited to accessible skin surfaces. RF current impedance is a function of tissue composition and attributes of skin tissue, including collagen density and integrity, hydration levels, and the like. Control of parameters such as distance between electrodes, and pulse length and frequency can be theoretically adjusted to optimize the effect and avoid safety concerns, but such adjustment Without the benefit of a device or other means for monitoring changes in tissue status, it is almost impossible. In the case of EM energy-based delivery, surface skin typically reaches the maximum treatment temperature, so temperature monitoring at the surface can prevent unwanted damage to the subepithelial tissue. . However, in the case of RF current, the subepithelial tissue actually reaches a higher treatment temperature than the surface skin, so that damage occurs in deeper tissue without measurable manifestation at the surface. There is a fear. Except for clinical settings that use highly functional instrumentation under the supervision of highly trained medical personnel, devices and regimens intended for personal use by consumers based on a single supply of RF current are: Therefore, in the absence of a suitable subepidermal monitoring mechanism, safety considerations at an effective treatment level continue to exist and are generally avoided. Handheld energy delivery devices that provide RF current as a single treatment modality and are intended for consumer use in the home are virtually unknown. One device, referred to as such (STOP ™, Ultragen Ltd) is commercially available to consumers for personal use in Europe, but the treatment tolerance of that device is desirable Objective evidence of clinical efficacy under controlled conditions is not available by setting it very low to avoid any damage.

  Therefore, the role of RF in skin treatment is substantially limited to auxiliary or preliminary functions combined with other thermal melting procedures. For example, in 2002, Bitter and Mulholland ( "Report of a new technique for enhanced non-invasive skin rejuvenation using a dual mode pulsed light and radio frequency energy sources: selective radiothermolysis", J.Cosmet Dermatol 2002; 1: 142~ 145) proposed a treatment protocol based on a combination of RF current and IPL and reported results based on 100 subjects' facial treatment, but the author was unable to disclose specific treatment design parameters It was. In that study, RF was reported to reinforce the effects of IPL treatment. Side effects included reports of cosmetically undesirable pigmentation effects, and consumer perception of pain was controlled by superficial cooling.

  RF has also been suggested and discussed as being useful for cosmetic skin treatment in conjunction with the application of targeted light energy. Overall, due to this treatment protocol design, RF is used supplementarily to light energy and applied according to some parameters of light energy. For example, Hammes et al. (“Electro-optical synergy (ELOS (trademark)), for non-active skin rejuvenation: a preliminary production study,” Journal of Europe, 70. Focus on the tuning pulse frequency between RF current and light energy, and that there is a synergy between these forms of energy that can allow the use of lower, less invasive levels of light energy. Further suggests that the side effects associated with a single RF application are reduced or avoided by this combined protocol. To do. Furthermore, regimens employing these devices include means for mechanically cooling the skin in response to or to prevent overheating.

  In another example, U.S. Patent Application Publication No. 2008/0033516 (A1) (Altshuller) is a "skin tissue" technique that involves a combination of heating the skin to a target depth and irradiating the target area using electromagnetic radiation. Disclosed is “temperature-controlled photobiological stimulation”. Altshuller is thought to increase ATP production, cell proliferation, and protein production, and to cause growth responses by inducing low-grade inflammatory responses, low level light, low level laser, monochromatic and quasi It refers to the existing technology of monochromatic photostimulation-based skin treatment methods, but reports inconsistent results and lack of confirmation of clinical efficacy. Altshuler assumes that application of thermal energy can enhance the photostimulation response. Altshuler teaches that a constant volume of skin thermotherapy can be achieved by any known source capable of raising the temperature of that volume, preferably from 37 ° to 45 ° C .; Illustrates heating with hot air, AC or DC current, use of a conductive heat source, ultrasonic or microwave radiation, or EM radiation of any suitable wavelength in the range of 380-2700 nm. However, all Altshuller embodiments rely on EM energy to achieve the desired treatment effect. Altshuller teaches that heat provides a synergistic enhancement of the desired effect of light stimulation, but heat in the absence of EM can lead to delayed repair of radiation-induced DNA damage, subsequent heat application. Skin treatment, suggesting that it may result in undesired biostimulation, such as the production of heat shock proteins that build resistance to and the modification of enzymatic processes, including those associated with the regeneration and repair of skin tissue As a modality, it generally teaches away from heat in the absence of light. Altshuller does not suggest a way to overcome the deficiencies associated with the inability to assess or monitor the condition of the subepidermal skin.

  Furthermore, consumer compliance is always a problem with currently available devices. Consumers have a limited amount of daily time for their beauty regimen. A device manufacturer wants to encourage consumers to use their device for an extended period of time to ensure that consumers get the most benefit, but the consumer is unlikely to comply . Therefore, there is a trade-off between recommending long-term use of the device and understanding that the amount of daily free time to use the device is limited to consumers. Exists.

  Consumer experience is also important when designing devices, compositions, and regimens. For example, sonography is a commonly performed procedure and provides significant medical benefits. However, gels used in sonographic procedures are thick and difficult to remove, causing consumer discomfort. Furthermore, many energy delivery devices heat up the outer skin too quickly or too hot, causing an unpleasant consumer experience.

US Patent Application Publication No. 2008/0033516 (A1)

Bitter and Mulholland ( "Report of a new technique for enhanced non-invasive skin rejuvenation using a dual mode pulsed light and radio frequency energy sources: selective radiothermolysis", J.Cosmet Dermatol 2002; 1: 142~145) Hammes et al. ("Electro-optical synergy (ELOS ™) for nonskinny skin rejuvenation, a preprominent prospective study 70, Journal of Europe 70, Academic.

  Therefore, there is a continuing need for methods that sufficiently improve the condition of the skin so as to avoid the need for more invasive procedures and the risks associated with those procedures. There is also a need for a better method of determining the effectiveness of energy delivery devices, and using that method to develop a more consumer-acceptable experience while improving the appearance of the skin. The desired result can be achieved.

  There remains a need in the art for safe and effective non-resective skin treatment and rejuvenation devices, therapies, and regimens suitable for personal use by consumers. Specifically, there is a need for a means of treating epidermal skin tissue by enhancing collagen synthesis and dermal remodeling without causing undesirable damage to the target tissue or surrounding tissue to be treated. It is left. There is a specific need in the art for a method to evaluate and monitor the effectiveness of RF current-based consumer performance treatments in order to optimize the administration of RF current with respect to providing the desired benefit, There also remains a need for an optimized RF current-based therapy that avoids the problems associated with EM-based therapy and does not rely on mechanical cooling associated with the procedure.

  Thus, by taking advantage of recent advances in genomics and proteomics biotechnology that allow accurate and complex feedback of cellular responses to manipulation of physical and chemical environments, we have To examine and determine genetic fingerprints related to the effect of RF currents electrically conducted to skin tissue, to provide the gene chip of the present invention, as well as safe and effective RF current based treatment regimens and treatment selection protocols Developed a method of adopting those gene chips. The inventors surprisingly found that optimization of RF current-based treatment provides benefits related to collagen synthesis and provides specific damage-inducing benefits that stimulate dermal remodeling, while It has been found that it avoids the problematic injury-induced inflammatory cytokine response. Even more surprising, the RF current-based treatment regime according to the present invention does not require supplemental energy delivery, and in particular, administration of electromagnetic (EM) energy is not in the electro-wavelength portion of the EM emission spectrum. It has been found that no mechanical surface cooling is required. The benefits of the present invention are therefore provided in the absence of known problems associated with the administration of EM energy to the skin.

  One embodiment of the present invention provides a gene that is regulated in mammalian skin in response to the generation of sufficient high frequency current within the tissue volume of mammalian skin to heat the tissue volume to a treatment temperature. Including a genetic panel. The genes of this panel are selected from Table 1 and / or Table 2 provided that at least one gene is selected from Table 3. Aspects include microarrays and gene signatures based on the microarrays.

  Another broad embodiment is directed to a method for providing benefits to mammalian skin, which benefits in the dermal layer of mammalian skin in the absence of an inflammatory cytokine response that damages the skin. Inducing collagen formation and / or remodeling of the dermis, wherein the method involves applying a high frequency current within a tissue volume of mammalian skin for a treatment cycle sufficient to heat the tissue volume to a treatment temperature. While avoiding up-regulation of the expression of the genes listed in Table 3.

  According to a further embodiment, a method for assessing the treatment effectiveness of an energy delivery device is provided. This energy delivery device is designed on the surface to provide benefits to the skin by heating the skin, treating the skin with the application of the energy delivery device, and extracting mRNA from the treated skin sample. It can be evaluated by a method comprising extracting and generating an expression profile for a gene panel according to the present invention. A facial skin treatment regimen is also effective in providing collagen and / or dermal remodeling benefits to mammalian skin without stimulating inflammatory cytokine responses that damage the skin, according to another embodiment. Can be screened or evaluated. The method comprises treating facial skin according to a treatment regimen, extracting mRNA from a sample of treated facial skin, creating a gene expression profile for a gene panel according to the invention, and generating the gene expression profile. When compared to the reference profile and the expression profile reflects the upregulation of the gene selected from Table 1 and / or Table 2 and the substantial lack of regulation of the gene selected from Table 3, Determining that the facial treatment regimen is effective. In certain aspects of the invention, cosmetically active substances and / or compositions can be similarly screened or evaluated for providing benefits according to the invention or for synergy with the treatment methods of the invention.

DETAILED DESCRIPTION While the specification concludes with claims that particularly point out and distinctly claim the invention, the invention is better understood from the following description taken in conjunction with the accompanying drawings. It is possible.
Schematic of spontaneous damage and repair cycle in human skin. Characteristics of human collagen. Characteristics of human collagen. Schematic of a biological model of the efficacy of RF current administration. 24 hour average fold change for organized genes. 24 hour average fold change for organized genes. 24 hour average fold change for exemplary genes. 24 hour average fold change for exemplary genes. Monthly average fold change for organized genes. Monthly average fold change for organized genes. Monthly average fold change for exemplary genes. Monthly average fold change for exemplary genes. Two graphs of RF simple heat transfer. Two graphs of RF simple heat transfer. Schematic of RF simple heat transfer. Schematic of the crease area of the corner of the eye around the consumer's eye. Region A of the consumer's skin. Region B of the consumer's skin. Region C of the consumer's skin. Region D of the consumer's skin. Region E of the consumer's skin. Region F of the consumer's skin.

  Unless otherwise defined, scientific and technical terms used in connection with the invention described herein shall have the meanings that are commonly understood by those of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. In general, the nomenclature and techniques used in connection with cell and tissue culture, molecular biology, and chemistry and hybridization of proteins and oligonucleotides or polynucleotides as described herein. Is well known in the art and commonly used. For example, standard techniques are used for the purification and preparation of genetic material (nucleic acid), chemical analysis, and oligonucleotide synthesis. Enzymatic reactions and purification techniques are performed according to the manufacturer's specifications or as commonly accomplished in the art or as described herein. The techniques and procedures described herein are generally in accordance with various general references and more specific examples that are cited and discussed in accordance with conventional methods well known in the art and throughout the specification. This is done as described in the various references. The nomenclature utilized in connection with the present specification, as well as the laboratory procedures and laboratory techniques described herein, are well known and commonly used in the art.

  Numerous specific details are set forth below for purposes of explanation and illustration of various aspects of the invention. However, those skilled in the art will appreciate that the present embodiments may be practiced without the use of these specific details. In other instances, well-known structures and devices are shown or discussed more generally for purposes of efficient illustration of the invention and to avoid obscuring aspects of the invention. It is done. It should be noted that there are a wide variety of alternative configurations, devices, software, and techniques to which the disclosed invention and embodiments describing the invention can be applied. The various embodiments and the full scope of the invention itself are not limited to the specifically described examples.

  Where a range of values is provided, unless otherwise specified by context, each intermediate value between the upper and lower limits of the range, up to one-tenth of the lower limit unit, and any within the specified range It will be understood that other defined or intermediate values are included within the scope of the present invention. The upper and lower limits of these smaller ranges can be individually included within that smaller range, and are also encompassed within the scope of the present invention, but any specifically within its stated range. Follow the excluded limits. Where the specified range includes one or both of the limits, ranges excluding either or both of those included limit values are also encompassed by the invention. Unless otherwise indicated, all numbers representing amounts of reactants, reaction conditions, etc. used in the specification and claims are understood to be modified in all cases by the term “about”. I want.

  It should be understood that the steps recited in any method claims attached herein can be performed in any order unless otherwise specified. For example, in the method claims describing step (a), step (b), and step (c), step (c) may be performed before step (a) and step (b), or It can be performed during the process. Furthermore, although the individual steps are described as separate steps, they can be performed during periods of partial or complete overlap.

  As used herein, “regulating skin condition” means improving skin condition and / or prophylactically regulating skin condition, eg, protecting tissue from ultraviolet radiation And regulating the signs of skin aging. As used herein, “improving the condition of mammalian skin” means bringing about a favorable change in the appearance and feel of tissue that is visually and / or tactilely perceptible. Conditions that can be adjusted and / or improved include, but are not limited to, one or more of the following: wrinkles and rough deep fine lines, fine lines, cracks, protrusions, and the appearance of large pores. Reducing; thickening the skin (e.g. building the epidermis and / or dermis and / or subdermal layer of the skin, to reduce skin, hair, or nail atrophy, Applicable to the stratum corneum); increasing the dermis-epidermal boundary (also known as interpapillary ridge); preventing loss of skin or hair elasticity, eg loss of functional skin elastin, damage And / or by inactivation resulting in conditions such as elastic fibrosis, sagging, loss of recoil from skin or hair deformation; reduction of cellulite; skin, hair, or Caused by changes in coloration, such as dark circles under the eyes, smudged state (eg due to uneven red coloration, eg due to rosacea), poor color, telangiectasia or spider angioma Discoloration, dryness, brittleness, and gray hair.

  As used herein, “indication of skin aging” includes visual and tactile perceptible manifestations on the outer surface and any macroscopic or microscopic effects of skin aging. For example, but not limited to. These symptoms may arise from the processes listed below, but are not limited to those processes: wrinkles and rough deep wrinkles, fine wrinkles, skin wrinkles, tears, protrusions, large pores, non-uniformity, or roughness, etc. Development of discontinuities in tissue structure; exfoliation; drying; loss of skin elasticity; discoloration (including dark circles under the eyes); smudged state; poor colour; poor pigmentation such as aging stains and buckwheat Excessive skin area; keratosis; abnormal differentiation; hyperkeratinization; elastic fibrosis; collagen degradation and other histological changes in the stratum corneum, dermis, epidermis, vasculature (eg, telangiectasia) Or spider angioma), and other tissue structure changes within the underlying tissue (eg, lipids and / or muscles), particularly in the underlying tissue proximate the skin.

  “Hyperpigmentation” as used herein refers to areas of skin where the pigmentation is greater than the pigmentation of adjacent skin areas (eg, pigmented spots, aging spots, etc.).

  As used herein, “personal care composition” means a composition suitable for topical application on mammalian skin. The personal care compositions described herein may contain one or more skin care actives. “Skin care active substance” or “active substance” as used herein provides, for example, a benefit or improvement to the skin in regulating the condition of the skin, and the condition of other mammalian skin Means a compound that helps by

  “Energy delivery device” as used herein means any device used to deliver energy to mammalian skin and / or hair. As used herein, “supplying energy” means that the surface and / or layer of the skin is exposed to energy emanating from the energy supply device, and that energy includes any desired hair shaft and / or hair follicle. It means that it can penetrate the skin layer.

  “Sustained level”, as used herein, means that the energy supplied by the device, ie, the energy output, is at an essentially constant level between the start-up of the device and the non-operation of the device. It means that it is maintained.

  “Pulsed”, as used herein, means that the energy output varies in a predictable manner between the start-up of the device and the non-operation of the device, and the higher output Periods (pulses) and lower power periods are characterized by alternating appearances. The start of the pulse may be abrupt or gradual. “Predictable” means that the pulse peak intensity, pulse shape, pulse duration, and time interval between pulses are substantially the same. The duration of the pulse and the time between pulses may vary.

  “Handheld” as used herein means that the device is of a weight and dimensions suitable for an average adult to comfortably hold.

  Human skin can be divided into two major structural layers, the epidermis and the subepidermal or underlying dermis. The epidermis with a stratum corneum serves as a biological barrier to the environment. Within the basal layer of the epidermis are pigmented cells called melanocytes, which are the main determinants of skin color.

  The underlying dermis provides the primary structural support for the skin. The lower dermis is mainly composed of an extracellular protein called collagen. Collagen is produced as a triple helix produced by fibroblasts and having three polypeptide chains connected by a thermolabile chemical bond and a thermostable chemical bond. When the collagen-containing tissue is heated, the modification of the physical properties of the protein matrix occurs at the characteristic temperature. The structural transition of collagen contraction and remodeling of the collagen matrix occurs with heat.

  Inside the skin, a certain amount of repair activity occurs and promotes continuous collagen production. During aging, the rate at which damage occurs may increase faster than repair activity, or damage may continue to occur at a substantially constant rate, but repair activity is delayed. In either case, the result is a reduction in collagen and manifestation of impaired appearance as a sign of aging, including the appearance of surface defects such as fine wrinkles, wrinkles, and hyperpigmentation. is there. The normal repair activity cycle and the resulting effects on appearance over time are shown in FIGS.

  In FIG. 1, the box represents the normal cause of collagen production. For many reasons, the increase in collagen in the skin and the rate at which the collagen is repaired and replaced contributes to a more complete, healthier, more attractive-looking skin. Continuous growth and dermal remodeling occurs naturally, but unfortunately these processes slow down as they age. Collagen is also produced when the skin is damaged, for example after inflammation or injury, for example by radiation from the sun. The circles in FIG. 1 represent some of the mechanisms by which collagen is formed and the ways in which collagen formation can be tracked. Only MMP and cytokine activity are two measurable quantities that serve to track the cycle of collagen repair and replenishment.

  FIG. 3 is a schematic diagram of a dual action biological model 20 for the efficacy of elure. The skin 21 is classified into three layers: a stratum corneum 30, an epidermis 32, and a dermis 34. Collagen remodeling and the production of new collagen occurs within the dermis layer. Normal injuries 22 to the skin 21 occur constantly and include normal aging, UV injuries, pH changes, chemical injuries and the like. Normal injury 22 results in damage to collagen 24. Similarly, normal low levels of inflammation 26 occur, resulting in cytokine, HSP, and HSF activity. Ultimately, repeated low level thermal energy produces an increase in MMP activity 36, resulting in the production of collagen fragments. Increased heat causes an upregulation 38 of cellular activity that causes collagen formation and repair. Both mechanism 36 and mechanism 38 result in the formation and remodeling of healthy collagen 28.

  Collagen is measured to have a melting temperature of up to 50 ° C., but the repair cycle can be modified and enhanced by the addition of heat at lower levels. Thermal cleavage of intramolecular hydrogen bond bridges is created by an equilibrium between cleavage events and relaxation events (hydrogen bond re-formation). No external force is required to generate this process. As a result, intermolecular stresses are created by thermal cleavage of intramolecular hydrogen bonds. The contraction of the tertiary structure of the bridging molecule creates a contraction vector between the initial molecules. The RF model heating curve is shown in FIG.

  The dermis structure is mainly composed of collagen 1. Collagen is expressed as pro-collagen, a single chain protein, by fibroblasts. After expression, procollagen is cut into collagen 1 and folded into a triple helical conformation called “tropocollagen”. RF model heating curves are shown in FIGS. 8A, 8B, and 9. FIG.

  The dermis structure is mainly composed of collagen 1 (50 in FIGS. 2A and 2B). Collagen is expressed as pro-collagen 52, a single chain protein, by fibroblasts. After expression, procollagen 52 is cut into collagen 1 (50) and folded into a triple helical conformation called “tropocollagen” 54. This process is illustrated in FIGS. 2A and 2B.

  Collagen cross-linking may be intramolecular (covalent or hydrogen bonding) or intermolecular (covalent or ionic). Collagen denaturation as a function of aging is due to thermal energy injury, mechanical injury, the effect of pH on collagenase and MMP rates, hydration status, and collagen fibrils “zipped” or “unzipped”. And the general disruption of the natural equilibrium of collagen fibrils, which can be easily digested by MMPs. These represent a number of injury types, but all are rate controlled by temperature. Furthermore, normal collagen turnover cycles can be adjusted within a temperature range of <37 ° C to 43 ° C.

  Collagen bond breakage also occurs at lower temperatures, albeit at a slower rate. Low levels of thermal cleavage are frequently associated with relaxation phenomena, where bonds are reshaped without a net change in molecular length.

  Dermal remodeling is a biophysical phenomenon that occurs at the cellular and molecular level. Molecular contraction or partial denaturation of collagen involves the application of an energy source that destabilizes the longitudinal axis of the molecule by breaking the heat labile bond of the triple helix. As a result, stress is created and breaks the matrix intermolecular bonds. This is essentially an extracellular process, but cell contraction requires a lag phase with respect to fibroblast migration and proliferation into the damaged area. The healing response generally involves an early inflammatory process, which consists of wetting by leukocytes that dispose of cellular debris. This process is followed by an increase in turnover with a final increase in the collagen available for deposition and / or deposition at the site of injury. Fibroblasts differentiate into contractile myofibroblasts, the source of contraction of the cell's soft tissue. Following cell contraction, collagen is placed as a static support matrix within the constricted soft tissue structure. The deposition and subsequent reconstruction of this new scar matrix provides a means for altering soft tissue consistency and geometry for aesthetic purposes.

  The application of thermal energy, causing damage and repairing the cascade to ultimately achieve an improvement in skin surface appearance, is known in the art, but is currently known in the art Accompanying defects. For example, laser delivery devices use specific wavelengths of light that penetrate the skin, bind to specific chromophores, and remove various colors and pigments from the skin through a process called selective photothermal melting. Lasers are large and expensive capital equipment that attacks only certain problems or colors in the skin, is prone to laser burns, scarring, can cause hyperpigmentation and / or reduced pigmentation, This can result in eye trauma to the user and patient. Intense broadband light systems emit light of multiple wavelengths, improve skin discoloration through selective photothermal melting, and improve skin texture non-specifically through skin heating. This system is also large and expensive, with improved skin texture and wrinkles, and there is also a risk of skin burns, reduced or hyperpigmentation, and scarring. In general, the application of electromagnetic energy is achieved across the epidermis with penetration limited by surface pigmentation factors and the composition of the dermis layer.

  High frequency technology is also known in the art of skin treatment. RF technology uses electrical current to heat the dermis and stimulate the production of collagen and elastin fibers that firm and tighten the skin. However, there are substantial drawbacks in the state of the art because treatment parameters cannot be optimized. Application of RF current creates a temperature gradient in the skin that is the opposite of other thermal energy delivery techniques. The administration of RF current is conducted between electrodes that are spaced apart at some distance on the skin. Current is conducted between the electrodes through the dermis, causing the dermis temperature to rise more rapidly than the temperature at the skin surface. Since most skin parameter measuring devices are designed for skin surface measurements, excessive heating of the dermis may be performed before it is recognized.

  Therefore, treatment with RF currents in the absence of thermal quenching, mechanical cooling, or other techniques designed to control internal heating is avoided in the art.

  However, taking advantage of recent advances in biotechnology, genomics and proteomics, we have developed a method to evaluate the effects of dermal administration of RF currents. Specifically, we screened a group of potential genes identified as being involved in the dermal collagen matrix, dermal inflammation and remodeling, and epidermal differentiation. Based on the resulting gene expression profile, analysis and scrutiny of differential signature modulation of potential genes when gene signatures and gene chip components are subject to specific RF current treatment parameters, conditions, and regimens Judged.

  The energy delivered to and / or into the skin layer can be in the form of RF energy, including, for example, radio frequency and microwaves. Exemplary RF energy devices are described in US Pat. Nos. 6,889,090, 6,702,808, 6,662,054, 5,569,242, and 5,755. 753, 6,241,753, 6,430,446, 6,350,276, 5,919,219, 5,660,836, Nos. 6,413,255, 6,228,078, 5,366,443, and 6,766,202.

  RF application and treatment regimens, including the administration of RF current through the dermis, can be designed for the first time to optimize the desired effects of RF treatment. Surprisingly, by analyzing and exploring gene expression data, potential gene datasets have increased dermal collagen, and the desired hormetic stress-induced dermal remodeling of the more problematic inflammatory cytokine damage response. It has been found that in optimizing a treatment regimen to be provided in the absence, it can be combined into three subsets with particular utility. To the best of our knowledge, this represents the first time genomics have been applied to cosmetic procedures that employ RF current technology. The expression profile provides a favorable hormetic stress that allows controlled and optimized administration to initiate and sustain the desired dermal remodeling while avoiding the traditional damage associated with undesirable biological effects. Clarify what will result. The present invention provides novel gene chips, gene signatures, screening methods, and optimizable regimens based on these findings.

  Genes considered as potential genes include genes associated with dermal matrix integrity (FBN1, FBLN1, TNXB, FN1, LOXL2, COL3A1, COL1A1, ELN, and LOXL1), genes associated with dermal inflammation initiation remodeling Genes (TIMP2, IL1A, TIMP1, TNF, MMP1, MMP9, MMP3, SOD2, and IL1B) and genes related to epidermal barrier function (KRT2, KRT6A, CLDN1, LOR, FLG, IVL, DRT10, AQP3, and KRT14) ). Subsets derived from the analysis of expression data for these genes include Table 1 (dermal marker), Table 2 (matrix remodeling marker, ie, the initiator of the preferred hormetic stress), and Table 3 (inflammatory cytokine response, ie, Marker of damage outside repair reaction).

  A gene expression profile provides information about the cellular response to a set of conditions. A gene includes instructions for making messenger RNA (mRNA). However, at any given point in time, each cell makes mRNA from only a portion of the genes carried by each cell. A gene is said to be “on” when it is used to produce mRNA, otherwise it is said to be “off”. The term “regulation” refers to causing a transcriptional state that is different from the regulatory state of a gene. For example, “up-regulation” may include simply turning on or may refer to increasing the transfer rate beyond a reference rate derived from control or reference conditions.

  In the expression profile, the relative amount of mRNA expressed under two or more experimental conditions is measured. Changes in the level of mRNA suggest an altered need for the protein encoded by the mRNA. For example, an increase in transcription of an enzyme catalyst or enzyme cofactor is observed in response to an increase in the concentration of the enzyme's substrate within the cellular environment.

  Overall, gene expression profiles include genes that demonstrate significant differences under changing experimental conditions. This is typically a subset of some data set that may include the entire genome. For one type of cell, a group of genes whose combination of expression patterns is uniquely characteristic for a given condition constitutes a gene signature for that condition. Genetic signatures can be used, for example, to select patients who can benefit from a particular treatment, or to design treatment protocols that maximize the desired signature.

  Gene expression profiles for potential gene datasets can be determined using microarrays. Exemplary cDNA microarrays are commercially available and are available from Agilent Technologies, Affymetrix Inc. (Santa Clara, Calif.), Nanogen (San Diego, Calif.), And Protogene Laboratories (Palo Alto, Calif.). Specific hybridization techniques that can be similarly practiced to generate expression profiles employed in the subject methods include US Pat. No. 5,143, the disclosure of which is incorporated herein by reference. , 854, 5,288,644, 5,324,633, 5,432,049, 5,470,710, 5,492,806, 5,503,980, 5,510,270, 5,525,464, 5,547,839, 5,580,732, 5,661, No. 028, No. 5,800,992, and WO 95/21265, No. 96/31622, No. 97/10365, No. 97/27317, EP No. 373203, and 7852 The techniques described are exemplified by No. 0. These techniques generally involve contacting an array of “probe” nucleic acids, including probes for each of the phenotyping genes whose expression is being assessed, with a target nucleic acid as described above. Contact is performed under hybridization conditions, eg, stringent hybridization conditions, and then unbound nucleic acid is removed. The resulting hybridized nucleic acid pattern provides information related to expression for each of the genes sought, which expression information typically indicates at what level the gene is expressed. This expression data, ie the expression profile, can be both qualitative and quantitative. Alternatively, the expression profile is determined by quantitative PCR or by other quantitative methods for measuring mRNA.

  One embodiment of the present invention provides a gene that is regulated in mammalian skin in response to the generation of sufficient high frequency current within the tissue volume of mammalian skin to heat the tissue volume to a treatment temperature. A gene panel is provided. At least one gene is selected from Table 1 or Table 2, and at least one gene is selected from Table 3. In a specific embodiment, the at least one gene is selected from each of the three tables. All gene panels according to the present invention are conceived to contain at least one gene from Table 3, which is indicated by the lack of a change in the expression of those genes to indicate a lack of a “malignant” damage response. Because. Probes are used to construct highly specific microarrays that have utility in designing, selecting, adapting, or monitoring treatment regimens for desired effects, or for validation of treatment regimens in subjects being treated. It can be designed to target each gene constituting the gene chip of the present invention. A microarray comprising a set of immobilized nucleic acid probes capable of hybridizing and detecting the genes constituting the gene panel according to the present invention is envisaged.

  As used herein, a “probe” is a naturally occurring or synthetically produced capable of specifically hybridizing with a nucleic acid having a sequence complementary to the probe. An oligonucleotide, polynucleotide, or DNA molecule that is either. The probe of the present invention specifically refers to an oligonucleotide attached to a solid support in a DNA microarray substrate. The probe can be either single stranded or double stranded. The probe typically contains 15 to 25 or more nucleotides, but may contain fewer nucleotides. The probes herein are selected to be complementary to the various strands of a particular target nucleic acid sequence, and thus specifically hybridize with the corresponding target strand under a set of predetermined conditions. It must be sufficiently complementary so that it is possible. Therefore, the probe sequence need not reflect the exact complementary sequence of the target. For example, non-complementary nucleotide fragments can be attached to the 5 'or 3' end of the probe and the remainder of the probe sequence can be complementary to the target strand. Alternatively, if the probe sequence has sufficient complementarity to the target nucleic acid sequence and specifically anneals to the target nucleic acid sequence, non-complementary bases or longer sequences can be introduced into the probe. Can be scattered.

  Another embodiment of the invention includes a method for providing benefits to mammalian skin. This benefit includes inducing collagen formation and / or dermal remodeling in the dermal layer of mammalian skin in the absence of an inflammatory cytokine response that damages the skin, the method comprising: Within the tissue volume of the skin, including generating a high frequency current for a sufficient treatment cycle to heat the tissue volume to the treatment temperature, while avoiding upregulation of the expression of the genes listed in Table 3. . The RF current can be generated multiple times within one treatment cycle.

  As used herein, terms such as “treat” and “treatment” generally refer to obtaining a desired cosmetic or aesthetic effect supported by a target biological response. . "Treatment" as used herein includes treatment in mammals, specifically humans, and (a) cosmetically undesirable skin conditions such as fine wrinkles, wrinkles, hyperpigmentation spots, And preventing or avoiding the development of other skin irregularities that result from aging or environmental aging, or impact on the skin, (b) preventing the appearance of cosmetically undesirable skin conditions, Remission or delay, (c) reverse or regress cosmetically undesirable skin conditions.

  One or more treatment cycles according to the present invention may be performed throughout the treatment period. Treatment cycles can be as short in duration as necessary to produce the desired response. In specific embodiments, the treatment cycle is about 1 minute or less, but in other specific embodiments, the treatment cycle is longer than about 1 minute. In other specific embodiments, the treatment cycle lasts from about 1 to about 6 minutes, and in other embodiments from about 2 to about 6 minutes. A treatment period according to the present invention comprises one or more treatment cycles, and in a specific embodiment is at least one week and comprises at least one treatment cycle. In another specific embodiment, the treatment period is from 1 week to 12 weeks. In a more specific embodiment, the treatment period is between 3 weeks and 8 weeks. In certain embodiments, each week of the treatment period comprises 1 to 6 treatment cycles, although it will be understood in the art that this may vary with the unique characteristics of the individual being treated.

  In general, administration of RF current produces a higher temperature in the dermis than the skin surface, but the temperature of the skin is measured on the surface in the most convenient and non-invasive manner. Thus, in certain embodiments, the treatment temperature, defined herein as the temperature of the tissue volume through which RF current is conducted, is less than about 45 ° C., the temperature of the skin surface above the tissue volume. Give rise to In a more specific embodiment, the treatment temperature produces a skin surface temperature above the tissue volume of about 37 ° C to about 43 ° C.

  A desired benefit according to the present invention is that mRNA is extracted from a sample taken from a tissue volume through which RF current is conducted and at least one gene selected from Table 1 and / or Table 2 and Table 3 Can be evaluated by determining the expression profile of a gene panel comprising at least one gene selected from The benefit is indicated by the expression profile reflecting the upregulation of the gene selected from Table 1 and / or Table 2 combined with substantially no change in the expression of the gene selected from Table 3 And instructing that benefits are being provided.

  Overall, gene regulation according to the present invention is reflected in the expression fold change data. There are many methods for the analysis of microarray-based experiments that identify genes that are differentially expressed between conditions, and the choice of that method can affect the set of genes identified. Known in the art. Magnification changes are believed to provide the most reproducible results. The fold change can be defined as the ratio between the average control observation and the average treatment observation (or the difference between the average log control data and the average log treatment data). A magnification change of 1 therefore represents no change over the control observation. A positive fold change indicates an increase in expression over a period of time, referred to herein as up-regulation, and a negative fold change is expressed over a period of time, referred to herein as down-regulation. Direct the decrease. The significance of fold change can be determined by conventional statistical methods.

  According to certain aspects of the invention, the benefits can be optimized by adjusting treatment temperature and / or other treatment parameters in response to the expression profile, from Table 1 and / or Table 2. While upregulation of the selected gene is maximized, the expression of the gene selected from Table 3 is maintained substantially unchanged. The treatment regimen of the present invention can include maintenance treatments, and benefits are maintained beyond the treatment period by one or more maintenance treatments, each maintenance treatment comprising at least one maintenance cycle.

  The skin treatment regimen of the present invention contemplates the application of thermal energy through electrically conducted RF current via a handheld RF current supply device. A preferred RF delivery device according to the present invention delivers RF as an electrically conducted current between two electrodes placed on the skin at some distance from each other. The delivery device that produces the benefits of the present invention is the absence of electromagnetic radiation at visible or infrared frequencies in the electromagnetic spectrum and the absence of a supplemental monochromatic or polychromatic light source directed towards the tissue volume. Underneath as well as in the absence of mechanical cooling, high frequency current is conducted through the tissue volume.

  Certain embodiments of the present invention provide the ability to evaluate the effectiveness of any energy supply device or combination of energy supply technologies in providing the benefits of the present invention. Therefore, a specific method for assessing the treatment effectiveness of an energy delivery device, designed to provide benefits to the skin by heating the skin, treats the skin by application of the energy delivery device. And extracting mRNA from the treated skin sample and generating an expression profile for the gene panel according to the present invention. As described in Example 3 below, an RF current supply device exists that includes parameter specification tolerances that can be dialed to produce an expression profile according to the present invention. A suitable device is manufactured by Syneron. An example of a device that is not designed to achieve the genomic reaction goals of the present invention is a handheld personal use RF delivery device manufactured by Ultragen Ltd, distributed in Europe under the trademark STOP ™. This device is also described in Example 3 for purposes of comparison, but cannot cause upregulation of the genes in Tables 1 and 2. This device specification is designed to accommodate a regimen based on the provision of substantially smaller thermal energy applied over a larger skin area, and the preferred homology achieved by the device and regimen according to the present invention. The level required to achieve a heterogeneous effect cannot be reached.

  According to another embodiment of the invention, a method for screening a facial skin treatment regimen is provided. This method is screened for effectiveness in providing collagen and / or dermal remodeling benefits to mammalian skin without stimulating inflammatory cytokine responses that damage the skin. The method generally includes treating facial skin according to a treatment regimen, extracting mRNA from a sample of treated facial skin, creating a gene expression profile for a gene panel according to the present invention, Comparing the gene expression profile with a reference profile, and the expression profile shows a substantial lack of up-regulation of genes selected from Table 1 and / or Table 2 and regulation of genes selected from Table 3 Determining if the facial treatment regimen is effective. In a specific embodiment, the facial skin treatment regimen includes using a high frequency current generating device to generate a pulsed high frequency current that passes through the first tissue volume of the facial skin over the treatment cycle. The treatment regimen may include moving the high frequency current generating device to generate a pulsed high frequency current passing through the second tissue volume of the facial skin during the treatment cycle.

  In the sorting method, the area of skin is selected for matching with the characteristics of the target treatment area, in particular the facial characteristics. Cosmetically vulnerable facial skin areas include the periorbital and peroral areas. Therefore, the facial skin around the ear, known as periauricular skin, is structurally similar to the target treatment area, but is easily visible in the case of wrinkles that can result from simple biopsy procedures. Not. Thus, in embodiments of the invention where a biopsy is required, the required sample is taken by biopsy of the treated periauronic skin, and the reference is by biopsy of the pre-treatment or non-treated perineural skin. Collected. In general, pre-treatment or untreated periauricular reference skin includes skin that is substantially adjacent to the treated periauricular skin. In a very specific embodiment, the expression profile of a gene selected from Table 1 and / or Table 2 reveals a fold change over reference that is statistically greater than 1.

  Obviously, the present invention can provide benefits to skin other than facial skin, however facial skin is most readily manifested in the effects of both aging and environmental aging and damage, and the body's It is highlighted in the illustrative example because it poses the greatest cosmetic treatment challenge in that it is initially thinner and more fragile than the other areas. Furthermore, facial skin is a very aesthetically important area for consumers of cosmetic treatment techniques.

  In certain embodiments, the benefits provided by application of RF current can be reinforced or enhanced by the application of a cosmetically active substance or composition to the facial skin in conjunction with the generation of pulsed radio frequency current. This active substance or composition may have an enhancing or synergistic effect on the gene expression profile according to the invention. Non-limiting examples of suitable cosmetically active substances include retinol propionate and its derivatives, caffeine, hyaluronic acid, and generally plant extracts.

  The sorting method according to the present invention includes substantially the same parameters as the treatment method according to the present invention with respect to the treatment cycle, treatment period, treatment temperature, and delivery device, and these parameters are adjusted to the desired sorting tolerance.

  According to a further embodiment of the present invention, a genetic signature of a differentially expressed gene suitable for identifying cosmetic skin benefits is provided. This benefit includes inducing collagen formation and / or dermal remodeling in the dermal layer of mammalian skin in the absence of an inflammatory cytokine response that damages the skin. , At least one gene selected from each of Table 2 and Table 3. In general, a gene signature is a subset of genes obtained from a data set of genes associated with a particular property, trait, or biological function. A gene signature can be obtained from all or part of a genetic data set, and a signature according to the invention can be from any number of genes from at least about two genes or up to a number comprising the entire data set. Information may be included. When using a subset of a data set, the subset can include genes that are up-regulated, genes that are down-regulated, genes that are not substantially regulated, or combinations thereof.

  The method of the present invention includes applying a first personal care composition, and optionally a second personal care composition, to an area of mammalian skin. The first and second personal care compositions can be in a variety of forms, including lotions, creams, serums, foams, gels, sprays, ointments, masks, sticks, moisturizers, patches, Examples include, but are not limited to, powders and / or wipes. In one embodiment, the first personal care composition is applied before and / or during the supply of energy. In an alternative embodiment, the second personal care composition is applied after application of the first composition and supply of energy. Optionally, the method of the present invention includes the step of applying a third personal care composition to the skin, the third composition including a conditioning agent. In one embodiment, the third personal care composition is applied prior to the application of the first personal care composition. Preferably, the third personal care composition is applied at least 24 hours prior to the delivery of energy. In alternative embodiments, the first personal care composition is applied twice a day and energy is supplied once a day, alternatively once a week, and alternatively once a month. In one embodiment, the first personal care composition is applied to the skin twice daily and energy is delivered to the skin once a week.

  The first, second, and third personal care compositions can include a variety of ingredients, non-limiting examples of which are found in The CTFA International Cosmetic Indicative and Handbook, Tenth Edition (2004). Can do.

  The composition of the present invention may comprise from 50% to 99.9% of a dermatologically acceptable carrier. The carrier of the present invention is in the form of an emulsion. As used herein, “emulsion” generally includes an aqueous phase and an oil phase. The oil can be derived from animals, plants, or petroleum, can be natural or synthetic, and can include silicone oils. Emulsion carriers include, but are not limited to, oil-in-water, water-in-oil, water-in-oil-in-water, and oil-in-water-in-silicone emulsions. In one embodiment, the dermatologically acceptable carrier comprises an oil-in-water emulsion, and alternatively a silicone-in-water emulsion. The emulsion may further comprise humectants such as glycerin and nonionic, cationic and / or anionic emulsifiers. Suitable emulsifiers are described, for example, in US Pat. Nos. 3,755,560 (Dickert et al.), 4,421,769 (Dixon et al.), And McCutcheon's Detergents and Emulsifiers, North American Edition, pages 317-324. (1986).

A wide range of amounts of the composition of the present invention can be employed to improve skin conditions. The amount of personal care composition applied to the skin can vary depending on the location on the body and the desired benefit. Exemplary amounts include ^{0.1mg / cm 2 ~40mg / cm 2} . The amount of One useful application is ^{0.5mg / cm 2 ~10mg / cm 2} .

  Temperature changes can be induced simultaneously in the skin or in a composition applied to the surface of the skin. This temperature change is independent of any temperature change induced by the supplied energy itself. For example, the skin can be heated before the supply of energy, or the skin can be cooled before, during and / or after the supply of energy.

  Using post-treatment gene panels and gene signatures and extensive consumer research, a regimen region was developed that optimizes the size of the treatment area, and an optimal treatment time was developed. Contrary to the teaching in the art, the use of a wide area device has been found not to be the best treatment method. If an excessively large area is treated, the first part may cool and return before the consumer sweeps over that part again. If an excessively small area is treated, the consumer runs the risk of overtreatment and unnecessary damage. Furthermore, the treatment itself is a task that requires concentration for the consumer, and dividing the treatment into modest amounts makes it easier for the consumer to comply with the regimen completely. Similarly, a personal care composition used with a device allows a consumer to apply the personal care composition to a small area to treat the area and then remove any remaining personal care composition. If it can, it is much more attractive to consumers. Placing the personal care composition over the entire face or even over half of the face can make the treatment experience uncomfortable. This also adversely affects consumer compliance.

  Referring now to FIG. 10, a wrinkle area 60 on the outer corner of the consumer 62 is identified by a dashed circle and occurs above and below the eye 64 adjacent to the outer corner of each eye. It is well known that the wrinkle area 60 of the corner of the eye tends to form wrinkles and fine wrinkles with human aging and is subject to environmental injury. FIG. 11 shows region A (70), and arrows within region A are shown to demonstrate an exemplary treatment path for a device (not shown). The number of treatments has been described above. FIG. 10 shows a region B (72) that is on the same eye of the consumer 62 and overlaps the crease area 60 of the corner of the eye. Exemplary arrows are shown to illustrate possible treatment paths. By defining region A (70) and region B (73) in this manner, the corner area 60 of the eye corner is treated with twice the length of the area A and region B that is not the corner of the eye corner. . This is an important finding because most of the fine wrinkles and wrinkles on the consumer's face that require treatment are in the wrinkle area of the corner of the eye.

  13 and 14 show a region C (74) and a region D (76) on the other eye 64 of the consumer 62. FIG. As expected, these treatment areas are substantially similar to Region A and Region B and are treated in the same manner.

  An alternative treatment protocol is shown in FIGS. 15 and 16, with region E (80) and region F (82) shown on consumer 62. FIG. These “C” shaped treatment areas treat the eye corner wrinkle area 60 of FIG. 10 only once for each pass, while this longer “C” shaped treatment area is used once more by the device. Allowing each area of the skin to be treated to cool slightly before returning to pass. Thus, although consumer comfort is improved, the crease area 60 in FIG. 10 is only treated during one treatment cycle.

  The second personal care composition can be used in conjunction with the methods described above, if desired. The second personal care composition can be used between successive treatment periods employing the first personal care composition and a thermal heating device. The second personal care composition preferably comprises at least one skin care active that is not present in the first personal care composition.

  The following examples are provided to illustrate specific features and advantages of various embodiments of the invention and should not be construed as limiting the scope of the invention.

Example 1 Treatment Protocol This example describes the treatment of the periauricular area of facial skin with the aim of performing genomic and histological assessments. All treatments are therefore administered by a technician within the clinical setting for control purposes. Treatment is administered 3 times a week for 6 weeks for 4 regions within the treatment area, 4 minutes per region for a total of 16 minutes. This protocol applies to this procedure only in the peri-ear area on one side, and applies the sham procedure consisting of gel and power-free device to the corresponding per-ear area in the other side. This is due to the split face / neck study in the design. The thermal imaging camera is used to ensure that the target skin surface temperature of 40-43 ° C. is reached and maintained within the first minute during each 4 minute treatment. Monitored at.

  The treatment site is centered and placed directly under the ear. The site is about 1.5 inches wide and about 4 inches long. If the area of skin to be treated is too dry or too hot, a separate gel can be applied. The subject's face and neck are clean and free of lotions, fragrances, and the like. The hair is clipped backwards from the treatment area.

Device action:
1. Apply one pump (about 1.4 g gel) to the treatment site and spread evenly without wiping. Additional gel pumps to the cheeks above the treatment area can be applied as needed during the treatment to cool the skin if the subject expresses that the skin becomes too hot. .
2. Switch on the device and set a 4-minute timer.
3. The applicator head of the device is applied to the skin and moved along the treatment site using back and forth motion and light contact. Maintain connectivity while manipulating the head of the device throughout the treatment area. Proper contact with the skin is indicated by a lens on the applicator that should continue to flash red to indicate contact.
4). Thermal camera images are monitored during the procedure. Within 30-60 seconds, the skin in the treatment area reaches an optimum temperature of 40-43 ° C. A color pattern on the thermal image indicates the temperature. In general, as the device warms, the applicator is moved quickly across the skin to avoid discomfort due to elevated temperatures. As the procedure continues, the movement is slowed to maintain an ideal temperature. The subject is warned to feel the device warm but not to burn the subject.
5. If the thermal image shows a reddish area, the temperature is above 43 ° C. The temperature can be increased on the cheek by increasing the treatment speed of the fore-and-aft movement, by slightly increasing the size of the treatment area, by alternating back-and-forth movement and the figure 8 pattern, or in preparation for this contingency event. It is controlled by adding more gel by bringing the applicator head part into contact with the attached separate gel.
6). If the treatment area cannot reach the optimum temperature, or if the treatment area cools, the back and forth movement should be slowed until most of the treatment area is at the target temperature.

Example 2: Biopsy Protocol This example is a relatively tolerant biopsy used to collect samples for evaluation and monitoring purposes according to certain aspects of the invention and support data disclosed herein. The inspection protocol will be described.

  A clinical biopsy study was performed on 30 adult female subjects using an RF current delivery device. Treatment was provided in the peri-ear (around the ear) area of the facial skin. Periauricular skin is known in the cosmetic art as a suitable periorbital substitute. Treatment was administered clinically using a Syneron V8 device (see Example 3) and a 4 minute heating profile as the target profile. Subjects received treatment 3 times a week for a total of 6 weeks. A biopsy was performed to examine the skin tissue structure as a function of treatment and create a genomics profile.

Biopsy technique:
One 4 mm biopsy was taken from the periauricular area from both the right and left sides of the neck (this area is located directly below the ear). Using a 30 gauge sterile needle, anesthetic containing 2% xylocaine with epinephrine is injected directly under the skin to be biopsied. After the subject indicates that the area is insensitive, an appropriately sized punch biopsy is collected using standard antibacterial techniques and subsequently sutured occluded. A 20% aqueous solution of aluminum chloride is used as necessary from the viewpoint of bioconstancy. Following the biopsy, the punch site is monitored for normal healing and the suture is removed after 7 days.

Sample handling after biopsy:
The biopsy tissue is divided into two separate samples, one for gene expression testing and the other for histological evaluation. The sample is equally divided in half by a line parallel to the line drawn from the stratum corneum to the dermis. Sample handling follows industry standards.

Example 3: mRNA extraction and analysis This example is an RNA biomarker that supports certain embodiments of the invention, including the discovery of biological models that guide gene chips, gene signatures, and RF delivery regimen design. The identification and analysis will be described.

  A 4 mm biopsy according to Example 2 was taken twice from both sides of the face / neck just below the ear as described above, once for 24 hours after the final treatment and once for 4 weeks after treatment.

  The biopsy sample was transferred to a 2 mL centrifuge tube containing 1.5 mL tissue storage reagent (RNAlater® solution, invitrogen, Life Technologies (Carlsbad, Calif.)). The tube was refrigerated overnight at 2-8 ° C. The storage reagent was removed and the samples were placed in a freezer at −80 ° C. until processed. Immediately prior to processing, samples were removed from the freezer and 1.5 mL of a single phase solution of phenol and guanidine isothiocyanate (TRIzol® reagent, Invitrogen, Life Technologies (Carlsbad, Calif.)) And one 3 mM tungsten carbide bead ( 3 mM tungsten carbide beads, Qiagen, catalog # 69997) were added to each tube. Samples were homogenized in a mixer mill (Qiagen Inc.) with 4 shakes at 30 / sec for 3 minutes with the adapter inverted after each round. Samples were centrifuged at 12,000 rpm for 10 minutes to remove necrotic tissue debris. The supernatant is then transferred to a pre-spun phase-locked gel heavy tube (Phase Lock gel heavy tubes, Eppendorf (New York, NY), catalog # 0032-005-152) and 300 μL of chloroform (Sigma) is added. The tube was shaken vigorously without vortexing excessively. The sample was centrifuged at 12,000 rpm for 10 minutes, after which the supernatant was transferred into a new 2 mL centrifuge tube.

  The binding state was adjusted by adding 800 μL of 70% ethanol, the tube was vortexed to mix and then spun briefly. 830 mL of sample was transferred to the RNeasy mini spin column in the vacuum manifold and vacuum was applied. The remaining sample (approximately 21.1 mm (830 mil)) was transferred onto the same RNeasy column and a vacuum was applied. 700 μL and 500 μL of buffer RW1 were continuously pipetted onto the RNeasy column, and contaminants were removed by applying a vacuum after each rinse.

  The RNeasy spin column was transferred to a new 0.05 mm (2 mil) collection tube, the tube was centrifuged at 14,000 rpm for 2 minutes, and the column was newly transferred into a 1.5 mL collection tube. Residual ethanol was aspirated from the inner ridge of each column. 30 μL of preheated Rnase free water was pipetted directly onto the RNeasy membrane, the membrane was incubated for 5 minutes and then centrifuged at 14,000 rpm for 2 minutes. The eluate was then collected to provide ready-to-use RNA in water.

  After separation, RNA yield was measured using Agilent Technologies, Inc. Determined using RNA 6000 Nano LabChip® kit # 5065- 4476 available from (Santa Clara, CA.). RNA was evaluated by a one-step RT-PCR method. For RT-PCR biomarker analysis, RNA was diluted to a final concentration of 5 ng / well.

  Purified RNA was converted to cDNA using an RT-PCR kit (QSScript ™ One-Step Fast MGB qRT-PCR kit, available from Quanta BioScience, Inc. (Gaithersburg, MD)). 500 nanograms of RNA was then mixed with the QScript enzyme / buffer mixture and subjected to a thermal cycler according to the kit instructions. 1 μL of the resulting cDNA was then mixed with Quanta Perfecta Master Mix and a custom array plate (Custom RT Profiler ™ PCR array, SABiosciences, Corp. (Federick) containing pre-verified primers for the following genes: , MD) available): KRT2, KRT6A, CLDN1, LOR, FLG, IVL, KRT10, AQP3, KRT14, FBN1, FBNL1, TNXB, FN1, LOXL2, COL3A1, COL1A1, ELN, LOXL1, TIMP2, IL1A, TIMP1, TNF, MMP1, MMP9, MMP3, SOD2, and IL1B. The array plate was then sealed and subjected to a thermal cycler (StepOnePlus ™ Real-Time PCR System Upgrade, Applied Biosystems, Inc. (Foster City, Calif.)).

  Data analysis was performed using data analysis software provided by SABiosciences (Frederick, MD.).

  Expression fold change data for the above 27 organized genes at the target time of 24 hours and 4 weeks are described in FIGS. 4A, 4B and FIGS. 6A, 6B, respectively. Genes associated with the dermal matrix are grouped as 90 and 91, genes associated with dermal inflammation reconstruction are grouped as 92 and 93, and genes associated with epidermal differentiation are grouped together as 94 and 95. The fold change data for an exemplary gene is collagen 1A1 in FIG. 5A, elastin in FIG. 5B, collagen 1A1 in FIG. 7A, and elastin in FIG. 7B. Note the co-regulation between many collagen 1A1 and elastin genes.

  The dimensions and values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Rather, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as “40 mm” shall mean “about 40 mm”.

  All documents cited herein, including any patents or patent applications that are cross-referenced or related, are wholly excluded unless expressly excluded or otherwise limited. Which is incorporated herein by reference. Citation of any document is not an admission that the document is prior art to all inventions disclosed or claimed herein, and that document alone or in any other way Neither is it acceptable to teach, suggest, or disclose any such invention in any combination with any of the above references. Further, any meaning or definition of a term in this document is assigned to that term in this document to the extent that it contradicts any meaning or definition of the same term in the document incorporated by reference. It shall conform to the meaning or definition.

  While specific embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.

  When the fold-change data is scrutinized and analyzed and grouped, surprisingly, if genes were isolated that showed very little regulation in response to treatment (IL1A, IL1B, and TNF), the genes are , Including inflammatory cytokine response genes, while genes associated with collagen synthesis and turnover and dermal remodeling were found to be upregulated under treatment conditions. The genes selected based on fold change data are therefore tabulated in Table 1, Table 2, and Table 3 below.

  Furthermore, it was surprisingly found that the favorable effect of the treatment regimen on gene expression, which was present at 24 months, was maintained almost undiminished at the off-target month. Maintenance treatment may be desirable to maintain the genomic effect over a longer period of time.

  The gene expression profile is expected to reflect downstream upregulation in the translation product. Although the observed fold changes may be relatively low, sustained up-regulation in the synthesis of certain proteins and enzymes over a long period of time has a substantially long-lasting cosmetic benefit to the skin. Considered to bring.

  Example 4 Personal Care Composition

  The following are exemplary personal care compositions for use with the devices of the present invention.

Claims (11)

  A gene panel comprising genes that are regulated in the mammalian skin in response to generation of sufficient high frequency current within the mammalian skin tissue volume to heat the tissue volume to a treatment temperature. A gene panel wherein at least one gene is selected from Table 1 or Table 2 and at least one gene is selected from Table 3.   The gene panel of claim 1, wherein at least one gene is selected from Table 1, at least one gene is selected from Table 2, and at least one gene is selected from Table 3.   A microarray comprising a set of immobilized nucleic acid probes capable of hybridizing and detecting the genes constituting the gene panel according to claim 1 or 2.   A method for assessing the treatment efficacy of an energy delivery device designed to provide benefits to the skin by heating the skin, said method comprising applying said energy delivery device 3. A method comprising treating skin, extracting mRNA from the treated skin sample, and generating an expression profile for the gene panel of claim 1 or claim 2.   A method for screening a facial skin treatment regimen for effectiveness in providing collagen and / or dermal remodeling benefits to mammalian skin without stimulating inflammatory cytokine responses that damage the skin. 3. The method comprises treating facial skin according to a treatment regimen, extracting mRNA from the treated facial skin sample, and generating a gene expression profile for the gene panel of claim 1 or claim 2. Comparing the gene expression profile with a reference profile, the expression profile being upregulated for a gene selected from Table 1 and / or Table 2, and the regulation of a gene selected from Table 3 Determining that the facial treatment regimen is effective when reflecting a general lack   The method of claim 5, wherein the facial skin treatment regimen includes generating a pulsed high frequency current through a first tissue volume of the facial skin over a treatment cycle using a high frequency current generating device.   The treatment regimen further comprises moving the high frequency current generating device to generate a pulsed high frequency current passing through a second tissue volume of the facial skin during the treatment cycle. the method of.   The facial skin comprises periauricular skin, the sample is taken by biopsy of treated periauronic skin, and the reference is taken by biopsy of pre-treatment or untreated perineural skin. Item 8. The method according to Item 7.   9. The method of claim 8, wherein the pre-treatment or non-treatment periauricular reference skin comprises skin that is substantially adjacent to the treated periauricular skin.   10. The method of claim 9, wherein the gene selected from Table 1 demonstrates a statistically greater fold increase in expression that exceeds the reference.   A genetic signature of a differentially expressed gene suitable for identifying cosmetic skin benefits, wherein the benefits are in the absence of an inflammatory cytokine response that damages the skin of mammalian skin Inducing collagen formation and / or remodeling of the dermis within the dermis layer, wherein the gene signature comprises at least one gene selected from each of Table 1, Table 2 and Table 3 .