COMPOSITIONS FOR PROMOTING SKIN REGENERATION, SKIN REJUVENATION, AND WOUND HEALING AND METHODS FOR PREPARATION AND USE THEREOF

Abstract

Compositions for promoting skin regeneration, skin rejuvenation, and wound healing that include tissues derived from the human placenta, such as the amniotic membrane, chorionic membrane, and umbilical cord. The placental tissues can be combined or compounded with aloe or allantoin or both to form topical treatment creams. Additionally, these compositions have therapeutic and cosmetic uses.

Claims

1-47. (canceled)

48. A composition for promoting skin regeneration, skin rejuvenation, and wound healing comprising: a micronized human placental powder comprising dried amnion and dried chorion present at an effective amount in the composition to exhibit bacteriostatic and/or bacteriocidal effects on at least one bacterium from E. coli, S. Aureus, P. aeruginosa, M. smegmatis, or Methicillin-Resistant Staphylococcus aureus (MRSA) after a twenty four hour time period of being contacted with the composition.

49. The composition of claim 48, further comprising Hepatocyte Growth Factor (HGF).

50. The composition of claim 49, wherein the composition exhibits bacteriostatic and/or bacteriocidal effects on at least two bacterium from E. coli, S. Aureus, P. aeruginosa, M. smegmatis, or Methicillin-Resistant Staphylococcus aureus (MRSA) after a twenty four hour time period of being contacted with the composition.

51. The composition of claim 50, wherein the composition exhibits bacteriostatic and/or bacteriocidal effects on at least three bacterium from E. coli, S. Aureus, P. aeruginosa, M. smegmatis, or Methicillin-Resistant Staphylococcus aureus (MRSA) after a twenty four hour time period of being contacted with the composition.

52. The composition of claim 51, wherein the composition exhibits bacteriostatic and/or bacteriocidal effects on each of E. coli, S. Aureus, P. aeruginosa, M. smegmatis, and Methicillin-Resistant Staphylococcus aureus (MRSA) after a twenty four hour time period of being contacted with the composition.

53. The composition of claim 48, wherein the composition dried amnion and dried chorion are present within the composition at an overall concentration of 20 wt % to 100 wt % of the composition.

54. The composition of claim 53, wherein the composition dried amnion and dried chorion within the composition at an overall concentration of 40 wt % to 100 wt % of the composition.

55. The composition of claim 53, further comprising at least one of aloe and allantoin.

56. The composition of claim 55, wherein the compositions comprises aloe and allantoin.

57. The composition of claim 56, wherein aloe is present in the composition at a concentration ranging from 0.75 μl to 2 μl per cc of the human placental powder and allantoin is present in the composition at an overall concentration of 0.5 wt % to 2.0 wt % of the composition.

58. The composition of claim 56, wherein the composition is configured for topical use.

59. The composition of claim 58, wherein the composition is a cream.

60. The composition of claim 52, wherein the composition dried amnion and dried chorion are present within the composition at an overall concentration of 20 wt % to 100 wt % of the composition.

61. The composition of claim 60, wherein the composition dried amnion and dried chorion within the composition at an overall concentration of 40 wt % to 100 wt % of the composition.

62. The composition of claim 61, further comprising at least one of aloe and allantoin.

63. The composition of claim 62, wherein the compositions comprises aloe and allantoin.

64. The composition of claim 63, wherein aloe is present in the composition at a concentration ranging from 0.75 μl to 2 μl per cc of the human placental powder and allantoin is present in the composition at an overall concentration of 0.5 wt % to 2.0 wt % of the composition.

65. The composition of claim 64, wherein the composition is configured for topical use.

66. The composition of claim 65, wherein the composition is a cream.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0040] A more complete understanding of the present invention may be obtained by references to the accompanying drawings when considered in conjunction with the subsequent detailed description. The embodiments illustrated in the drawings are intended only to exemplify the invention and should not be construed as limiting the invention to the illustrated embodiments.

[0041] FIG. 1 is a micrograph showing freeze-dried placental tissue including amnion, chorion, and umbilical cord tissue.

[0042] FIGS. 2A-C are micrographs showing different views of micronized placental tissue, including amnion, chorion, and umbilical cord tissue.

[0043] FIGS. 3A-H are photographs showing different samples of micronized amnion, chorion and umbilical cord tissue. Each sample includes an optimal grinding time. This data is also shown in Table 1.

[0044] FIG. 4 is a graph showing the tissue weight prior to grinding and the grind weight after grinding. This data is for the micronized tissues shown in FIGS. 3A-H.

[0045] FIG. 5 is a graph showing % tissue loss due to the grinding process for micronized tissues shown in FIGS. 3A-H.

[0046] FIG. 6 is a graph showing Hepatocyte Growth Factor (HGF) concentration in umbilical cord tissue.

[0047] FIG. 7 is representative photographs showing zones of inhibition for gram-positive and gram-negative bacteria. These “zones of inhibition” are indicative of antibacterial activity by showing where the composition has prevented/slowed or killed bacterial growth.

[0048] FIG. 8 is a photograph showing establishment of optimal aloe concentration in the compositions. Data summarized in Table 2.

[0049] FIG. 9 shows photographs of representative placental tissue and aloe-based topical creams of various consistencies.

[0050] FIG. 10 is a graph showing average diameters of zones of inhibition of treated bacterial species against creams of varying amnion/chorion membrane AC concentrations (Table 3).

[0051] FIGS. 11A-D show results of a disk diffusion assay with Escherichia coli. FIGS. 11A-C are agar plates showing zones of inhibition obtained using several or all creams of Samples 1-8 (Table 3). FIG. 11A: Samples 1 (100%), 2(0%), positive control (30 μg tetracycline), and negative control (water). FIG. 11B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (5%). FIG. 11C: Samples 7 (3%) and 8 (1%). FIG. 11D is a bar graph showing average diameters of zone of inhibition for the assay with Escherichia coli.

[0052] FIGS. 12A-D show results of a disk diffusion assay with Staphylococcus aureus. FIGS. 12A-C are agar plates showing zones of inhibition obtained using several or all creams of Samples 1-8 (Table 3). FIG. 12A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 12B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (5%). FIG. 12C: Samples 7 (3%) and 8 (1%). FIG. 12D is a bar graph showing average zone of inhibition diameters for the assay with Staphylococcus aureus.

[0053] FIGS. 13A-D show results of a disk diffusion assay with Pseudomonas aeruginosa (Micrococcus pyocyaneus). FIGS. 13A-C are agar plates showing zones of inhibition obtained using several or all creams of Samples 1-8 (Table 3). FIG. 3A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 13B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (5%). FIG. 13C: Samples 7 (3%) and 8 (1%). FIG. 13D is a bar graph showing average zone of inhibition diameters for the assay with Pseudomonas aeruginosa.

[0054] FIGS. 14A-D show results of a disk diffusion assay with Mycobacterium smegmatis (commonly used as a surrogate model for Mycobacterium tuberculosis). FIGS. 14A-C are agar plates showing zones of inhibition obtained using several or all creams of Samples 1-8 (Table 3). FIG. 14A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 14B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (5%). FIG. 14C: Samples 7 (3%) and 8 (1%). FIG. 14D is a bar graph showing average diameters of zones of inhibition for the assay with Mycobacterium smegmatis.

[0055] FIGS. 15A-D show results of a disk diffusion assay with Methicillin-Resistant Staphylococcus aureus (MRSA). FIGS. 15A-C are agar plates showing zones of inhibition obtained using several or all creams of Samples 1-8 (Table 3). FIG. 15A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 15B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (5%). FIG. 15C: Samples 7 (3%) and 8 (1%). FIG. 15D is a bar graph showing average diameters of zones of inhibition for the assay with MRSA.

[0056] FIG. 16 shows representative experiments of the testing mode for bacterial inhibition. The agar plates exhibit variations in the modes of inhibition, bactericidal (cidal) or bacteriostatic activity (static), obtained using several or all creams of Samples 1-8 (Table 3).

[0057] FIGS. 17A-C show results of an experiment evaluating mode of bacterial inhibition, bactericidal (cidal) or bacteriostatic (static), when testing MRSA. The agar plates of FIGS. 17A-B exhibit variations in the modes of inhibition of MRSA obtained using several or all creams of Samples 1-8 (Table 3). FIG. 17C is a bar graph showing average diameters of zones of inhibition and modes of inhibition obtained when testing MRSA.

[0058] FIGS. 18A-C show results of an experiment evaluating mode of bacterial inhibition, bactericidal (cidal) or bacteriostatic (static), when testing with Mycobacterium smegmatis. The agar plates of FIGS. 18A-B exhibit variations in the modes of inhibition of with Mycobacterium smegmatis obtained using several or all creams of Samples 1-8 (Table 3). FIG. 18C is a bar graph showing average diameters of zones of inhibition and modes of inhibition obtained when testing Mycobacterium smegmatis.

[0059] FIGS. 19A-C show results of an experiment evaluating mode of bacterial inhibition, bactericidal (cidal) or bacteriostatic (static), when testing with Pseudomonas aeruginosa. The agar plates of FIGS. 19A-B exhibit variations in the modes of inhibition of with Pseudomonas aeruginosa obtained using several or all creams of Samples 1-8 (Table 3). FIG. 19C is a bar graph showing average diameters of zones of inhibition and modes of inhibition obtained when testing Pseudomonas aeruginosa.

DETAILED DESCRIPTION OF THE INVENTION

[0060] For the purpose of promoting an understanding of the principles of the invention, reference will now be made to embodiments illustrated herein and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended. Any alterations and further modification in the described compositions, powders, creams, formulations, cosmetics, methods, and kits along with any further application of the principles of the invention as described herein, are contemplated as would normally occur to one skilled in the art to which the invention relates.

[0061] The inventive compositions are both therapeutically and cosmetically applicable and can be used for promoting skin regeneration and wound healing or skin rejuvenation. The compositions include tissues derived from the placenta, such as the amniotic membrane, chorion and umbilical cord. The placental tissues are micronized and combined with at least one of aloe and allantoin to form topical treatment creams. The compositions are specifically formulated as a cream or alternatively, the micronized placental tissues can be added to commercially available preparations to form compounded, topical treatment creams.

Placental Powder

[0062] Placental powder (FIG. 1 and FIGS. 2A-C) is a primary component of the inventive composition for maximal delivery of growth factors. This component is prepared by subjecting freeze-dried or dehydrated amnion and/or chorion and/or umbilical cord (FIG. 1) for reduction of particle size, preferably, but not limited to, micronization at subzero temperatures (FIGS. 2A-C, FIGS. 3A-H, FIG. 4, and FIG. 5).

TABLE-US-00001 TABLE 1 Amnion/ Chorion UC Time of Run Lot (g) (g) Speed/sec (min) A 0.18 0.54 30 2:00 B 0.18 0.54 30 2:30 C 0.18 0.54 30 3:00 D 0.18 0.54 30 1:00 E 0.75 30 2:30 F 0.75 30 3:00 G 0.75 30 3:30 H 0.75 30 4:00

[0063] Placental powders have known therapeutic efficacy in promotion of skin regeneration % rejuvenation and wound healing. The elution of HGF from the tissue over 24 hours shows that the micronized placental powder is able to locally deliver this regenerative growth factor (FIG. 6). The half-life of HGF is very short in the vascular system, approximately 3 hours. The duration of the HGF for 24 hours, at least, in the local environment points to a therapeutic advantage. In addition, the micronized particulate exhibits anti-microbial properties (FIG. 7).

Aloe

[0064] The addition of aloe to the placental tissue preparation is not intended to enhance the anti-microbial properties shown in FIG. 7, but to facilitate the delivery of anti-microbial properties of the placental tissue preparation to the tissues. For example, as demonstrated herein, the optimal amount of aloe added ranges from about 0.75 μl to about 2 μl per cc of placenta (FIG. 8). The term “about” in this context refers to amounts of aloe near to close to the disclosed range of 0.75 μl to about 2 μl that allows the composition to still reasonably achieve the described properties and/or function.

TABLE-US-00002 TABLE 2 Amount Mixed Time of Run Aloe Added Lot (cc) (min) (μl) B 2.0 2:30  500 B 2.0 2:30 1000 B 2.0 2:30 1500 B 2.0 2:30 2000 B 2.0 2:30 2500 B 2.0 2:30 3000 E 2.0 2:30 2250 F 2.0 3:00 2250 G 2.0 3:30 2250 H 2.0 4:00 2500

Examples One-Four: Compositions

Example One

[0065] Placental tissues, specifically amnion, chorion and/or umbilical cord are obtained from Cesarean section deliveries following informed consent of the mother. The maternal donors are screened for infectious disease with a comprehensive questionnaire (DRAI and AATB) and serology to exclude HIV, hepatitis B, hepatitis C, syphilis, and Zika virus. The processed tissues are then freeze dried or dehydrated and cryogenically ground. All donated tissues are processed aseptically and terminally sterilized using e-beam radiation. The fine powder of micronized tissue is then added to an aloe and allantoin preparation and mixed to form a cream for topical application (FIG. 9).

Example Two

[0066] Placental tissues, specifically amnion, chorion and/or umbilical cord are obtained from Cesarean section deliveries following informed consent of the mother. The maternal donors are screened for infectious disease with a comprehensive questionnaire (DRAI and AATB) and serology to exclude HIV, hepatitis B, hepatitis C, syphilis, and Zika virus. All donated tissues are processed aseptically. The processed tissues are then freeze dried or dehydrated and cryogenically ground. The fine powder of micronized tissue is then combined with aloe and mixed to form a cream for topical application.

Example Three

[0067] Placental tissues, specifically amnion, chorion and/or umbilical cord are obtained from Cesarean section deliveries following informed consent of the mother. The maternal donors are screened for infectious disease with a comprehensive questionnaire (DRAI and AATB) and serology to exclude HIV, hepatitis B, hepatitis C, syphilis, and Zika virus. The processed tissues are then freeze dried or dehydrated, cryogenically ground, and packaged. All donated tissues are processed aseptically and terminally sterilized using e-beam radiation. Separately, aloe or a combination of aloe and allantoin at the optimal ratio is packaged. The fine powder of micronized tissue and aloe-based diluent is provided at the optimal ratio for the desired consistency, depending on intended application. Instructions are included for the processor (e.g. nurse, preparation and distribution center) and/or the end user (e.g. nurse, physician, consumer) to prepare the final composition, which includes combination of the powder and diluent to form a cream for topical application.

Example Four

[0068] Placental tissues, specifically amnion, chorion and/or umbilical cord are obtained from Cesarean section deliveries following informed consent of the mother. The maternal donors are screened for infectious disease with a comprehensive questionnaire (DRAI and AATB) and serology to exclude HIV, hepatitis B, hepatitis C, syphilis, and Zika virus. The processed tissues are then freeze dried or dehydrated and cryogenically ground. All donated tissues are processed aseptically and terminally sterilized using e-beam radiation. Allantoin is added at the optimal ratio and the mixture is packaged. Separately, aloe or a combination of aloe and a second diluent at the optimal ratio, is packaged. The fine powder and aloe-based diluent is provided at the optimal ratio for desired consistency, depending on intended application. Instructions are included for the processor (e.g. nurse, preparation and distribution center) and/or the end user (e.g. nurse, physician, consumer) to prepare the final composition, which includes combination of the powder and diluent to form a cream for topical application.

Example Five: Examining Antibacterial Properties of Placental Tissue Components

[0069] As the use of antibiotics has increased across the globe, the rate of antibiotic resistance has grown leading to a pressure to develop new antibiotics. Placental tissue is known to exhibit antimicrobial activity against bacteria common to public areas and medical facilities, making it a viable candidate for antibiotic use. Further, antimicrobial peptides have been reported to be released within placental tissue layers, for example human beta-defensin (41). However, there are no antibacterial products currently on the market that take advantage of these properties of placental tissue. With this knowledge in hand, various components of placental tissue were tested to determine the portions that exhibited antibacterial activity. Dried placental tissue samples were micronized and rehydrated. The goal of this experimental study was to take the described products and test them on multiple bacterial species, some common to human skin, found in hospital/surgical environments. The described products (creams) are contemplated for use during surgical procedures to limit chances of bacterial infection. The bacterial species tested were Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Mycobacterium smegmatis, and Methicillin-Resistant Staphylococcus aureus (MRSA). A disk diffusion method was used. Bacteria were inoculated into nutrient broth, incubated overnight at 37° C., and then diluted in water to match the turbidity of a McFarland Standard. The bacteria were spread on Mueller-Hinton Agar plates. Disks with 20 uL of the hydrated tissue were placed on the inoculated plates and incubated at 37° C. overnight. Analysis of the plates showed that many of the placental tissues had a zone of inhibition around the disks, which indicated antibacterial activity (FIG. 10). The determination of whether the components exhibited bactericidal or bacteriostatic properties was seen. Bacteria within the zones of inhibition were collected, streaked onto nutrient agar plates and incubated overnight at 37° C. The analysis showed that most of the tissue components had a bacteriostatic mode of inhibition, which indicates its potential in applications intended to promote healing.

Cream Formulations

[0070] Creams were formulated as described above through processes of micronization and rehydration. Eight samples (Table 3) were formulated having different ratios of amnion/chorion membrane (AC Placental) and umbilical cord/amnion tissue (UC:A).

TABLE-US-00003 TABLE 3 50:50 AC Placental UC:A Cream Sample (%) (%) 1 100  0 2  0 100 3  40  60 4  20  80 5  10  90 6  5  95 7  3  97 8  1  99

Disk Diffusion Assay

[0071] A disk diffusion assay was performed to see how the different components of the placental tissue might lead to differences in antimicrobial activity against the bacteria through analysis of the zone of inhibitions produced after the assay. Overnight cultures were made for each bacterial species and were incubated at 37° C. in a shaking incubator. The bacteria were then diluted in sterile water to match a McFarland standard and spread on MHA plates. 20 μl of each cream sample was applied on 6 mm diameter disks and aseptically placed onto the swabbed plates. A Tetracycline BBL™ Sensi-Disc™ Antimicrobial Susceptibility Test Disk was used as a positive control and distilled water was used as a negative control. 0% AC cream contained Warren Laboratories Georges Aloe Vera. The plates were incubated at 37° C. Diameters were measured after incubation.

Results of Disk Diffusion Assay

[0072] Escherichia coli(FIGS. 11A-D, Table 3)

[0073] Creams composed of more AC placental powder exhibited greater antibacterial activity (large disk diameter) and creams composed of 20% and 10% AC placental powder showed similar activity (similar disk diameters). FIG. 11A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 11B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (5%). Creams composed of the least amount of AC placental powder showed no antibacterial activity (no zone of inhibitions). FIG. 11C: Samples 7 (3%) and 8 (1%). FIG. 11D is a bar graph showing average diameter of zones of inhibition for the assay with Escherichia coli. Notably, there is no significant difference in the Z of I when AC component is reduced from 100% to 40%. (+) bar represents the tetracycline positive control.

Staphylococcus aureus (FIGS. 12A-D, Table 3)

[0074] Creams composed of more AC placental powder exhibited greater antibacterial activity (large disk diameter). All creams exhibited antibacterial activity, even at the lowest concentration of 1% (Sample 8). FIG. 12A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 12B: Samples 3 (40%), 4 (20%), S (10%), and 6 (5%). FIG. 12C: Samples 7 (3%) and 8 (1%). FIG. 12D is a bar graph showing average diameter of zones of inhibition for the assay with Staphylococcus aureus. (+) bar represents the tetracycline positive control.

Pseudomonas aeruginosa (Micrococcus pyocyaneus) (FIGS. 13A-D, Table 3)

[0075] Creams composed of more AC placental powder exhibited greater antibacterial activity (large disk diameter). A significant drop in the zones of inhibition was observed between creams composed of 40% (Sample 3) AC placental powder and 10% (Sample 4) AC placental powder. There was no significant difference in the zones of inhibition between creams composed of 100% AC placental powder and positive control. FIG. 13A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 13B: Samples 3 (40%), 4 (10%), 5 (10%), and 6 (5%). FIG. 13C: Samples 7 (3%) and 8 (1%). FIG. 13D is a bar graph showing average diameters of zones of inhibition for the assay with Pseudomonas aeruginosa. (+) bar represents the tetracycline positive control.

Mycobacterium smegmatis (FIGS. 14A-D, Table 3)

[0076] Creams composed of more AC placental powder exhibited greater antibacterial activity (large disk diameter). There was no significant difference in the zones of inhibition between creams composed of 100% or 40% AC placental powder and positive control. Creams composed of 1% (Sample 8) AC placental powder showed no antibacterial activity. FIG. 14A: Samples 1 (100%), 2(0%), positive control (30 μg tetracycline), and negative control (water). FIG. 14B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (5%). FIG. 14C: Samples 7 (3%) and 8 (1%). FIG. 14D is a bar graph showing average diameters of zones of inhibition for the assay with Mycobacteriun smegmatis. (+) bar represents the tetracycline positive control.

Methicillin-Resistant Staphylococcus aureus (MRSA) (FIGS. 15A-D, Table 3)

[0077] Creams composed of more AC placental powder exhibited greater antibacterial activity (large disk diameter). Creams composed of 100% (Sample 1), 40% (Sample 3), 20% (Sample 4), and 10% (Sample 5) AC placental powder had a larger zone of inhibition than the positive control with 100% (Sample 1) and 40% (Sample 3) being significantly higher. Creams composed of 1% (Sample 8) AC placental powder showed no antibacterial activity. FIG. 15A: Samples 1 (100%), 2 (0%), positive control (30 μg tetracycline), and negative control (water). FIG. 15B: Samples 3 (40%), 4 (20%), 5 (10%), and 6 (S %). FIG. 15C: Samples 7 (3%) and 8 (1%). FIG. 15D is a bar graph showing average diameters of zones of inhibition for the assay with MRSA. (+) bar represents the tetracycline positive control.

Test for Bactericidal and Bacteriostatic Activity

[0078] The bactericidal and bacteriostatic activity was measured by swabbing the zone of inhibition on the plates followed by swabbing on fresh nutrient agar plates and incubating the newly swabbed plates overnight at 37° C. (FIG. 16).

[0079] The results showed variation in the modes of inhibition of the creams. All creams exhibited bactericidal activity at 100% and 40% concentrations.

[0080] MRSA exhibited bactericidal activity at concentrations of 100%, 40%, 20%, and 10% of AC placental powder and bacteriostatic activity at concentrations of 5% and 3% AC placental powder (FIGS. 17A-C).

[0081] Mycobacterium smegmatis showed results similar to that of MRSA by exhibiting bactericidal activity at concentrations of 100%, 40%, 20%, and 10% of AC placental powder and bacteriostatic activity at concentrations of 5% and 3% AC placental powder (FIGS. 18A-C).

[0082] Pseudomonas aeruginosa (Micrococcus pyocyaneus) exhibited bactericidal activity at a concentration of 40% AC placental powder and bacteriostatic activity at concentrations of 100% and 20% AC placental powder (FIGS. 19A-C).

CONCLUSION

[0083] The invention described and exemplified herein represents a new treatment composition for promoting skin regeneration or rejuvenation and wound healing. These novel compositions exhibit increased clinical efficacy and decreased costs as compared to conventional regenerative medical techniques.

[0084] All patents and publications mentioned in this specification are indicative of the levels of those skilled in the art to which the invention pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. It is to be understood that while a certain form of the invention is illustrated, it is not intended to be limited to the specific form or arrangement herein described and shown. It will be apparent to those skilled in the art that various changes may be made without departing from the scope of the invention and the invention is not to be considered limited to what is shown and described in the specification. One skilled in the art will readily appreciate that the present invention is well adapted to carry out the objectives and obtain the ends and advantages mentioned, as well as those inherent therein. The compositions, formulations, creams, cosmetics, powders, kits, methods, procedures, and techniques described herein are presently representative of the preferred embodiments, are intended to be exemplary and are not intended as limitations on the scope. Changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention. Although the invention has been described in connection with specific, preferred embodiments, it should be understood that the invention as ultimately claimed should not be unduly limited to such specific embodiments. Indeed various modifications of the described modes for carrying out the invention which are obvious to those skilled in the art are intended to be within the scope of the invention.

REFERENCES

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