Dermal Fx (Hypoallergenic, Non-comedogenic and Nonirritant)

Study of the safety and hydrating properties of a cosmetic cream

Jose Guerrosantos1, MD, Ferando Guerrossantos1, MD, Eriberto Orellana Morales1, MD 1. Clinica Doctores Guerrerosantos, Instituto Jalisciense de Cirugia Reconstructiva, Jalisco, Guadalajara, Mexico Abel Chajchir2, MD, Irina Chajchir2, MD, Pankaj Modi3*, PhD,MD 2. Centro MedicoBarrancas, Cirugia Plastica Estetica, Buenos Aires, Argentina 3. Dr. Pankaj Modi, PhD, MD., DPM Therapeutics Corp,

Abstract;

Objective; To study Dermal Fx cream formulation in 120 to 150 Healthy Subjects for Hypoallergenic, Non-comedogenic and Non-irritant properties.

Design; This open, randomized, intra-individual study was conducted in accordance with the principles of current international ethical standards originating from the Declaration of Helsinki and in compliance with local regulatory requirements. The following instruments were used to assess skin conditions after the treatment with DermalFx;

Corneometer® (to evaluate water content of the stratum corneum) corneometer is capable of measuring the amount of moisture in the skin.

• Dermal Torque Meter® (to evaluate skin elasticity and moisturisation) cutometer and the twistometer evaluate skin tone and elasticity.
• EP3 Evaporimeter® (to measure rate of water loss across the epidermis)
• Sebumeter® (to measure sebum on skin) The sebumeter or "seb-u-tape" measures the amount of oil on the skin's surface.
• Minolta Chromameter® (to measure colour)
• Image Analysis equipment (to assess the surface of the skin) Skin prints
make it possible to obtain quantitative and qualitative measurements. This is the technique used to measure the number and depth of wrinkles.
• Profilometry for skin smoothness, anti-wrinkle, anti-ageing and fine line reductions

expression in the face and neck. Hyperfunctional facial lines are distressing to
patients because they often are misinterpreted as anger, fear, fatigue,
melancholia, and aging. They represent a prevalent clinical scenario. Patients
often look to aesthetics medicine for resolution.

Skin moisturizers are used to help prevent skin from becoming dry or to return dry skin to its hydrated condition. Several compounds are known to act as skin moisturizers or humectants. Among these are glycerol (glycerin), urea,
petrolatum (petroleum jelly), polyethylene glycols (PEGs), lanolins (wool wax),
lactic acid (lactate), and ceramides. In skin care products, these ingredients are formulated together with a large variety of other ingredients used for maximizing efficacy, stability and shelf life. For skin care products, the subjective impressions of the user are particularly important. Thus, ingredients for improving subjective properties such asstickiness/greasiness, distribution/spreading, absorption, odor, or color are also added. Typically, skin care products are comprised of about 20-50 different ingredients, since it is the experience of the personal skilled in the art that favorable results may not be achieved by formulations with fewer ingredients.

The ingredients of skin care formulations are generally combined to form an
emulsion, since they consist of hydrophobic and hydrophilic ingredients. In
principle, two basic types of emulsions are possible. In an oil-in-water (O/W)
emulsion, the oil phase forms the discontinuous phase, whereas in a water-in-oil (W/O) emulsion, the water phase represents the discontinuous phase.

Most skin care products are formulated as O/W emulsions. O/W emulsions are usually preferred for cosmetics, due to their favorable cosmetic properties that are pleasing for consumers. An O/W emulsion however represents a trade off in favor of such user-pleasing properties, since O/W-emulsions are generally more effective than W/O emulsions in treating or ameliorating dry skin conditions.

A common problem associated with skin care products is that they cause adverse effects in consumers or patients. Such unwanted effects include irritation (subjective and objective), allergic contact dermatitis, photocontact dermatitis, and immediate contact reactions ( "Dry Skin and Moisturizers: Chemistry & Function ", p. 403, Eds. Loden & Maibach, 2000). The most frequently observed side effect is allergic contact dermatitis. This reaction is usually caused by fragrances and preservatives present in these products. Another origin of allergic reactions are moisturizers such as lanolin and its derivatives, emulsifiers such as cocamidopropyl betaine, and emollients.

The novel formulation (Dermal Fx) provides a hypoallergenic and non-irritant cosmetic skin care formulation to improve the texture [or soften the appearance] of hyperfunctional facial rhytids (wrinkles). The oil phase contains cetearyl octanoate, dicaprylyl ether, caprylic / capric triglyceride, polyglyceryl- 3-diisostearate and polyglyceryl-2-dipolyhydroxystearate.

All Dermal Fx (DPM Therapeutics) formulations comprise an oil phase dispersed in a water phase. In this embodiment, the oil phase may comprise cetearyl octanoate, dicaprylyl ether, caprylic / capric triglyceride, polyglyceryl-3-diisostearate and polyglyceryl-2-dipolyhydroxystearate. In general, the ingredients used for the dermatologic formulations of the invention are of high
quality and purity.

The skin care formulations of Dermal Fx are free of ingredients with a known allergenic potential, thus being particularly suitable and useful for consumers with dry, sensitive, wrinkled or aging skin.

The primary objective of the present formulation (Dermal Fx) is to solve the problems associated with available and known skin care products by providing a hypoallergenic and non-irritant skin care formulation. Aside from being hypoallergenic and non-irritant, the formulation of the novel and stable product effectively moisturizes dry, sensitive skin and is pleasing to consumers seeking quality cosmetic care.

Dermal Fx is a non-animal, stabilized Sodium Hyaluronate gel with powerful hydrating properties. Injectable Hyaluronic Acid derivatives have also been developed for soft tissue augmentation, and are currently used in South America, USA, Europe and Canada for the correction of moderate facial wrinkles and folds and for lip enhancement. As with the Sodium Hyaluronate contained in Dermal Fx products, they confer less risk of immunogenicity; unlike collagen, hyaluronic acid is chemically identical across all species.

Stabilization (cross-linking) of the hyaluronic acid molecule is achieved through minimal chemical modification, thereby improving its resistance to enzymatic degradation within the dermis without compromising its biocompatibility. Because Dermal Fx Sodium Hyaluronate is derived from a non-animal source (it is produced from bacteria by a bio-fermentation process), there is no likelihood of contamination with antigenic proteins during manufacture; consequently, the risk of hypersensitivity reactions and transmission of potentially harmful infectious agents is reduced. Preliminary clinical findings indicate that Dermal Fx is well tolerated and provides a good cosmetic effect when used on facial wrinkles and folds. This controlled study of Dermal Fx was conducted to evaluate its tolerability as well as cosmetic effectiveness and persistence over a 6-month period in subjects requiring cosmetic care of facial wrinkles and folds.

Application
Dermal Fx is a cosmetic product manufactured by DPM Therapeutics Corp, in Canada. It is a non-comedogenic oil-inwater emulsion primarily marketed for moisturizing and improving the appearance of facial wrinkles. The subjects were instructed by our clinicians how to use the product. It was then applied daily for four weeks by the subjects twice daily (once in the morning after their shower and once at the bed time). The use of this novel cream result in a convincing effect on the subjects’ skin wrinkles by quickly making them younger looking. Subjects were followed for at least four weeks for a possible change in appearance or any adverse effects.

Treatment
Dermal Fx is a cosmetic (anti aging, anti wrinkle) product manufactured by DPM Therapeutics Corp, in Canada. It is a non-comedogenic oil-in-water emulsion primarily marketed for facial wrinkles correction. The patients were instructed by our clinicians how to use the product. It was then applied daily for four weeks by the patients twice daily (once in the morning after their shower and once at the bed time). The use of this novel cream result in a convincing effect on the patients' skin wrinkles by removing them quickly and making them look years younger. Patients were followed for at least 4 weeks for a possible flare-up or any adverse effects.

Since the investigation was performed during the time interval between the subjects' first visit and the termination of applications after four weeks, the resulting effects on these subjects were fairly constant throughout the observation period.

Subjective, objective, visual and instrumental procedures were used to evaluate all categories of products. Environmentally controlled rooms offer a wide range of options. Skin bio-engineering measurements were typically performed at 55% (+/- 5%) relative humidity and 20°C (+/- 2°C) temperature. In assessing skin condition, factors such as hydration, trans-epidermal water loss and oiliness were
routinely measured. Instrumental measurements form a significant part of these evaluations and routine use was made of:

• Corneometer® to evaluate water content of the stratum corneum, i.e., it is
  capable of measuring the amount of moisture in the skin.
  
• Dermal Torque Meter® (to evaluate skin elasticity and moisturisation). The
  cutometer and the twistometer evaluate skin tone and elasticity.
• EP3 Evaporimeter® (to measure rate of water loss across the epidermis)
• Sebumeter® or "seb-u-tape" (to measure sebum/oil on the skin's surface)
• Minolta Chromameter® (to measure color)
• Image Analysis equipment (to assess the surface of the skin). Skin prints
  make it possible to obtain quantitative and qualitative measurements of the
  number and depth of wrinkles.
• Profilometry for skin smoothness, wrinkles and fine lines

Barrier Function
An Evaporimeter® was employed to measure the Trans Epidermal Water Loss
(TEWL). This is a useful aid in the determination of barrier function,
moisturization and skin.

Evaluation Criteria
All criteria were assessed at baseline, before
any product application, and after 15 days of product use. Skin
discomfort signs (redness, dryness, roughness, desquamation, or
any other sign) and symptoms (stinging, itching, burning,
discomfort/tightness, or any other symptom) were evaluated
according to a 5- point scale (0 = none, 1 = mild, 2 = moderate, 3
= severe, 4 = very severe). The appraisal of the moisturizing cream
effect was complemented by objective measurements on the
subject's cheeks evaluating electrical capacitance and TEWL, as well
as the D-Squame[R] test. The electrical capacitance was measured
by using a Corneometer CM 820 (Courage & Khazaka electronic
GmbH, Koln, Germany) the measurements of which are expressed in
arbitrary units. This is an automated instrument used to measure
the amount of moisture in the outer layer of the skin (skin
hydration) and the ability of the skin to retain moisture (skin
moisture capacity). (11) TEWL, expressed in g/[m.sup.2]/h, was
measured with a Tewameter TM 210 (Courage & Khazaka electronic
GmbH, Cologne, Germany), which is an indicator of water vapor
pressure gradient between the boundary layer of the skin surface
and the ambient air. Since the skin barrier regulates the rate of
water loss from the body, the rate of TEWL is a measure of the
condition of the skin barrier. When skin is damaged, its barrier
function is impaired resulting in high water loss. D-Squame tapes
are adhesive coated discs intended for the measurement of skin
desquamation by sampling corneocytes. After staining, the samples
were assessed visually in a medical viewer and a special system for
the measurement of optical transmission of the tapes was used.
At the end of the study, subjects completed a satisfaction
questionnaire including items regarding the effect of the moisturizing
cream on the skin, their purchase intent and a global score (from 1
meaning bad to 10 meaning excellent).

Statistical Analyses
Individual results obtained from the objective assessments before and after
application of Cetaphil® Moisturizing cream were analyzed using the Wilcoxon
Signed-Rank Test. Clinical signs and symptoms were descriptively summarized.
The cream was tested on human volunteers as follows to determine the safety
and efficacy in the control of cutaneous pruritus.

Sensitization and Irritation Risk (HRIPT-Test)

A human repeated insult patch test (HRIPT. Made from our proprietary Dermal Fx formulation consisting hyaluronic acid, collagen (organic vegetable based) and elastin was carried out as a 40-days randomized, controlled study to assess the sensitizing and irritating potential of the dermatological skin care formulations of the Dermal Fx. The study was started with 85 healthy volunteers of either sex between 18 and 65, all enrolled volunteers completed the study without any major side reactions and health issues.

In the induction phase (day 1 - 30), the volunteers were subjected to 15 occlusive treatments for 24 hours with 75 Nl of the test formulation and applied to their backs on days 1, 3, 5, 8, 10, 12, 15, 17, 19, 31, 23, 25, 27, 30. Untreated test fields without application of a test formulation were used as a concurrent control. Skin reactions were evaluated 24 hours (1/2 hour after removal of test chambers; on days 2, 4, 6, 9, 11, 13, 16, 18, 20, 22, 24, 26, 28, 30; fifteen readings) and 48 hours after the application of occlusive treatments on the backs of the volunteers (1/2 hour after removal of test chambers on days 3, 5, 10, 12, 17,19; six readings) by a visual 6-point-score.

Visual 6-point-score

Negative/ no reaction 0
Equivocal erythema 0.5
Erythema 1
Erythema and induration (edema) 2
Erythema, induration (edema) and vesicles 3
Erythema, induration (edema) and bullae and/or spreading 4
The mean cumulative irritation index (M.C.I.I.) was calculated for the 24 hours
readings (M.C.I.I./ 24 h.) and for the 48 hours readings (M.C.I.I./ 48 h.).

After the treatment-free interval (30 days; days 31-45; no treatment), an
occlusive challenge application of the test formulations on yet untreated test sites on the backs of the volunteers was done for 24 hours. The test formulations were evaluated 24 h, 48 h, 72 h and 96 h after the application by the same visual 6-point score (challenge phase; 5 days, days 36-45).

Results:

Sensitization: No case of sensitization occurred in both ITT- and VC-population.
Irritation: No irritation potential observed in comparison to the empty untreated test field could be observed in both ITT- and VC-population. Furthermore, no adverse events or serious adverse event related to the test formulations was observed during the course of the study.
Sesitization Effect; none (98.3%), 2 subjects report minor tingling effect
Irritation Effect; none (100%)

Irritation Potential Study;

The primary objective of this study was to evaluate the cutaneous tolerability of
the skin care formulations of the invention over a 8-weeks period when used in
the intended way (application twice daily to face or body, respectively) in the
intended patient/consumer group having dry and sensitive skin. The secondary
objectives of this study were to evaluate the irritation potential of the test
formulation when used around the eyes, to evaluate the hydrating effect of all
test formulations as measured by corneometry, to evaluate the volunteer's
general cosmetic acceptance of all test formulations, and finally to evaluate the cosmetic acceptance of the test formulation as make-up foundations.
The study was designed as a single center, randomized study; with
interindividual comparison of treatments between the test formulation and
comparative commercially available products for facial application and between test formulation and comparative commercially available products for body application, and was performed for 56 consecutive days.

Healthy volunteers of either sex between 18 and 65 years old meeting specific
inclusion/ exclusion criteria with dry skin proven by low corneometer values (=70) were chosen for this study. All volunteers in all treatment groups treated face and body twice daily at home over 56 consecutive days with the respective test formulations.

For the measurement of the hydrating effects of the test formulations for facial application, corneometry of both cheek bones was performed on days 1, 28 and 56. For the test formulation for body application, corneometry of both anterior lower legs was performed on screen and on days 1, 28 and 56.

A clinical assessment was performed at screen and on days 1, 8, 15, 30, 45 and 56 using a visual 5-point score (no / doubtful / slight / moderate / severe signs of irritation, erythema and/or scaling). Out of the results of the visual 5-point score an irritation product profile was created with four irritation groups being the primary variable of this study. Subjective symptoms were asked on days 1, 8, 15, 30, 45 and 56 using a 5-point score for subjective evaluation (no / doubtful / slight / moderate / severe sensation of stinging / burning / pruritus with specification in case of any positive answer). Additionally, an ophthalmological examination investigating fore- and background of the eyes as well as the conjunctival mucosa and subjective ophthalmological symptoms using a 5-point score for subjective ophthalmological symptoms took place on days 1, 8, 15, 30, 45 and
56.

Additionally, a questionnaire concerning the sensorial profile and overall
cosmetic acceptance (parameters: distribution/spreading, absorption, feeling on the skin, stickiness, greasiness, oily shine, odor, and color; evaluation on a 5-point scale) was filled in by the volunteers on days 1, 8, 15, 30, 45 and 56.

Results:
Subject satisfaction questionnaire.
(% of subjects) (% of subjects)
1. Skin comfort is very good or good 97.3 3.7
2. The product softens skin* 96.7 3.3
3. The product firms up skin* 95.2 4.8
4. The product hydrates skin* 100
5. The product is not sticky* 96.7 3.3
6. The product doesn't sting eyes* 100
7. Product's smell is very 90 10
pleasant or pleasant
8. The product provides a soothing 96.0 4.0
sensation during application*
9. Intent to buy 93.4 6.4 (cost factor)
10. Global score 9.6/10
*completely agree or agree (yes or perhaps (may be yes)
Overall opinion excellent very good good acceptable
Cosmetic qualities 52% 48%
Distribution 68% 22% 10%
Spreading 70% 30%
Absorption 78% 22%
Feeling on the skin 56% 24% 11% 9%
Stickiness NO STICKINESS OBSERVED 100%
Greasiness NO GREASINESS OBSERVED 100%
Oily shine NO Oily Shine Observed 100%
Odor Very Lightly Fragrant 100%
Color of Skin 47% 33% 20% (Improved markedly)

As shown above, the subjects clearly recognized the superior fluidity and quicker absorption rate of Dermal Fx. The subjects also perceived a significant reduction in stickiness on the skin as well as a significantly softer skin feeling

Conclusions

A skin efficacy test, conducted showed that daily use of Dermal Fx helps improve
skin hydration. As shown, Dermal Fx showed a 92 % increase in skin hydration
when compared to untreated skin. The formulation found to be Hypoallergenic
and Non-Comedogenic and non-irritant.

* Provides lightweight moisturization for a cashmere soft feel
Dermal Fx offers a pleasantly soft, non-oily skin sensation. When rubbed between
the fingers, Dermal Fx has satiny skin feel. Once dry, it leaves the skin feeling
cashmere soft.

* Maintains skin's natural water balance for a more radiant appearance
Dermal Fx provides advanced protection against moisture loss by acting as a
barrier to help maintain the skin's natural moisture level. Increased hydration
helps skin appear more radiant and supple.

* Conditions skin to a silky smooth finish with enhanced tone
Dermal Fx conditions skin without build-up, making it ideal for everyday use. Its
conditioning properties result in easy to apply, silky smooth skin.

* The skin care formulations of the Dermal Fx are useful to moisturize dry,
sensitive skin and protect the skin's moisture barrier. They also exhibit
favorable sensorial properties.

References

1. Dry Skin and Moisturizers: Chemistry & Function ", p. 403, Eds. Loden &
Maibach, 2000
2. Bikowski J. The use of therapeutic moisturizers in various dermatologic
disorders. Cutis 2001;68(suppl):3-11. Review.

Cetaphil ® is a trademark of Galderma Laboratories, L.P. United States