CLINICAL TRIAL ON THE EFFICACY OF A NEW ANTI-ACNE REGIMEN IN THE TREATMENT OF MILD TO MODERATE ACNE VULAGARIS

The Clinical Trial was conducted at the Philippine Dermatologist Research and Testing Foundation that conducts research on topical medicines.

Objectives:
A. To determine the efficacy of CRC anti-acne skin regimen to help dry and prevent eruption of new acne.
B. To monitor the development of signs and symptoms of adverse reactions of the regimen when applied daily over the duration of the study. (Tolerability)
Test Regimen: Mask Cleanser and Acne Cream (CRC)
Panelist: 30 panelist ages 16-35 with mild to moderate acne.

CONCLUSION

Acne regimen consisting of Mask Cleanser and Acne Cream (CRC) is effective in the treatment of mild to moderate acne, with significant improvement from week 1 to week 4.
It is safe regimen and was well tolerated by the panelists suffering from mild to moderate acne.

For the reduction of acne comedones, there was a significant improvement after week 1 (40%) compared to the baseline data. After week 2 (58%) week 3 (68%) and week 4 (72 %).
This improvement is very satisfactory, considering that comedone extraction was not done.

Among the 3 types of acne lesions, the pustular lesions responded best to the acne regimen. As early as Week 1 there was already 70% improvement and at Week 2 and 3, the improvement increased significantly to 94% and 92% with 95% improvement at the end of Week 4.

Nodulo-cystic acne eruptions improved significantly from week 1 to week 4, 45%, 87%, 84% and 87% respectively.

Pustular and nodulo-cystic lesions do not usually improve with topical treatment alone. In clinical practice, patients suffering from pustular and nodulo-cystic type of acne are also given oral antibiotics in addition.

There was a significant improvement in the reduction of acne lesions as a whole
(44%) as early as week 1 compared to the baseline data. Week 2 (62 %), Week 3 (70%) and Week 4 (76%). Data analysis used was T-Test paired samples.

PICTURES OF SOME PATIENTS BEFORE AND AFTER CRC ACNE TREATMENT
STUDY ON CRC PHYTIC TREATMENT CONCENTRATE*


This study was conducted to determine if exposure to the Phytic Treatment Concentrate can increase cell proliferation in the Mat-Tek EpiDerm
skin model and also assesses the any tissue damage and changes in tissue viability after exposure to the test materials using standard H&E stain techniques and the metabolic dye MTT.

Test materials:
1.CRC Phytic Treatment Concentrate
2. Glycolic Acid 50 % Solution

For each test material the percent viability value for each replicate was plotted on the y-axis versus the exposure time on the x-axis.

Histology were done on tissues (H&E staining and Ki-67 staining) analysis.

Results:
The level of cytotoxicity was determined in tissues after15 and 30 minutes of exposure to CRC Phytic Treatment Concentrate and after 15 minutes to Glycolic Acid 50% solution.

One H&E stain was prepared from one tissue in each set. One Ki-67 (Mib-1) immunostain was prepared from another tissue in each set including negative controls.

Mitoses in H&E stained issues were counted at 40X magnification and expressed as number per high power field (HPF). The epidermal thickness was measured using a micrometer ocular at 40X magnification, measuring from the base of the basal layer (equivalent of the basement membrane) to the bottom of the stratum granulosum.

After exposing tissue to Phytic Treatment Concentrate for 15 minutes the tissue shows all layers of normal skin, including a basal layer, spinous layer, granular layer and cornified layer. No abnormal or pathological processes are identified.

After exposure for 30 minutes using the same material the tissue apparently represents epidermis, but there is widespread accumulation of granular material in both the basal and granular layer. The material appears, at least in part, to be situated intracellularly. The cells of the granular layer are partially torn apart. The cornified layer shows some parakeratosis.

After exposuing tissue to Glycolic Acid 50% solution for 30 minutes the same changes as described for 30 minutes of exposure to Phytic Treatment Concentrate are present.

Based on the results of the MTT assay, Phytic Treatment Concentrate is non-cytotoxic after exposure for 15 minutes but is cytotoxic after exposure for 30 minutes. Glycolic Acid 50% solution is cytotoxic after exposure for 15 minutes.

Based on the results of the histology the tissue exposed to Phytic Treatment Concentrate for 15 minutes appeared normal with no abnormal or pathological processes identified. Mib-1 (Ki-67) stain is positive for cell proliferation. The tissue exposed to Phytic Treatment Concentrate for 30 minutes and Glycolic Acid 50% for 15 minutes show widespread accumulation of granular material in both the basal and granular layer. The cells of the granular layer are partially torn apart and the cornified layer shows some parkeratosis. The Mib-1 stain is negative. This is evidence of tissue damage.

PICTURES

Study of Tyrosinase Inhibitory Activity for Phytic Treatment Concentrate*

Concerning functional cosmetic, tyrosinase-inhibitory activity and superoxide scavenging activity have become very interesting research targets. Tyrosinase plays an important role in the formation of melanin, because it facilitates melanization by catalyzing reaction from tyrosine to dopa and from dopa to dopaquinone, and probably from dihydroxy indole to indole-5, 6-quinone. Superoxides have some relationship with skin inflammation, cancer and aging. Therefore, the development of superoxide scavengers and tyrosinase inhibitors will contribute to the prevention of skin pigmentation and aging, and to the maintenance of skin health. The inhibition ratio against tysosinase activity will be calculated by the measurement of optical density at 475nm with a routine method.

The mixture containing different concentrations of Kojic acid or the single concentration of Phytic Treatment Concentrate were incubated with the reaction mixture at 37 Degrees C for 20 minutes and absorbance was recorded at 475 nm.

The inhibitory effect of the sample on the tyrosinase activity is represented as inhibition % compared with control. The relative inhibitory activity of the product and the means of the results are displayed as % vehicle control (lacking an inhibitor) as a function of inhibitory concentration (100 % represents 0% inhibitor).

The process of melanin biosynthesis a determinant of skin color, and its disturbance plays an important role in local hyperpigmentation such as melasma, and lentigo. Therefore, tyrosinase is a useful target in the fields of hyperpigmentation and cosmetic materials.

Phytic Treatment Concentrate exhibited significant inhibitory effects on mushroom tyrosinase at more than 150 ug/ml. The product exhibited an IC50 values of 207 ug/ml on tyrosinase. Therefore, Phytic Treatment Concentrate exhibited similar order of magnitude (Y x10 ug/ml) in the level of potency with Kojic acid used as a positive control. From the fact that the activity of Phytic Treatment Concentrate is comparative order of magnitude with that of Kojic acid, therefore, this compound may serve in functional skin care products for whitening and photoprotective functions.