1. INTRODUCTION:

Recent advances in the cosmetic industry combined with the growth of Asian markets, including the Chinese and Southeast Asian markets, have had a significant technical and economic impact¹. The domestic cosmetic industry has been strategically widening market access to other Asian countries and has been trying to gain insights into the culture, social atmosphere, economy, and religion of different countries to plan commercialization directions^1.2.

Current trends in the development of cosmetic products involve the development of eco-friendly products using natural materials and traditional herbal medicines that are targeted for South Korea, China, and Japan. In South Korea, cosmetic products are being actively developed using natural materials and traditional herbal medicines, and their competitiveness in the domestic market has been strengthened by taking advantage of the highly developed media and culture contents in South^3.4. Furthermore, Korea and China have consistently benefited from the development of products on the basis of the theories of Korean and Chinese medicines, which essentially share the same origin, and the strategic marketing of these products. Therefore, it has become important to consider the traditional usage of medicinal herbs and to demonstrate the modern scientific basis of oriental medicine for industrial and economical purposes in the cosmetic industry^5.6.

Skin is a large organ that accounts for 16% of the total body weight. It consists of the epidermis, dermis, and subcutaneous tissue, and has various functions including prevention of the entry of foreign substances, sensation, body temperature control, excretion, and absorption^7.8. The dermis, which exists between the epidermis and fat layers, consists of fibrous connective tissue composed of fibrous proteins such as collagen, elastin, and reticulin, protects the body from external injury, and is closely associated with joint movement and local joint flexibility⁹. Changes in elastin and hyaluronic acid contents within the dermis are an important factor associated with healthy skin and are closely associated with wrinkle development due to aging and moisture loss¹⁰.

Wrinkles develop when the water content of the corneometer decreases due to external stress factors such as ultraviolet rays, dry atmosphere, and physical and chemical stimuli. Reduced water content of the corneometer causes it to expand and the epidermis to contract, and eventually brings about quantitative and qualitative changes in collagen and elastin contents in the dermis. This ultimately reduces the overall flexibility and elasticity of the skin and promotes wrinkle formation¹¹. Accordingly, materials and natural products that can reduce wrinkles and increase skin elasticity by increasing dermal compactness have consistently been developed and sought¹².

According to the theories of traditional Korean medicine, herbal medicines have various tastes, exert certain effects on internal organs, and control the physiological functions of certain internal organs; therefore, they can be used to treat diseases and maintain good health¹³. We selected 3 herbal medicines, Coptischinensis,Cimicifugaheracleifolia,and Poriacocos (Schw.), that are associated with skin health according to the theories of Korean medicine, and investigated their effects on dermal compactness. This clinical assessment on dermal compactness was approved by the institutional review board (IRB) and conducted in accordance with a guideline for clinical assessment of cosmetic products released by the Ministry of Food and Drug Safety. We have reported the results of this study, as they were meaningful.

2. MATERIALS AND METHODS:

2.1. Experiment materials:

The 3 medicinal herbs, Coptischinensis, Cimicifugaheracleifolia, and Poriacocos, were purchased from Puremind Co. (Yongchoen, Korea). The samples were stored in a common laboratory of BIT College of Convergence at Seowon University. One hundred grams (100 g) of each medicinal herb were macerated in 600 mL of 70% (v/v) ethanol solvent once every 24 hours up to a total of 3 times in 72 hours. The solution was filtered with filter paper and collected in a bottle. The solution was then run in a rotary evaporator and freeze-dried for 3 days. The extraction yield of the mixed extract was 11.5%. The freeze-dried sample was diluted in glycerin and used in a clinical test at a concentration of 1%.

2.2. Participant selection:

The present study was approved by the IRB of Semyung University Oriental Medicine Hospital=(IRB No. SMCTC-49-15-01), and was conducted between May 28, 2015 and June 26, 2015. Subjects aged 35-55 years who satisfied the inclusion criteria and did not meet the exclusion criteria were included. For the measurement of various parameters of the test sample, the participants were informed about the research procedure by our researchers through verbal and written communication. The participants began their participation after submitting their consent form. A total of 22 women who satisfied the inclusion criteria were finally selected. The effectiveness of the mixed extract at improving dermal compactness was assessed, and a skin irritation test was performed in accordance with the guidelines for clinical assessment of cosmetic products released by the Ministry of Food and Drug Safety.

2.3. Skin irritation test:

Skin irritation symptoms such as erythema, swelling, itchiness, and burning sensation were assessed and graded.

2.4. Test method:

The anti-wrinkle effects of the extract were assessed on the eye rims. The extract was always applied under the same experimental conditions in a clean and dry state after the patients rested in an air-conditioned room for at least 30 min (22±2℃, R.H. 40~60%). A cutometer (MPA 580, Courage and Khazaka Electronic Co., Germany) was used to measure wrinkle improvements. This optical measuring system measures the amount of skin drawn into the probe by negative pressure. An infrared light is passed through the skin, and the ratio of the intensity of light passing through the skin to the intensity of the light that does not pass through the skin is calculated to determine skin elasticity. An arbitrary unit (A.U.) was used as the unit of measurement.

2.5. Test schedule and procedure:

2.5.1. First visit (0 days after the test initiation):

Participants were selected according to the inclusion and exclusion criteria. Following an assessment of skin conditions, skin elasticity was assessed using a cutometer.

2.5.2. Second visit (2 weeks after the test initiation):

Skin irritation test and skin elasticity assessments using a cutometer were performed after the participants used the test sample for 2 weeks according to the instructions.

2.5.3. Third visit (at 4 weeks after the test initiation):

Skin irritation test and skin elasticity assessments using a cutometer were performed after the participants used the test sample for 4 weeks according to the instructions. The effectiveness of the test sample was also assessed through a questionnaire survey.

2.6. Statistical analysis:

Changes in facial skin elasticity before the extract application, and at 2 and 4 weeks after the application were statistically analyzed by performing a paired t-test using SPSS 18.0software. The level of statistical significance was set at p<0.05.

3. RESULTS AND DISCUSSION:

3.1. Participant composition:

The participants consisted of women aged 37-54 years (mean 44.3 years). The proportions of women in their 30s, 40s and 50s were 13.6%, 77.3%, and 9.1%, respectively. A total of 22 subjects participated in the test, and none of the subjects dropped out or quit participating during the research period[Table 1].

Table 1 : Construction of participants

Age	Number	Dropout	Number of completion
30’s	3	0	3
40’s	17	0	17
50’s	2	0	2
Total	22	0	22

3.2. Skin irritation test:

No abnormal skin reactions against the test sample were observed by the examiners and the participants. While some natural products have shown to cause skin irritation, the mixed extract of Coptischinensis, Cimicifugaheracleifolia, and Poriacocos was shown to be safe when applied topically.

3.3. Facial skin elasticity:

The effectiveness of the test sample at preventing facial wrinkles was subjectively and clinically assessed in accordance with the guideline for clinical assessment of cosmetic products released by the Ministry of Food and Drugs. The sample was applied onto participants’ eye rims twice a day. Skin elasticity was assessed before the application of the sample (0 week), and at 2 and 4 weeks after the application. Skin elasticity results are shown in [Table 2 and Fig. 1]. The mean skin elasticity was 0.787 A.U. before the application, 0.887 A.U. at 2 weeks after the application, and 0.873 A.U. at 4 weeks after the application. A significant difference in skin elasticity was observed between 2 weeks and 4 weeks after the application; the sample significantly improved skin elasticity (p<0.05).

Table 2 : Changes of skin dermal compactness values (Unit: A.U.)

Average	Week 0			Week 2			Week 4
Average	0.787	±	0.056	0.887	±	0.076^*	0.873	±	0.062^*

Values are mean ± SD (n=22). Week 0: just before application with sample, Week 2: two weeks after application with sample, Week 4: four weeks after application with sample. ^*p<0.05.

Figure 1.Changes of skin dermal elasticity values.

Bars represent mean ± SD (n=22). Week 0: just before application with sample, Week 2: two weeks after application with sample, Week 4: four weeks after application with sample. The values of week 2 and week 4 are significantly different from the value of week 0 by paired t-test (*p<0.05).

3.4. Changes in skin elasticity:

Changes in skin elasticity were measured by subtracting the value of skin elasticity measured before the application from that of skin elasticity measured after the application [Table 3]. The value of skin elasticity significantly increased at 2 and 4 weeks after the application (p<0.05), indicating that the mixed extract of the 3 medicinal herbs significantly improved skin elasticity.

Table 3 : Changes of skin dermal compactness values (Unit: A.U.)

Average	Week 2 - Week 0			Week 4 - Week 0
Average	0.100	±	0.070^*	0.086	±	0.064^*

Values are mean ± SD (n=22). Week 0: just before application with sample, Week 2: two weeks after application with sample, Week 4: four weeks after application with sample. The changed value at week 4 is significantly different from the changed value at week 0 by paired t-test (*p<0.05).

3.5. Rate of improvement in skin elasticity:

The rate of improvement in skin elasticity (%) was calculated by measuring changes in skin elasticity at the site of extract application [Table 4, Fig. 2]. Statistical analysis on the rate of improvement in skin elasticity (%) showed a more significant improvement at 4 weeks after the application (1.99%) than at 2 weeks after the application (1.42%) (p<0.05).

The results of the clinical assessment showed that the mixed extract of the 3 medicinal herbs created on the basis of the theories of Korean medicine improved facial wrinkles. Korean medicine is based on the theories of oriental medicine in which the body is perceived as a whole entity based on the traditional medical theory among internal organs, and maintenance of good health and treatment of diseases are achieved by promoting balance and harmony within the body¹⁴. Coptischinensis, Cimicifugaheracleifolia,and Poriacocoswere selected for this study as they are suggested to act on the heart, lungs, and spleen to promote maintenance of healthy skin and hair according to oriental medicine. Coptischinensisis the rhizome of a perineal herb that belongs to the Ranunculaceae family. It has a bitter taste, acts on the heart, and treats inflammatory disorders caused by heat¹⁵. Coptischinensishas significant anti-inflammatory and skin regeneration effects¹⁶. Therefore, it appears that Coptischinensisalso has skin regeneration and anti-wrinkle effects. Cimicifugaheracleifoliais the rhizome of a perennial herb that belongs to the Ranunculaceae family. It tastes spicy, acts on the lungs, and treats skin diseases caused by heat toxins¹⁷. It is also known to inhibit collagenase synthesis and promote hyaluronic acid synthesis¹⁸. These physiological activities of Cimicifugaheracleifolia may have certain effects that reduce facial wrinkles and promote healthy skin. A Poria coco (Schw) is the sclerotium of a fungus that belongs to the Polyporaceae family. It has a sweet taste and acts on the spleen to promote the flow of Qi blood, thereby increasing the flow of nutrients into the skin¹⁹. A Poria coco has been found to have antioxidant, anti-wrinkle, and anti-inflammatory effects according to a previous study²⁰. As can be seen, the anti-wrinkle effects of the 3 medicinal herbs selected for this study can be predicted based on the results of previous literatures.

In this study, we created a prescription of medicinal herbs on the basis of the theories of traditional Korean medicine and assessed the anti-wrinkle effects of 3 medicinal herbs using a scientific experimental method. The results of this study suggest that an individual medicine, as well as a combination of various medicines, based on the theories of traditional oriental medicine, have certain effects on skin health.

Table 4 : Improvement of skin dermal compactness values (Unit: A.U.)

Recovery rate	Week 2			Week 4
Recovery rate	12.936	±	9.631	11.267	±	8.662

Values are mean ± SD (n=22). Week 2: two weeks after application with sample, Week 4: four weeks after application with sample. The changed rate(%) at week 4 is significantly different from the changed rate(%) at week 2 by paired t-test (*p<0.05).

Figure 2.Comparison in the improvement of skin dermal compactness values.

Values are mean ± SD (n=22). Week 2: two weeks after application with sample, Week 4: four weeks after application with sample.

4.CONCLUSION:

In the present study, the effectiveness of a mixed extract of Coptischinensis, Cimicifugaheracleifolia, and Poriacocos (Schw.) at improving facial wrinkles was clinically assessed following the approval of the IRB. Improvement in skin elasticity was significantly higher at 2 weeks after the extract application (12.936%) than at 4 weeks after the application (11.267%) (p<0.05). These results confirmed that a mixed extract of Coptischinensis, Cimicifugaheracleifolia, and Poriacocos (Schw.) can inhibit wrinkle formation caused by aging and can reduce existing wrinkles. They also suggest the potential of the 3 medicinal herbs, Coptischinensis, Cimicifugaheracleifolia, and Poriacocos (Schw.) as raw materials for the development of cosmetic products and topical dermatologic drugs.

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http://e-ajbc.org/journal/view.php?number=847

Received on 10.12.2017 Modified on 11.02.2018

Research J. Pharm. and Tech 2018; 11(7): 3031-3035.

DOI: 10.5958/0974-360X.2018.00558.9

Anti-wrinkle Effects of Mixed Extracts of Three Medicinal Herbs