Effects of Oral Supplementation of Trichovitals^TM  on Human Skin, Hair and Nail Physiology

Chandramohan Marimuthu*, Jayaganesh Murugan, Srisunmathi Ravichandran, Sruthi Sukumar

Microcore Research Laboratories India Pvt Ltd., 9th km, 30 Feet Road, 204 - A Poondurai Main Road, Checkmedu, Erode - 638115, Tamil Nadu.

*Corresponding Author E-mail: info@microcoreresearch.com

ABSTRACT:

In this study, Microcore’s oral supplements of Trichovitals^TM contains Collagen type (I,III,V,X), elastin, ellagic acid, Sabbery b glucogallin, Biotin, Curcumin C3 reduct ODN and Murraya koenigii. ESM ingestion is associated with significant impact on hair thickness, reduction in hair breakage and improvement in hair growth.  This study leads to evaluate the impacts of a nutritional intervention as compared to control group (without oral supplements intake) on the appearances of hair, skin, and nails in healthy middle-aged adults. Two pilot studies were run to assess Trichovitals^TM on skin parameter. 1. Self-assessment questionnaire intended to perform an assessment on skin, hair, and nail health after 12 weeks of treatment for a group of 25 female volunteers 2. Evaluated the efficacy of Trichovitals^TM effect of 5-week treatment on hydration of skin by corneometry, on elasticity with the cutometer, and on pigmentation with themexameteron for a group of 7 female volunteers, Microscopic studies of hair strength and nail growth. In study 1-Participants were predominantly satisfied with the effects obtained on general face (100% volunteers satisfied) and (94% volunteers satisfied) body skin condition and skin properties (100% volunteers satisfied) with facial skin softness, 95% ith facial skin hydration, 63% of facial skin brightness and 89% with body skin hydration and significant effects on hair damages (100% volunteers satisfied), hair falls (95% volunteers satisfied) and hair growth (96% volunteers satisfied) and nail appearance (80% volunteers satisfied) and improvement in brittle nails. In this open-label 5 weeks study, Oral Microcore’s Trichovitals^TM ingestion was associated with a significant improvement in skin appearance such as increase skin hydration and skin elasticity, reduces skin pigmentation after 5 weeks treatment and facial skin tone, with significant impact on hair thickness, reduces split ends, hair loss, hair breakage and improvement in hair growth after 5 weeks treatment, increase of 80% nail growth rate and a decrease of 42% brittle nails after 6 weeks. Microcore’s Trichovitals^TM achieved significant change improvement in skin, hair and nail appearance (thickness/density) than control group (without oral supplements intake) by week 12 of the study. From the result analysis, Microcore’s Trichovitals^TM is deemed to be one of the most effective anti-aging products in cosmetic sector and overall physical appearance. Oral supplementation of 500mg/day of Microcore’s Trichovitals^TM for 12 weeks is associated with significant improvement in the appearance of facial skin, hair and nail.  Increases in skin hydration and skin elasticity with reduced pigmentation on skin were observed after 5 weeks treatment, which is a very important marker for skin aging. Also significant impact on hair thickness reduces split ends, hair loss, hair breakage and improvement in hair growth and healthy development of nails after 5 weeks treatment.

KEYWORDS: Collagen, Eggshell, Oral supplement, Pilot study, Skin, Hair, Nail.

1. INTRODUCTION:

In the human body, skin related concerns hair loss and nail health are faced by everyone these days. To be able to provide effective treatment for skin related problems, hair loss, hair fall and nail health. Skin is the largest organ and is involved in several important functions. The two main layers that make up the skin are the outer layer, or epidermis, and the inner layer, or dermis. The epidermis is composed of a stratified squamous epithelium that overlies a basal layer composed of columnar cells arranged perpendicularly (proliferating and differentiated keratinocytes). The dermis is composed of connective tissue consisting of diverse extracellular matrix components, including collagen and elastin fibers and glycosaminoglycans (GAGs), (Shaw G et al., 2017), which are synthesized by dermal fibroblasts. Underlying these layers is subcutaneous fat tissue. Hair loss in women is often an overlooked and underappreciated condition that affects women almost more frequently than it affects men. 40% of people with hair loss are women and 40% of women have visible hair loss by age of 40 in USA. Hair loss may begin in women as early and increases significantly with age. In one case study, 6% of women aged under 50 years were diagnosed as having female pattern hair loss, increasing to 38% in subjects aged 70 years and over (M. P. Birch et al., 2001). Signs of biotin deficiency include skin rashes, hair loss, and brittle nails (Zempleni J et al., 2009), (Mock D M et al., 2010). Therefore, biotin supplements are often promoted for hair, skin, and nail health.

In the effort to halt or slow aging, consumers often turn to interventions that may include cosmetic dermatology, over-the-counter products and dietary supplements. It has been estimated that in 2017, sales of products and services in this category hit $5.16 billion dollars.1 According to Nielson, in 2018, Americans spent $12 billion dollars on beauty and personal care products, with hair, skin, and nails falling under this category (Bagchi D et al., 2002). While there are many products and services available to address skin and hair issues, the efficacy of many of these products and services has been questioned. Therefore, due to the extensive cost to the consumer and the need for claim substantiation of such products, evaluation of efficacy is warranted.

It is known that the appearance of the hair, skin, and nails changes as part of the aging process. This is due to normal intrinsic and extrinsic aging as well as to environment exposures such as UV sunlight.

For example, the collagen density of the dermis decreases with age, which leads to a reduction in dermal thickness.

Similar changes also impact hair and hair appearance (shine, thickness, fullness, etc), and more specifically, molecular alterations lead to structural modifications of the hair fiber, decrease in melanin production, and lengthening of the telogen phase of the hair cycle, and thus altered hair appearance and health. The appearance of the nails also can change due to aging, including alterations in nail thickness, contour, surface, and color. Kompel AJ et al., (2019). Therefore, approaches to offset or even reverse these changes are desirable to consumers.

Currently, many medical treatments are available to men that are too dangerous for women or are unsafe for use in women of child-bearing age, such as finasteride. Trichovitals^TM is an dietary supplement which is specifically designed for skin, hair and nail related problems. Trichovitals^TM is a proprietary blend of Collagen type (I,III,V,X), elastin, ellagic acid, Sabbery b glucogallin, Biotin, Curcumin C3 Reduct^® ODN and Murraya koenigii which provides essential nutrients to nourish naturally to improve the appearance of skin, treats wrinkles, thinning hair, protects the hair damages, prevents hair falls, promotes hair growth and gives healthy nails. It does not contain hormones or other drugs.

Eggshell membrane, an organic substance, is known to increase cellular activity, increase collagen production, slow skin aging. Chicken egg shell and membrane are byproducts of eggs. The egg shell membranes are true source of valuable bioactive materials, including collagen that has many applications in medical, health and cosmetics. Eggshell membranes are an abundant raw material that are a novel source for naturally occurring bioactive compounds such as glucosamine (Picard J et al., 1973) chondroitin sulfate (Baker J R and Balch D A, 1962), hyaluronic acid (Long F D et al., 2005), fibrous proteins such as collagen Type I (Wong M et al., 1984) and sulfur-rich proteins (Tsai et al., 2006) and other components including lysozyme, ovotransferrin, ovocalixin and desmosine and isodesmosin. Glycosaminoglycan and proteins  essential for maintaining healthy joint and connective tissues.

Collagen Type (1,V,X) is the most important protein produced by the human body, it is mainly formed by the amino acid glycine (33%), proline and hydroxyproline (22%) (primary structure) in a triplex helix which is formed by three α chains. Elastin is a protein critical to skin, cardiovascular, cartilage and spinal column health. As its name suggests, elastin gives tissue the elastic tension and ability to resume their shape after stretching. Desmosine and isodesmosine - two little known amino acids that is responsible for elastin’s elastic and rubbery properties.

Chondroitin sulfate is a sulfated glycosaminoglycan (GAG) composed of a chain of alternating sugars (N-acetylgalactosamine and glucuronic acid). It is usually found attached to proteins as part of a proteoglycan. Chondroitin sulfate is an important structural component of cartilage and provides much of its resistance to compression.

The eggshell membrane contains a number of bioactive components that may impact the physiology and presentation of the hair, skin, or nails. For example, the eggshell membrane is rich in glycosaminoglycans (GAGs). GAGs are highly negative charged polysaccharides that appear to influence biomineralization. GAGs are complex molecules and can be part of larger molecules. One such example is proteoglycans, proteoglycans are glycosylated molecules where one or more specific glycosaminoglycan (GAG) and/or O- and N-linked oligosaccharides are attached to a core protein (Liu Z, Sun X, Cai C, He W, Zhang et al,.2016) There are several types of GAGs which are typically sulfated. These include chondroitin sulfate/dermatan sulfate (CS/DS), keratan sulfate (KS), and heparan sulfate (HS)/heparin. There are four specific GAG types in the chicken eggshell, hyaluronan, keratan sulfate, chondroitin sulfate, and heparan sulfate. Hyaluronan is the most prevalent GAG in eggshell membranes.

Eggshell membrane contains hyaluronan, thus also contains the GAG hyaluronic acid (HA). HA production occurs in the skin and has been shown to stimulate internal production of HA with oral intake. Keratan sulfate (KS) is also a natural GAG occurring in the hydrolyzed eggshell membrane. (Oe M, Sakei S, Yoshida H, et al.2017) In the body, KS may influence wound healing, especially that of the skin. Chondroitin sulfate (CS), another GAG, has also been tested for its ability to impact skin health.

Ellagic acid, also known as 2,3,7,8-tetrahydroxy(1)benzopyrano(5,4,3-CDE)(1)benzopyran-5,10-dione, is a well-known molecule belonging to the group of polyphenols. Ellagic Acid Pomegranate has the highest concentration of ellagic acid. Apart from the antioxidant activity, ellagic acid has skin whitening effect and also reduces UV-induced skin pigmentation, which is due to inhibition of tyrosinase activity by binding to copper at its active site (Shimogaki H et al., 2000). Ellagic acid has excellent hair growth promoting activity by enlarging the follicular size and prolongation of anagen (growing) phase. Ellagic acid can also be used in antidandruff shampoo preparations (Derkx T et al., 2011).  Formulation containing ellagic acid can be used as antipollution cosmetic agent, which protects the skin from detrimental effect of pollution. Ellagic acid-containing formulation has been reported for treating graying of the hair (Commo S, 2013).

Saberry more commonly known as Amla extract. It contains a minimum of 10% ß-glucogallin and 50% gallates. It protects the skin from the damaging effect of free radicals and heavy metals induced oxidative stress. Elevates the mitochondrial activity of human skin fibroblast and promotes the production of procollagen (J Varani  et al., 2000). It is an anti-aging ingredient. Also ß-glucogallin (an amla fruit extract) is a powerful antioxidant molecule and rejuvenator which prevents hair loss.

Biotin is a water-soluble vitamin and an essential coenzyme for several important enzymes. The use of these agents for hair loss is based on the observation (D’Amato P 2016). Biotin deficiency leads to dry, irritated skin and cracking of nails. One study in women who had brittle nails or splitting of the nails (Onychoschizia) noted that oral supplementation of biotin at 2.5mg over the course of at least six months increased nail thickness by 25% (reaching normal/healthy control values) (Colombo V E  etal., 1990). About half the group with brittle nails also experienced reductions in nail splitting. All nails from biotin-treated individuals showed some improvement when assessed by electron microscopy. Biotin protects the skin from acne, fungal infections, rashes, severe dryness and cracking of nails.

Curcumin, the active ingredient of turmeric, has been used for centuries as an anti-inflammatory agent (Laura V et al., 2019). It helps to treat acne and lighten the pigmentation. It is a powerful antioxidant which reduces hair loss, thinning of hair, reduces hair loss through breakage.

Murraya Koenigii is long known for its properties to strengthen hair roots. Promotes hair growth, prevents hair thinning and hair loss.

Oral supplementation of Trichovitals^TM has been linked with positive improvements in skin elasticity and appearance of the skin. It was the intent of this study to evaluate whether oral supplementation with a specific combination of Collagen type (I,III,V,X), elastin, ellagic acid, Sabbery b glucogallin, Biotin, Curcumin C3 reduct ODN and Murraya koenigii that contains a number of bioactive compounds that improves appearance, health of hair, skin, and nails. The aim of this study is to evaluate the impacts of a nutritional intervention on the appearances of hair, skin and nails in healthy middle-aged adults and to study the effectiveness of Trichovitals^TM biophysical parameters on skin, hair, nail, such as elasticity, pigmentation, and hydration, related to cutaneous aging, hair volume, hair thickness, hair loss, nail hardness, nail growth and nail irregularity.

2. MATERIALS AND METHODS:

2.1 Composition:

Composition for Microcore’s Trichovitals^TM was shown in the Table 1.

Table 1: Composition for Microcore’s Trichovitals^TM

S. No	Contents	Quantity
1.	Collagen (I,III,V,X) and Elastin (Desmosin + Isodesmosin)	455 mg
2.	Ellagic Acid (90%) (An extract from pomegranate fruit)	5 mg
3.	Sabbery (10%) b glucogallin (An extract from Amla fruit)	5 mg
4.	Biotin (a water soluble vitamin)	25 mg
5.	Curcumin C3 reduct ODN	5 mg
6.	Murraya Koenigii (An extract from curry leaf)	5 mg

2.2 Food supplement under investigation:

The dietary supplement of Microcore’s Trichovitals^TM used in this study contains Muttaijow Complex ^TM (Natural Egg Membrane Concentrate) consisting of egg membranes separated from eggshells by a patented process. Samples of Muttaijow Complex^TM were later retrieved for composition analyses followed by George W and Latimer JR AOAC official method, 2019. Compositional analysis of eggshell membranes has identified a high content of protein (collagen type I-V-X, elastin, keratin)and moderate quantities of GAGs (glucosamine, chondroitin sulfate, HA) (Picard, Paul-Gardais, and Vedel, 1973).We have confirmed the presence of the major components collagen, elastin, HA, Glycoaminoglycans in Muttaijow Complex ^TM composition.

2.3. Pilot study level - I

2.3.1 Selection of participants:

The study was conducted in Tamil Nadu Southern part of India at Erode district. The study enrolled a group of 25 adults, healthy volunteers of female. 40% of volunteers were in the age range of 30–45 years; 27% were 46–60 years; and 33% were over 60 years old.

2.3.2 Study design:

Participants took 500 mg of Microcore’s Trichovitals^TM daily for 12 weeks. Products were taken orally at home according to the instructions given by the investigator. The study consisted of a baseline and a follow-up visit at the end of treatment. Participants completed a self-assessment questionnaire regarding their experience with the regimen at 12 weeks. The questionnaire comprised three sections intended to perform an assessment on skin, hair, and nail health. The first part consisted of nine questions related to skin health. The second part included seven questions related to hair health, and the final section comprised four questions about nail health. Each question has five possible answers with an assigned score: (70-100%) great improvement, (30-60%) mild improvement, (0%-10%) no change. Finally, we calculated the percentage of volunteers giving a specific answer (Asserin J, et al.,2015)

2.4 Pilot study - II

2.4.1 Selection of participants:

The study was conducted in Tamil Nadu Southern part of India at Thiruchirapalli district. The study enrolled a group of 7 volunteers were in the age range of 25–60 years. Participants were treated daily over a 5-week period with oral supplementation of 500 mg of Microcore’s Trichovitals^TM for skin, hair and nail testing parameters.

2.4.2 Study Design:

Skin parameter testing:

Participants had the following assessments at each visit: skin hydration, skin pigmentation, and skin elasticity. Measurements were performed with the multifunctional skin analyzer MSD800 (Courage and Khazaka, Cologne, Germany), which allows measurement of skin parameters using different probes such as a corneometer, acutometer, and amexameter. The corneometer assesses skin surface hydration by electrical capacitance, which is the most common method used to determine the skin surface moisture (Proksch E et al., 2014). It measures the reactive capacitance of skin, using the stratum corneum as a dielectric membrane. Measurements are arbitrarily expressed as indices of hydration (0–99), which increase with increasing skin hydration. A score less than 30 implies very dry skin; a score of 30–60 indicates dry skin; and a score over 60 implies sufficiently moisturized skin. The mexameter measures the level of melanin, or pigmentation, and erythema, or redness, by reflection. Measurements are arbitrarily expressed as indices of pigmentation. Skin elasticity was performed with the cutometer, based on the well-established suction method. The resistance of the skin to be sucked up by negative pressure (firmness) and its ability to return to its original position (elasticity) are displayed as curves. To analyze skin elasticity, R₂ value (gross elasticity: resistance vs. ability of returning) was recorded; R₂ increases with increasing skin elasticity. The participants compliance (dosage and way of intake) and tolerance toward the products were checked weekly.

Hair Parameter Testing:

Daily scalp hair counts can be useful to the physician to help quantify how much the patient is losing and make sure that this is not more than the physiologic hair loss. It is said that it is normal to lose up to 100 hairs per day (Shapiro J et al., 2002). Patients are instructed to collect hairs shed in one day, count them and place them in plastic bags for every week of interval. All shed hairs in the shower or sink or on the brush are collected. If the patient is losing approximately more than 100 hairs per day, then the hair should be examined microscopically to detect any

abnormalities.

Nail Parameter Testing:

The properties of nails like nail growth, brittleness, flexibility and cracked nails were measured. After the oral consumption of Microcore’s  Trichovitals^TM over 5 week period, the growth of nail and its properties were studied after every week  interval and the brittleness of nails were also tested. (Zague V, et al.,2011) The subjects received a form in which they should record daily how many times their nails had been cracked or chipped on each hand.

3. RESULT AND DISCUSSION:

3.1 Compositional analysis of 500 mg of Microcore’s Trichovitals^TM

Muttaijow Complex^TM :

Typical analysis of Muttaijow Complex^TM resulted Solubility (20 %), Fat (0 %), Carbohydrate (0 %) and Ash (0 %) was shown in the Table 2. Amino acid composition of Muttaijow Complex ^TM for Lysine (2.98%), Tryptophan (2.71%), Leucine (3.96%), Aspartic acid (7.05%), Proline (8.25%), Isoleucine (2.12%), Threonine (4.67%), Glycine (4.01%), Histidine (2.91%), Arginine (5.36%), Tyrosine (1.34%), Glutamic acid (8.25%), Cystine (6.14%), Alanine (2.05%), Methionine (2.85%), Valine (5.15%), Phenylalanine (1.49%), Serine (4.3) were shown in the Table 3. Constituents of Muttaijow Complex ^TM resulted 96% protein, 17% Collagen, 27% Elastin (16.2% Desmosine, and 10.8% Iso desmosine), 28.3% Chondroitin sulphate, 20.5. Glycosaminoglycan and 5% Hyaluronic acid was shown in the Table 4. Collagen and elastin taken for formulation of Microcore’s Trichovitals^TM.

Table 2: Typical analysis of Muttaijow Complex^TM

S. No	Sample	% Muttaijow Complex ^TM
1	Solubility	40
2	Fat	0
3	Carbohydrate	0
4	Ash	0

Table 3: Typical Amino acid composition of Muttaijow Complex^TM

S. No	Aminoacid	% Muttaijow Complex ^TM
1	Lysine	2.98
2	Tryptophan	2.71
3	Leucine	3.96
4	Aspartic acid	7.05
5	Proline	8.25
6	Isoleucine	2.12
7	Threonine	4.67
8	Glycine	4.01
9	Histidine	2.91
10	Arginine	5.36
11	Tyrosine	1.34
12	Glutamic acid	8.25
13	Cystine	6.14
14	Alanine	2.05
15	Methionine	2.85
16	Valine	5.15
17	Phenylalanine	1.49
18	Serine	4.3

Table 4: Typical constituents of Muttaijow Complex^TM

S. No	Sample	% Muttaijow Complex ^TM
1	Protein	96
2	Collagen (Collagen Type (I,III,V, IX)	17
3	Elastin	27
4	Desmosine	16.2
5	Isodesmosine	10.8
6	Chondroitin sulfate	28.83
7	Glycosaminoglycan	20.15
8	Hyaluronic acid	5

Compositional analysis of other ingredients such as Ellagic acid, Sabbery, Biotin, Curcumin and Murraya koenigii analysed by George W and Latimer JR AOAC official method, 2019.

3.2 Pilot study-I:

Self-assessment questionnaire responses were analyzed, and results are shown Table 5 below. The majority of the participants observed improvement on general face and body skin condition. Of these, 39% and 50% noticed great improvement in general face and body skin condition, respectively. Volunteers’ self-assessment of specific skin properties showed variable percentages, with 33% to 100% of participants observing an improvement. The parameters that received the most positive evaluation (over 50% of participants manifested some positive change) were, in this order; facial skin softness (100%), facial skin hydration (95%), body skin hydration (89%), facial skin brightness (83%), and smoothness of facial skin (72%). Of these, facial skin softness (85%) and body skin hydration (80%) showed great improvement. The effects on hair and nail general condition were valued positively by 67% and 50% of volunteers showed great improvement, respectively. The most outstanding qualities according to volunteers’ self-assessment were skin brightness (63%) showed mild improvement above 25 years and nail hardness (61%) showed mild improvement above 45 years.

Table 5: Effects of Microcore’s Trichovitals^TM according to participants’ self- assessment questionnaires

S. No	Parameters	% of Volunteers satisfaction
		25% adults of 25-30 years (%)	40% of 30-45 years (%)	27% of 45-60 years (%)	33% of over 60 years (%)
1	Skin Property
	Facial Skin Hydration	95	95	94	92
	Facial Skin Brightness	63	62	60	60
	Smoothness of the Facial skin	79	77	75	72
	Facial skin softness	100	100	99	98
	General improvement of facial skin	98	98	95	93
	Body skin hydration	89	88	85	82
	General improvement of body skin condition	85	84	83	81
2	Hair Property
	Hair volume	96	93	92	91
	Hair thickness	97	96	95	92
	Prevent hair loss	95	94	93	85
	General improvement of hair condition	100	95	94	93
3	Nail Property
	Nail hardness	75	71	69	64
	Nail faster growth	80	75	73	65
	Decrease of nail irregularity	82	82	79	77
	General improvement of nail condition	82	80	65	63

[(71-100%) great improvement, (21-70%) mild improvement, (0%-20%) no change]

3.3 Pilot Study-II:

3.3.1 Skin biophysical parameters:

The overwhelming success of Microcore’s Trichovitals^TM treatment, among volunteers participating in the study, led us to conduct a second study aimed at measuring skin parameters such as skin hydration, pigmentation, and elasticity. In total, seven participants completed the study. There were no discomfort or adverse reactions reported. These pilot studies set out to explore the effect of Microcore’s Trichovitals^TM, a dietary supplement consisting of egg membranes, on skin biophysical parameters related to cutaneous aging. Increases in skin hydration and skin elasticity with reduced pigmentation on skin were observed after 5 weeks treatment of Microcore’s Trichovitals^TM supplements than control groups (without intake of oral supplement). Our results are in agreement with previous investigations in which oral administration of collagen-derived products has shown improvement in skin elasticity (Proksch E et al., 2014). The improvement observed in skin elasticity suggests a long-lasting positive physiological effect, in contract to effects of topical skin-care products, which increase skin elasticity predominantly by enhancing epidermal hydration. Moreover, treatment with eggshells has previously shown a positive effect on skin parameters such as pigmentation and hydration. From the results oral intake of 500 mg/day Microcore’s Trichovitals^TM shows significant improvement from initial to after 5 weeks of treatment and also a significant decrease in pigmentation of skin was shown in the Table 6 and Graph 1.

Table 6: Effect of Microcore’s Trichovitals^TM in skin parameters

S. No	Skin Parameters	Time (Weeks)
		Control	Initial	Week 1	Week 2	Week 3	Week 4	Week 5
1	Skin hydration	51.4	51.4	52.7	53.5	55.7	59.9	77.4
2	Skin pigmentation	25.6	25.6	23.8	20.6	18.5	16.7	15.1
3	Skin elasticity	71.4	71.4	66.7	68.9	69.5	72.5	80.5

Graph 1: Effect of Microcore’s Trichovitals^TM on skin biophysical parameters

3.3.2 Hair biophysical parameters:

500 mg of oral supplement of Microcore’s  Trichovitals^TM treated with split ends, hair loss, hair breakage, strong hair scalp improvement . Before and After 5 weeks of treatment (Shapiro J.et al.,2002), there is an significant impact on participants with overwhelming feedback. Improvement in thickened hair growth with reduced split ends, reduction in hair loss and increased in hair strength was shown in the Table 7 and Graph 2. In Figure 1, the split ends of hair was measured under the microscope before and after oral supplementation of Microcore’s Trichovitals^TM.

Table 7: Effect of Microcore’s Trichovitals^TM in hair parameters

S. No	Hair Parameters	Time (Weeks)
		Control	Initial	Week 1	Week 2	Week 3	Week 4	Week 5
1	Hair loss	94.5	94.5	85.2	71.8	65.34	50.6	30.3
2	Hair split ends	24.6	24.6	23.8	20.6	19.2	18.9	16.2

Table 8: Effect of Microcore’s Trichovitals^TM in nail parameters

S. No	Nail Parameters	Time (Weeks)
		Control	Initial	Week 1	Week 2	Week 3	Week 4	Week 5
1	Nail  growth(cm)	1.1	1.1	1.2	1.4	1.5	1.59	1.7

Graph 2: Effect of Microcore’s Trichovitals^TM on Hair biophysical parameters

Figure 1:  Microscopic view of hair at 4X of before and after Treatment with Microcore Trichovitals^TM

3.3.3 Nail biophysical parameters:

500 mg of oral supplement of Microcore’s Trichovitals^TM treated with participants. Before and after 5 week of treatment, increase of 80% nail growth rate and a decrease of 42% brittle nails within 4 weeks were observed in the Table 8 and Graph 3.

Graph 3: Effect of Microcore’s Trichovitals^TM on Nail biophysical parameters

Before starting the treatment, the participant’s frequency of broken nails was on average 10 times/mo. It significantly decreased to 6 times/mo after 5 weeks of treatment, representing a reduction of 60%. The ingestion of Microcore’s Trichovitals^TM led to a global clinical improvement in brittle nails, a significant increase in nail growth rate, and a decrease in the frequency of cracked or chipped nails in the present trial after 5 weeks of treatment.

4. CONCLUSIONS:

A pilot studies on oral supplementation of 500 mg/day of Microcore’s Trichovitals^TM for 12 weeks has been conducted to investigate the effect of on skin, hair and nail health by self-assessment questionnaires and their properties are studied. The following results were observed from the Pilot studies. In the study 1, self-assessment questionnaire are investigated to check the effect of Trichovitals on skin, hair, and nails. It was observed that participants were predominantly satisfied with the beneficial effects on skin health. Volunteers shown 100% satisfied on general face and 94% satisfied on body skin condition and 100% volunteers satisfied skin properties with facial skin softness, 95% with facial skin hydration, 63% of facial skin brightness and 80% with body skin hydration. Around 79% of volunteers shown a satisfactory result on body skin smoothness.  Almost 95% of people were shown a good development in hair volume, thickness and overall hair richness. 100% satisfied on significant effects on hair damages, 95% satisfied on hair falls and 96% satisfied on hair growth. There was also a significant improvement in nail and the volunteers of 70-80% gave development of clean nail and reduced white and gray lines. From the pilot study no 2 it was noted that the significant improvement in skin elasticity was absorbed after the 5-week with Microcore’s Trichovitals^TM treatment. Collagen type-1 is being a key molecule of skin as it is found in Microcore’s Trichovitals^TM and that plays an important role in maintaining its structure and hydration.  Thus the study concludes that oral intake of Microcore’s Trichovitals^TM led to a significant increase  in skin hydration and skin elasticity with reduced pigmentation on skin were observed after 5 weeks treatment, which is a very important marker for skin aging. Microcore’s Trichovitals^TM also shows significant impact on hair thickness, reduces split ends, hair loss, hair breakage and improvement in hair growth. Above 90% of hair loss has been reduced after 5 week of oral intake of Trichovitals. Improvement in nail physiology with increase of 80% nail growth rate and a decrease of 42% brittle nails within 5 weeks. Thus Microcore’s Trichovitals^TM achieving a significant change in skin, hair, nails than control group (without oral supplements intake). From the result analysis, Microcore’s Trichovitals^TM is deemed to be one of the most effective anti-aging products in cosmetic sector. Collagen and elastin present in Muttaijow Complex^TM of Microcore’s Trichovitals^TM gives natural glow in the skin and have proven a good dermatologic effects. Oral supplementation of 500mg/day of Microcore’s Trichovitals^TM 12 weeks is associated with significant improvement in the appearance of facial skin, hair, nail growth and improvement of brittle nails in conjunction with a notable decrease in the frequency of broken nails.

5. REFERENCES:

1.   George W and Latimer JR (2019), “Official method of analysis of AOAC” by AOAC international 21st Edition.

2.   Kompel AJ et al. (2019), “Intra-articular Corticosteroid Injections in the Hip and Knee: Perhaps Not as Safe as We Thought?” Radiology 190341.

3.   Oe M, Sakei S, Yoshida H, et al (2019), “Oral hyaluronan relieves wrinkles:a double blinded, placebo-controlled study over a 12-week period” Lin Cosmet Invest Dermatol vol-10  pp. 267-273.

4.   Laura V, Mattia F, Roberta G, Federico I, Emi D, Chiara T, Luca B and Elena C (2019), “Potential of Curcumin in Skin Disorders” Nutrients vol.11(9) pp.2169.

5.   Shaw G et al. (2017), “Vitamin C-enriched gelatin supplementation before intermittent activity augments collagen synthesis.” Am J Clin Nutr vol-105(1) pp.136-143.

6.   Liu Z, Sun X, Cai C, He W, Zhang F, Linhardt RJ (2016), “Characteristics of glycoaminoglycans in chicken eggshells and the influence of disaccharide composition on eggshell properties” Poult Sci. vol.95: pp.2879-2888.

7.   D’Amato P (2016), “Biotin: 5 healthy benefits of this B vitamin. Chatelaine”

8.   Asserin J, Lati E, Shioya T, Prawitt J (2015), “The effect of oral collagen peptide supplementation on skin moisture and dermal collagen network: evidence from an ex-vivo model and randomized, placebo controlled clinical trials” J Cosmet Dermatol vol-14 pp.291-301.

9.   Proksch E, Segger D, Degwert J, Schunck M, Zague V, Oesser S (2014), “Oral supplementation of specific collagen peptides has beneficial effects on human skin physiology: a double-blind, placebo-controlled study” Skin Pharmacol Physiol vol-27(1) pp.47–55.

10. Commo S (2013), “Use of ellagic acid for treating graying hair” Patent EP 1870081 B1.

11. Zague V, De FV, Rosa C, Jaeger RG (2011), “Collagen hydrolysate intake increases skin collagen expression and suppresses matrix metalloproteinase 2 activity” J Med Food vol-14 pp.618-624.

12. Derkx T, Richet L (2011), “Composition comprising ellagic acid and a particular cationic surfactant, and cosmetic use thereof” Patent WO 2011107432 A2.

13. Mock D M (2010), “Biotin. In: Coates PM, Betz JM, Blackman MR, et al., eds. Encyclopedia of Dietary Supplements” 2nd ed. London and New York: Informa Healthcare pp.43-51.

14. Zempleni J, Wijeratne SSK, Hassan YI (2009), “Biotin. Biofactors” vol-35 pp.36-46.

15. Tsai W.T., Yang J.M., Lai C.W., Cheng Y.H., Lin C.C., Yeh C.W (2006),“Characterization and adsorption properties of eggshells and eggshell membrane” Bioresource Technology pp. 488-493.

16. Long F.D., Adams R.G., DeVore D.P (2005), “Preparation of hyaluronic acid from eggshell membrane. USA Patent 6946, 551.

17. Bagchi D, Misner B, Bagchi M, Kothari SC., Downs BW., Fafard RD., Preuss HG (2002), “Effects of orally administered undenatured type II collagen against arthritic inflammatory diseases: A mechanistic exploration” Int J Clin Pharmacol Res vol-22(3- 4) pp.101-10.

18. Shapiro J (2002), “Assessment of the patient with alopecia. In: Hair Loss., editor. Principle of Diagnosis and Management of alopecia” 1st ed. London: Martin Dunitz Ltd.

19. M. P. Birch, J. F. Messenger, and A. G. Messenger (2001), “Hair density, hair diameter and the prevalence of female pattern hair loss,” British Journal of Dermatology, vol. 144, no. 2, pp. 297–304.

20. J Varani, R L Warner, M Gharaee-Kermani, S H Phan, S Kang, J H Chung, Z Q Wang, S C Datta, G J Fisher, J J Voorhees (2000), “Vitamin A antagonizes decreased cell growth and elevated collagen-degrading matrix metalloproteinases and stimulates collagen accumulation in naturally aged human skin” J Invest Dermatol vol-114(3) pp.480-6.

21. Shimogaki H, Tanaka Y, Tamai H, Masuda M (2000), “In vitro and in vivo evaluation of ellagic acid on melanogenesis inhibition” Int J Cosmet Sci vol-22 pp.291–303.

22. Colombo V E, Gerber F, Bronhofer M, Floersheim G L (1990), “Treatment of brittle fingernails and onychoschizia with biotin: scanning electron microscopy” J Am Acad Dermato vol-23 pp.1127-32.

23. Wong M., Hendrix M.J.C., Von der Mark K, (1984), “Collagen in the egg shell membranes of the hen” Dev. Biol vol-104 pp. 28-36.

24. Picard J, A. Paul-Gardais, Vedel M. (1973), “Sulfated glycoproteins from egg shell membranes and hen oviduct. Isolation and characterization of sulfated glycopeptides”, Biochim. Biophys. Actavol- 320 , pp. 427-441.

25. Baker J.R., Balch D.A (1962), “A study of the organic material of hen’s-egg shell” Biochem. J pp. 352-361.

Received on 15.02.2021            Modified on 11.03.2021

Accepted on 28.03.2021              ©A&V Publications all right reserved

Research J. Topical and Cosmetic Sci. 2021; 12(1):51-58.

DOI: 10.52711/2321-5844.2021.00008