Hair Regrowth

Serum, Shampoo, and Conditioner


Pure Guild Hair Regrowth Serum, Shampoo, and Conditioner set the new benchmark for molecular purity in topical treatments:

  • Pure Guild extracts pristine active compounds mechanically, using no industrial solvents or damaging heat, creating a standard of purity by which other products can be measured.
  • Pure Guild employs only the active ingredients proven most effective in rigorous clinical trials, creating a standard of cosmeceutical performance.

Pristine Mechanical Extraction

Active ingredients in Pure Guild Hair Regrowth Serum, Shampoo, and Conditioner maintain their molecular integrity and full spectrum of biological activities because no solvents are used and no heat is applied to extract them.

Pure Guild employs only gentle mechanical compression over time to render highly effective compounds from potent raw materials. Although costly, this process yields a superior molecule, while other organic brands use chemical solvents like hexane or ether, which adulterate the final product, and heat distillation, which inhibits therapeutic properties.

Super-premium Pure Guild cosmeceuticals contain no sodium lauryl sulfate or other detergents. They are strictly hypoallergenic, non-irritating, and never tested on animals.


Latest BioTechnical Science In Hair

Pure Guild Hair Regrowth Serum, Shampoo, and Conditioner incorporate only the active ingredients proven most effective via clinical trials. Dramatically, one extract of the botanical Lupinus albus, native to the Mediterranean region, has shown in multiple scientific studies to control testosterone and boost the density, activity, and blood supply of delicate hair follicles:

  • After 86 days of clinical treatment, Lupinus albus extract increased the density of hair by 12 percent, the number of follicles in anagen growth phase by 17 percent, and the ratio of active to dormant follicles by an astonishing 118 percent.
  • L albus extract inhibited the enzyme 5α-reductase by 18 to 31 percent — more than the oral drug finasteride — and reduced formation of the dihydrotestosterone that would cause hair follicles to miniaturize and eventually to stop producing hair.
  • L albus stimulated vascular endothelial growth factor by 17 to 30 percent and stimulated the development of new vascular branching to deliver essential nutrients to growing hair.


Hair follicle in longitudinal section


Hair follicle in cross section

Hair Growth

STATE OF THE SCIENCE

Hair is a social marker, an important component of attraction, and the loss of hair is associated with a loss of power. Male pattern balding starts at an early age, between 15 and 20, and may progress to total baldness very rapidly. To arrest and delay significant loss, specialists recommend prevention at the first sight of symptoms.

The leader in cosmeceutical science, Pure Guild researches hair pathology in depth and offers the most effective topical treatments to combat balding. Currently, attention is focused on active ingredients that affect four factors of the hair cycle: epithelial cells, peripheral nervous system, immune system, and vascular system. Pure Guild has developed today’s state-of-the-science treatment for hair regrowth, Lupinus albus extract, derived from the white lupine native to the Mediterranean. Rich in peptides, trace elements, and vitamins, the extract has been proven in clinical trials.

HAIR CYCLE

Lupinus albus extract performs simultaneously on three fronts of the hair cycle:

  • Enzymatic: L albus restores hormonal balance by inhibiting activity of the enzyme 5α-reductase II.
  • Metabolic: L albus stimulates metabolic activity in hair-follicle cells, favoring keratinization and hair growth.
  • Vascular: L albus improves vascularization of the follicle, facilitating nutrient supply to construct new hair.

Tested in vivo, the active botanical extract deployed in Pure Guild Hair Regrowth Serum, Shampoo, and Conditioner is proven to favor hair growth and return hair density to normal levels. In each hair cycle, a follicle undergoes three phases: anagen, catagen, and telogen.

ANAGEN PHASE

The active phase of hair growth lasts for three to five years. Normally, 80–85 percent of follicles are in anagen phase. Each can yield about 20 terminal hairs. Divided into six stages, anagen requires intense metabolic activity in the hair bulb:

Stage I: The follicle is regenerated from a bulge — believed to be stem cells — located where the sebaceous gland opens onto the shaft.
Stage II: Envelopment of the dermal papilla forms the bulb and its matrix.
Stage III: The bulb increases in size. Mitotic activity becomes intense. Signs of hair and epithelial-sheath differentiation appear. Melanocyte activity begins.
Stage IV: Keratinization of hair takes place.
Stage V: The hair grows flush with the epidermal surface.
Stage VI: Visible hair emerges.

CATAGEN PHASE

In this phase — about three weeks — the follicle regresses. Cells become keratinized. Mitotic activity stops. The follicle takes the form of a club.

TELOGEN PHASE

The quiescent phase lasts about three months. The hair remains anchored in the follicle and continues to rise. At the end, the hair falls, and a new cycle starts. During the hyperproliferative anagen phase, the follicle requires a particular physical, biochemical, and psychological balance to develop normally. Any stressors can thrust anagen follicles back into telogen phase, a condition called alopecia or baldness. There are three types:

  • Traumatic alopecia,
  • Alopecia due to disease, and
  • Androgenetic alopecia.

To restart the hair cycle, a treatment must reactivate the anagen growth phase by acting simultaneously on three factors:

  • Hormone balance,
  • Vascular system, and
  • Cell metabolism.

HORMONE BALANCE

Tested at 0.25% on normal human fibroblasts, Lupinus albus extract inhibits by 18 percent the activity of 5α-reductase II, an enzyme that enables conversion of testosterone into the 5α-dihydrotestosterone that causes male pattern baldness.

Androgens play a fundamental role in this phenomenon, characterized by dwindling hair diameter, a process called miniaturization. In the course of androgenetic alopecia, follicles experience increased activity from the enzyme 5α-reductase II and higher levels of the hormone 5α-dihydrotestosterone (5α-DHT).

The 5α-DHT appears to be the most important factor in miniaturization. It binds to androgen receptors and forms a hormone/receptor complex that activates the genes responsible for hair shedding.

VASCULAR SYSTEM

Tested at 0.5%, Lupinus albus extract increases the synthesis of vascular endothelial growth factor (VEGF) by 30 percent. L albus extract makes the existing vascular system of the hair follicle denser to facilitate better nutrient supply to cells.

Vascularization of a follicle is essential for development and growth, especially in anagen phase. Growth factors, cytokines, and other molecules are involved. One essential growth factor is VEGF, known to stimulate angiogenesis and vascular permeability. VEGF acts directly on dermal ridges and stimulates formation of a robust vascular system in proximity to follicles, fulfilling their nutritional needs. Levels are lower in individuals suffering from androgenetic alopecia, resulting in less-than-adequate vascularization.

CELL METABOLISM

During anagen phase, cellular activity is intense. Cells divide continuously to manufacture hair strands. After differentiation, fibers accumulate keratin, hardening and leading to the disintegration of nuclei. A growing hair is prima facie evidence of cellular multiplication. Pressure pushes the hair out through the surface.

Lupinus albus extract provides peptides, zinc, and iron to stimulate the metabolic activity of hair-bulb cells and boost keratinocyte differentiation, as seen in the 44-percent increase in expression of mRNA coding for type 1 transglutaminase (L albus extract at 1%).


Efficacy

GROWTH FACTOR

In clinical studies, Lupinus albus extract improves vascularization of the hair follicle, a process essential for the manufacture of hair. Tested at 0.5%, it leads to a 30 percent increase in synthesis of vascular endothelial growth factor (VEGF), required to formulate the vascular system.

The aim of this study was to determine the capacity of L albus extract to stimulate synthesis of the growth factor and to enable new vascularization in proximity to dependent hair follicles, thus favoring the essential nutrient supply required.

Day 1: Keratinocytes were inoculated in six-well plates at 100,000 cells per well.

Day 2: Keratinocytes were treated with L albus extract at 0.25% and 0.50%, followed by incubation for 72 hours.

Day 5: After incubation, the cell supernatants were recovered. VEGF was assayed with an enzyme-linked immunosorbent assay kit.


Agent tested
L albus extract 0.25%
L albus extract 0.50%
VEGF synthesis vs. control
+17%
+30%

Table 1. Effect of Lupinus albus on synthesis of VEGF

ANGIOGENESIS

In further studies, Lupinus albus extract at 0.25%, 0.50%, and 1.00% increased formation of new vessels, called ramifications, from the existing circulatory system. L albus extract favors angiogenesis, thus facilitates the supply of oxygen and nutrients essential for cells during intensive activity.

The aim of this study was to determine the capacity of L albus to stimulate angiogenesis, the process of forming new vessels.

When endothelial cells are cultivated in vitro, they create new vasculature by migration, forming continuous tubules under the influence of various angionenic factors. It is thus possible to obtain a hair-follicle structure with ramifications.

Endothelial cells were visualized, and ramifications were quantified by immunolabeling, using an antibody-to-platelet-endothelial-cell-adhesion molecule (PECAM-1), a marker of these cells.

Cells were cultured with L albus at 0.25%, 0.50%, and 1.00% for 12 days. Incubation was at 37°C in a 5% CO2 atmosphere. Quantification was done with an Olympus IX70 microscope coupled to a Visiolab 2000 image-analysis system. Results are shown in Tables 2 and 3.


Agent tested
Control
Length of tubular structures
1290 µm
L albus extract 0.25% 1395 µm (+8%)
L albus extract 0.50% 2231 µm (+73%)
L albus extract 1.00% 2530 µm (+96%)

Table 2. Effect of Lupinus albus on tubular structures


Agent tested
Control
Number of ramifications
1
L albus extract 0.25% 4
L albus extract 0.50% 8
L albus extract 1.00% 9

Table 3. Effect of Lupinus albus on ramifications

5A-REDUCTASE ACTIVITY

In enzymatic studies, Lupinus albus extract tested at 0.25% on human fibroblasts showed an 18 percent inhibiting effect against 5α-reductase II activity — comparable to the oral drug finasteride.

The aim of this study was to determine the effect of L albus extract on the enzyme 5α-reductase II, which catalyses the hydroxylation of testosterone into 5α-dihydrotestosterone, known to cause alopecia.

Confluent fibroblasts were preincubated for two hours in the extract at 0.25% and 2.50%, and in a reference compound, finasteride at 3 ng/ml. After 25.5 hours of treatment, cells and supernatants were recovered. Homogenates were extracted by dichloromethane. After evaporation, dry residues were dissolved in methanol. Deposits were carried out on silica gel plates. Non-radiolabeled standards, testosterone, 5α-dihydrotestosterone, and androstenedione were spotted on each. Plates were scanned, and standards were revealed by spraying with a sulfuric acid at 5%, then heating at 100°C for 10 minutes. Proteins were assayed.

The biotransformation of testosterone was seen by the formation of several metabolites of which two were 5α-DHT and androstenedione. The formation of 5α-DHT reflects the activity of 5α-reductase II. Measuring the percentage of transformation made it possible to evaluate the enzymatic activity. The percentages of testosterone metabolism into 5α-DHT were calculated, and 5α-reductase II activity was expressed as pmole of 5α-DHT formed/h/mg of cellular proteins.


Agent tested

Control
Finasteride (3 ng/ml)
L albus extract 0.25%
L albus extract 2.50%
5α-reductase as pmol
of 5α-DHT formed/h/mg proteins

11.3
9.1 (–21%)
9.3 (–18%)
7.7 (–31%)

Table 4. Inhibition of 5α-reductase with Lupinus albus

CELL METABOLISM

In studies of metabolic activity, Lupinus albus extract tested at 1% favored the expression of mRNA coding for type 1 transglutaminase, a marker of keratinocyte differentiation. By boosting metabolism, especially important in the anagen phase, L albus stimulates hair growth.

The aim of this study was to determine the effect of L albus on metabolic activity in cultured keratinocytes. It measured the expression of mRNA of the differentiation marker, type 1 transglutaminase.

Human keratinocytes were incubated in a medium for 48 hours at 37°C in an atmosphere containing 5% CO2 in the presence of L albus extract at 0.5% and 1.0%. After incubation, cells were recovered and total RNA was extracted. The RNA was subjected to reverse transcriptase, and the DNA obtained was analyzed with PCR, using oligonucleotides complementary to a sequence coding for the genes of the proteins studied. B-actin mRNA was also analyzed in each experimental condition.

The intensity of amplicon bands, which are pieces of DNA formed as products of amplification events, on agarose gels was quantified by image analysis. Results are expressed as a ratio of type 1 transglutaminase mRNA to the control, shown in Table 5.


Agent tested
L albus extract 0.50%
L albus extract 1.00%
MRNA vs. control
21% ± 7%
44% ± 8%

Table 5. Metabolic activity of keratinocytes after treatment with Lupinus albus extract

ANTI-HAIR-LOSS EFFECT

In vivo, after 12 weeks of treating with Lupinus albus extract, both hair density and anagen/telogen ratio increased significantly, proving the ingredient’s capability.

The aim of this study was to determine the action of topical L albus, tested at 10% on human subjects, to reduce hair loss. The study was conducted on 16 healthy male volunteers between 18 and 65 years of age, with slight to moderate baldness. No volunteer had treated before.

A 10% solution was applied once a day for 12 consecutive weeks. To evaluate efficacy, the phototrichogram is a standardized macrophotographic method.

A 1 cm² region was delimited in the right temporal region of each volunteer. Photos were taken on days 1 and 3 with a video-microscope at 25x magnification. Photos were taken again on days 42, 44, 84, and 86. Enumeration of hairs on the macrophotos enabled evaluation of:

  • Hair density
  • Number of anagen hairs — those growing during the two-day intervals — and
  • Number of hairs in telogen phase — those that stopped growing or fell out.

Analysis of the phototrichograms enabled comparisons at different points in time. Typical density for a subject free of alopecia is 200–400 hairs per cm². In balding subjects, however, density decreases considerably to 80 hairs/cm² or less.

Based on the number of hair shafts in anagen and telogen phases, it is possible to calculate the anagen/telogen ratio, normally 4 or 5. If less than 1.5, the scalp is pathologic. The ratio is used to assess efficacy of anti-hair-loss treatments.

  • Day 1: Temporal site was delimited, shaving hair down to emergence, and macrophotography was done.
  • Days 1–3: No hair treatment or shampoo was applied.
  • Day 3: New macrophotography was done, and volunteers received product.
  • Days 3–42: Product was applied each day.
  • Day 42: Temporal site was delimited, shaving hair down to emergence, and macrophotography was done. Acceptability and compliance were verified.
  • Days 42–44: No hair treatment or shampoo was applied.
  • Day 44: New macrophotography was done, and the second bottle of product was given. Acceptability and compliance were verified.
  • Days 44–84: Product was applied each day.
  • Day 84: Temporal site was delimited, shaving hair down to emergence, and macrophotography was done. Acceptability and compliance were verified.
  • Days 84–86: No hair treatment or shampoo was applied.
  • Day 86: Macrophotography and evaluation of cosmetic acceptability were done. Compliance was verified.

Analysis evaluated these parameters:

  • Total hair density,
  • Anagen-phase hair density, and
  • Anagen/telogen ratio.

Results are shown in Tables 6 and 7.
After 44 days of application:

  • Total hair density moderated,
  • Density of hair in the anagen phase remained stable, and
  • Anagen/telogen ratio grew 85.8 percent.

After 86 days of application:

  • Hair density increased by 11.9 percent,
  • Anagen density increased 17.1 percent, and
  • Anagen/telogen ratio grew 117.7 percent.

Day
Day 0
Day 44
Day 86
Mean number of hairs/cm²
110.4
106.2
123.6

Table 6. Effect of Lupinus albus extract on hair density
Day
Day 0
Day 44
Day 86
Mean anagen follicles/cm²
103.4
103.4
121.1

Table 7. Effect of Lupinus albus on follicles in anagen

 

Safety Tests

HYPOALLERGENIC, NON-TOXIC

Tests
Determination of tolerance after application under a
    bandage for 48 hours
Determination of sensitizing potential in volunteers with
    normal skin
Mutagenicity tests
Phototoxicity tests
Results

non-irritant

hypoallergenic
non-mutagenic
non-phototoxic

 

Directions

Serum: Part hair and apply 15 pumps onto the area affected by hair loss or thinning. Rub the solution into the scalp and allow it to dry. Wait at least three hours before washing hair, swimming, or engaging in strenuous exercise. For best results, apply twice daily without skipping.

Shampoo: Massage the product into the scalp and hair thoroughly and leave it on for one minute before rinsing. Application could be repeated on hair that needs additional cleansing. For best results, follow with Pure Guild Hair Regrowth Conditioner.

Conditioner: After washing, massage the conditioner thoroughly into hair and scalp and leave it on for five minutes before rinsing. For best results, follow with Pure Guild Hair Regrowth Serum.

 

Ingredients

Serum
Primary ingredient: Lupinus albus extract
Inactive ingredients: deionized water, glycerol (vegetable origin), xanthan gum, isostearyl lactate, phenoxyethanol

Shampoo
Primary ingredient: Lupinus albus extract
Inactive ingredients: Aloe vera gel, trimethylglycine (sugar beet), glycerin, seaweed extract, witch hazel extract, chamomile extract, arnica blossom extract, phenoxyethanol, sodium hyaluronate, ginseng extract, tocopherol, essential oils, glycolic acid, sodium chloride, potassium sorbate, sodium benzoate, citric acid

Conditioner
Primary ingredient: Lupinus albus extract
Inactive ingredients: Aloe vera gel, lavender extract, algae extract, jasmine extract, glycerin, cetyl alcohol, stearalkonium chloride, stearyl alcohol, glycol stearate (vegetable origin), seaweed extract, Spirulina extract, chamomile extract, almond oil, keratin oil, D-panthenol, retinyl palmitate, olive butter, phenoxyethanol, mango butter, ginseng extract, natural essential oils, tocopherol, potasium sorbate, sodium benzoate, citric acid