Targeting the Hair Follicle Basement Membrane to Improve Growth Conditions In vitro

Jan 1, 2011 | Contact Author | By: C. Gondran, C. Meyrignac, A. Perrin and N. Domloge, ISP Vincience, Global Skin Research Center; P. Mouser; and C. Dal Farra, ISP Corporate Research Center
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Title: Targeting the Hair Follicle Basement Membrane to Improve Growth Conditions In vitro
hair folliclex integrinsx cell communicationx hair growthx
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Keywords: hair follicle | integrins | cell communication | hair growth

Abstract: Signal transductions mediated by integrins are shown to be involved in hair growth and development. Here, the authors investigate the effect of a botanical corn extract on the expression of different integrins via in vitro measurements of hair shaft elongation. Scalp biopsies show the daily application of the active may improve hair elongation.

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C. Gondran et al, Targeting the Hair Follicle Basement Membrane to Improve Growth Conditions In vitro, Cosmet & Toil 126(1) 50 (2011)

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Hair is an essential element of physical appearance, well-being and attractiveness. The hair follicle (HF) is the hair shaft-producing unit that has been described as a complex and complete organ; indeed, it is characterized by continuous cycling with successive phases of growth (anagen), illustrated by the intense activity of hair matrix keratinocytes. The growth stage is followed by a complete regression of the lower segment of follicle and bulb (catagen) and relative quiescence (telogen), resulting in hair shedding (exogen). At the end of telogen, the HF is reactivated by intrafollicular and extrafollicular signals in order to re-initiate a new cycle.

This intense cellular activity is the result of complex interactions taking place in different compartments of the HF. In particular, the concentric organization of the HF implicates regulated interactions between the epithelial and connective tissue sheath. These epithelial-mesenchymal interactions have been shown to play a central role in HF activity and involve components of the HF basement membrane, which is located between the connective tissue sheath that surrounds the HF and the basal layer of the outer root sheath (ORS). This basement membrane also continues at the level of the dermal papilla, which is composed of mesenchymal tissue and considered a conductor of hair regeneration and growth.

In the skin, the basement membrane has been well-described as far as its molecular organization and role in maintaining epidermal-dermal cohesion. However, its role in relation to hair growth remains incompletely understood. The aim of the present study was to characterize the main components of the HF basement membrane by immunohistology to evaluate the modulation of their synthesis by a compound designed to boost cell energy levels.

Integrins

Integrins are transmembrane adhesion proteins that are involved in cell-cell and cell-matrix interactions and exert signalling functions. In the HF, they play key roles in interactions between the outermost cell layer of the ORS and the basement membrane. In particular, b1 integrin mediated signaling has been implicated in hair growth control. Moreover, b1 and a6 integrin subunits have been identified as epidermal stem cell markers.

With respect to their role in cellmatrix adhesion, a6b4 and a3b1 integrins have been described as main receptors for laminin-332 and laminin-511, respectively, which regulate hair growth. In particular, laminin-511, through interaction with its b1 integrin receptor, has been shown to promote dermal papilla development. Moreover, blocking laminin-511 has been shown to provoke alopecia on human scalp grafts.

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Figure 1. Longitudinal section of hair follicle in a biopsy of scalp skin

Figure 1. Longitudinal section of hair follicle in a biopsy of scalp skin

These epithelial-mesenchymal interactions have been shown to play a central role in HF activity and involve components of the HF basement membrane,3 which is located between the connective tissue sheath that surrounds the HF and the basal layer of the outer root sheath (ORS) as seen in Figure 1.

Figure 2. Immunostaining of a3, a6 and b1 integrins on human scalp biopsies (nonspecific staining of hair shaft)

Figure 2. Immunostaining of a3, a6 and b1 integrins on human scalp biopsies (nonspecific staining of hair shaft)

The staining of the different integrin subunits (a3, a6 and b1) was localized in the outermost cell layer of outer root sheath (see Figure 2).

Figure 3. Immunostaining of laminin-332 and laminin-511 on human scalp biopsies

Figure 3. Immunostaining of laminin-332 and laminin-511 on human scalp biopsies

Moreover, the staining of laminin-332 and laminin-511 in the basement membrane was more intense with the active than in the control (see Figure 3).

Figure 4. Immunostaining of collagen IV on human scalp biopsies (nonspecific staining of hair shaft)

Figure 4. Immunostaining of collagen IV on human scalp biopsies (nonspecific staining of hair shaft)

The effect of the active’s application to the basement membrane zone was also observed on collagen IV expression, which was boosted by the treatment (see Figure 4).

Figure 5. Immunostaining of fibronectin on human scalp biopsies

Figure 5. Immunostaining of fibronectin on human scalp biopsies

Furthermore, the staining of fibronectin, visualized in the dermal sheath, was enhanced by application of the active (see Figure 5).

Figure 6. Immunostaining of Ki67 (green) and p63 (red) on human scalp biopsies (nonspecific staining of hair shaft)

Figure 6. Immunostaining of Ki67 (green) and p63 (red) on human scalp biopsies (nonspecific staining of hair shaft)

Results indicated the active positively modulated the expression of these two markers, favoring the activity of hair growth and regeneration (see Figure 6).

Figure 7. Photographs of scalp skin biopsies showing hair elongation at day 0, day 7 and day 14

Figure 7. Photographs of scalp skin biopsies showing hair elongation at day 0, day 7 and day 14

It is interesting to note that this ex vivo model maintained the vitality of scalp biopsies in vitro and provided the required conditions to actually enable the measurement of hair shaft elongation (see Figure 7).

Figure 8. Hair shaft elongation measured on human scalp biopsies; * = significant with student’s t-test, p < 0.05.

Figure 8. Hair shaft elongation measured on human scalp biopsies; * = significant with student’s t-test, p < 0.05.

Application of both the active and placebo resulted in enhancement of the hair shaft length on biopsies treated daily; although the active produced 71% greater growth than the placebo at day 8 and 110% greater growth than the placebo at day 14 (see Figure 8).

Footnotes (CT1101 C. Gondron)

a William’s E medium was obtained from Lonza.
b Insulin was obtained from Sigma.
c Hydrocortisone also was obtained from Sigma.
d Primocin is a product of Invivogen, San Diego, CA, USA.
e OCT media was obtained from CellPath, UK.
f Alexa Fluor 488 is a product of InVitrogen.
g Fluoromount-G was obtained from Electron Microscopy Sciences.
h The Nikon Eclipse E600 microscope was used for this assessment.
j The Nikon DXM1200F camera with ACT-1C software was used for this assessment.
k The Vivacam camera is manufactured by Lucid, USA.
m Image Pro analysis software is manufactured by Media Cybernetics.

Footnotes (CT1101 C. Gondron)

a William’s E medium was obtained from Lonza.
b Insulin was obtained from Sigma.
c Hydrocortisone also was obtained from Sigma.
d Primocin is a product of Invivogen, San Diego, CA, USA.
e OCT media was obtained from CellPath, UK.
f Alexa Fluor 488 is a product of InVitrogen.
g Fluoromount-G was obtained from Electron Microscopy Sciences.
h The Nikon Eclipse E600 microscope was used for this assessment.
j The Nikon DXM1200F camera with ACT-1C software was used for this assessment.
k The Vivacam camera is manufactured by Lucid, USA.
m Image Pro analysis software is manufactured by Media Cybernetics.

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