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Promotive Effect of Topical Ketoconazole, Minoxidil, and Minoxidil with Tretinoin on Hair Growth in Male Mice

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Recently topical use of 2% Ketoconazole solution has been reported to have a therapeutic effect on androgenic alopecia. Minoxidil is a vasodilatory medication used primarily as antihypertensive drug. It was discovered to have the side effect of hair growth and reversing baldness. Tretinoin is commonly used topically for acne treatment and in the treatment of photoaging. It is used by some as hair loss treatment. Objective. To compare the stimulatory effect of Ketoconazole, Minoxidil, and Minoxidil with Tretinoin on hair growth in a mouse model. Materials and Methods. Coat hairs on the dorsal skin of seven weeks old male mice were gently clipped and then stained by using commercial dye. These mice were divided into four groups each of five treated with topical application of ethanol 95%, Ketoconazole solution 2%, Minoxidil solution 5%, and Minoxidil with Tretinoin solution 0.1%, respectively. The drugs were applied once daily for three weeks, the clipped area was photographed, and the ratio of regrown coat area was calculated. Results. The results demonstrated that Ketoconazole, Minoxidil, and Minoxidil with Tretinoin had a significant stimulatory effect on hair growth compared with the control group and Minoxidil was the most effective drug among them.
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Research Article
Promotive Effect of Topical Ketoconazole, Minoxidil, and
Minoxidil with Tretinoin on Hair Growth in Male Mice
Muhsin A. Aldhalimi,1Najah R. Hadi,2and Fadaa A. Ghafil2
1Department of Dermatology, Kufa College of Medicine, Kufa, Iraq
2Department of Pharmacology and erapeutics, Kufa College of Medicine, Kufa, Iraq
Correspondence should be addressed to Najah R. Hadi; drnajahhadi@yahoo.com
Received  December ; Accepted  January ; Published  March 
Academic Editors: R. urmond, R. Villalobos-Molina, and S.-N. Wu
Copyright ©  Muhsin A. Aldhalimi et al. is is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly
cited.
Recently topical use of % Ketoconazole solution has been reported to have a therapeutic eect on androgenic alopecia. Minoxidil
is a vasodilatory medication used primarily as antihypertensive drug. It was discovered to have the side eect of hair growth and
reversing baldness. Tretinoin is commonly used topically for acne treatment and in the treatment of photoaging. It is used by some
as hair loss treatment. Objective. To compare the stimulatory eect of Ketoconazole, Minoxidil,and Minoxidil with Tretinoin onhair
growth in a mouse model. Materials and Methods. Coat hairs on the dorsal skin of seven weeks old male mice were gently clipped
and then stained by using commercial dye. ese mice were divided into four groups each of ve treated with topical application
of ethanol %, Ketoconazole solution %, Minoxidil solution %, and Minoxidil with Tretinoin solution .%, respectively. e
drugswereappliedoncedailyforthreeweeks,theclippedareawasphotographed,andtheratioofregrowncoatareawascalculated.
Results. e results demonstrated that Ketoconazole, Minoxidil, and Minoxidil with Tretinoin had a signicant stimulatory eect
on hair growth compared with the control group and Minoxidil was the most eective drug among them.
1. Introduction
Androgenic alopecia is a partial or complete loss of hair that
occurs in a progressive pattern in genetically predisposed
individuals []. A variety of genetic and environmental factors
likely play a role in androgenic alopecia, and most of con-
tributing factors remain unknown.
e age at onset of androgenic alopecia diers but occurs
usually in mid-twenties. e prevalence and severity of
androgenic alopecia increase directly with age []. e basis
of androgenic alopecia is a progressive decrease in the density
of terminal hairs and concurrent increase in density of vellus
hairs []. In eect, terminal hairs are turned o and are
transformed into vellus hairs, and this eect is due to hair
follicle miniaturization [], which is associated with substan-
tial reduction in hair diameter. Although the mechanisms of
these changes have not been established denitively, male pat-
ternbaldnessisknowntodependonandrogens,inparticular
the dihydrotestosterone DHT []. DHT is synthesized from
testosterone by 𝛼-reductase enzyme type- and type-, and
these enzymes are found on the nuclear membranes [].
Ketoconazole (KCZ) is an imidazole antifungal agent.
Longtermuseof%KCZwasreportedtobeeectiveagainst
androgenic alopecia in patients without seborrheic dermatitis
and dandru []. Minoxidil is a vasodilatory medication used
primarily as antihypertensive drug. Minoxidil is a potassium
channel agonist that has the chemical structure of nitric oxide
(NO),abloodvesseldilator,andmaybeanitricoxideagonist.
is appears to explain its vasodilatory eect []butmayalso
be linked to Minoxidil’s ability to stimulate hair growth and
treat hair loss.
Tretinoin is the acid form of vitamin A, also known as all
transretinoic acid (ATRA), helps normalize hyperkeratiniza-
tion, and has demonstrated signicant anti-inammatory
eects.Itisusedbysomeashairlosstreatment[].
2. Materials and Methods
2.1. Animals. e study was conducted on  mature male
Albino-Webster mice, which were housed in the air-con-
ditioned animal house of College of Medicine, University of
Hindawi Publishing Corporation
ISRN Pharmacology
Volume 2014, Article ID 575423, 5 pages
http://dx.doi.org/10.1155/2014/575423
ISRN Pharmacology
F : Mice aer clipping of the coat hair.
F : Mice aer staining of the dorsal coat hair.
Kufa, with standard animal diet and water. eir ages ranged
fromtoweeks,andtheirweightsrangedfromtogm.
e design of the study was approved by the local ethical
committee in the College of Medicine, University of Kufa.
2.2. Methods
2.2.1. Clipping of the Coat Hair of Mice. One day before
the experiment, the mice were anesthetized with diethyl
ether, and the coat hair on the dorsal skin was gently
clipped using an electric shaver to avoid injury. e skin
surface of the clipped area of each mouse was observed on
the day when the materials were to be applied to have a
pinkish color, suggesting that it was in the resting phase [].
Mice were randomly chosen and put into four groups, each
group included ve mice, and they had been photographed
(Figure ).
2.2.2. Staining. e denuded area of the dorsal skin of each
mouse was stained by using commercial dye (Homann,
, ) [,]; then it was washed by using alcohol,
staining done to nd the ratio of area showing hair regrowth
(area of white color) to the ratio of area denuded of hair (area
of black color). Aer staining of mice, also they had been
photographed (Figure ).
2.2.3. Study Groups. Group I was considered as a control
groupandtreatedwiththevehiclesolution(%),ethanol
alcohol, . mL applied by a micropipette to the denuded skin
T : Mean ratio of white color (area showing hair regrowth) to
the ratio of black color (area denuded of hair) aer  days of treat-
ment with KCZ solution %, Minoxidil solution %, and Minoxidil
solution % with Tretinoin solution .%.
Groups Mean ratio of color dierence 𝑃value
Control 1.49 ± 0.17 N.S.
Ketoconazole 15.98 ± 3.42 <.
Minoxidil 39.53 ± 8.42 <.
Minoxidil and Tretinoin 15.70 ± 4.71 <.
All data expressed as mean ±SEM.
and then spread by means of swab once daily  days/weak for
 weeks.
GroupIIwastreatedwithKetoconazolesolution%,
. mL applied to the denuded skin by a micropipette once
daily for  weeks.
Group III was treated with Minoxidil solution %, . mL
applied to the denuded skin by a micropipette once daily for
 weeks.
Group IV was treated with equal amounts of Minoxidil
solution % + Tretinoin .%, . mL of each of them applied
by a micropipette once daily for  weeks.
e mice were macroscopically observed and photogra-
phed every week until the st day.
Photographic Data Analysis. Photographic data were analyzed
by a special computer program called Photoshop Visual
Basic- program. is program calculated the ratio of the area
showing hair regrowth (which was represented by the area of
white color) to the ratio of the area denuded of hair (which
was represented by the area of black color), and this was done
for each mouse in the four groups.
2.3. Histological Sections. Aer completing the treatment
for three weeks, histological sections were obtained. ese
sections were examined microscopically for the hair follicle
number and diameter,in control and treated groups.
2.4. Statistical Analysis. Statistical analysis was done by using
one-way ANOVA test with post hoc test at level of signif-
icance 𝛼 = 0.05 to compare between control and treated
groups and then performing multiple comparisons between
the treated groups.
Chi-squaretesthadbeenusedtocomparebetweenthe
proportion of histological changes in various groups.
3. Results
ere was a signicant hair growth (𝑃value <.) in the
groups treated with KCZ, Minoxidil, and Minoxidil with
Tretinoin as compared with control group (Tables and and
Figures ,,and ).
e histological examination of the specimens showed no
signicant increase in the number of hair follicles (𝑃value >
.) in all treatment groups, while hair follicle diameter has
ISRN Pharmacology
F : Mice treated with Ketoconazole showing signicant hair
growth aer  weeks of treatment.
T:Comparisonofhairgrowthamongthetreatedgroupsaer
 days of treatment with KCZ solution %, Minoxidil solution %,
and Minoxidil solution % with Tretinoin solution .%.
Groups Mean dierence 𝑃value
KCZ-Minoxidil −23.54 ± 7.24 <.
KCZ-Minoxidil and Tretinoin 0.28 ± 7.24 N.S.
Minoxidil-Minoxidil and Tretinoin 28.82 ± 7.24 <.
All data expressed as mean ±SEM.
T : e mean number of hair follicles in control and treated
groups examined under high power eld per mm aer  days
of treatment with KCZ solution %, Minoxidil solution %, and
Minoxidil solution % with Tretinoin solution .%. is table shows
that hair follicle number increased insignicantly (𝑃value >.) in
all treatment groups.
Groups Mean number of hair
follicles/ mm 𝑃value
Control 5.200 ± 1.92
KCZ 9.600 ± 3.20 N.S.
Minoxidil 13.600 ± 2.50 N.S.
Minoxidil and Tretinoin 8.200 ± 2.58 N.S.
All data expressed as mean ±SD.
T : e mean diameter (in micrometer) of hair follicles in
control and treated groups aer  days of treatment with KCZ
solution %, Minoxidil solution %, and Minoxidil solution % with
Tretinoin solution .%.
Groups Mean diameter of hair
follicles (in 𝜇m) 𝑃value
Control 1.53 ± 0.09 N.S.
KCZ 2.71 ± 0.18 <.
Minoxidil 3.72 ± 0.20 <.
Minoxidil and Tretinoin 3.46 ± 0.18 <.
All data expressed as mean ±SEM.
been increased signicantly (𝑃value <.) in all treatment
groups (Tables ,,andand Figures ,,,and).
F : Mice treated with Minoxidil showing signicant hair
growth aer  weeks of treatment.
F : Mice treated with Minoxidil and Tretinoin showing sig-
nicant hair growth aer  weeks of treatment.
F : Mice of control group aer  weeks of topical alcohol
treatment showing insignicant hair growth.
F : Histological section from mouse skin in control group
(×) showing normal skin layers and hair follicles.
ISRN Pharmacology
T : Comparison of the diameter (in 𝜇m) of hair follicles among
the treated groups aer  days of treatment with KCZ solution %,
Minoxidil solution %, and Minoxidil solution % with Retin—A
solution .%. 𝑁=5in each group.
Groups Mean dierence 𝑃value
KCZ-Minoxidil −1.01 ± 0.24 N.S.
KCZ-Minoxidil and Tretinoin −0.75 ± 0.27 N.S.
Minoxidil-Minoxidil and Tretinoin 0.26 ± 0.28 N.S.
All data expressed as mean ±SEM.
F : Histological section from the skin of mouse treated with
Ketoconazole (×) showing normal skin layers and increase in hair
follicle diameter.
4. Discussion
is study demonstrated that topical Ketoconazole stimulates
hair growth signicantly. is result is in agreement with
Pi´
erard-Franchimont et al. [], Khandpur et al. [], Jiang et
al. [], Inui and Itami [],andHugoPerez[].
e ecacy of % KCZ shampoo in androgenic alopecia
patients appeared to stem from its antiandrogenic properties
[,]. However, our study demonstrated that topical KCZ
was eective on the androgen-insensitive coat hairs of mice,
so KCZ behaved as an androgen-independent biological res-
ponse modier. However, its action on hair growth and hair
follicle diameter is less than that of Minoxidil.
ere was a signicant increase in hair growth in the
group treated with topical Minoxidil solution. is result is
in agreement with Olsen et al. [], Weiss et al. [], and Mori
and Uno [].
Minoxidil induces rapid relaxation of vascular smooth
muscle induced by its sulphated metabolite, Minoxidil sul-
phate []. e conversion of Minoxidil to Minoxidil sulphate
is catalysed by sulphotransferase enzyme, which was initially
demonstrated in rat liver []andhassincebeenfoundin
human liver [], platelets [], and mouse vibrissae follicles
[].
In scalp skin of stump-tailed macaque, sulphotransferase
activity is largely localized in the hair follicle []. Bio-
chemical evidence for Minoxidil sulphation by two phenol
sulphotransferases has been found in human scalp skin [].
ere are individual variations in scalp sulphotransferase
activity and this correlates with the level in platelets []. In a
clinical setting scalp sulphotransferase activity was higher in
menwhorespondedtoMinoxidilcomparedwiththosewho
did not respond [].
F : Histological section from the skin of mouse treated with
Minoxidil (×) showing normal skin layers and increase in hair
follicle diameter.
F : Histological section from the skin of mouse treated with
Minoxidil and Tretinoin (×) showing normal skin layers and
increase in hair follicle diameter.
In this study there was a signicant hair growth in the
group treated with Minoxidil and Tretinoin, and this result
is in agreement with Bazzano et al. [,].
Tretinoin is known to alter cell proliferation and dier-
entiation and may promote vascular proliferation, and these
actions may be important to hair growth and so aect hair
follicle during the various growth and regression phases.
Preliminary studies of cRABP levels in whole scalp skin
ofhumansubjectswithmalepatternbaldnessindicatedthat,
in the scalp areas not normally aected by alopecia, levels
of cRABPs were higher than in areas with alopecia, and the
levelsofcRABPsinwholeskinwereincreasedbytopical
application of retinoic acid [].
It has been found that combining high concentration of
retinoic acid with Minoxidil causes less elongation than at low
concentration suggested that retinoic acid might increase the
tissue concentration of Minoxidil in hair follicles [].
Tretinoin was reported to increase percutaneous absorp-
tion of Minoxidil by increasing the stratum corneum per-
meability []. Our study demonstrated that Tretinoin in
combination with Minoxidil caused a signicant increase
in hair growth and a signicant increase in hair follicle
diameter. However, in this study the results obtained from
the combination of Minoxidil with low dose Tretinoin are
less than that of Minoxidil alone and this warrants further
studies to evaluate the role of Tretinoin in combination with
Minoxidil in the treatment of hair loss.
ISRN Pharmacology
Conflict of Interests
e authors declare that there is no conict of interests
regarding the publication of this paper.
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... Also, in animal studies, topical application of 2% KCZ solution stimulated the telogen-to-anagen hair conversion and positive hair growth in C3H/HeN mice (Jiang et al. 2005). Subsequently in 2014, Aldhalimi et al. have confirmed the significant trichogenic effects of topical KCZ in mice at the histopathological level with increased follicular diameter and new hair growth (Aldhalimi et al. 2014). ...
... The most common proposed mechanism of action for topical KCZ in androgenetic alopecia (AGA) is via its known anti-androgenic properties (Hugo Perez 2004). However, Aldhalimi et al. demonstrated that it was also effective on the androgen-insensitive coat hairs of mice McNemar test **Highly significant (P < 0.01) χ 2 Chi square test, NS non-significant (P > 0.05), P 1 baseline versus after 2 months, P 2 baseline versus after 4 months, P 3 baseline versus after 6 months (Aldhalimi et al. 2014). This finding indicates that topical KCZ may act through androgen-dependent as well as androgen-independent pathway, which warrants further investigations. ...
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Background Female pattern hair loss (FPHL) is a common non-scaring alopecia. Topical minoxidil is the only approved treatment for FPHL; however, it has frequent side effects. Ketoconazole is an antifungal with anti-androgenic properties. Methods The aim of this work was to evaluate the safety and possible trichogenic effects of ketoconazole in FPHL. Patients were randomly assigned into Group A with 20 patients who received 2% topical minoxidil solution and Group B involving 20 patients who were treated by 2% topical ketoconazole preparation. All patients were treated for 6 months and evaluated clinically plus via trichoscopy. Side effects were recorded, and patients’ satisfaction was measured. Results Regarding hair growth, a significant difference was detected between baseline and at 4th and 6th months in Group A. While in Group B, the improvement was delayed until the 6th month. Only 10% of Group B reported side effects. Patients’ satisfaction did not differ between the two groups. Conclusions Topical ketoconazole mediates a trichogenic effect in FPHL with few side effects. However, ketoconazole showed a delayed improvement versus minoxidil at similar concentrations. Further studies should evaluate the efficacy of higher concentrations of ketoconazole solutions for FPHL which is a promising therapy. Trial registration IRB approval status: reviewed and approved by IRB of Faculty of Medicine, Zagazig University; approval no. #3642
... One hundred women completed the study, with a mean reduction in hair shedding score (range 1-6) of 2.3 at 6 months and 2.6 at 12 months. [11] In addition, a recent retrospective review showed that low-dose minoxidil (2.5-5 mg once daily), given either as monotherapy or as an additional therapy for 6-12 months, was both effective and safe in 41 male patients with AGA [12,13]. Therianou et al. also reported the effectiveness of using oral minoxidil in patients allergic to topical minoxidil [14]. ...
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Introduction: Oral minoxidil is an antihypertensive vasodilator known to stimulate hair growth. The use of low-dose oral minoxidil for the treatment of male androgenetic alopecia (AGA) is receiving increasing attention. The aim of this study was to evaluate the efficacy and safety of oral minoxidil for the treatment of male AGA. Methods: This was an open-label, prospective, single-arm study. Thirty men aged 24-59 years with AGA types III vertex to V were treated with oral minoxidil 5 mg once daily for 24 weeks. Efficacy was evaluated by hair counts, hair diameter measurements, photographic assessment, and self-administered questionnaire. The safety of the treatment was closely monitored by means of physical examinations and laboratory investigations. Results: There was a significant increase in total hair counts from baseline at weeks 12 (mean change + 26, range 182.5-208.5 hairs/cm2) and 24 (mean change + 35.1, range 182.5-217.6 hairs/cm2) (both p = 0.007). Photographic assessment of the vertex area by an expert panel revealed 100% improvement (score > + 1), with 43% of patients showing excellent improvement (score + 3, 71-100% increase). The frontal area also showed a significant response but less than that of the vertex area. Common side effects were hypertrichosis (93% of patients) and pedal edema (10%). No serious cardiovascular adverse events and abnormal laboratory findings were observed. Conclusion: Oral minoxidil 5 mg once daily effectively increased hair growth in our male patients with AGA and had a good safety profile in healthy subjects. However, oral minoxidil should be used carefully with men who have severe hypertension and increased risk for cardiovascular events.
... Ratios of hair regrowth to denuded area were significantly greater in the ketoconazole group than the ethanol control group, with a ratio 0.61 (p = 0.006) on the 21st day. Similarly, a prospective murine study by Aldhalimi, Hadi, and Ghafil (2014) separated 20 shaved mice into four groups for 3 weeks of treatment with: (1) ketoconazole 2% solution, ...
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Androgenetic alopecia (AGA) is common and associated with significant psychosocial distress. Treatment options are needed for patients that do not adequately respond to first line treatments of finasteride or minoxidil. Topical ketoconazole has been proposed as a promising treatment. The goal of this systematic review was to evaluate the efficacy of topical ketoconazole in the treatment of AGA. A systematic literature search was conducted within the MEDLINE database using the key terms "ketoconazole" and "alopecia." Forty-seven papers were screened for inclusion, of which 9 were assessed for eligibility. Seven articles were included in the qualitative synthesis, including 2 animal studies (total of 40 participants) and 5 human studies (total of 318 participants). Murine studies demonstrated a significant increase in mean ratio of hair regrowth to denuded area in the ketoconazole treatment groups compared to controls. Human studies reported increased hair shaft diameter following ketoconazole use. One study reported a significant increase in pilary index (percent anagen phase x diameter) following treatment. Studies also demonstrated clinical improvement of AGA based on photographic assessment and subjective evaluation. Topical ketoconazole is a promising adjunctive or alternative therapy in the treatment of AGA. Randomized controlled trials are needed. This article is protected by copyright. All rights reserved.
... Male 5.5-week C57BL/6 mice were purchased from Hamamatsu (Shizuoka). Two d before the experiment, hair was clipped from the dorsal surface of each mouse with an electric shaver under anesthesia, without causing damage or injury to the skin 76 . To determine the optimal time interval between doses of MSC-EVs, we assayed MSC-EV retention in mice (n = 3) with DiD-labeled MSC-EVs (MSC-EVs/DiD) by fluorescence imaging on an IVIS Lumina III In Vivo Imaging System (PerkinElmer). ...
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Hair loss is a common medical problem. In this study, we investigated the proliferation, migration, and growth factor expression of human dermal papilla (DP) cells in the presence or absence of treatment with mesenchymal stem cell extracellular vesicles (MSC-EVs). In addition, we tested the efficacy of MSC-EV treatment on hair growth in an animal model. MSC-EV treatment increased DP cell proliferation and migration, and elevated the levels of Bcl-2, phosphorylated Akt and ERK. In addition; DP cells treated with MSC-EVs displayed increased expression and secretion of VEGF and IGF-1. Intradermal injection of MSC-EVs into C57BL/6 mice promoted the conversion from telogen to anagen and increased expression of wnt3a, wnt5a and versican was demonstrated. The first time our results suggest that MSC-EVs have a potential to activate DP cells, prolonged survival, induce growth factor activation in vitro, and promotes hair growth in vivo.
... Topical prototypic retinoid ATRA (tretinoin) alone and in combination with 0.5% minoxidil has been shown to be effective for the promotion of hair growth in vitro [91,92] and in mice [93]. Tretinoin can promote and regulate cell proliferation and differentiation in the epithelium and may promote vascular proliferation to prolong the anagen phase of the hair cycle [91]. ...
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Introduction: Treatments for androgenetic alopecia constitute a multi-billion-dollar industry, however, currently available therapeutic options have variable efficacy. Consequently, in recent years small biotechnology companies and academic research laboratories have begun to investigate new or improved treatment methods. Research and development approaches include improved formulations and modes of application for current drugs, new drug development, development of cell-based treatments, and medical devices for modulation of hair growth. Areas covered: Here we review the essential pathways of androgenetic alopecia pathogenesis and collate the current and emerging therapeutic strategies using journal publications databases and clinical trials databases to gather information about active research on new treatments. Expert opinion: We propose that topically applied medications, or intra-dermal injected or implanted materials, are preferable treatment modalities, minimizing side effect risks as compared to systemically applied treatments. Evidence in support of new treatments is limited. However, we suggest therapeutics which reverse the androgen-driven inhibition of hair follicle signaling pathways, such as prostaglandin analogs and antagonists, platelet-rich plasma (PRP), promotion of skin angiogenesis and perfusion, introduction of progenitor cells for hair regeneration, and more effective ways of transplanting hair, are the likely near future direction of androgenetic alopecia treatment development.
Chapter
Circulating androgens can profoundly influence target cutaneous structures such as the pilosebaceous unit (hair follicle and sebaceous gland), and play a central role in the pathogenesis of cutaneous disorders of hyperandrogenism (CHA) which include androgenetic alopecia (AGA), acne vulgaris, and hirsutism. Therapeutically, two main categories of drugs affect androgens and their activity. The first are ‘antiandrogens’ which are agents that block the androgen receptor (AR) and include spironolactone, flutamide, and cyproterone acetate. The second are ‘androgen inhibitors,’ which block androgen synthesis, and include finasteride and dutasteride (these are specific 5-α reductase inhibitors, that inhibit formation of dihydrotestosterone [DHT]), as well as leuprolide, (which is a gonadotropin-releasing hormone (GnRH) agonist, and works at the level of the ovary and pituitary). In this chapter ‘antiandrogens’ is the term used for specific AR blockers, and the term ‘androgen inhibitors’ will be used to distinguish those compounds that work by suppressing or inhibiting the formation of DHT. Other agents such as progestins and combined oral contraceptives (COC) are both antiandrogens and androgen inhibitors. The various drugs discussed in this chapter are categorized by their most important clinical mode of action. This chapter provides an overview of various antiandrogens (drugs that block the AR) and androgen inhibitors (enzyme inhibitors and other mechanisms). How these specific compounds work, their approved and off-label indications, dosing, and adverse effects will be presented. Particular emphasis is given to spironolactone, finasteride, dutasteride, and oral contraceptives.
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In this research, we examined the effect of rosuvastatin calcium-loaded nanoparticles on the hair growth–promoting activity on Albino rats. Nanoparticles were prepared using 2:1 weight ratio of drug to methyl-β-cyclodextrin with 10, 20, and 30% stabilizers (phospholipid, polyvinyl pyrrolidone K30, and Compritol 888 ATO) using nanospray dryer. Subsequently, the prepared nanoparticles were evaluated for their process yield, particle size, polydispersity index, zeta potential, and in vitro drug release as well as in vivo studies. The dried nanoparticles showed process yield values up to 84% with particle size values ranging from 218 to 6258 nm, polydispersity index values ranging from 0.32 to 0.99, and zeta potential values ranging from − 6.1 to − 11.9 mV. Combination of methyl-β-cyclodextrin with 10% polyvinyl pyrrolidone K30 accomplished nanoparticles with the lowest particle size (218 nm) and polydispersity index (0.32) values. These nanoparticles had suitable process yield value (70.5%) and were able to retard drug release. The hair growth–promoting activity for the selected nanoparticles revealed the highest hair length values in Albino rats after 14 days of the hair growth study compared with non-medicated nanoparticles, nanoparticles’ physical mixture, rosuvastatin solution, and marketed minoxidil preparation groups as well as the control group. The immunohistochemistry images for both selected nanoparticles and marketed minoxidil groups showed a significant increase in the diameter of hair follicle and percent area fraction of cytokeratin 19 in the outer root sheath of hair follicle compared with other tested groups. Rosuvastatin nanoparticles prepared by nanospray drying technique could be a good competitor to minoxidil for hair growth–promoting activity. Graphical abstract
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Introduction: Although labelling changes and market withdrawal have been implemented for oral ketoconazole (KTZ) due to serious adverse effects, topical KTZ is generally thought to be effective and safe for the treatment of superficial fungal infections. New dermatologic indications for the use of topical KTZ have arisen such as onychomycosis, blepharitis and hair loss. This article aims to review the literature on topical KTZ's efficacy and adverse effects, as well as provide an overview on current insights regarding its mechanism of action and upcoming developments. Methods: A PubMed search was done to include randomized-controlled trials focusing on the use of topical KTZ on human subjects. Results: Forty studies with 4566 patients were included in this review. Topical KTZ is clinically effective for the treatment of Malassezia-related conditions such as seborrheic dermatitis and pityriasis versicolor with a reported efficacy of 63-90% and 71-89%, respectively. Discussion: Topical KTZ demonstrates high clinical efficacy for Malassezia-related conditions. More efficacious alternatives are now available for Tinea and Candida. Although topical KTZ is safe, clinicians should be aware that allergic contact dermatitis may occur. Further studies should be completed to investigate the use of topical KTZ for hair loss and inflammatory dermatoses.
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OVOL1 and OVOL2, ubiquitously conserved genes encoding C2H2 zinc finger transcription factors in mammals, control epithelial cell proliferation, and differentiation, including those in skin. OVOL1 and OVOL2 expression is coordinately mediated via the Wnt signaling pathway, and OVOL1 negatively regulates OVOL2 expression in a transcriptional manner. Our previous study of OVOL1 expression in human skin revealed that OVOL1 is preferentially expressed in the inner root sheath of the hair follicle. Therefore, we hypothesized that the OVOL1-OVOL2 axis is involved in normal and neoplastic follicular differentiation. Immunohistochemical analysis showed that OVOL1 and OVOL2 were strongly expressed in a mutually exclusive manner in the cytoplasm of inner root sheath cells and matrix cells, respectively, in normal follicles. OVOL2 was also expressed in pilomatricoma, with only partial expression of OVOL1. Cultured human keratinocytes expressed OVOL1 and OVOL2 on both the mRNA and protein levels. The expression of OVOL2 was higher in keratinocytes transfected with siRNA of OVOL1. Ketoconazole, a hair growth stimulant, up-regulated the expression of OVOL1 but did not affect OVOL2 expression. These results indicated that the OVOL1-OVOL2 axis may actively contribute to cell differentiation and proliferation in the hair bulb, suggesting that the OVOL1 and OVOL2 may be therapeutic targets of hair disorders, including alopecia, and play important roles in the tumorigenesis of pilomatricoma.
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Background. We examined the efficacy of a new regimen to treat AGA, with attention to male patients who are atopic. Objective. To assess the efficacy of a four-part regimen for the treatment of AGA in atopic and nonatopic patients. NuH Hair is a novel topical combination of finasteride, dutasteride, and minoxidil, which is blended in a hypoallergenic lotion. The other three components included Rogaine foam, Propecia, and ketoconazole shampoo. Methods. A prospective pilot study was conducted in 15 patients. All patients were assessed for the presence of atopy. Each patient served as their own control. All patients were treated specifically with NuH Hair and were given the option to add any of the other components of the protocol to their regimen. Photographs were taken of each patient's scalp at months 0, 1, 3, 6, and 9. Results. All 15 patients demonstrated significant growth of hair. In those patients who utilized all 4 components, significant growth was achieved in as little as 30 days. In those patients who choose only to utilize NuH Hair, significant growth was demonstrated after 3 months. Conclusion. Aggressively treating AGA achieves significant and rapid growth of new hair. This is effective in atopic and nonatopic male patients.
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Nineteen healthy male volunteers completed a three-way, randomized, crossover study to determine the effect of the synthetic retinoid, tretinoin, on percutaneous absorption of minoxidil. Subjects received, for 20 days, twice-daily administrations of 1 ml of an aqueous 2% topical minoxidil solution either alone, with once-daily applications of a 0.05% tretinoin cream, or with once-daily applications of a vehicle control cream. When minoxidil was coadministered with tretinoin cream, minoxidil absorption was increased nearly threefold, compared with a 1.3-fold increase in absorption observed with coadministration of vehicle control cream. Transepidermal water loss measurements, which are sensitive to changes in stratum corneum function, were also significantly increased with tretinoin. No treatment-related changes in stratum corneum thickness were observed on the basis of skin biopsy analysis. The findings indicate that percutaneous minoxidil absorption is enhanced by tretinoin as a result of increased stratum corneum permeability.
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Androgenetic alopecia is characterised by progressive, patterned hair loss from the scalp. Recently the pathogenesis and genetic basis of the hair loss have been better understood, as has the distress experienced by men who have lost their hair. There have also been breakthroughs in the treatment of androgenetic alopecia.The transition of some terminal hairs into vellus hairs is a universal physiological secondary sexual characteristic.1 Androgenetic alopecia becomes a medical problem only when the hair loss is subjectively seen as excessive, premature, and distressing.The prerequisites for premature androgenetic alopecia are a genetic predisposition and sufficient circulating androgens.2 Eunuchs do not go bald.3 Every white man possesses the autosomal inherited predisposition,4 and 96% lose hair to some degree,5 but because of the variabity of gene expression far fewer have appreciable premature hair loss. Summary points Androgenetic alopecia is a specific type of hair loss mediated by systemic androgens and genetic factorsRecent advances in understanding of the biology of hair follicles have shed light on the pathogenesis of androgenetic alopeciaThough most men learn to deal with their androgenetic alopecia without it impairing their psychosocial functioning, some men tolerate hair loss poorly and have a negative overall body image and diminished quality of lifeSafe and effective treatments are currently available for androgenetic alopecia, but advice and counselling remain the most important aspects of management
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Minoxidil is an antihypertensive agent and hair growth promoter that is metabolized by sulfation to the active compound, minoxidil sulfate. Thermostable phenol sulfotransferase (TS PST or P-PST) was initially thought to catalyze the reaction, and the enzyme was designated minoxidil sulfotransferase (MNX-ST). Information about human ST activities toward minoxidil would be useful in developing the capacity to predict individual responses to minoxidil based on tissue levels of STs. Therefore, human STs were studied from platelet homogenates, partially purified platelets, scalp skin high speed supernatants and COS-1 cell cDNA expressed preparations using a radiochemical enzymatic assay with minoxidil as the substrate. Studies showed the presence of TS PST, TL (thermolabile) PST and MNX-ST activities in human scalp skin. Biochemical properties and correlation studies suggested that in addition to TS PST, the TL PST activity, another ST activity or both were involved in the reaction. Partially purified human platelet TL PST tested with minoxidil and dopamine showed identical thermal stabilities and similar responses to the inhibitors 2,6-dichloro-4-nitrophenol (DCNP) and NaCl. To characterize the activity of TL PST toward minoxidil, several biochemical properties of the enzyme expressed from a human liver cDNA clone were investigated. When assayed with minoxidil and dopamine, thermal stabilities of the expressed enzyme were identical and IC50 values for the inhibitors DCNP and NaCl were similar. It was also demonstrated that cDNA encoded human liver dehydroepiandrosterone sulfotransferase and estrogen sulfotransferase contributed to the sulfation of minoxidil. The results confirm that at least four human STs contribute to minoxidil sulfation. MNX-ST activity represents a combination of ST activities. The data indicate that multiple ST activities should be taken into account in attempts to predict the regulation of minoxidil sulfation and individual responses to minoxidil.
Article
An important step in understanding minoxidil's mechanism of action on hair follicles was to determine the drug's active form. We used organ-cultured vibrissa follicles to test whether it is minoxidil or its sulfated metabolite, minoxidil sulfate, that stimulates hair growth. Follicles from neonatal mice were cultured with or without drugs and effects were assessed by measuring incorporation of radiolabeled cysteine in hair shafts of the treated follicles. Assays of minoxidil sulfotransferase activity indicated that vibrissae follicles metabolize minoxidil to minoxidil sulfate. Dose-response studies showed that minoxidil sulfate is 14 times more potent than minoxidil in stimulating cysteine incorporation in cultured follicles. Three drugs that block production of intrafollicular minoxidil sulfate were tested for their effects on drug-induced hair growth. Diethylcarbamazine proved to be a noncompetitive inhibitor of sulfotransferase and prevented hair growth stimulation by minoxidil but not by minoxidil sulfate. Inhibiting the formation of intracellular PAPS with chlorate also blocked the action of minoxidil but not of minoxidil sulfate. Acetaminophen, a potent sulfate scavenger blocked cysteine incorporation by minoxidil. It also blocked follicular stimulation by minoxidil sulfate apparently by directly removing the sulfate from the drug. Experiments with U-51,607, a potent minoxidil analog that also forms a sulfated metabolite, showed that its activity was inhibited by both chlorate and diethylcarbamazine. These studies show that sulfation is a critical step for hair-growth effects of minoxidil and that it is the sulfated metabolite that directly affects hair follicles.
Article
The N,O-sulfate of minoxidil (Mnx) is the active agent in producing the vasodilation and the hair-growth stimulating responses observed with Mnx treatment. In this report, Mnx sulfation activity was assayed in cytosol prepared from several normal human livers, and Mnx sulfation was shown to correlate significantly with the activity of the phenol-sulfating form of phenol sulfotransferase (P-PST) activity in the same livers. No correlation was observed between Mnx sulfation and the dopamine or dehydroepiandrosterone (DHEA) sulfotransferase activities present in human liver. Mnx sulfation also copurified with P-PST activity during the purification of P-PST from human liver. During the purification procedure, Mnx and p-nitrophenol sulfotransferase (P-PST) activities were resolved from the dopamine and DHEA sulfation activities catalyzed by the monoamine-sulfating form of phenol sulfotransferase (M-PST) and DHEA sulfotransferase respectively. Also, purified DHEA sulfotransferase was not capable of sulfating Mnx, and no data were obtained to indicate that Mnx is a substrate for M-PST. p-Nitrophenol, a substrate for P-PST, was demonstrated to be a competitive inhibitor of Mnx sulfation catalyzed by purified P-PST when Mnx was the variable substrate. These results indicate that Mnx is sulfated and, therefore, bioactivated by P-PST in human liver.