Canine Noninflammatory Alopecia: A Comprehensive Evaluation of Common and Distinguishing Histological Characteristics

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DOI: 10.1111/j.1365-3164.2012.01049.

Canine noninflammatory alopecia: a comprehensive


evaluation of common and distinguishing histological
characteristics
Tabitha Müntener*, Gertraud Schuepbach-Regula†, Linda Frank‡, Silvia Rüfenacht§ and
Monika M. Welle*¶
*Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, Länggassstrasse 122, CH-3012 Bern, Switzerland
†Veterinary Public Health Institute, Vetsuisse Faculty, University of Bern, Schwarzenburgstrasse 155, CH-3097 Liebefeld, Switzerland
‡College of Veterinary Medicine, University of Tennessee, C247 Veterinary Medical Center, Knoxville, TN 37996-4544, USA
§Dermavet, Muhenstrasse 56, CH-5036 Oberentfelden, Switzerland
¶DermFocus, Vetsuisse Faculty, University of Berne, Switzerland
Correspondence: Monika M. Welle, Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, Länggassstrasse 122, CH-3012 Bern,
Switzerland. E-mail: monika.welle@vetsuisse.unibe.ch

Background – Noninflammatory alopecia is a frequent problem in dogs, and the pathogenesis is still unclear.
Objective – The objective of this study was a comparative histological description of skin biopsies from dogs
with different alopecic disorders and control dogs matched for coat type, season and disease duration.
Animals – Twenty-one cases of alopecia X in plush-coated dogs, 12 cases of recurrent flank alopecia, three
cases of hyperestrogenism, 15 cases of hyperadrenocorticism, 12 cases of hypothyroidism and 12 cases of pri-
mary alopecic disorders of unknown cause were evaluated. The controls were biopsies from 38 dogs of different
coat types.
Methods – We evaluated five serial sections of each biopsy histologically and immunohistologically to compare
the histological findings within the disease groups and with the control.
Results – All the dogs with hair cycle disorders had a significant increase in the number of hairless hair follicles,
which we assigned to kenogen. In addition, dogs with alopecia X had the lowest percentage of anagen follicles
and the highest percentage of telogen follicles.
Conclusions – The marked increase in kenogen follicles is a strong indication that the induction of the new ana-
gen phase is impaired in hair cycle disorders. The findings in dogs with alopecia X further suggest that premature
catagen is also involved in the pathogenesis. Further work to investigate the stem cell compartment and possible
initiating factors for the different cycle phases is required to elucidate the exact pathogenesis.

Histological features commonly reported in primary


Introduction
alopecic disorders include predominance of telogen folli-
Noninflammatory alopecia is a frequent problem in dogs; cles, hairless telogen follicles,1,8 follicular atrophy,1 follicu-
however, the pathogenesis of this group of disorders is lar dysplasia,9 so-called ‘flame follicles’ with excessive
still unclear, and diagnosis is based on the history, clinical trichilemmal keratinization1 and dystrophic follicles in
findings, histopathology of skin biopsies and laboratory cyclical flank alopecia.6
findings. Histopathology findings in skin biopsies, how- All of these HC disorders are associated with arrest or
ever, are often unspecific and do not discriminate impaired promotion of the HC. During the normal HC, the
between different hair cycle (HC) disorders. Potential hair follicle (HF) undergoes recurrent phases of controlled
diagnoses include alopecia X, hyperadrenocorticism, hypo- growth (anagen), regression of the lower follicle (catagen)
thyroidism, hyperestrogenism and recurrent flank alope- and relative quiescence (telogen). After a variable time in
cia, as well as primary alopecic disorders of unknown telogen, initiation of a new anagen phase occurs through
cause in nonplush-coated dogs (i.e. dogs without endocri- activation of stem cells. The release and shedding of
nological abnormalities that present with bilateral sym- hair has been described as a distinct phase named exo-
metrical alopecia and histological changes supporting a gen.10,11 Depending on the hair type and the species,
hair cycle disorder).1–7 shedding can occur at any point in the HC (Figure 1).11,12
For example, mice shed their overfur hairs in anagen,
Accepted 27 February 2012
while their underfur hairs are shed in anagen or subse-
Sources of Funding: The study was supported by an ESVD quent telogen.10,12 According to our observations, dogs
research grant and the Specialization committee (SPEZKO) of the shed their old club hair during the following anagen IV
Vetsuisse Faculty of the University of Bern. phase at the latest. Kenogen is a recently introduced term
Conflict of Interest: No conflicts of interest have been declared. that applies to HFs that have lost their hair fibre (exogen)
ª 2012 The Authors. Veterinary Dermatology
206 ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.
Histology of canine noninflammatory alopecia

coat types. We were not, however, able to obtain control biopsies


from dogs with long hair and dense undercoat or with an anagen-
dominated hair cycle, and there was only one biopsy from a plush-
coated type breed. Details about the control dogs are presented in
Table 2.
The biopsies were embedded in paraffin, sectioned at 4 lm and
stained with haematoxylin and eosin. In addition, five serial sections
of each biopsy were cut and stained immunohistochemically for
laminin (Dako, Baar, Switzerland), vimentin (Dako) and CAM5.2 (anti-
cytokeratin) (Becton Dickinson, Allschwil, Switzerland), and the
histochemical stains Sacpic and Masson Fontana, as previously
described.16
The biopsies were read blinded, and either a minimum of 50 or,
where fewer, all the hair follicles were assessed. Hair follicles were
assigned to a specific cycle stage: haired telogen, kenogen (formerly
hairless telogen), early anagen (anagen I–IIIb), late anagen (ana-
gen IIIc–VI), early catagen, late catagen and not determinable.16 The
presence of atrophy, dystrophy, excessive trichilemmal keratiniza-
tion, infundibular hyperkeratosis (not present, mild, moderate or
severe) and follicular casts (not present or present) was also
recorded. Other parameters included the following: the length of the
biopsy section; the depth of the dermis at its thinnest point; the num-
ber of epidermal cell layers (1–2, 2–3 and >3); epidermal pigmenta-
tion (not present, multifocal mild pigmentation and multifocal marked
or diffuse hyperpigmentation); epidermal orthokeratotic hyperkerato-
Figure 1. The hair cycle. After the telogen stage, the hair follicle
sis (not present, mild and moderate); the size of the sebaceous
may either lose its hair shaft and rest in kenogen for an indefinite
glands (normal, atrophic and hyperplastic); and the presence of
time period or may progress directly to the subsequent anagen
dermal inflammation. The morphological criteria are described in
stage. Exogen, the release of the old club hair and its shedding, can
Figures 2–7.
occur at any time during the subsequent hair cycle but takes place
All statistical analyses were performed with the statistical soft-
most often during anagen.
ware NCSS 2007 (NCSS, Kaysville, UT, USA). Mean values were
used where more than one biopsy was evaluated. Comparison of the
outcome measures between controls and cases, age and sex
and remain empty for a certain time before anagen is groups, diagnosis groups, coat types and duration of disease were
reinitiated.13 The percentage of kenogen follicles is made using the appropriate ANOVAs, Kruskal–Wallis tests, or chi-
increased in androgenetic alopecia in humans.13 squared tests. Control samples were also evaluated for different
Most HFs in primary alopecia are said to be in telogen, sample times following euthanasia. Values of P < 0.05 were consid-
ered statistically significant.
have an increased number of catagen stages and show
secondary atrophy. It has been suggested that the pre-
dominance of telogen is associated with lack of anagen Results
induction, impaired anagen promotion or premature cat-
Evaluation of hair cycle stages in control dogs
agen induction.14 Lack of anagen induction could mean
The distribution of the different hair follicle cycle stages in
that the signals that induce germ cells to develop into
the 38 control dogs (Tables 3, 4, and 5) was similar to that
follicular keratinocytes or that the stem cell precursors
reported for beagle dogs.16 The percentage of not deter-
themselves are not fully functional. Hair follicles may also
minable follicles was 27.2% for our control dogs versus
be forced into premature catagen.
34.7% in the beagles. Telogen (telogen and kenogen)
The objectives of our study were to determine whether
follicles comprised 30.6%, anagen follicles 39.9% and
distinct histological features can be used to discriminate
catagen follicles 2.4% versus 27.6, 30.1 and 7.6%,
between alopecic disorders, and if the predominant HC
respectively, in the beagles.16 In control dogs with a
stage indicates the time point of HC interruption in alope-
normal coat, the percentage of haired telogen follicles
cic disorders. This would allow further insight into the
was higher than in control dogs with either a fine, long
pathological mechanisms involved in HC arrest and be
coat or a wire-haired coat (Table 4), but no differences
helpful for future investigations.
were seen for other hair follicle stages. There was also a
trend for plush-coated dogs to have a higher hair follicle
Materials and methods density than dogs with other coat types (10.9 versus 4.9–
6.8 follicles ⁄ mm of epidermal width, respectively).
Biopsies from 76 dogs with alopecia were evaluated. Histopatho-
logical findings, detailed signalment, a complete clinical history and
laboratory test results were used to achieve a final diagnosis of Comparison between control dogs and all alopecic
alopecia X in plush-coated dogs (n = 21; 15 were described in a pre- dogs
vious study15), alopecia of unknown cause in nonplush-coated The comparison of the histological parameters between
breeds (n = 12), recurrent flank alopecia (n = 13; one provided by the control dogs and alopecic dogs is presented in
Joan Rest, Focus Histopathology, Warrington, UK), hyperestroge-
Table 3. The percentage of not determinable follicles was
nism (n = 3), hyperadrenocorticism (n = 15) and hypothyroidism
lower in skin biopsies from alopecic dogs compared with
(n = 12). The signalment, duration of clinical signs and time of the
year when the dogs were biopsied are presented in Table 1. Post- control animals. In alopecic dogs, there was a significant
mortem control biopsies were collected from the shoulder and thigh increase in kenogen follicles with a significant decrease in
of 38 dogs without skin problems of different breeds with different anagen (both early and later stages) and catagen follicles
ª 2012 The Authors. Veterinary Dermatology
ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44. 207
Müntener et al.

Table 1. Case summary of dogs with primary alopecia


Case Coat Age Season Duration since
number Disease Breed type (years) Sex of biopsy onset of alopecia
1* Alopecia X Pomeranian 4 3.5 MC 2 4
2* Alopecia X Pomeranian 4 3.5 MC 2 4
3* Alopecia X Pomeranian 4 1.5 M 2 3
4* Alopecia X Pomeranian 4 4 MC 3 4
5* Alopecia X Pomeranian 4 6 M 3 3
6* Alopecia X Pomeranian 4 4 FS 3 4
7* Alopecia X Pomeranian 4 6.5 FS 3 4
8* Alopecia X Pomeranian 4 4 M 3 3
9* Alopecia X Pomeranian 4 6.5 MC 3 4
10* Alopecia X Pomeranian 4 2.5 MC 3 4
11* Alopecia X Pomeranian 4 3.5 FS 3 2
12* Alopecia X Pomeranian 4 3 FS 4 3
13* Alopecia X Pomeranian 4 2 M 4 3
14* Alopecia X Pomeranian 4 4 MC 4 3
15* Alopecia X Pomeranian 4 1.5 M 4 3
16* Alopecia X Pomeranian 4 10 FS 1 3
17 Alopecia X Pomeranian 4 1.5 M 3 4
18 Alopecia X Siberian husky 4 5 M 1 4
19 Alopecia X Miniature Pomeranian 4 2.5 M 1 2
20 Alopecia X Miniature Pomeranian 4 5 M 2 1
21 Alopecia X Miniature Pomeranian 4 6 FS 4 2
22 Alopecia of unknown cause Bernese mountain dog mix 3 9 M 3 —
23 Alopecia of unknown cause Scottish terrier 5 5 M 3 3
24 Alopecia of unknown cause Poodle 7 15 M 1 4
25 Alopecia of unknown cause Yorkshire terrier 3 10 MC 2 —
26 Alopecia of unknown cause Dachshund, longhaired 3 9 M 1 4
27 Alopecia of unknown cause Toy poodle 7 6 M 2 3
28 Alopecia of unknown cause Maltese 3 7 M 2 —
29 Alopecia of unknown cause Labrador · Appenzell mountain dog 1 10 FS 3 —
30 Alopecia of unknown cause Maltese 3 8 M 1 —
31 Alopecia of unknown cause Landser · Newfoundland 6 4 M 3 2
32 Alopecia of unknown cause Papillon 3 8 M 4 1
33 Alopecia of unknown cause Mixed breed - 3 F 4 3
34 RFA Weimaraner 2 4 F 2 4
35 RFA Boxer 2 Adult F 4 2
36† RFA Miniature dachshund, wirehaired 5 2 MC 3 —
37 RFA Airedale terrier 5 4 MC 1 —
38 RFA Pumi mix 7 6 MC 1 —
39 RFA Dogue de Bordeaux 2 5 F 1 2
40 RFA Labrador retriever 1 2 FS 2 2
41 RFA Airedale terrier 5 2 FS 4 1
42 RFA Labrador retriever 1 5 MC 1 4
43 RFA German wirehaired pointer 5 3 FS 1 2
44 RFA Shih tzu 6 10 FS 2 —
45 RFA Rhodesian ridgeback 2 4 F 2 1
46 RFA Airedale terrier 5 10 FS 4 —
47 Hyperestrogenism Belgian Groenendael 6 12 M 4 —
48 Hyperestrogenism Pinscher mix 2 11 F 3 4
49 Hyperestrogenism Schnauzer · poodle 7 12 M 1 1
50 Hyperadrenocorticism Yorkshire terrier 3 9 MC 1 —
51 Hyperadrenocorticism Boxer 2 7 M 1 —
52 Hyperadrenocorticism Cairn terrier 5 9 F 2 —
53 Hyperadrenocorticism Fox terrier 5 5 MC 4 1
54 Hyperadrenocorticism Old German shepherd dog 6 8 FS 4 4
55 Hyperadrenocorticism Husky mix 1 6 FS 1 1
56 Hyperadrenocorticism Briard · Bergamasco shepherd 6 9 M 1 2
57 Hyperadrenocorticism Bichon frisé 3 12 M 2 1
58 Hyperadrenocorticism Boston terrier 2 9 FS 3 —
59 Hyperadrenocorticism Dachshund, shorthaired 2 5 M 4 4
60 Hyperadrenocorticism Boxer · pitbull 2 6 FS 3 3
61 Hyperadrenocorticism Bichon frisé 3 10 FS 4 —
62 Hyperadrenocorticism Coton de Tuléar 6 12 M 1 4
63 Hyperadrenocorticism Miniature Pomeranian 4 5 F 1 2
64 Hyperadrenocorticism Boxer 2 9.5 M 4 1
65 Hypothyroidism White Swiss shepherd 1 8 M 1 —
66 Hypothyroidism Doberman 2 9 M 4 1
67 Hypothyroidism Old German sheepdog 6 11 FS 3 1

ª 2012 The Authors. Veterinary Dermatology


208 ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.
Histology of canine noninflammatory alopecia

Table 1. (Continued)
Case Coat Age Season Duration since
number Disease Breed type (years) Sex of biopsy onset of alopecia
68 Hypothyroidism Border collie 1 6 FS 3 1
69 Hypothyroidism American Staffordshire terrier 2 4 M 4 1
70 Hypothyroidism Rhodesian ridgeback 2 7 FS 1 2
71 Hypothyroidism Bergamasco shepherd 6 11.5 M 2 1
72 Hypothyroidism Pyrenean shepherd 6 8 M 4 4
73 Hypothyroidism Poodle 7 4 MC 2 4
74 Hypothyroidism Doberman 2 4 FS 2 —
75 Hypothyroidism Bearded collie 6 13 FS 2 —
76 Hypothyroidism Labrador retriever 1 10 M 3 1
Abbreviations: F, female; FS, female spayed; M, male; MC, male castrated; and RFA, recurrent flank alopecia. Coat types: 1, normal; 2, short fine;
3, long fine; 4, plush coated; 5, wire haired; 6, long, dense undercoat; and 7, anagen dominated. Duration since onset of alopecia: 1, up to
3 months; 2, 3–6 months; 3, 6–12 months; and 4, over 12 months. Season of biopsy: 1, spring (March–May); 2, summer (June–August); 3,
autumn (September–November); and 4, winter (December–February).
*Cases of alopecia X in plush-coated dogs that were previously characterized.15
†Case provided by Joan Rest (Focus – Veterinary Histopathology International).

Table 2. Case summary of control dogs


Control number Breed Coat type Age (years) Sex Season of biopsy
1 Great Dane 2 8 F 4
2 Mixed breed; short, fine coat 2 12 F 4
3 Boxer 2 1 M 4
4 Bernese mountain dog mix 3 9 F 4
5 Golden retriever 1 8 F 4
6 Borzoi 3 6 FS 4
7 Belgian Malinois 1 4 M 4
8 Terrier 5 12 F 4
9 German shepherd 1 M 3
10 Appenzell mountain dog · Border collie 1 15 FS 4
11 Field spaniel 3 9 MC 4
12 Cocker spaniel 3 11 FS 4
13 Dachshund, longhaired 3 7 M 4
14 Cairn terrier 5 14 FS 4
15 German shepherd 1 7 M 4
16 Greater Swiss mountain dog 1 9.5 M 4
17 West Highland white terrier 5 14.75 FS 4
18 American cocker spaniel 3 11 M 3
19 West Highland white terrier 5 8.5 M 3
20 German shepherd · Rottweiler 1 8 F 3
21 German shepherd 1 0.3 M 3
22 Flat-coated retriever 3 11 MC 3
23 Golden retriever 1 7 M 3
24 Cairn terrier 5 7 FS 3
25 French bulldog 2 0.6 M 3
26 Bernese mountain dog 3 0.5 M 3
27 Great Dane 2 4 FS 3
28 Terrier mix 5 Adult — 3
29 Golden retriever 1 6.5 MC 3
30 West Highland white terrier 5 Adult — 3
31 Malamute mix 1 12 FS 3
32 Cocker spaniel 3 6 MC 3
33 Mixed breed; short, fine coat 2 8 MC 4
34 French bulldog 2 5.5 FS 4
35 Eurasian (spitz) 4 10.5 FS 4
36 Bruno Jura hound 2 Adult M 4
37 English cocker spaniel 3 12 F 1
38 Terrier 5 2 FS 1
Abbreviations: F, female; FS, female spayed; M, male; and MC, male castrated. Coat types: 1, normal; 2, short fine; 3, long fine; 4, plush coated;
and 5, wire haired. Season of biopsy: 1, spring (March–May); 2, summer (June–August); 3, autumn (September–November); and 4, winter
(December–February).

compared with control dogs. The number of telogen folli- nization, follicular atrophy or follicular dystrophy was sig-
cles did not differ between control and alopecic dogs. The nificantly increased in alopecic dogs. In both control dogs
number of hair follicles with excessive trichilemmal kerati- (94.5%) and alopecic dogs (99.3%), the majority of the
ª 2012 The Authors. Veterinary Dermatology
ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44. 209
Müntener et al.

(a) (b) (c) (d)

1
2

(e) (f) (g) (h)


1
1
2

3
1
2
3

Figure 2. Representative photomicrographs of hair cycle stages. (a) Telogen follicle containing a hair shaft (thick arrow). Note the position of the
dermal papilla (arrow 1) directly adjacent to the club hair without trailing connective tissue sheath cells and situated at the border of subcutaneous
fat and dermis. The club hair is anchored in trichilemmal keratin, and no inner root sheath is present. (b) Multiple kenogen follicles (thick arrows).
These are also located in the dermis but do not contain hair shafts. (c) Two hair follicles in early anagen (thick arrows). While the right one is in ana-
gen I, with a thickened and prolonged keratinocyte strand between the dermal papilla and the club hair, the left one is in anagen II, characterized
by the partial enclosure of the dermal papilla by matrical cells. Arrow 1 indicates a secondary follicle in telogen. Note that the two early anagen folli-
cles still contain their old club hairs (arrow 2). (d) Several hair follicles in late anagen in which their dermal papillae have reached the deepest posi-
tion in the subcutaneous fat tissue and the hair shaft is well developed (thick arrows). (e) Example of a hair follicle in early catagen (thick arrow).
The follicle is still located deep in the subcutaneous fat tissue. Its dermal papilla is wide and has almost achieved the typical onion-shape of cat-
agen III (arrow 1). Note also the papillary stalk of fibroblasts between the dermal papilla and the connective tissue sheath (arrow 2). The neighbour-
ing follicle is in late anagen (arrow 3). (f) A hair follicle in late catagen (thick arrow). The hair shaft has developed into a club hair (arrow 1) and the
epithelial strand is well developed (arrow 2). Even though the follicle is still surrounded by subcutaneous fat tissue, its upward movement is obvi-
ous by the trailing connective tissue sheath cells (arrow 3). (g) Examples of tangential follicular sections, where it cannot be determined whether
they are in late anagen or early catagen (thick arrows). They are completely surrounded by subcutaneous fat tissue and contain a well-developed
hair shaft. (h) Examples of follicles for which the cycle stage was not determined (thick arrows). They are located in the dermis and thus cannot be
assigned to either anagen or catagen. In addition, tangential sections of telogen follicles, which can be recognized by the trichilemmal keratin sur-
rounding the hair shaft, can be seen (arrows 1). They could also be in late catagen, but as catagen is of short duration, the likelihood that they are
in telogen is much higher. Thus, they were counted as telogen with hair shaft. Also visible in this image are epithelial structures of kenogen follicles
(arrows 2). They are high up in the dermis and have no hair shaft. Haematoxylin and eosin stain; scale bars represent 100 lm.

atrophic follicles were in kenogen, with the remainder in disorder is presented in Table 5. As only three dogs with
telogen. hyperestrogenism were available, they were not included
Epidermal hyperkeratosis did not differentiate control in the statistical analyses. In dogs with alopecia X, the
dogs from dogs with hair cycle disorders. In contrast, number of telogen follicles with a hair was significantly
infundibular hyperkeratosis and follicular casts were sig- higher than in the control dogs and in dogs with other
nificantly more frequent in alopecic dogs. Epidermal alopecic disorders (although it was similar in the three dogs
pigmentation also differed between control and alopecic with hyperestrogenism). The percentage of anagen folli-
dogs. While multifocal mild epidermal pigmentation was cles was highest in dogs with recurrent flank alopecia or
significantly more common in control dogs, multifocal hypothyroidism, although it was significantly lower in
marked or diffuse hyperpigmentation was significantly alopecic dogs than in the control animals. The proportion of
more common in alopecic dogs. hair follicles with excessive trichilemmal keratinization in
dogs with hyperadrenocorticism was at least double that
Comparison between control dogs and dogs with in other alopecic dogs. Severe infundibular hyperkeratosis
specific alopecic disorders was most obvious in dogs with recurrent flank alopecia.
The comparison of the histological parameters between Epidermal pigmentation was significantly increased in
the control dogs and dogs assigned to a specific alopecic all alopecic disorders, but was most pronounced in
ª 2012 The Authors. Veterinary Dermatology
210 ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.
Histology of canine noninflammatory alopecia

(a) (b) (c)

(d) (e) (f)

1 2

Figure 3. Representative photomicrographs of hair follicles displaying excessive trichilemmal keratinization, follicular atrophy or follicular dystro-
phy. (a) Example of a telogen follicle containing an excessive amount of trichilemmal keratin, which protrudes, flame like, into the outer root sheath
(thick arrow). Additionally, this follicle is dystrophic, with an abnormal shape and a thickened connective tissue sheath. Note also the extremely
large and misshapen dermal papilla (arrow 1). (b) Three examples of kenogen follicles with an excessive amount of trichilemmal keratin that pro-
trudes, flame like, into the outer root sheath (thick arrows). They have no hair shaft and are assigned to kenogen by the position of their dermal
papilla (border pf subcutis and dermis) as well as the absence of trailing connective tissue sheath cells, which would categorize them as catagen.
Arrow 1 indicates a telogen follicle that contains a hair shaft. (c) Examples of atrophic kenogen follicles, at most two cell layers across (thick
arrows). (d) Cross-sections of atrophic telogen follicles (thick arrows). Note the adjacent kenogen follicles of normal size, characterized by the pres-
ence of trichilemmal keratin but no hair (arrows 1). (e) Atrophic kenogen follicles (arrows 1) as well as a mildly dystrophic kenogen follicle with
abnormal shape and a thickened connective tissue sheath (arrows 2). The absence of a hair shaft was confirmed by serial sections. (f) Example of
a dystrophic kenogen follicle displaying a bizarre shape and a thickened connective tissue sheath (thick arrow). Haematoxylin and eosin stain. Scale
bars represent 100 lm.

dogs with recurrent flank alopecia and in the three parameters and the dermal thickness (Table 6). There
dogs with hyperestrogenism. Epidermal thickness, der- were few histological differences that discriminated
mal thickness and size of the sebaceous glands were between the different diseases. Dogs with alopecia X
not significantly decreased in dogs with hyperadreno- had significantly fewer anagen follicles (early and late
corticism in comparison to the other alopecic condi- anagen) and significantly more telogen follicles contain-
tions. However, the dermis was significantly thinner in ing a hair than dogs with other hair cycle disorders.
dogs with alopecia X and significantly thicker in dogs Another consistent finding was that dogs with hypo-
with hypothyroidism. More dogs with hypothyroidism thyroidism had a significantly thicker dermis. Dogs with
had dermal inflammation, but the difference was not recurrent flank alopecia had a significantly lower num-
significant. ber of telogen follicles containing a hair than dogs with
alopecic disorders of unknown cause, hyperadreno-
Comparison between the specific alopecic disorders corticism and hypothyroidism. Excessive trichilemmal
When comparing the specific hair cycle disorders, we keratinization was significantly less common in dogs
statistically evaluated only the hair cycle-specific with hypothyroidism than in dogs with alopecia X or
ª 2012 The Authors. Veterinary Dermatology
ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44. 211
Müntener et al.

(a) (b) (c)

(d) (e) (f)

Figure 4. Photomicrographs of follicles with variable infundibular hyperkeratosis. (a) Normal follicular infundibulum, which is not dilated and con-
tains a normal amount of infundibular keratin (thick arrow). (b) Mild infundibular hyperkeratosis (thick arrows). (c) Moderate infundibular hyperkera-
tosis (thick arrows). (d) Severe infundibular hyperkeratosis (thick arrow). (e) Example of a follicular infundibulum typically seen in cases of
recurrent flank alopecia (thick arrow). In addition to the severe infundibular hyperkeratosis, the infundibulum is pear shaped (wider at the bottom
than at the top) and the infundibular keratin extends into the lumina of the secondary hair follicles (arrows 1). (f) Comedone, characterized by an
extremely dilated infundibulum filled with keratin (thick arrow). Haematoxylin and eosin stain. Scale bars represent 100 lm.

hyperadrenocorticism. Dogs with alopecic disorders of alopecia showed significantly more (P < 0.05) multifocal
unknown cause and hyperadrenocorticism had a signi- marked or diffuse hyperpigmentation than dogs with
ficantly increased percentage of atrophic follicles in alopecia X.
comparison to dogs with recurrent flank alopecia or There were no significant differences in epidermal
hypothyroidism. thickness between dogs with the alopecic disorders and
Plush-coated dogs with alopecia X had significantly control dogs. However, the category of three or more
greater density of hair follicles (10 follicles ⁄ mm; P < 0.05) epidermal layers was significantly more common
than all the other disease groups except hypothyroidism. (P < 0.05) in dogs with recurrent flank alopecia and
The other disease groups consisted predominantly of hypothyroidism than in dogs with alopecia X, and it was
dogs with other coat types and had on average 4.2– also significantly more common in dogs with hypo-
5.8 follicles ⁄ mm of epidermal width, which is a similar thyroidism than in those with hyperadrenocorticism
density to the control dogs. (P < 0.05).
Dogs with alopecia X showed significantly more
(P < 0.05) multifocal mild epidermal pigmentation than Comparison of hair cycle parameters between
dogs with recurrent flank alopecia and hypothyroidism, control dogs of different coat types
and dogs with hyperadrenocorticism had more than those We were not able to compare data from plush-coated
with recurrent flank alopecia. Dogs with recurrent flank dogs, long-haired dogs and dogs with anagen-dominated
ª 2012 The Authors. Veterinary Dermatology
212 ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.
Histology of canine noninflammatory alopecia

(a)

(b)

Figure 5. Photomicrograph of follicular casts characterized by follic-


ular hyperkeratosis that projects as a cast over the epidermal surface
(thick arrows). Haematoxylin and eosin stain. Scale bars represent
100 lm. (c)

hair cycles owing to low numbers. Analysis of the other


coat types revealed only few statistically significant differ-
ences (Table 4). Control dogs with a normal coat had
significantly more (P < 0.05) follicles in telogen than dogs
with a long, fine coat or a wire-haired coat. While the total
number of anagen hair follicles did not differ significantly
between dogs with a short, fine or a long, fine coat, dogs
with a short, fine coat had significantly more follicles in
early anagen and less in late anagen than dogs with a
long, fine coat (P < 0.05). Dogs with a wire-haired coat
had significantly more follicles in catagen than dogs with
a long, fine coat and more follicles in early catagen than Figure 6. Photomicrographs of the epidermis showing the grading
dogs with a long, fine or a short, fine coat (P < 0.05). applied to epidermal pigmentation. (a) No epidermal pigmentation.
Dogs with a wire-haired coat had a significantly thinner (b) Multifocal mild epidermal pigmentation characterized by scat-
tered small aggregates of melanin. (c) Diffuse hyperpigmentation,
(P < 0.05) dermis than the dogs with normal, or short,
located mainly in the basal layer but extending into the superficial
fine or long, fine coats. layers. Masson Fontana stain. Scale bars represent 100 lm.

Comparison of hair cycle parameters between


control dogs and alopecic dogs with specific coat cantly increased in alopecic dogs with a long, fine coat
types (Table 4).
It was not possible to include dogs with alopecia X in the
analysis because there were insufficient control dogs. Comparison of hair cycle stages and histological
Alopecic dogs had a significant increase in the number of parameters with disease duration
kenogen follicles compared with control dogs with the The dogs with alopecia were grouped according to
same coat type. In addition, the number of anagen and, in the duration of their condition, as up to 3 months,
most groups, catagen follicles was decreased. In most 3–6 months, 6–12 months and over 12 months (Table 7).
groups, there was a significant increase in the number of The duration was not known for all cases, however, and
follicles with excessive trichilemmal keratinization and the small group sizes for each condition precluded any
dystrophic follicles, but follicular atrophy was only signifi- analysis (data not shown). Combing the conditions did not
ª 2012 The Authors. Veterinary Dermatology
ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44. 213
Müntener et al.

(a) (b)

(c) (d)

Figure 7. Photomicrographs showing the grading applied to sebaceous glands. (a) and (b) The variability in sebaceous gland size in the control
dogs. Even though they may be small and inconspicuous, they are usually present with every follicular complex (thick arrows). (c) Examples of
atrophic sebaceous glands (thick arrows). In addition to being very small and inconspicuous, they are missing from most follicular complexes (see
the follicular complex on the right side of the picture). (d) Example of hyperplastic sebaceous glands in a dog with recurrent flank alopecia showing
a marked increase of follicular lobuli dispersed around the follicular complex (thick arrows). Haematoxylin and eosin stain. Scale bars represent
100 lm.

yield any clear trends, except for dogs with cyclical flank winter had significantly fewer anagen (specifically late
alopecia, in which the percentage of telogen follicles anagen) follicles, but there were no other significant
decreased and the percentage of anagen follicles findings.
increased with disease duration.
Comparison of hair cycle stages and histological
Comparison of hair cycle stages and histological parameters with age and sex
parameters with time of sampling In dogs with alopecia X, neutered female dogs had signi-
Analysis was only possible between biopsies taken ficantly fewer telogen follicles and significantly more
in the autumn (September–November) and winter kenogen and atrophic follicles than male dogs. No other
(December–February) because too few dogs were significant findings were observed in the alopecic or
sampled at other times of the year. Biopsies taken in control groups.
ª 2012 The Authors. Veterinary Dermatology
214 ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.
Histology of canine noninflammatory alopecia

Table 3. Percentage distribution of different hair cycle stages and other histological parameters and their statistical significance in alopecic disor-
ders
Alopecia Recurrent
Control All alopecic X-like flank Hyperoestro- Hyperadreno- Hypo-
dogs cases Alopecia disease alopecia genism corticism thyroidism
(n = 38) (n = 76) X (n = 21) (n = 13) (n = 12) (n = 3) (n = 15) (n = 12)
Dermis (mm)† 1.09 0.99 0.65** 0.92 1.12 0.87 1 1.55**
ND total (%) 27.22 17.54** 23.54 18.35* 12.43** 19.72 10.14** 19.97*
ND with hair (%) 25.73 12.33** 14.55** 14.55* 7.77** 17.18 7.71** 15.14*
ND without hair (%) 1.49 5.2* 8.94* 3.8* 4.66* 2.53 2.44 4.83*
Tel with hair (%) 16.54 18.65 31.23** 16.47 4.2** 31.5 15.27 14.45
Kenogen (%) 14.01 54.2** 42.59** 54.73** 66.23** 43.35 64.98** 51.17**
Ana total (%) 39.85 5.79** 0.81** 4.56** 12.61** 2.55 6.08** 9.48**
Ana ND (%) 0.1 0.1 0.08 0 0.48 0 0 0.03
Ana early (%) 8.59 3.08** 0.28** 2.49** 6.78 1.12 2.69** 5.93*
Ana late (%) 31.16 2.61** 0.45** 2.07** 5.35** 1.43 3.4** 3.52**
Ana-Cat (%) 0 3.5** 1.8** 5.52** 4.16** 2.87 3.21** 4.15**
Cat total (%) 2.37 0.33** 0.07** 0.38* 0.37** 0 0.31** 0.78*
Cat early (%) 0.77 0.1** 0** 0.08* 0.22* 0 0* 0.32
Cat late (%) 1.63 0.23** 0.07** 0.3* 0.15* 0 0.31* 0.46*
ETK (%) 0.6 8.57** 9.3** 6.56* 4.47* 7.17 17.49** 2.79
FA (%) 5.31 20.02** 19.05* 27.24** 12.42 10.7 28.24** 13.49
DF (%) 0.33 10.18** 6.96** 12.72** 5.74** 10.2 16** 9.94**
Abbreviations: Ana, anagen; Ana-Cat, not determinable if late anagen or early catagen; Cat, catagen; DF, dystrophic follicles (the given percentage
applies to the total number of assessed hair follicles); ETK, excessive trichilemmal keratinization (the given percentage applies to the total number
of assessed hair follicles); FA, follicular atrophy (the given percentage applies to telogen without hair follicles); ND, not determinable; and Tel, telo-
gen. No statistical significance is indicated for cases of hyperestrogenism due to the small group size (three dogs).
*P < 0.05, **P < 0.001, statistically significant difference from control group.
†The values given are mean values of the dermis thickness of all dogs in the respective group.

Follicular atrophy was mainly found in kenogen follicles;


Discussion
only 5.6 and 0.7% of the atrophic follicles in control and
These findings show that the histological diagnosis of alopecic dogs, respectively, contained a hair shaft. This
specific hair cycle disorders remains a challenge, and no indicates that follicular atrophy occurs after telogen, and
features consistently differentiate these conditions. Nev- that the appropriate stimuli and ⁄ or hair follicle stem or
ertheless, our data do highlight some discriminative fea- progenitor cells are lacking. In rodents, hair follicle down-
tures that may become useful in histological analysis. A growth is initiated by stem cells activated by signals from
striking feature of all these hair cycle disorders was the dermal papilla.19 Miniaturization of hair follicles in
a 3- to 4.5-fold increase in the number of kenogen hair male androgenic alopecia, furthermore, is associated with
follicles. Kenogen has been described in humans and a lack of CD200-rich and CD34-positive hair follicle pro-
denotes that the club hair is shed directly after telogen genitor cells.20 Follicular stem cells are present in the
and the remaining empty follicle rests in a lag phase17 outer root sheath of the canine hair follicle,21 but their role
without coincident entry of the hair follicle into a new ana- in follicular atrophy has not been studied.
gen phase.12,13,18 Hairless follicles have been previously Other histological features that differentiated control
observed in dogs, and it has been suggested that their dogs from alopecic dogs included excessive trichilemmal
increased number is the only true indicator of alopecia keratinization, follicular dystrophy, infundibular hyperkera-
(Bob Dunstan, personal communication). These follicles tosis, follicular casts and epidermal pigmentation. This is
were termed ‘hairless telogen’ because trichilemmal in agreement with earlier reports.1 Hyperkeratosis, which
keratin can be seen in the lumen. However, we suggest has been described as a histological feature of hair cycle
using the term ‘kenogen’ in veterinary medicine, as in disorders, was inconsistent in our study, as a third of the
human dermatology. Our findings, together with those control dogs had mild hyperkeratosis and a quarter of
from other studies16 (Bob Dunstan, personal communica- the alopecic dogs had no hyperkeratosis. However, the
tion), suggest that kenogen follicles may account for degree of hyperkeratosis in biopsy samples might have
up to 20% of the follicular stages in healthy hair coats. been influenced by preparation of the biopsy site. In
However, an increased number clearly indicates hypotri- contrast to others,14,22,23 we did not find an increase in
chosis or alopecia. Based on the fact that all our alopecic catagen follicles in dogs with hair cycle disorders. One
dogs, irrespective of the cause, had a significantly reason might be that different criteria were used to iden-
increased number of kenogen follicles, we conclude that tify the follicular stages. Our criteria were based on a pre-
anagen induction is impaired in hair cycle disorders. This viously published guide16 adapted from the classification
is in contrast to earlier work suggesting that there is a lack scheme for murine hair follicles.24 This emphasized the
of anagen initiation in hyperadrenocorticism, impaired depth of the follicle and dermal papilla. A follicle with its
anagen promotion in hypothyroidism, and premature dermal papilla in the dermis or at the subcutis–dermis
catagen induction or impaired catagen propagation in border without trailing connective tissue sheath cells
alopecia X and recurrent flank alopecia.14 was classified as telogen, even if distorted or with
ª 2012 The Authors. Veterinary Dermatology
ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44. 215
216
Table 4. Percentage distribution of different hair cycle stages and other histological parameters and their statistical significance in alopecic disorders in regard to coat type
Müntener et al.

Coat: long, Coat: anagen


Coat: normal Coat: short, fine Coat: long, fine Coat: plush coated Coat: wire haired dense undercoat dominated

C A C A C A C A C A C A C A
(n = 8) (n = 6) (n = 8) (n = 13) (n = 13) (n = 10) (n = 1) (n = 23) (n = 8) (n = 8) (n = 0) (n = 10) (n = 0) (n = 5)
Dermis (mm)‡‡ 1.31§ 1.36 1.25** 1.3 1.05†† 0.75† 1.11 0.64 0.76§**†† 1.1† — 1.28 — 1.04
ND total (%) 24.93 15.89 26.84 14.85† 25.5 18.19 25.85 22.31 32.86 9.82* — 16 — 20.02
ND with hair (%) 23.46 11.86† 25.36 8.97† 23.83 13.79† 24.95 13.86 31.56 8.04* — 14.15 — 15.1
ND without hair (%) 1.46 4.03† 1.48 5.88 1.67 4.4† 0.9 8.4 1.31 1.78 — 1.85 — 4.92
Tel with hair (%) 20.73‡§ 16.67 24.14 14.47 11.9‡ 12.37 11.6 30.14 12.93§ 12.42 — 17.6 — 7.34
Kenogen (%) 12.15 49.6* 12.68 60.14* 15.91 59.83* 31.25 44.51 11.98 65.99* — 55.96 — 51.18
Ana total (%) 40.02 13.31† 33.3 7.5* 45.43 5.7* 29.5 1.13 38.45 8.14* — 6.75 — 7.65
Ana ND (%) 0.46 0.05 0 0.28 0 0 0 0.07 0 0.25 — 0 — 0
Ana early (%) 8.46 7.05 12.58¶ 5.88† 6.38¶ 1.97† 8.05 0.26 8.4 4.7 — 3.27 — 2.47
Ana late (%) 31.11 6.2† 20.72¶ 1.33† 39.05¶ 3.73* 21.45 0.8 30.05 3.19* — 3.48 — 5.18
Ana-Cat (%) 0 3.24† 0 2.64† 0 3.8† 0 1.8 0 3.64† — 3.49 — 12.68
Cat total (%) 2.18 1.3 3.04 0.39† 1.25†† 0.11† 1.8 0.14 3.79†† 0* — 0.2 — 1.12
Cat early (%) 0.69 0.28 0.33** 0.08 0.35†† 0.11 0.9 0 1.96**†† 0† — 0.2 — 0.34
Cat late (%) 1.49 1.03 2.72 0.31† 0.98 0† 0.9 0.14 1.83 0† — 0 — 0.78
ETK (%) 0.89 3.13 0 5.53* 0.85 8.67† 0.9 11.46 0.46 8.33† — 10.84 — 7.09
FA (%) 3.89 16.61 3.13 13.86 8.45 30.82* 8.95 18.95 3.34 15.5 — 23.82 — 20.71
DF (%) 0.46 7.45† 0 8.93* 0.35 14.3* 0.9 8.32 0.44 11.32† — 15.71 — 5.42
Abbreviations: A, alopecic; Ana, anagen; Ana-Cat, not determinable if late anagen or early catagen; Cat, catagen; C, Controls; DF, dystrophic follicles (the given percentage applies to the total number of assessed hair
follicles); ETK, excessive trichilemmal keratinization (the given percentage applies to the total number of assessed hair follicles); FA, follicular atrophy (the given percentage applies to telogen without hair follicles); ND,
not determinable; and Tel, telogen. Examples of coat types: normal, German shepherd; short, fine, boxer; long, fine, Bernese mountain dog; plush coated, Pomeranian; wire haired, Airedale terrier; long, dense under-
coat, Bergamasco shepherd; and anagen dominated, poodle.
*Statistically significant difference from control group of same coat type, P < 0.001.
†Statistically significant difference from control group of same coat type, P < 0.05.
‡Statistically significant difference between coat type normal versus long, fine, P < 0.05.
§Statistically significant difference between coat type normal versus wire-haired, P < 0.05.
¶Statistically significant difference between coat type short, fine versus long, fine, P < 0.05.
**Statistically significant difference between coat type short, fine versus wire-haired, P < 0.05.
††Statistically significant difference between coat type long, fine versus wire-haired, P < 0.05.
For the plush-coated breeds, no statistical significance is indicated owing to the small number of control dogs (n = 1). For the breeds with long hair and dense undercoat and the anagen-dominated dogs, no statistical
significance can be indicated owing to the absence of controls.
‡‡The given values are mean values of the dermis thickness of all dogs in the respective group.

ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.


ª 2012 The Authors. Veterinary Dermatology
Table 5. Percentage and number of cases evaluated and their statistical significance for definite histological parameters between control dogs and cases of different diagnosis groups
All alopecic Alopecia Recurrent
Control dogs cases Alopecia X X-like disease flank alopecia
(n = 38) (n = 76) (n = 21) (n = 13) (n = 12) Hyperoestrogenism Hyperadrenocorticism Hypothyroidism
[% and (n)] [% and (n)] [% and (n)] [% and (n)] [% and (n)] (n = 3) [% and (n)] (n = 15) [% and (n)] (n = 12) [% and (n)]

ª 2012 The Authors. Veterinary Dermatology


No hyperkeratosis 60.5 (23) 23.7 (18)* 9.5 (2)* 15.4 (2) 41.7 (5) 66.7 (2) 26.7 (4) 25 (3)
Mild hyperkeratosis 34.2 (13) 69.7 (53) 71.4 (15) 76.9 (10) 58.3 (7) 33.3 (1) 73.3 (11) 75 (9)
Moderate hyperkeratosis 5.3 (2) 6.6 (5) 19 (4) 7.7 (1) 0 0 0 0
No infundibular hyperkeratosis 76.3 (29) 1.3 (1)** 0** 0* 0* 33.3 (1) 0* 0*
Mild infundibular hyperkeratosis 21.1 (8) 18.4 (14) 19 (4) 15.4 (2) 8.3 (1) 0 13.3 (2) 41.7 (5)
Moderate infundibular hyperkeratosis 2.6 (1) 46.1 (35)** 38.1 (8)* 69.2 (9)** 25 (3)* 66.7 (2) 60 (9)** 33.3 (4)*
Severe infundibular hyperkeratosis 0 34.2 (26)** 42.9 (9)** 15.4 (2)* 66.7 (8)** 0 26.7 (4)* 25 (3)*

ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.


No follicular casts 97.4 (37) 26.3 (20)** 23.8 (5)* 23.1 (3)* 33.3 (4) 33.3 (1) 26.7 (4)* 25 (3)*
Follicular casts present 2.6 (1) 73.7 (56)** 76.2 (16)** 76.9 (10)** 66.7 (8)** 66.7 (2) 73.3 (11)** 75 (9)**
1–2 epidermal cell layers 60.5 (23) 51.3 (39) 61.9 (13) 69.2 (9) 41.7 (5) 0 53.3 (8) 33.3 (4)
2–3 epidermal cell layers 31.6 (12) 36.8 (28) 38.1 (8) 30.8 (4) 33.3 (4) 33.3 (1) 46.7 (7) 33.3 (4)
>3 epidermal cell layers 7.9 (3) 11.8 (9) 0 0 25 (3) 66.7 (2) 0 33.3 (4)
Normal sebaceous glands 89.5 (34) 78.9 (60) 71.4 (15) 84.6 (11) 83.3 (10) 100 (3) 86.7 (13) 66.7 (8)
Atrophy of sebaceous glands 10.5 (4) 17.1 (13) 28.6 (6) 15.4 (2) 0 0 13.3 (2) 25 (3)
Hyperplasia of sebaceous glands 0 3.9 (3) 0 0 16.7 (2) 0 0 8.3 (1)
No dermal inflammation 81.6 (31) 89.5 (68) 85.7 (18) 92.3 (12) 100 (12) 100 (3) 100 (15) 66.7 (8)
Dermal inflammation 18.4 (7) 10.5 (8) 14.3 (3) 7.7 (1) 0 0 0 33.3 (4)
No epidermal pigmentation 34.2 (13) 14.5 (11) 14.3 (3) 15.4 (2) 0* 0 20 (3) 25 (3)
Multifocal mild epidermal pigmentation 63.2 (24) 31.6 (24)* 61.9 (13) 30.8 (4) 0* 0 40 (6) 8.3 (1)*
Multifocal marked or diffuse hyperpigmentation 2.6 (1) 53.9 (41)* 23.8 (5)* 53.8 (7)* 100 (12)** 100 (3) 40 (6)* 66.7 (8)*
Alopecia X like, alopecia of unknown cause in nonplush coated breeds. No statistical significance is indicated for cases of hyperestrogenism owing to the small group size (three dogs).
*P < 0.05, **P < 0.001, statistically significant difference from control group.

217
Histology of canine noninflammatory alopecia
218
Table 6. Statistical comparison of different hair cycle disorders in regard to different hair cycle stages and other histological parameters
Alopecia
Alopecia X versus Alopecia Alopecia Alopecia X-like Alopecia X-like Recurrent flank Hyperadreno-
Müntener et al.

X versus recurrent X versus X versus disease versus disease versus Alopecia X-like alopecia versus Recurrent flank corticism
Alopecia X-like flank hyperadre- hypo- recurrent flank hyperadreno- disease versus hyperadreno- alopecia versus versus hypo-
disease alopecia nocorticism thyroidism alopecia corticism hypothyroidism corticism hypothyroidism thyroidism
Dermis (mm)† n.s. S** (0.7 S* (0.7 S** (0.7 n.s. n.s. S* (0.9 versus 1.6) n.s. S* (1.1 S* (1 versus 1.6)
versus 1.1) versus 1) versus 1.55) versus 1.6)
ND total (%) n.s. S* (23.5 S** (23.5 n.s. n.s. S* (18.4 versus 10.1) n.s. n.s. S* (12.4 S* (10.1 versus 20)
versus 12.4) versus 10.1) versus 20)
ND with hair (%) n.s. S* (14.6 S* (14.6 n.s. n.s. S* (14.6 versus 7.7) n.s. n.s. S* (7.8 S* (7.7 versus 15.1)
versus 7.8) versus 7.7) versus 15.1)
ND without hair (%) n.s. n.s. S* (8.9 n.s. n.s. n.s. n.s. n.s. n.s. n.s.
versus 2.44)
Tel with hair (%) S* (31.2 S** (31.2 S* (31.2 S* (31.2 S* (16.5 n.s. n.s. S* (4.2 versus S* (4.2 n.s.
versus 16.5) versus 4.2) versus 15.3) versus 15.4) versus 4.2) 15.3) versus 14.5)
Kenogen (%) n.s. S** (42.6 S** (42.6 n.s. n.s. n.s. n.s. n.s. n.s. n.s.
versus 66.2) versus 65)
Ana total (%) S** (0.8 S* (0.8 S** (0.8 S** (0.8 n.s. n.s. n.s. n.s. n.s. n.s.
versus 4.6) versus 12.6) versus 6.1) versus 9.5)
Ana ND (%) n.s. n.s. n.s. n.s. n.s. — n.s. n.s. n.s. n.s.
Ana early (%) S* (0.3 S* (0.3 S* (0.3 S** (0.3 n.s. n.s. n.s. n.s. n.s. n.s.
versus 2.5) versus 6.8) versus 2.7) versus 5.9)
Ana late (%) S** (0.5 S* (0.5 S* (0.5 S* (0.5 n.s. n.s. n.s. n.s. n.s. n.s.
versus 2) versus 5.4) versus 3.4) versus 3.5)
Ana-Cat (%) n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.
Cat total (%) n.s. n.s. n.s. S* (0.1 n.s. n.s. n.s. n.s. n.s. n.s.
versus 0.8)
Cat early (%) n.s. n.s. n.s. S* (0 n.s. n.s. n.s. n.s. n.s. S* (0 versus 0.3)
versus 0.3)
Cat late (%) n.s. n.s. n.s. S* (0.1 n.s. n.s. n.s. n.s. n.s. n.s.
versus 0.5)
ETK (%) n.s. n.s. n.s. S* (9.3 n.s. n.s. n.s. n.s. n.s. S* (17.5 versus 2.8)
versus 2.8)
FA (%) n.s. n.s. n.s. n.s. S* (27.2 n.s. S* (27.2 S* (12.4 versus 28.2) n.s. S* (28.2 versus 13.5)
versus 12.4) versus 13.5)
DF (%) n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.
Abbreviations: alopecia X like, alopecia of unknown cause in nonplush-coated breeds; Ana, anagen; Ana-Cat, not determinable if late anagen or early catagen; Cat, catagen; DF, dystrophic follicles (the given percent-
age applies to the total number of assessed hair follicles); ETK, excessive trichilemmal keratinization (the given percentage applies to the total number of assessed hair follicles); FA, follicular atrophy (the given per-
centage applies to telogen without hair follicles); ND, not determinable; n.s., no statistically significant difference; S, statistically significant difference; and Tel, telogen. No statistical significance is indicated for cases
of hyperestrogenism owing to the small group size (three dogs).
*P < 0.05, **P < 0.001.
†The given values are mean values of the dermis thickness of all dogs in the respective group.

ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.


ª 2012 The Authors. Veterinary Dermatology
Histology of canine noninflammatory alopecia

Table 7. Percentage distribution of different hair cycle stages and larity between dogs with alopecia X and hyperestroge-
other histological parameters and their statistical significance in nism may involve the estrogen receptor, which inhibits
alopecic dogs in regard to disease duration
anagen and has been investigated in alopecia X.4,25,26
Duration Duration Duration Duration Our findings in alopecia X in plush-coated breeds
up to 3–6 6–12 over were similar to those previously reported, and included
3 months months months 12 months
hyperkeratosis, follicular keratosis, excessive trichilem-
(n = 14) (n = 11) (n = 12) (n = 20)
mal keratinization, epidermal atrophy, few small anagen
Dermis (mm)‡ 1.22* 1.11 0.77* 0.88 bulbs, epidermal hyperpigmentation, melanin aggregates
ND total (%) 17.3 16.62 19.48 22.15
within follicular keratin, follicular atrophy and more telo-
ND with hair (%) 12.67 11.03 14.08 14.18
ND without hair (%) 4.63 5.59 5.4 7.92
gen follicles, with most telogen follicles retaining their
Tel with hair (%) 18.1 19.18 24.43 21.69 hair shaft.1,15,16 We also found a thinner dermis, but this
Kenogen (%) 57.66 54.31 52.39 45.08 should be interpreted with caution because the apparent
Ana total (%) 4.66* 5.8 1.78* 6.11 thickness of the dermis depends on how vertically the
Ana ND (%) 0.02 0.34 0 0.09 biopsy is cut.
Ana early (%) 2.14 3.56 0.96 2.57 Dogs with hypothyroidism had a significantly thicker
Ana late (%) 2.51 1.9 0.82† 3.45†
epidermis and dermis than the control animals and other
Ana-Cat (%) 1.77 3.59 1.77 4.77
Cat total (%) 0.5 0.5 0.16 0.25
disease groups, and fewer atrophic follicles than the
Cat early (%) 0.2 0.1 0 0.1 other disease groups. Thus, even accounting for variation
Cat late (%) 0.3 0.4 0.16 0.15 in dermal thickness by biopsy location or orientation,
ETK (%) 11.89 6.66 12.22† 7.02† these criteria could help differentiate hypothyroidism. It
FA (%) 20.01 16.56 18.92 17.45 has been reported that telogen follicles, often hairless,
DF (%) 13.28 8.44 7.56 6.76 predominate in hypothyroidism,8,23 whereas we found
Abbreviations: Ana, anagen; Ana-Cat, not determinable if late anagen that kenogen rather than telogen follicles were most fre-
or early catagen; Cat, catagen; DF, dystrophic follicles (the given per- quently associated with all the alopecic disorders. Thyroid
centage applies to the total number of assessed hair follicles); ETK, hormones prolong anagen duration in humans,27 and
excessive trichilemmal keratinization (the given percentage applies
dermal papilla fibroblasts are a direct target of thyroid-
to the total number of assessed hair follicles); FA, follicular atrophy
(the given percentage applies to telogen without hair follicles); ND, stimulating hormone.28 Experimentally, hypothyroid bea-
not determinable; and Tel, telogen. There are no statistically signifi- gles exhibit fewer hair shafts in the follicles and a failure
cant differences between duration groups ‘up to 3 months’ and to initiate anagen, which was corrected by thyroid hor-
‘3–6 months’, as well as between groups ‘up to 3 months’ and ‘over mone replacement therapy.8
12 months’, ‘3–6 months’ and ‘6–12 months’, and ‘3–6 months’ and Recurrent flank alopecia has been associated with the
‘over 12 months’.
following characteristics: follicular atrophy and infundibu-
*Statistically significant difference between duration groups ‘up to
3 months’ and ‘6–12 months’, P < 0.05.
lar hyperkeratosis, with keratin extending into the opening
†Statistically significant difference between duration groups ‘6–12’ of primary and secondary follicles; occasional distorted
and ‘over 12 months’, P < 0.05. follicles; telogen follicles narrowed to a thin cord of cells;
‡The values given are mean values of the dermis thickness of all pigmentation of the epidermis, outer root sheath and
dogs in the respective group. sebaceous glands; and pigmentary incontinence around
the deep hair follicle.6,29 In our cases, recurrent flank alo-
trichilemmal keratin at the level of the sebaceous glands pecia was mainly distinguished by marked infundibular
or above, rather than catagen. Serial sections were very hyperkeratosis, rare telogen follicles, frequent kenogen
helpful for the allocation of the stages, as reported else- and atrophic follicles, and epidermal hyperplasia and pig-
where.16 Moreover, our finding that telogen follicles were mentation. Pear-shaped severe infundibular hyperkerato-
no more frequent in alopecic dogs than in control dogs, sis, with keratin extending into the openings follicle of the
with the exception of alopecia X and hyperestrogenism, secondary follicles (Figure 4e), appeared to be character-
supports our criteria. Otherwise, an increased number of istic of recurrent flank alopecia and was rare in other alo-
catagen follicles should result in an increased number of pecic conditions. The number of anagen follicles was
telogen follicles, because catagen inevitably progresses higher than in the other alopecic diseases, which is possi-
into telogen. bly associated with the recurrent nature of the disease,
We could not reliably distinguish histologically between although the frequency of the different follicular stages
the different alopecic disorders. Alopecia X was associ- may differ when the alopecia is developing or resolving.
ated with fewer anagen and kenogen follicles (although Hyperadrenocorticism is reported to have predomi-
they were still three times more frequent than in the nance of hairless, atrophic telogen follicles,1 with atrophic
control dogs) and more telogen follicles than the other sebaceous glands, epidermis and infundibular epithe-
conditions. A similar picture was seen in dogs with hyper- lium.23 Comedones and calcinosis cutis are not always
estrogenism, but more dogs need to be studied to con- present but are typical.1,4 We saw the hairless (kenogen)
firm this. These and other findings suggest that catagen follicles, which were often atrophic, as well as a moder-
is induced prematurely,14 resulting in fewer anagen and ate to severe infundibular hyperkeratosis, but atrophy of
more telogen follicles. We did not see more catagen folli- the epidermis, dermis and sebaceous glands was not
cles, but this might be due to the short duration of cat- clearly reflected in our cases. Most of the dogs we evalu-
agen. It is also possible that these findings are normal for ated, with the exception of those with hyperestrogenism
plush-coated breeds, because a comparison with healthy or hypothyroidism, had a thin epidermis, but we could not
plush-coated dogs was not performed. The possible simi- easily differentiate sebaceous glands in affected dogs
ª 2012 The Authors. Veterinary Dermatology
ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44. 219
Müntener et al.

from those in the control dogs. We assessed the seba- 4. Frank LA. Comparative dermatology – canine endocrine derma-
ceous glands as normal if they were present with most toses. Clin Dermatol 2006; 24: 317–325.
5. Scott-Moncrieff JCR, Guptill-Yoran L. Hypothyroidism. In:
follicular complexes, even if small and inconspicuous. We
Ettinger SJ, Feldman EC, eds. Textbook of Veterinary Internal
regarded them as atrophic if they were missing from Medicine, 6th edn. St Louis, MO: Elsevier-Saunders, 2005;
most complexes, with the remaining ones small and 1535–1544.
inconspicuous. This strict definition may reflect the differ- 6. Gross TL, Ihrke PJ, Walder EJ et al. Dysplastic diseases of the
ences from the reported findings. Glucocorticoids induce adnexa. In: Skin Diseases of the Dog and Cat. Clinical and Histo-
catagen in mice.30 In addition, human hair follicles display pathologic Diagnosis, 2nd edn. Oxford: Blackwell, 2005; 518–
a functional equivalent of the hypothalamic–pituitary– 536.
7. Daminet S, Paradis M. Evaluation of thyroid function in dogs suf-
adrenal axis and synthesize cortisol upon stimulation
fering from recurrent flank alopecia. Can Vet J 2000; 41: 699–
by adrenocorticotrophic hormone and corticotrophin- 703.
releasing hormone.31 Corticotrophin-releasing hormone 8. Credille KM, Slater MR, Moriello KA et al. The effects of thyroid
inhibits hair shaft elongation and hair keratinocyte prolifer- hormones on the skin of beagle dogs. J Vet Intern Med 2001;
ation, and stimulates catagen induction and hair keratino- 15: 539–546.
cyte apoptosis.31 The number of catagen follicles was 9. Rothstein E, Scott DW, Miller WH Jr et al. A retrospective study
not, however, increased compared with biopsies from of dysplastic hair follicles and abnormal melanization in dogs with
follicular dysplasia syndromes or endocrine skin diseases. Vet
control dogss or from dogs with other alopecic disorders,
Dermatol 1998; 9: 235–241.
and the effects of cortisol, adrenocorticotrophic hormone 10. Milner Y, Sudnik J, Filippi M et al. Exogen, shedding phase of
and corticotrophin-releasing hormone in dogs need fur- the hair growth cycle: characterization of a mouse model. J In-
ther study. vestig Dermatol 2002; 119: 639–644.
The control dogs with a normal coat had significantly 11. Stenn K. Exogen is an active, separately controlled phase of the
more telogen follicles than dogs with long and fine or hair growth cycle. J Am Acad Dermatol 2005; 52: 374–375.
12. Higgins CA, Westgate GE, Jahoda CAB. From telogen to exo-
wire-haired coats. In addition, dogs with long, fine hair
gen: mechanisms underlying formation and subsequent loss of
had more follicles in late anagen. As late anagen is the the hair club fiber. J Investig Dermatol 2009; 129: 2100–2108.
phase of continued hair growth, this seems to reflect the 13. Rebora A, Guarrera M. Kenogen, a new phase of the hair cycle?
coat type. In winter, irrespective of the coat type, dogs Dermatology 2002; 205: 108–110.
had a significantly lower percentage of late anagen folli- 14. Cerundolo R, Mecklenburg L. Clinical and histological aspects of
cles in comparison to biopsies taken in the autumn. This non-inflammatory alopecia in dogs. Proceedings of the 22nd
finding reflects the longer coat during winter and the need Annual Congress of ESVD-ECVD, Mainz, 2007; 25–32.
15. Frank LA, Donnell RL, Kania SA. Oestrogen receptor evaluation
to save energy at this time of the year.
in Pomeranian dogs with hair cycle arrest (alopecia X) on melato-
In summary, all the hair cycle disorders in this study nin supplementation. Vet Dermatol 2006; 17: 252–258.
were associated with an increase in the prevalence of 16. Müntener T, Doherr MG, Guscetti F et al. The canine hair cycle –
kenogen follicles, which were often atrophic. This indi- a guide for the assessment of morphological and immunohisto-
cates that induction of new anagen growth is impaired. chemical criteria. Vet Dermatol 2011; 22: 383–395.
Only dogs with alopecia X or hyperestrogenism differed; 17. Sinclair R. Male pattern alopecia. BMJ 1998; 317: 865–869.
they showed the lowest percentage of anagen follicles 18. Rebora A, Guarrera M. Teloptosis and kenogen: two new con-
cepts in human trichology. Arch Dermatol 2004; 140: 619–
and the highest percentage of telogen follicles, suggest-
620.
ing that premature catagen induction may be an underly- 19. Hsu YC, Pasolli HA, Fuchs E. Dynamics between stem cells,
ing pathogenetic factor. Further work to investigate the niche and progeny in the hair follicle. Cell 2011; 144: 92–105.
stem cell compartment and possible initiating factors for 20. Garza LA, Yang CC, Zhao T et al. Bald scalp in men with androge-
the different cycle phases is now required to elucidate netic alopecia retains hair follicle stem cells but lacks CD200-rich
the exact pathogenesis of these disorders. and CD34-positive hair follicle progenitor cells. J Clin Invest
2011; 121: 613–622.
21. Kobayashi T, Shimizu A, Nishifuji K et al. Canine hair-follicle
Acknowledgements keratinocytes enriched with bulge cells have the highly prolifera-
tive characteristics of stem cells. Vet Dermatol 2009; 20: 338–
We thank all the submitting veterinarians for the addi- 346.
tional data they provided for their cases; and Manuela Bo- 22. Ebling FJ, Hale PA. Hormones and hair growth. In: Goldsmith
zzo, Eveline Rohrer and Erika Garchi for their excellent LA, ed. Biochemistry and Physiology of the Skin, 2nd edn. New
technical support. York, NY: Oxford University Press, 1983; 522–546.
23. Gross TL, Ihrke PJ. The histologic analysis of endocrine-related
alopecia in the dog. In: von Tscharner C, Halliwell REW, eds.
References Advances in Veterinary Dermatology, vol 1. London, UK: Baillière
Tindall, 1990; 77–88.
1. Gross TL, Ihrke PJ, Walder EJ et al. Atrophic diseases of the 24. Müller-Röver S, Handjiski B, van der Veen C et al. A comprehen-
adnexa. In: Skin Diseases of the Dog and Cat. Clinical and sive guide for the accurate classification of murine hair follicles in
Histopathologic Diagnosis, 2nd edn. Oxford: Blackwell, 2005; distinct hair cycle stages. J Investig Dermatol 2001; 117: 3–15.
480–517. 25. Frank LA. Oestrogen receptor antagonist and hair regrowth in
2. Cerundolo R, Lloyd DH, Vaessen MMAR et al. Alopecia in pom- dogs with hair cycle arrest (alopecia X). Vet Dermatol 2007; 18:
eranians and miniature poodles in association with high urinary 63–66.
corticoid:creatinine ratios and resistance to glucocorticoid feed- 26. Oh H, Smart RC. An estrogen receptor pathway regulates the
back. Vet Rec 2007; 160: 393–397. telogen–anagen hair follicle transition and influences epidermal
3. Leone F, Cerundolo R, Vercelli A et al. The use of trilostane for cell proliferation. Proc Nat Acad Sci USA 1996; 93: 12525–
the treatment of alopecia X in Alaskan malamutes. J Am Anim 12530.
Hosp Assoc 2005; 41: 336–342.

ª 2012 The Authors. Veterinary Dermatology


220 ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.
Histology of canine noninflammatory alopecia

27. Van Beek N, Bodo E, Kromminga A et al. Thyroid hormones 30. Paus R, Handjiski B, Czarnetzki BM et al. A murine model for
directly alter human hair follicle functions: anagen prolongation inducing and manipulating hair follicle regression (catagen):
and stimulation of both hair matrix keratinocyte proliferation and effects of dexamethasone and cyclosporin A. J Investig Derma-
hair pigmentation. J Clin Endocrinol Metab 2008; 93: 4381–4388. tol 1994; 103: 143–147.
28. Bodo E, Kromminga A, Biro T et al. Human female hair follicles 31. Ito N, Ito T, Kromminga A et al. Human hair follicles display a
are a direct, nonclassical target for thyroid-stimulating hormone. functional equivalent of the hypothalamic-pituitary-adrenal
J Investig Dermatol 2009; 129: 1126–1139. (HPA) axis and synthesize cortisol. FASEB J 2005; 19: 1332–
29. Miller MA, Dunstan RW. Seasonal flank alopecia in boxers and 1334.
Airedale terriers: 24 cases (1985–1992). J Am Vet Med Assoc
1993; 203: 1567–1572.

Résumé
Contexte – L’alopécie non-inflammatoire chez le chien est un problème fréquent dont la pathogénie reste
obscure.
Objectif – L’objectif de cette étude est de réaliser une description histopathologique comparative de biop-
sies cutanées de chiens atteints de différentes alopécies et de chiens contrôles associés en fonction du
type de leur pelage, de la saison et de la durée de la maladie.
Animaux – Vingt et un cas d’alopécie X chez des chiens à pelage pelucheux, 12 cas d’alopécie récidivante
des flancs, 3 cas d’hyperoestrogénisme, 15 cas d’hyperadrénocorticisme, 12 cas d’hypothyroı̈die et 12 cas
d’alopécies primaires de cause inconnue ont été évalués. Les contrôles étaient des biopsies de 38 chiens
de différents types de pelage.
Méthodes – Nous avons évalués cinq séries de chaque biopsie d’un point de vue histopathologique et
immunohistologique afin de comparer les résultats histologiques entre les groupes de maladie et les con-
trôles.
Résultats – Tous les chiens atteints de troubles du cycle pilaire avaient une augmentation significative du
nombre de follicule pileux sans poils, que nous avons identifié comme kénogène. De plus, les chiens
atteints d’alopécie X avaient un pourcentage plus faible de follicules anagènes et le pourcentage le plus
élevé de follicules en phase télogène.
Conclusions – L’augmentation marquée des follicules kénogènes constitue une forte indication que
l’induction de la nouvelle phase anagène est altérée dans les troubles du cycle pilaire. Les résultats obte-
nus chez les chiens atteints d’alopécie X suggèrent en outre que la phase catagène prématurée est égale-
ment impliquée dans la pathogénie. Un approfondissement des études sur le compartiment des cellules
souches et les possibles facteurs responsables pour les différentes phases du cycle est nécessaire afin
d’en élucider la pathogénie exacte.

Resumen
Introducción – la alopecia no-inflamatoria es un problema frecuente en perros y la patogenia sigue siendo
confusa.
Objetivo – El objetivo de este estudio era una descripción histológica comparativa de las biopsias de piel
de perros con diversos desórdenes alopécicos y perros control apareados por capa similar, por muestras
en la misma estación y por duración de la enfermedad.
Animales – Se evaluaron veintiún casos de alopecia X en perros con capa de pelo densa, 12 casos de alo-
pecia recurrente del flanco, 3 casos de hiperestrogenismo, 15 casos de hiperadrenocorticismo, 12 casos
de hipotiroidismo, y 12 casos de desórdenes alopécicos primarios de causa desconocida. Los controles
fueron biopsias tomadas de 38 perros con diversos tipos de capa.
Métodos – Evaluamos cinco secciones seriadas de cada biopsia por histologı́a e inmunohistologı́a para
comparar los resultados histológicos dentro de los grupos de enfermedad y control.
Resultados – Todos los perros con desórdenes del ciclo del pelo tenı́an un aumento significativo en el
número de folı́culos pilosos sin pelo, que asignamos a la fase quenógena. Además, los perros con la alope-
cia X tenı́an el porcentaje más bajo de folı́culos pilosos en fase anágena y el porcentaje más alto de los folı́-
culos en fase telógena.
Conclusiones – El aumento marcado en folı́culos en fase quenógena es indicativo de que la inducción de
la nueva fase anágena en el pelo está deteriorada en desórdenes del ciclo folicular. Los resultados en per-
ros con la alopecia X sugieren además que una fase catágena prematura está implicada en la patogénesis.
Para aclarar la patogenia de estos desordenes se necesitan nuevos estudios que investiguen el comparti-
mento de células madre y los factores que inician las diversas fases del ciclo folicular.

Zusammenfassung
Hintergrund – Eine nicht entzündliche Alopezie ist ein häufiges Problem bei Hunden, wobei die Pathoge-
nese noch immer unklar ist.
Ziele – Das Ziel dieser Studie war eine vergleichende histologische Beschreibung von Hautbiopsien von
Hunden mit unterschiedlichen alopezischen Erkrankungen und Kontrollhunden, die bezüglich Felltyp,
Jahreszeit und Dauer der Erkrankung mit den Patienten abgestimmt waren.
Tiere – Einundzwanzig Fälle von Alopezia X bei Hunden mit einem Plüschfell, 12 Fälle von wiederkehrender
Flankenalopezie, 3 Fälle von Hyperöstrogenismus, 15 Fälle von Hyperadrenocortizismus, 12 Fälle einer

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ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44. 221
Müntener et al.

Hypothyreose und 12 Fälle einer primären alopezischen Erkrankung unbekannter Ursache wurden eva-
luiert. Die Kontrollproben kamen von 38 Hunden mit unterschiedlichen Haarkleidtypen.
Methoden – Wir untersuchten fünf aufeinander folgende Schnitte einer jeden Biopsie histologisch und
immunhistologisch, um die histologischen Befunde innerhalb der Gruppen mit unterschiedlichem Kran-
kheitsbild und den Kontrollen zu vergleichen.
Ergebnisse – Alle Hunde mit Erkrankungen des Wachstumszyklus der Haare zeigten eine signifikante
Zunahme der Anzahl der haarlosen Haarfollikel, was wir als kenogen bezeichneten. Zusätzlich hatten
Hunde mit Alopezia X den niedrigsten Prozentsatz an anagenen Haarfollikeln und den höchsten Prozent-
satz an telogenen Follikeln.
Schlussfolgerung – Die deutliche Zunahme an kenogenen Follikeln ist ein deutlicher Hinweis darauf, dass
die Induktion der neuen anagenen Phase bei Fehlsteuerungen des Haarzyklus beeinträchtigt ist. Die Ergeb-
nisse bei Hunden mit Alopezia X weisen weiters darauf hin, dass eine frühzeitige Katagen-Phase in der
Pathogenese ebenfalls eine Rolle spielt. Weitere Studien sollten Stammzellen und mögliche Auslösefakto-
ren für die verschiedenen Zyklusphasen untersuchen, um die genaue Pathogenese aufzuklären.

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e44 ª 2012 ESVD and ACVD, Veterinary Dermatology, 23, 206–e44.

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