Original Article

Toxicologic Pathology
2016, Vol. 44(5) 636-654

Normal Anatomy, Histology, and ª The Author(s) 2016

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Spontaneous Pathology of the Nasal DOI: 10.1177/0192623315626523
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Cavity of the Cynomolgus Monkey
(Macaca fascicularis)

Ronnie Chamanza1,2, Ian Taylor3, Michela Gregori1, Colin Hill3,

Mark Swan3, Joel Goodchild3, Kane Goodchild3, Jane Schofield3,
Mark Aldous3, and Vasanthi Mowat1

Abstract
The evaluation of inhalation studies in monkeys is often hampered by the scarcity of published information on the relevant nasal
anatomy and pathology. We examined nasal cavities of 114 control cynomolgus monkeys from 11 inhalation studies evaluated 2008
to 2013, in order to characterize and document the anatomic features and spontaneous pathology. Compared to other laboratory
animals, the cynomolgus monkey has a relatively simple nose with 2 unbranched, dorsoventrally stacked turbinates, large maxillary
sinuses, and a nasal septum that continues into the nasopharynx. The vomeronasal organ is absent, but nasopalatine ducts are
present. Microscopically, the nasal epithelium is thicker than that in rodents, and the respiratory (RE) and transitional epithelium
(TE) rest on a thick basal lamina. Generally, squamous epithelia and TE line the vestibule, RE, the main chamber and nasopharynx,
olfactory epithelium, a small caudodorsal region, while TE is observed intermittently along the passages. Relatively high incidences
of spontaneous pathology findings, some resembling induced lesions, were observed and included inflammation, luminal exudate,
scabs, squamous and respiratory metaplasia or hyperplasia, mucous cell hyperplasia/metaplasia, and olfactory degeneration. Regions
of epithelial transition were the most affected. This information is considered helpful in the histopathology evaluation and inter-
pretation of inhalation studies in monkeys.

Keywords
nasal anatomy, cynomolgus monkey, Macaca fascicularis, nonhuman primate, inhalation, spontaneous pathology, toxicity studies

Introduction pathology findings is of particular importance when evaluating

Inhalation studies are designed to assess drug or chemical-
induced respiratory effects in animals in order to predict the
potential risk to human health. For biological pharmaceuticals
that are to be given via the inhalation route, nonhuman primates
may be the only pharmacologically relevant species due to their
close phylogenic relationship to man. For this reason, inhala-
tion studies are often performed in monkeys, with a detailed
evaluation of the nasal cavity. However, the evaluation of the
monkey nose is often hampered by the scarcity of published
information on the gross anatomy, normal histology, and back-
ground pathology of the nasal cavities in this species (Renne et
al. 2007). A working knowledge of the comparative anatomy
and histology of the nasal cavity of the laboratory test species is
required for a proper evaluation of toxic effects in the nasal
cavity and the extrapolation of the data to human risk assess-
ment (Chamanza et al. 2015; Harkema, Carey, and Wagner
2006). In addition, information on the range and incidences
of the more commonly occurring background or incidental
inhalation studies, since some of them may be confused with
induced lesions.
For the cynomolgus monkey (Macaca fascicularis), which
is the most widely used purpose-bred nonhuman primate in
preclinical safety testing, no published information on these
parameters is readily available. This study was therefore
designed to investigate, describe, and document the character-
istic gross anatomical features, morphology and distribution of
the epithelium, and the nature and incidence of spontaneous
1
Huntingdon Life Sciences, Huntingdon, Cambridgeshire, United Kingdom
2
Syngenta Limited, Jeallot’s Hill International Research Centre, Bracknell,
Berkshire, United Kingdom
3
Huntingdon Life Sciences, Eye Research Centre, Eye, Suffolk, United Kingdom
Corresponding Author:
Ronnie Chamanza, Syngenta Limited, Jeallot’s Hill International Research
Centre, Bracknell, Berkshire, RG42 6EY, United Kingdom.
Email: chamanza@aol.com; ronnie.chamanza@syngenta.com
Chamanza et al.
pathological findings in the nasal cavity of young control cyno-
molgus monkeys in order to produce readily available refer-
ence material.
Materials and Method
The Animals
Nasal cavities were obtained from 114 control cynomolgus
monkeys (57 animals per sex) from 11 inhalation toxicity stud-
ies of 4- to 13-week duration, conducted between 2008 and
2013. All animals used in these studies were of Vietnamese
origin, purpose bred for laboratory use, and obtained from
accredited suppliers. They were all juvenile monkeys between
27 and 42 months of age (with a full set of deciduous teeth and
1 or 2 permanent erupted molars or incisors) at the start of the
studies and weighed between 2.5 and 3.5 kg.
Study animals were housed in groups of 1 or 3 animals of
the same sex and dose group in custom-designed U.K. Home
Office–compliant primate cages (chapter 14, section 21, U.K.
Animals [Scientific Procedures] Act of 1986). The temperature
and humidity were automatically controlled at 21 C + 4 C and
55% + 10%, respectively, with a minimum of 15 air changes
per hour. An automatic light cycle of 7:00 a.m. to 7:00 p.m.
(20-hr cycle) was maintained. Each individual gang pen had
drinking water and food hoppers, and animals were fed a com-
mercial primate diet (Mazuri diet, Special Diet Service Ltd,
Witham, Essex, England). Twice weekly fruit supplements
were also offered to all animals. Animals received an appro-
priate veterinary examination, which included serological tests
to confirm that they were free of routinely tested infectious
agents. They were confirmed free of respiratory infections and
the following viral diseases during quarantine: simian immu-
nodeficiency virus, Cercopithecine herpesvirus 1 (B virus),
simian retroviruses type D, rabies, simian T-cell leukemia
virus, measles, and filoviruses. Tuberculin tests; other bacter-
iology screening tests for Shigella, Yersinia, and Salmonella;
and parasitological screening (including malaria smears) were
carried out on arrival at the test facility.
All studies were conducted in accordance with the U.K.
Animals (Scientific Procedures) Act 1986, which conforms to
the European Convention for the Protection of Vertebrate Ani-
mals Used for Experimental and Other Scientific Purposes
(Strasbourg, Council of Europe).
Pathology Procedures
A detailed necropsy examination was performed on all animals
following euthanasia by intravenous injection with sodium
pentobarbitone and exsanguination. Soon after euthanasia, the
heads of the monkeys were removed from the body and the
nasal passages flushed with 10% neutral buffered formalin
(NBF) retrograde through the pharynx. After flushing the nasal
cavities, the brain, mandible, eyes, integument, and muscula-
ture were removed from the head, which was then immersed in
a large volume of NBF.
637
Nasal Cavity Trimming of Routine Sections
After removing the lower jaw and the soft tissues, and prior to
decalcification, the remaining upper part of the head was fixed
to a clamp and serially trimmed transversely, from the vesti-
bule to the nasopharynx, using a water-cooled diamond-bladed
band saw. A total of 5 to 6 coronal (transverse) sections were
cut in a rostro-caudal direction, using dental and palatal land-
marks (Figure 1A), to produce a minimum of 4 tissue slices
(levels 1–4). Each slice was then placed with the rostral face
down in a cassette.
Cut 1. Rostral vestibule (not examined on most studies)—
Immediately caudal to the second incisor or at the level
of the canine or the first palatal ridge.
Cut 2. Level 1—Immediately rostral to the first premolar
(or the first deciduous molar) and between the first and
second palatal ridges.
Cut 3. Discarded—Immediately rostral to the second pre-
molar (or the second deciduous molar) and between
the 3rd and 4th palatal ridges.
Cut 4. Level 2—Immediately rostral to the first (perma-
nent) molar and between the 5th and 6th palatal ridges.
Cut 5. Level 3—Immediately caudal to the first (perma-
nent) molar.
Cut 6. Level 4—Caudal to the second (permanent) molar
or the mesiodistal diameter of a molar.
Nasopharynx—Caudal to the last molar and through the
soft palate.
The trimmed coronal nasal sections were decalcified using
the Ultrasonic Decalcifying Automate (Medite1 Medizintech-
nik GmbH, Burgdorf—Germany) and the Medite USE 33 dec-
alcifying method before embedding. After decalcification,
tissues were embedded in paraffin wax, sectioned to a 3- to
4-mm thickness, and stained with hematoxylin and eosin. They
were examined histopathologically and the findings entered
directly into a computerized database (Pristima Path/Tox,
Xybion Medical Systems Corporation, Morris Plains, NJ).
Study materials including histological incidence tables,
individual animal data listings, and glass slides were retrieved
from the archives and analyzed for the normal histology and
spontaneous pathology of the nasal cavity. In addition, a his-
torical control data search was performed for pathology find-
ings recorded under the nasal cavity. Data were available from
114 control animals that had been sham dosed with air, saline,
or lactose vehicle via the nose-only method of inhalation
exposure.
Nasal Cavity Trimming of Longitudinal Sections
Further investigations of the gross anatomy of the nasal cavity
and its associated structures were carried out on 4 heads
obtained from control animals on noninhalation studies, from
638 Toxicologic Pathology 44(5)

Figure 1. Gross sections of the nasal cavity (NC) of young cynomolgus monkeys (Macaca fascicularis) of 29 to 33 months of age and the landmarks
used for nasal trimming. (A) Hard palate from a young female cynomolgus monkey, showing dental structures and palatal ridges used as landmarks
for the trimming of coronal sections of the NC. The incisive papilla where nasopalatine ducts (NPDs) exit is shown (arrow). (B) and (C)
Parasagittal sections of the NC of young female cynomolgus monkeys showing the vestibule (V), vestibular wings (arrow and arrowhead),
maxilloturbinates (MTs), ethmoturbinates (EMTs), NPDs, nasolacrimal ducts (NLDs), the nasopharynx (NP), the ventral meatus (VM), and the
nasopharyngeal septum(NPS). (D)–(F) Coronal (transverse) sections of the NC of a juvenile male cynomolgus monkey (note that some teeth are
Chamanza et al.
which at least 3 mid- or parasagittal sections were obtained
(Figure 2B and C). The sections were obtained by cutting long-
itudinally (along the long axis of the head) through the right
nasal fossae from the nares to the pharynx and exposing the
lateral meatus and turbinates.
Results
Gross and Subgross Anatomic Features on Parasagittal
Sections (Figure 1B and C)
The gross examination of the cynomolgus monkey nose shows
considerable variability in the size and shape of the main nasal
structures. The main interanimal differences are mainly in the
shape of the turbinates and the overall length of the nasal
cavity, and they do not appear to be sex- or age dependent
(animals were of a relatively similar age). However, despite
this, the general layout of nasal features is relatively uniform
between individual animals. Parasagittal sections reveal that,
from rostral to caudal, the nasal cavity of the cynomolgus
monkey can be roughly divided into a vestibule, a main nasal
chamber with 2 dorsoventrally stacked turbinates, and a
nasopharynx that is divided by a membranous median septum
(Figure 1B and C). The vestibule is relatively large and empty
(in comparison to that of rodents and dogs), and there are no
well-developed atrioturbinates or alar folds, although a dorso-
lateral vestibular (alar) wing that forms an atrioturbinates-like
projection is present (Figure 1B, C, and D). The vestibule is
elevated above the floor of the main nasal chamber due to the
prominence of the separate premaxilla. This allows the ventral
floor of the main nasal cavity to continue forward (rostroven-
tral) below the floor of the vestibule as the incisive canal or
nasopalatine duct (NPD; Figure 1B and D). The NPDs (or
Stenson’s ducts) open into the oral cavity at the incisive papil-
lae (Figure 1A).
In the nasal chamber proper, the dorsally located ethmotur-
binates (EMTs) can be easily identified by their bright yellow
color, which is often retained even after prolonged fixation
(Figure 1B and C). They appear as long, horizontal finger-
like projections that are attached only at the dorsocaudal end
of the nasal cavity (Figure 1B and C). The rostral ends are free
on 3 surfaces, for as much as a rostral 3rd of the length of the
turbinates. Interanimal variations in the shape of the ventral
edge of the EMTs may be appreciated on parasagittal sections,
but they are more conspicuous on coronal sections (Figure 2A–
F). The variations in the shape of the EMTs represent the
greatest interindividual differences in the gross anatomy of the
cynomolgus monkey nose and do not seem to be age or sex
related. Below the EMTs are the ventrally located pair of max-
illoturbinates (MTs), which span about two-thirds of the nasal
cavity caudorostrally. The MTs (which correspond to the
Figure 1. (continued) yet to erupt). (D) Rostral V showing NPDs (arrowheads) and how the lateral nasal cartilages or vestibular wings (arrows)
create a crude dorsolateral meatus. (E) Rostral NC at level 2 showing MTs, unattached EMTs, nasal septum (S)(SP), and the dorsolateral ridge
(arrowheads). (F). Section of the NC at level 3, showing MTs, EMTs, NLD, SP, and maxillary sinuses (MS). The NLDs lie lateral to the EMTs at this
level and the septal cartilage is reduced to small piece in the middle of the septum.
639
human inferior turbinates) are the rostral-most turbinates by
only a very small margin in monkeys and widen to blend with
the lateral walls both caudally and ventrally (Figure 1B and C).
The shape of their ventral surface also shows some slight var-
iation on longitudinal sections, but as with EMTs, the intera-
nimal variations are more apparent on coronal sections. There
are no analogous structures to the superior nasal turbinates of
humans and nasoturbinates are absent, although a small bulge
or nasal ridge formed by a small separate bone is often
observed on the dorsolateral wall, at the level of the nasal
atrium (Figure 3B). In some animals, a poorly defined vestigial
structure is occasionally observed attached to the dorsal surface
of the EMT.
Gross and Subgross Anatomic Features on Coronal
Sections
The main anatomic features on the monkey nasal sections pro-
duced by the method described above (Figure 1A) are summar-
ized in Table 1. Minor variations between sections taken at the
same level were relatively common and were attributed to a
combination of interindividual variability in monkeys and tan-
gential sectioning. However, in general, all coronal sections
(Figure 1D–F) reveal a relatively simple nose that is divided
into 2 separate and symmetrical passages by a median nasal
septum, which is mostly cartilaginous rostrally and bony caud-
ally. The nasal septum divides the nasal cavity from the naris to
the choana and past the caudal edge of the vomer into the
nasopharynx, where it continues thereafter as a membranous
median septum that divides the nasopharynx into 2 equal
halves, for as much as two-thirds of its length. A more detailed
description of the toxicologically relevant gross and subgross
anatomic features on the nasal sections from rostral to caudal is
given subsequently.
The coronal section of the vestibule rostral to level 1 reveals
how the dorsal and lateral cartilaginous projections of the ves-
tibular wing create a crude dorsolateral meatus of varying sizes
(Figure 1C). Caudally, level 1 shows the caudal vestibule or the
entrance to the main nasal chamber, which is often wide and
empty and devoid of any projections. A nasal ostium as found
in humans is not present at the entrance of the main chamber,
but a slight narrowing of the nasal passage is achieved rostrally
by a bulging of the septum in the middorsal 3rd (Figures 1E and
3A). This midseptal bulge or tubercle is often present from the
vestibule to a level just rostral to the tip of the MTs and is
limited to the mucosa, without involvement of the cartilage.
Level 2, 3, and 4 sections expose the main nasal chamber
and the nasal turbinates. Dorsally at level 2, which is taken
slightly caudal to the septal bulge but before the attachment
of the EMTs, the dorsolateral wall exhibits a small ridge which
is a frequent site of spontaneous pathological changes (Figures
640 Toxicologic Pathology 44(5)

Figure 2. Subgross coronal sections of the ventral nasal meatus of young cynomolgus monkeys (Macaca fascicularis) of 36 to 40 months of age.
(A)–(C) Interanimal variations in the size and shape of ethmoturbinates (EMTs) and maxilloturbinates (MTs) in young female cynomolgus monkeys
at level 2 of the nasal cavity (NC), which shows the exit of the nasolacrimal ducts (NLD; arrow) in the ventral meatus (VM). Note how at this level
the SP is mostly composed of cartilage. Foci of cartilage (arrowhead) and large venous sinuses present in the MTs are also shown. (D) and (E)
Interanimal variations in the ventral extension of EMTs in relation to MTs in young female monkeys at level 3 of the NC, which shows the
maximum diameter of the maxillary sinus (MS). Note that at this level, the MTs arise from a separate secondary bone that divides the nose
into the nasal meatus and the MS. (F) Caudal VM of a young male cynomolgus monkey at level 4, showing that the MS and the NC are
separated only by a mucous membrane (arrow) at this level. All images, hematoxylin and eosin, 1 original objective. SP ¼ nasal septum,
NALT ¼ nasal-associated lymphoid tissue.
Chamanza et al.
Figure 2. (continued).
3B and 4H). This dorsolateral ridge continues caudally and
may be present on some level 3 sections. Ventrally, there are
no vomeronasal organs (VNO) or vestiges of them at the floor
of the septum, and only 1 paranasal sinus, the maxillary sinus
(MS), is present within the maxilla (Figures 1F and 2D–F) at
levels 3 and 4. The relatively large (compared to dogs and
rodents) MS has a roughly spherical to polygonal shape, and
its floor lies above the roots of the permanent molar teeth while
the roof occupies most of the orbital floor (Figure 1F).
The 2 nasal turbinates present in the monkey can be viewed
on all coronary sections passing through the main nasal cham-
ber (levels 2, 3, and 4). They appear as 2 pairs of simple and
unbranched structures that project ventromedially and dorsally
into the meatus from the maxilla and ethmoid bone, respec-
tively (Figures 2 and 3), leaving very little space for free air
passage between them and the nasal septum, particularly in the
dorsal meatus. Although the MTs appear to arise from the
maxilla, subgross histological sections at level 3 reveal that
they do not actually arise from the maxilla itself but from a
separate, secondary membrane bone that divides the nose into 2
compartments, the nasal meatus and the MS (Figures 1F and
2D). Caudally at level 4, this bony partition is largely absent on
the lateral wall of the nasal cavity, so that the MS is separated
from the nasal cavity only by a mucous membrane, and the MT
has no central bony core (Figures 2F and 3D). Slight interanimal
variation may be observed in the shape of the ventral surfaces
of the MTs, but the fist-shaped or slightly curved presentation
is by far the most common presentation (Figure 2A–F).
Although the EMTs are also present on all sections cutting
through the main nasal chamber (levels 2, 3, and 4), at level 2,
only the unattached rostral ends, which often float free within
the nasal meatus as triangular- or rectangular-shaped structures
(Figures 1E and 3B), are present. The interanimal variations in
the shape of the ventral surfaces of the EMTs can be appre-
ciated at this level and at level 3 (Figure 2A–C), and these
641
generally show that the long and pointed tip (Figures 1F and
2C–E) is the most common presentation, while the blunt or
rounded surface (Figure 2A), or midway in between the two
(Figure 2B), are not uncommon.
The largest profile of the EMTs is present on level 3 sec-
tions. This level shows that the medial and lateral surfaces of
the EMTs have more constant presentations that roughly follow
the contours of the septum and the lateral meatus, respectively
(Figures 1F, 2B and C, and 3D and E). Therefore, the medial
surface of the EMTs consistently appears straight, mirroring the
perpendicular shape of the nasal septum, while the lateral surface
has a curved appearance that conforms to the curved contours of
the lateral nasal wall (Figures 2B and C and 3E), with very little
interanimal variation. Besides the variations in the shape, the
dorsoventral relationship of the 2 turbinates also differs slightly
among individuals, with some EMTs extending further ventrally
to overlap or reach the same ventral limits as the MTs (Figure 2E).
Level 4 sections reveal that the monkey’s nasopharynx lies
caudal and not ventral to EMTs; therefore, a transverse lamina
that divides the EMTs from the nasopharyngeal passageway in
other laboratory animals is absent (Figure 3F and G). Near this
caudal edge of the nasal cavity, septal deviations may be
observed at the bone–cartilage interface, where the septal carti-
lage is reduced to its smallest size in the middle of the septum
(Figure 3E and F). They are differentiated from artifactual dam-
age by the presence of some tissue reaction, including fibrosis
and hemorrhage on histology (Figure 3E). A septal window as
seen in rodents is not present at this or any other level.
The main gross anatomic differences between the nasal cav-
ities of the cynomolgus monkey and humans can therefore be
summarized as the presence of just 1 paranasal sinus (no frontal,
sphenoid, or ethmoid sinuses), 2 instead of 3 turbinates, a
slightly elevated vestibule, the presence of NPDs with no ves-
tiges of the VNO, and the presence of a median septum that
continues to about two-thirds of the nasopharynx, in monkeys.
642 Toxicologic Pathology 44(5)

Figure 3. Subgross coronal sections of the dorsal nasal meatus and the nasopharynx (NP) from young cynomolgus monkeys (Macaca fascicularis)
of 33–42 months of age. (A) Caudal nasal vestibule of a young female cynomolgus monkeys at level 1 showing the septal bulge and the distribution
of squamous (SE), transitional (TE), and respiratory epithelium (RE) in the lateral and septal (SP) walls. Inflammation is present in the lateral wall at
the SE and TE junctions (arrowhead). (B) Dorsolateral nasal ridge at level 2 of the nasal cavity (NC) of a young female cynomolgus monkey. The
ridge is formed by a small separate bone (arrowhead) and is a region of frequent inflammatory lesions. Dorsal to the ridge at this level is a region of
a poorly defined epithelium that resembles both respiratory and olfactory epithelia (OE; arrow). The unattached ethmoturbinates (EMTs) are
lined by RE. (C) and (D) Caudal NC of young male cynomolgus monkeys at level 4, showing the dorsal and ventral limits of the OE (arrowheads)
on the EMTs and septum (SP) and the location of the nasolacrimal ducts (NLD) dorsal to the NC at this level. Ventrally at this level, the maxillary
sinus (MS) and the NC are separated only by a mucous membrane while the maxilloturbinates (MTs) are not anchored by a bony core. (E) EMTs in
a young female cynomolgus monkey at level 3, showing regions of frequent background changes (arrowheads) and that the medial and lateral
surfaces roughly follow the contours of the adjacent nasal walls. Septal deviation with soft tissue reaction and swelling at the ends of the septal
cartilage (SPC) are present (arrow). (F) Level 4 of the NC of a young male cynomolgus monkey showing the caudal edge of the nasal meatus and
Chamanza et al. 643

Figure 3. (continued) the beginning of the NP, where the SPC is reduced to its smallest size and is a frequent site for septal deviations. (G) The
NP of a young female cynomolgus monkey showing septal tonsillar lymphoid tissue. (H) Level 2 of the NC of a young female cynomolgus monkey
showing a rostral dorsal meatus lined by nonolfactory RE and some nasal-associated lymphoid tissue. The distal tips of EMT at this level are a
frequent site of spontaneous pathology and are lined by TE and RE. All images hematoxylin and eosin, 1 original objective.
644 Toxicologic Pathology 44(5)

Table 1. Main Anatomical Features and Epithelial Types Present on Various Coronary Sectional Levels of the Nasal Cavity of Young Cynomolgus
Monkeys (Macaca fascicularis).

Level Main Anatomic Features Epithelial Types

Rostral vestibule V-shaped meatuses Mostly SE

Dorsal and lateral alar cartilages present RE or TE may be observed ventrally on tangential sections
Nasopalatine ducts may be present ventrally
below the floor of the vestibule
Mild midseptal bulge
Septum completely cartilaginous
Level 1 Caudal vestibule, wide, empty, and smooth walled SE mainly in the dorsal regions
Thick midseptal bulge TE mainly in the mid to ventral regions
Septum completely cartilaginous RE mainly in the ventral regions
Level 2 Unattached ethmoturbinates present dorsally Mostly RE throughout
Dorsolateral ridge present Small areas of TE may be present on angular surfaces of
Maxilloturbinates attached to maxilla ethmoturbinates
Septum completely or largely cartilaginous On more caudal sections (characterized by a partly bony septum),
(middle eight-tenths) dorsal region may be lined by epithelium between RE and OE
Profile or exit of nasolacrimal duct present
ventrally
Level 3 Ethmoturbinates attached dorsally, shape conforms OE present dorsally
to contours of nasal walls RE present ventrally
Maxillary sinus present A poorly defined epithelium resembling both OE and RE intervenes
Maxilloturbinates attached to separate bone in between
dividing nasal cavity from maxillary sinus Small areas of TE may be present on ventral and angular surfaces
Nasolacrimal duct present lateral to of ethmoturbinates
ethmoturbinates
Dorsolateral ridge may still be present
Septal cartilage reduced to small area in the middle,
septal deviations may be present
Level 4 Ethmoturbinates attached dorsally, show straight and OE present dorsally and lines largest surface area
narrower profile RE present ventrally
Maxillary sinus present Small areas of TE may be present on ventral and angular surfaces
Maxilloturbinates attached to membranous of ethmoturbinates
partition between nasal cavity and maxillary sinus
Nasolacrimal duct present dorsally and prominent
Septal cartilage reduced to very small area in the
middle, deviations may be present

Note. OE ¼ olfactory epithelium; RE ¼ respiratory epithelium; SE ¼ squamous epithelium; TE ¼ transitional epithelium.

Histology The SE covering the monkey vestibule is a nonkeratinized-

or lightly keratinized-stratified epithelium composed of basal
The general distribution of epithelial types at each level of the
cells lying on a barely visible basal lamina and 5 to 8 layers of
monkey nose is summarized in Table 1. The epithelial distri-
more flattened cells toward the surface. In general, there is a
bution at each level can also be influenced by the interanimal
rostral to caudal decrease in the thickness of the SE, so that the
variations and tangential sectioning. However, as in other
thinnest SE, with keratohyaline granules and pigmented kera-
laboratory animals, 4 main types of epithelial populations,
tinocytes, is observed in the dorsal regions, where it extends
squamous (SE), transitional (TE), respiratory (RE), and olfac-
more caudal than in the ventral regions. The thickest SE is
tory epithelium (OE), line the vestibule, nasal chamber, and
present in the most rostral parts and on the midseptal or mid-
turbinates in the cynomolgus monkey.
lateral regions of the vestibule where it abruptly transitions to
The rostral vestibule (which is not always examined) is
TE. This abrupt transition from SE to TE also marks the sudden
completely lined by a stratified SE, but further inward at level
appearance of a slightly thicker basal lamina lying beneath the
1, the SE often becomes restricted to the dorsal and dorsolat-
TE (Figure 4B and C).
eral regions (Figure 3A). The relatively thin SE present in the
The nasal TE of the cynomolgus monkey is a stratified,
dorsal regions at this level gives way to a TE mainly in the
nonciliated cuboidal to low columnar epithelium of 4 to 6 cell
midseptal, midlateral, and ventrolateral regions (Figure 3A).
thickness (Figure 4A–C), which is mostly devoid of goblet
However, in some individuals, the ventral parts of the vestibule
cells. It is intermittently present over a relatively wide long-
at level 1 may also be lined by a SE that is often thicker than that
itudinal (rostrocaudal) area that extends from as far forward as
lining the dorsal regions, possibly due to oblique sectioning.
the vestibule at level 1 to as far back as the middle or rostral 3rd
Chamanza et al. 645

Figure 4. Normal histological appearance of the nasal epithelium in young cynomolgus monkeys (Macaca fascicularis) of 36 to 42 months of age.
(A) Transitional (TE) and respiratory epithelium (RE) on the distal ethmoturbinates at level 2 of a male monkey, showing increased goblet cell
density with pseudoglands or intraepithelial crypts formation (arrowheads). Hematoxylin and eosin, 4 original objective. (B) TE on the septal
bulge at level 1, in a male monkey. Hematoxylin and eosin, 20 original objective. (C) Gradual transition from TE to RE in the rostral lateral meatus
at level 2 of the nasal cavity (NC) of a female monkey. There is increased goblet cell density and thickness of the basal lamina (arrow and
arrowhead) in the RE. Hematoxylin and eosin, 10 original objective. (C, insert) Gradual transition from TE to RE in the rostral lateral meatus at
level 2 of the NC of a female monkey. Higher magnification, hematoxylin and eosin, 20 original objective. (D) The ventromedial surface of the
maxilloturbinate of a control female monkey at level 3 of the NC showing a tall, densely ciliated, RE with the highest density of goblet cells in the
nose. Submucosal NALT and seromucous glands (SSG), obscured the cartilage and arrowhead are also present. Hematoxylin and eosin, 4
original objective. (D, insert) A higher magnification of 4-D, showing a tall, densely ciliated, RE lying on the thickest basal lamina (arrowheads) of the
whole NC. Hematoxylin and eosin, 20 original objective. (E) and (F) Caudal lateral meatus of a young control male monkey at level 3 showing RE
with a relatively high goblet cell density and a thick basal lamina (arrowheads). Hematoxylin and eosin, 20 original objective. (G) Ventral maxillary
sinus (MS) of a young control male monkey at level 3, showing an increase in goblet cells and the accumulation of intraluminal mucus. Hematoxylin
and eosin, 4 original objective. (G, insert) The lateral NC and the adjacent medial surface of the MS of a young control female monkey at level 3,
showing the huge difference in the morphology of their respective RE, and the corpora amylacea (arrowhead) in the mucosa of the MS. Besides its
characteristic low profile, the RE lining the MS also shows no thick or visible basal lamina. Hematoxylin and eosin, 20 original objective. (H) Septal
olfactory epithelium at level 4 of a young control male monkey, showing nuclei of cell types arranged in a roughly lamina pattern from the apical
surface to the basal lamina, the sustentacular cell (arrowhead), mature and immature olfactory sensory neurons, and globose and horizontal basal
cells (thick arrows). Bowman’s glands and Bowman’s glands/duct cells are also shown. Hematoxylin and eosin, 20 original objective.

of the nasal cavity at levels 2 and 3. The most commonly less frequently, the ventral vestibule. Septal TE is most com-
affected areas are the septum and lateral meatus of the vestibule monly observed in the middle regions, along the septal bulge at
(Figure 4B and C), the angular surfaces and rostral ends of level 1, where it is usually at its thickest (Figure 4B), while
turbinates (Figure 4A), much of the rostral nasal septum, and turbinate TE is most commonly observed on the 2 or more
646
Figure 4. (continued).
angular surfaces of the unattached turbinates at level 2 (Figures
3B and 4A) or on smaller areas of the ventral and medial sur-
faces of the caudal EMTs (Figure 3E) at level 3. On the MTs, it is
often restricted to a small area at the rostral limits of the turbi-
nate, which is usually not present on any of the sections.
The distribution of TE on the EMTs suggests that in the
cynomolgus monkey, TE, or a TE-like nonciliated epithelium
with a low profile and scant goblet cells, may be encountered in
certain areas of the main nasal chamber away from the rostral
region. This TE-like epithelium may also be observed on the
dorsolateral ridge, dorsal to the nasal atrium (Figures 1E and 3B),
and just before the transition from RE to OE. Since these regions
are frequently associated with inflammatory changes in the
lamina propria (as regions of high particle impaction), it is often
difficult to determine whether the morphology of this epithelium
represents a normal appearance or a metaplastic change.
The nasal RE in the cynomolgus monkey is a tall, pseudos-
tratified ciliated columnar epithelium that lines the largest sur-
face area in the nasal cavity and is therefore present on all 4
levels (levels 1–4). It lies on a microscopically visible and
strikingly thick basal lamina (Figure 4C–F), which appears as
Toxicologic Pathology 44(5)
a thick amorphous eosinophilic structure that often obscures
cellular detail on tangential sections (Figures 4F and 5H). The
basal lamina of the RE is much thicker than that beneath the TE
and can therefore be used to demarcate the start of RE at the
transition from TE (Figure 4C) or the end of the RE at the RE/
OE transition zone. The transition from TE to RE is otherwise
gradual and almost imperceptible, although the scarcity of gob-
let (mucous) and ciliated cells may be used to identify the TE.
The RE is also on average taller than the TE.
The RE is composed of basal cells, ciliated columnar cells,
goblet cells, and nonciliated columnar cells. Goblet cells are
abundant in the RE lining both the nasal meatus and the MS. In
the MS, they are more concentrated on the ventral aspect where
mucus often collects (Figure 4G). The highest density of goblet
cells however is found on the medial wall of the MTs, which
also has the tallest and most densely ciliated RE, and the thick-
est basal lamina (Figure 4D). Goblet cells are also abundant in
the RE lining the caudal ventral meatus, the caudal nasal sep-
tum, and the nasopharynx. There is therefore a general increase
in the density of goblet cells from rostral to caudal, particularly
on the septum. Increased goblet cell density with pseudogland
Chamanza et al.
formation and mucous cell metaplasia of the submucosal
glands are a common feature on the MTs and the septum.
In terms of epithelial height, the RE lining the ventral and
the ventrolateral meatus (Figure 4G), and to some extent the
septum, is of a relatively lower height than that found on the
turbinates and the lateral and dorsal walls, while that covering
the MS shows the lowest profile (Figure 4G; less than half the
height of the RE on the MTs).
The monkey OE lines only a small part of the caudodorsal
meatus and the dorsal attachments of EMTs. Therefore, on
nasal sections produced by the method described in this article
(Figure 1A), the rostral dorsal meatus and the unattached EMTs
at level 2 are usually not lined by OE (Figure 3H), and the
neuroepithelium first appears on the level 3 section. However,
serial trimming of further coronal sections and examination of
parasagittal sections show that the most rostral limits of the OE
lie somewhere in between levels 2 and 3; and at this level, OE
lines just the dorsal meatus and approximately a dorsal 5th of
the septum. It first appears on the EMTs at their attachments to
the dorsal meatus and lines the medial surface first, covering
approximately nine-tenths of the dorsomedial surface, in addi-
tion to the dorsal half of the septum (Figure 3C and D). The
lateral surface of the EMTs (dorsal 3rd) and a small part of the
dorsolateral wall are lined by OE only in the caudal most
regions (Figure 3C) at level 4. Thus, at any given level, the
OE lines larger areas of the medial surfaces of the EMTs and
the nasal septum, than it does the lateral aspects of the EMTs
and the lateral nasal wall. There is therefore on level 3 or 4
sections (mainly level 3), a dorsolateral angle formed by the
lateral surface of the EMTs and the lateral wall that is never
fully lined by OE, but mainly by a poorly defined epithelium
that resembles both RE and OE, and is often a site of epithelial
inflammation or vacuolation (Figure 6D).
In general, the transition from RE to OE is not well defined,
and some clusters of ciliated RE cells may be found within the
OE, making it difficult to distinguish such normal features from
respiratory metaplasia of the OE (Figure 6E and F). Since the
basal lamina of the OE is not readily visible, the absence of a
thickened basal lamina and the presence of Bowman’s glands
and nerve bundles are helpful in identifying metaplastic OE.
The OE of the cynomolgus monkey is a very tall pseudos-
tratified columnar epithelium of up to twice the height of the
RE. It is composed of 5 cell types whose nuclei are lined in a
roughly lamina pattern in the following order: from the apical
surface to the basal lamina, the sustentacular cell, mature and
immature olfactory sensory neurons (OSNs), and globose and
horizontal basal cells, respectively (Figure 4H). In addition,
Bowman’s glands/ducts cells extend through the epithelium
from the lamina propria to the apical surface, constituting the
6th cell type. There are no goblet cells and the mucus covering
the luminal surface of the OE is produced and secreted by the
subepithelial Bowman’s glands.
Three main types of submucosal glands are present as in
other animals: serous glands, seromucous glands, and Bow-
man’s glands of the olfactory mucosa. However, the lateral
nasal (Steno’s) glands (of the dog and rodents) are not present.
647
Submucosal glands are more abundant in the rostral septum, lat-
eral walls, and the MTs. Rostral septal glands have ducts that
extended rostrally to open in the vestibule, while a few glands
are also present in the submucosa of the vestibule itself. Besides
the large number of submucosal glands, the submucosa of the
MTs also typically contains large and abundant venous sinuses
and foci of cartilage as normal features (Figures 2A and 4D). In
the olfactory mucosa, dark submucosal pigment is often observed
affecting the mucous glands, sustentacular cells, connective tis-
sue cells, nerve bundles, and the Bowman’s gland epithelium
(Figure 6H). This pigment is believed to be responsible for the
grossly visible yellow color of the olfactory mucosa or EMTs.
Most of the nasopharynx is lined by a low RE, but a TE is
present at the level of the exits of the Eustachian tubes. The
septal wall of the nasopharynx contains large amounts of none-
ncapsulated tonsillar lymphoid tissue (Figure 3G) analogous to
the Waldeyer’s ring in man. The tonsillar tissue and nasal
associated lymphoid tissue (NALT) are also often present in
the lateral walls of the nasopharynx. The NALT in the cyno-
molgus monkey is not confined to a particular region but can be
found anywhere throughout the nasal cavity, including the ven-
tral meatus (Figures 2D and 7D), dorsal meatus (Figure 3H),
MTs (Figure 2D), or the vestibule (Figure 7C). The NALT of
the vestibule often causes partial occlusion of the nasal pas-
sages in some animals (Figure 7C).
Background Pathology
The most common spontaneous pathology findings observed in
the nasal cavity of young cynomolgus monkeys are presented
in Tables 2 to 5. The findings were recorded on the basis of the
anatomic (e.g., ventrolateral meatus) and functional (epithelial
type) rather than the section level, in order to avoid repetition.
The most common findings were inflammatory lesions, which
showed a greater predilection for the rostral nasal cavity, the
dorsolateral ridge (Figure 3B), the ventral and medial surfaces
of the EMTs, and other regions of epithelial transition. They
were invariably associated with other degenerative, reparative,
and adaptive changes in the nasal epithelium, which were often
not recorded separately. These included squamous or transi-
tional cell hyperplasia (Figure 5A), RE attenuation and squa-
mous metaplasia (Figure 5D, E, and G), mucous cell
hyperplasia/metaplasia (Figure 5C), OE degeneration/atrophy
(Figure 6A–C), OE vacuolation (Figure 6D), RE hyperplasia
(Figure 5H), and olfactory epithelial hyperplasia (Figure 6E).
The inflammatory lesions reached high incidences of up to
100% of the animals in the studies where they occurred. How-
ever, the prevalence over the 11 studies (data not shown) was
not as high as the total incidences within individual studies.
The severity grades ranged from minimal to moderate.
Lesions of the SE
In a large number of the animals, the midlateral or midseptal
regions of the vestibule or at the transition from SE to TE at
level 1 were covered with scabs or focal inflammatory exudate
648 Toxicologic Pathology 44(5)

Figure 5. Spontaneous findings of the squamous (SE), transitional (TE), and respiratory epithelium (RE) in the NC of young cynomolgus monkeys
(Macaca fascicularis) of 27 to 42 months of age. (A) Rostral vestibule of a young control female monkey, showing inflammation and hyperplasia of
the SE with rete pegs formation. Hematoxylin and eosin, 20 original objective. (B) Vestibule of a young control male monkey at level 1, showing
inflammation and scabs on the lateral wall at the SE-TE epithelial junction. Hematoxylin and eosin, 10 original objective. (C) The rostral nasal
septum (SP) of a young control male monkey at level 1, showing mucous cell hyperplasia/metaplasia of the TE with pseudogland formation
(arrowhead and insert) and hyperplasia of the SE. Hematoxylin and eosin, 10 original objective (insert) and hematoxylin and eosin, 20 original
objective. (D) Degeneration and attenuation of the RE with minimal early SE metaplasia in the lateral wall of the rostral nasal cavity (NC) of a
young control female monkey at level 2. Hematoxylin and eosin, 10 original objective. (E) Inflammation and attenuation of the RE on the medial
surface of the ethmoturbinate at level 2 of the NC of a young control male monkey. Hematoxylin and eosin, 20 original objective. (F)
Inflammation and glandular microabscess formation in the RE that lies adjacent to the olfactory epithelium on the caudodorsal SP at level 3 of
a young control male monkey. Hematoxylin and eosin, 20 original objective. (G) Early SE metaplasia or attenuation of the RE, in the lateral wall of
the rostral NC at level 2 of a male monkey. Note the much thickened basement membrane that identifies this as the RE (arrowheads).
Hematoxylin and eosin, 20 original objective. (H) RE hyperplasia with rosette formation and marked thickening of the basal membrane (arrows)
in the caudal dorsolateral angle at level 3 of the NC of a young control female monkey. Hematoxylin and eosin, 10 original objective.

(Figures 3A and 5B). The lesion at this site was one of the main Lesions of the TE and RE
causes of high incidences of inflammatory lesions in studies
The most common findings in the TE and RE were inflamma-
and was considered to be associated with nose picking. Asso-
tion, squamous metaplasia, mucous cell hyperplasia/metapla-
ciated findings included mixed inflammatory cell infiltrates in
sia, and respiratory hyperplasia. Rostrally, inflammation was
the lamina propria, squamous cell hyperplasia with rete peg
most commonly observed in the regions of transition from TE
formation (Figure 5A), and goblet cell metaplasia in the SE/
to RE in the midseptum or the septal bulge, the midlateral
TE transitional zone.
Chamanza et al.
Figure 5. (continued).
region, and the rostral ends of the turbinates (Figure 5E). Caud-
ally, inflammation of the RE most commonly occurred in the
dorsolateral ridge at level 2 or 3, the regions of transition from
RE to OE in the dorsolateral angle or dorsal septum (Figures 3B
and 5F), and the ventral/ventromedial surfaces of the EMTs.
Inflammation in the dorsolateral ridge and the regions of RE/
OE interface was often associated with intraepithelial micro-
abscesses and cysts (Figure 5F), respiratory hyperplasia with
rosette formation and further thickening of the basal lamina
(Figures 5H and 6F), and epithelial vacuolation (Figure 6D).
Squamous metaplasia (Figure 5G) also occurred com-
monly in the regions of transition from TE to RE in the mid-
lateral and midseptal regions of the rostral nasal cavity or
vestibule, the rostral ends of the turbinates, or on other angu-
lar surfaces of the EMTs. In its earliest form, minimal squa-
mous metaplasia was characterized by subtle degenerative
changes in the RE such as disorganization and attenuation
of the epithelium with superficial cells showing an altered
649
polarity toward a horizontal orientation, loss of cilia, and a
reduction in the number of goblet cells (Figure 5D and E). As
a result, the RE initially resembled the cuboidal TE lying
adjacent to it, giving the impression of an extension of the
TE to regions normally occupied by RE. Therefore, some
degenerative changes of the RE or early forms of squamous
metaplasia in the regions of TE/RE transition were either
missed or recorded as TE extension or hyperplasia. Attenua-
tion of the TE and/or RE was also often accompanied by
inflammatory cell infiltrations into the lamina propria (Figure
5D and E). Since this finding was considered a precursor to
squamous metaplasia, minimal grades of squamous metapla-
sia or inflammatory cell infiltrates were usually diagnosed.
Goblet cell hyperplasia and metaplasia were often combined
as a single term, but in other studies, a distinction was made
between the 2 terms. In the latter cases, goblet cell metaplasia
was recorded in the TE/RE zones on the septum, where it was
characterized by an increase in the number of goblet cells in an
650 Toxicologic Pathology 44(5)

Figure 6. Spontaneous findings of the olfactory epithelium (OE) in the nasal cavity (NC) of young control cynomolgus monkeys (Macaca
fascicularis) of 27 to 42 months of age. (A) OE inflammation and degeneration in the dorsolateral meatus of a young control female monkey at
level 4. Hematoxylin and eosin, 20 original objective. (B) OE degeneration, with cell-specific necrosis of olfactory sensory neurons, and sparing
of sustentacular cells, in the dorsal meatus at level 4 of the NC of a young female monkey. Minimal inflammation is also present. Hematoxylin and
eosin, 20 original objective. (C, insert). OE degeneration with respiratory epithelial metaplasia and deposition of amyloid-like eosinophilic
material in the dorsolateral meatus of a young control female monkey at level 4. Hematoxylin and eosin, 20 original objective. (D) OE
vacuolation at the respiratory–olfactory epithelial border on the dorsal nasal septum of a young female monkey at level 4. Minimal inflammation
is also present. Hematoxylin and eosin, 20 original objective. (E) OE hyperplasia with rosette formation and respiratory metaplasia, at the
respiratory–olfactory epithelial border, in the dorsolateral angle of the caudal NC of a young female monkey at level 3. Hematoxylin and eosin,
20 original objective. (F) Respiratory metaplasia/hyperplasia with hyperplasia of Bowman’s glands, rosette formation, and deposition of amyloid-
like material at the respiratory–olfactory epithelial border in the dorsolateral angle of the caudal NC at level 3 of a young male monkey.
Hematoxylin and eosin, 20 original objective. (G) Inflammation at the border between the respiratory epithelium (RE) and OE on the
dorsolateral ridge of the caudal NC at level 3 in a young male monkey. Note the thick basal lamina (arrowhead) beneath the RE. Hematoxylin
and eosin, 20 original objective. (H) Pigment in the olfactory mucosa of the caudodorsal meatus at level 4 of the NC in a young control female
monkey. Hematoxylin and eosin, 20 original objective.

epithelium that normally contains scant goblet cells (Figure
5C). Goblet cell metaplasia of the TE was often associated with
acute inflammation and influx of neutrophils into the lamina
propria, epithelium, or lumen. Goblet cell hyperplasia on the
other hand was used to describe an increase in the density of
goblet cells in regions which normally have a high goblet cell
density, such as the MT, MS (Figure 4D and G), or the caudal
ventrolateral meatus (Figure 4E and F). Goblet cell hyperpla-
sia/metaplasia was characterized by mucosal folding and pseu-
doglands or intraepithelial crypts formation (Figure 5C and F).
Rupture of hypertrophic goblet cells in these regions resulted in
intraepithelial mucus-filled cysts (Figure 5F).
Chamanza et al. 651

Figure 6. (continued).

Lesions of the OE present in the lumen (Figure 6A) and were often used to differ-
entiate genuine OE degeneration from artifactual thinning of
As with the other epithelial types, inflammatory lesions were
the epithelium. More chronic changes were associated with
the most commonly observed changes in the OE, followed by
basal cell hyperplasia, rosette formation, and/or respiratory
degeneration/atrophy and respiratory metaplasia. Inflammation
metaplasia (Figure 6C, E, and f).
and degenerative changes often occurred together in the olfac-
Other miscellaneous findings included the frequently
tory mucosa and were usually characterized by varying grades
observed corpora amylacea in the wall of the MS (Figures
of polymorphonuclear or mixed cell infiltration into the lamina
4G and 7B), submucosal glandular metaplasia, and dilatation
propria or intraepithelial (Figure 6A–E), luminal inflammatory
of submucosal glands of the MS (Figure 7A), NALT hyperpla-
exudate (Figure 6A), OE degeneration, (Figure 6B and C), and
sia (Figure 7C and D), OE pigment, and eosinophilic inclusions
the intraepithelial elaboration of an amyloid-like eosinophilic
in the RE. Corpora amylacea started as mineral deposition in
material (Figure 6B and C). The earliest degenerative change
the glands of the MS and was present in almost every animal.
observed in the OE was atrophy/disorganization, which was
characterized by the disruption of the laminar pattern of the
nuclei of the OSNs (Figure 6B) and a vacuolated appearance of
the epithelium caused by a nonspecific loss of either OSNss or Discussion
sustentacular cells. This was followed by thinning or attenua- The nose is a complex organ with many functions that include
tion of the mucosa. An eosinophilic proteinaceous material or olfaction, warming, humidifying, and filtering of the inspired
cellular debris overlying the attenuated mucosa were usually air (Negus 1958). Its anatomy varies greatly between
652 Toxicologic Pathology 44(5)

Figure 7. Miscellaneous background findings in the nasal cavity (NC) of young control cynomolgus monkeys (Macaca fascicularis) of 27 to 42
months of age. (A) Dilatation of submucosal glands in the maxillary sinus (MS) of a young male monkey at level 3 of the NC. Hematoxylin and eosin,
20 original objective. (B) Corpora amylacea in the subepithelium of the MS of a young male monkey at level 3 of the NC. Hematoxylin and eosin,
20 original objective. (C) Rostral vestibule of a young female monkey showing nasal associated lymphoid tissue (NALT) hyperplasia, with partial
occlusion of the nasal passages. Hematoxylin and eosin, 4 original objective. (D) Multifocal NALT hyperplasia in the nasal cavity of a young male
monkey at level 2 of the nasal cavity. Hematoxylin and eosin, 1 original objective.

laboratory animals and man. The anatomical differences may nasal walls and presence or absence of accessory structures,
affect the occurrence and distribution of lesions following may influence nasal airflow and therefore species-specific
exposure to inhaled nasal toxicants. Differences in the gross uptake and deposition of inhaled material. In addition, inter-
anatomy, such as the turbinate structure, folds, or grooves on species variations in the morphological composition and
Chamanza et al. 653

Table 2. Incidences of Pathological Findings in the Transitional Table 5. Incidences of Pathological Findings in the Olfactory
Epithelium of the Nasal Cavity in 114 Control Cynomolgus Monkeys Epithelium of the Nasal Cavity in 114 Young Control Cynomolgus
(Macaca fascicularis) from 11 Inhalation Studies of 4- to 13-Week Monkeys (Macaca Fascicularis) from 11 Inhalation Studies of 4- to
Duration. 13-Week Duration.

Males (57) Females (57) Males (57) Females (57)

Findings Incidence Percentage Incidence Percentage Findings Incidence Percentage Incidence Percentage

Inflammation, transitional 4 7.02 5 8.77 Inflammation/infiltrates, 10 17.54 9 15.79

epithelium lamina propria
Hyperplasia/metaplasia, 2 3.51 2 3.51 Inflammation 1 1.75 3 5.26
mucous cell Degeneration/atrophy/ 1 1.75 3 5.26
Hyperplasia, transitional 2 3.51 1 1.75 disorganization
epithelium Metaplasia, respiratory 1 1.75 3 5.26
Cyst(s) in lamina propria 1 1.75 0 0 epithelium
Inflammatory infiltrates, 1 1.75 0 0 Eosinophilic inclusions 0 0 3 5.26
lamina propria Brown pigment, lamina 1 1.75 0 0
propria
Rosette formation 1 1.75 0 0
Intraepithelial cysts 0 0 1 1.75
Table 3. Incidences of Pathological Findings in the Respiratory
Erosions 0 0 1 1.75
Epithelium of the Nasal Cavity in 114 Control Cynomolgus Monkeys
(Macaca fascicularis) from 11 Inhalation Studies of 4- to 13-Week
Duration.
distribution of the nasal epithelium may influence the local
Males (57) Females (57) tissue susceptibility resulting in lesions with a species-
specific distribution pattern. The species- and site-specific
Findings Incidence Percentage Incidence Percentage responses to inhaled compounds may in turn affect the inter-
Inflammation/infiltrates, 13 22.81 14 24.56 pretation of results from inhalation toxicity studies and the
lamina propria extrapolation of data from experimental animal studies to
Inflammation 5 8.77 7 12.28 human risk assessment.
Hyperplasia, respiratory 4 7.02 2 3.51 We have described the normal anatomy and background
epithelium pathology of the nasal cavity of the cynomolgus monkey,
Hyperplasia/metaplasia, 3 5.26 4 7.02
including the characteristic gross features and the interanimal
mucous cell
Squamous metaplasia 3 5.26 4 7.02 variations, the light microscopic characteristics and distribu-
Pseudogland formation 3 5.26 3 5.26 tion of the epithelial types, and the nature and incidences of
Eosinophilic inclusions 0 0 1 1.75 spontaneous pathological changes. The results indicate that,
compared to other laboratory animals, the nasal gross anatomy
of the cynomolgus monkey is more similar to that of humans
(DeSesso 1993; Lucas 1932; Negus 1958). The main gross
Table 4. Incidences of Pathological Findings in the Olfactory anatomic differences to dogs and rodents are that the monkey
Epithelium of the Nasal Cavity in 114 Control Cynomolgus Monkeys
nose shows the wide interanimal variations in the shape and
(Macaca fascicularis) from 11 Inhalation Studies of 4- to 13-Week
Duration. size; fewer and less complex turbinates; and an absence of a
vomeronasal organ, a transverse lamina and olfactory recess,
Males (57) Females (57) Steno’s glands, and vestibular alar folds or atrioturbinates.
This description of the anatomy of the cynomolgus monkey
Findings Incidence Percentage Incidence Percentage
nose is closely similar to that described for other macaque
Inflammation/infiltrates, 10 17.54 9 15.79 monkeys in the literature (Harkema, Plopper, Hyde, Wilson,
lamina propria et al. 1987; Kepler et al. 1998; Lucas 1932). Any minor differ-
Inflammation 1 1.75 3 5.26 ences between this study and previous studies could be attrib-
Degeneration/atrophy/ 1 1.75 3 5.26 uted to species differences. For instance, in the rhesus
disorganization
Metaplasia, respiratory 1 1.75 3 5.26
macaque, variations in the shape of the turbinates were
epithelium observed on the medial surface of the EMTs, while the MTs
Eosinophilic inclusions 0 0 3 5.26 were reported to be more uniform (Lucas 1932). In this study,
Brown pigment, lamina 1 1.75 0 0 the biggest variations in the shape of the turbinates were
propria observed on the ventral surface of both turbinates. The medial
Rosette formation 1 1.75 0 0 and lateral surfaces were uniform and generally followed the
Intraepithelial cysts 0 0 1 1.75 contours of the septum and the lateral wall, respectively. Spe-
Erosions 0 0 1 1.75
cies differences between the rhesus and the cynomolgus
654
macaques could therefore account for these minor differences
in the description of the gross anatomy.
Similarly, the histological appearance and distribution of the
nasal epithelium is also consistent with what has been reported
for other macaque monkeys in the literature (Harkema, Plop-
per, Hyde, Wilson, et al. 1987; Monticello et al. 1989). The
study also confirms that, as with other macaque monkeys, the
morphology and distribution of the nasal epithelium in the
cynomolgus monkey is closer to that of humans than that of
rodents and dogs (Chamanza et al. 2015; DeSesso 1993;
Halama et al. 1990; Harkema, Plopper, Hyde, Wilson, et al.
1987). Briefly, the major histological differences between
monkeys and rodents include a generally thicker epithelium
of all subtypes in monkeys, a wider area lined by TE, and a
much reduced area lined by OE.
One of the main objectives of this study was to provide
readily accessible reference material on the common back-
ground changes in the nasal cavity of control cynomolgus mon-
keys, which could be used as an aid when evaluating inhalation
studies. This included describing the findings, the incidences
and the most affected regions, and therefore the possible
regions of toxicological significance in the cynomolgus mon-
key nose. The results indicate that the rostral nasal cavity, areas
of particular impaction (bulges and ridges), and areas of epithe-
lial transition were the most affected regions and these
included:
(a) the midlateral region within the vestibule or rostral
nasal cavity,
(b) the midseptal regions or septal bulge in the rostral
nasal cavity,
(c) the dorsolateral ridge (or vestige of the nasoturbinate)
that lies dorsal to the nasal atrium,
(d) the rostral margins and medial or ventral surface of the
EMTs, and
(e) the region of transition from RE to OE in the dorso-
lateral angle or on the adjacent septum.
These regions are largely consistent with those known to be
affected by inhaled irritants such as ozone (Harkema, Plopper,
Hyde, St. George, et al. 1987) and formaldehyde (Monticello
et al. 1989) in monkeys, and the spontaneous pathology
changes observed in these regions were also similar to those
induced by nasal irritants. Since the lesion distribution patterns
associated with nasal irritants are mostly attributable to
regional deposition as a result of airflow patterns (Morgan and
Monticello 1990) and an inherent susceptibility of the epithe-
lium, such as the reduced mucosubstances in the TE (Harkema,
Carey, and Wagner 2006), this suggests that nasal airflow char-
acteristics and tissue susceptibility may also play a role in the
distribution of spontaneous lesions in the nasal cavity of the
cynomolgus monkey. It also suggests that, although the exact
etiology of the spontaneous nasal lesions in juvenile monkeys
is not known, exposure to environmental factors such as smog
and other impurities in the urban air could be the contributing
factors. Ozone, which causes similar nasal responses to those
Toxicologic Pathology 44(5)
observed in our study (inflammation, epithelial hyperplasia,
and mucous cell hyperplasia of the TE) in both laboratory
animals and humans (Nikasinovic, Momas, and Seta 2003) is
the principal oxidant pollutant in smog. Differentiating these
background changes from induced lesions can be a challenge.
Author Contribution
Authors contributed to conception or design (RC); data acquisition,
analysis, or interpretation (RC, IT, MG, CH, MS, JG, KG, MA, JS,
VM); drafting the manuscript (RC); and critically revising the manu-
script (RC, IT, MG, CH, MS, JG, KG, MA, JS, VM). All authors gave
final approval and agreed to be accountable for all aspects of work in
ensuring that questions relating to the accuracy or integrity of any part
of the work are appropriately investigated and resolved.
Declaration of Conflicting Interests
The author(s) declared no potential conflicts of interest with respect to
the research, authorship, and/or publication of this article.
Funding
The author(s) received no financial support for the research, author-
ship, and/or publication of this article.
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