Endocrine Care

doi: 10.4183/aeb.2010.335

RETICULAR FIBERS NETWORK PARTIALLY DISCRIMINATES

BETWEEN NORMAL HYPOPHYSIS AND PITUITARY
ADENOMAS
R.A. Ceausu1, B. Balinisteanu1, A.M. Cimpean*1, P.N. Gaje1, C. Capatina2,
M. Gheorghiu2, V. Ciubotaru3, M. Coculescu2, M. Raica1
1
2

Dpt. Histology, V. Babes University of Medicine and Pharmacy -, Timisoara,

Dpt. Endocrinology, "C. Davila" University of Medicine and Pharmacy Bucharest,
3 Bagdasar-Arseni Hospital for Neurosurgery, Bucharest, Romania

Abstract
Background. Usually, silver stain is needed to differentiate between normal or
hyperplastic hypophysis and pituitary adenomas. Many papers reported the lack of reticular
fibers network as mandatory for pituitary adenoma diagnosis.
Aim. Differences between the architecture of reticular fibers in normal pituitary and
pituitary adenomas.
Methods. Gordon- Sweet silver staining of pituitary specimens, prelevated during
pituitary surgery performed in 138 patients with the endocrine and imagistic diagnosis of
pituitary macro-adenomas.
Results. Pituitary specimens of pituitary adenomas was confirmed in 133 cases; 3
specimens were with normal pituitary tissue, 1 with pituitary hyperplasia, 1 with pituitary
apoplexy. Twelve of 133 pituitary adenomas specimens were associated also with normal
pituitary tissue. There was a loss of acinar network of reticular fibres in 115 cases, but the
present study describes the persistence of reticular fibers networks in 18 (13.53%) of 133
pituitary adenomas. We identified five distribution patterns of remanent reticular fibers network
in pituitary adenomas using silver staining.
Conclusion. Persistence of reticular fibers in a small number of pituitary adenomas
could be a possible pitfall in the discrimination between the normal hypophysis and pituitary
adenoma.
Key words: hypophysis, pituitary adenomas, reticular fibres, silver staining.

INTRODUCTION
Named by Van Gehuchten l`organ enigmatique, one century ago (1),
hypophysis keeps this controversial features both for normal and pathological
*Correspondence to: Anca Cimpean, MD, Victor Babes University of Medicine and Pharmacy Histology, Piata Eftimie Murgu 2, Timisoara, Timis 300041, Email: ancacimpean1972@yahoo.com
Acta Endocrinologica (Buc), vol. VI, no. 3, p. 335-341, 2010
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conditions (2). Pituitary adenomas are benign epithelial tumors arising in the
anterior part of the pituitary gland and consisting of adenohypophyseal cells.
Pituitary adenomas are the most common lesions in the sella region, representing
75% of sellar lesions (3). They are revealed in 20-25% of autopsy-obtained
pituitaries, performed for other diseases, and have to be differentiated by various
non-neoplastic tumor-like lesions (2). Pituitary adenomas were classified by
WHO according to staining affinities of the cell cytoplasm, size, endocrine activity,
histologic characteristics, hormone production and contents, ultrastructural features,
(granularity of the cell cytoplasm, cellular composition), cytogenesis and growth
pattern (4). From the first morphologic classification proposed by Cushing until
now, classifications of pituitary adenomas have aimed to combine the morphologic
aspects of these tumors with clinical aspects. Previously, pituitary adenomas were
separated on the grounds of staining characteristic of the cell cytoplasm into
acidophilic, basophilic and chromophobic tumors. On the basis of their size,
pituitary adenomas can be divided into microadenomas and macroadenomas.
Modern pathological classifications are based on immunohistochemistry and other
molecular methods (4, 5).
Pituitary hyperplasia is a condition difficult to diagnose. There are reports of
patients with enlarged pituitary gland submitted by mistake to surgery in the best
European centers, as pituitary adenomas, although the pathology proved to be
physiological pituitary hyperplasia (6). Pituitary hyperplasia is defined as a nonneoplastic increase of one or more functionally distinct types of pituitary cells (7, 8).
Any cell population within the pituitary gland can undergo hyperplasia and sometime
adaptive transdifferentiation (9). This process can be physiological (e.g. lactotrophs
hyperplasia in pregnancy) or in pathological conditions. Some types of the pituitary
hyperplasia - e.g. thyrotrophs in primary mixedema - could be largely enough to
compress adjacent structures and mimic a neoplasic process, and when hyperplasia is
prolonged it may progress to adenoma, at least in rodents (10-12).
The Gordon-Sweet silver stain identifies reticular fibers network in various
tissues. For pituitary gland specimens, this method is applied to highlight the presence
of reticular fibers network around nested pituitary cells (acini) in normal hypophysis
and hyperplasia and lack of it in pituitary adenomas. The absence of acinar reticular
fibers in adenomas is considered by many authors as mandatory for their differentiation
from normal or hyperplastic pituitary gland (2).
In contrast with many papers that reported the complete loss of acinar reticular
fibers in pituitary adenomas, in the present study we describe five patterns of reticular
fibers remanents in adenomatous pituitary gland compared with normal pituitary gland.

MATERIALS AND METHODS

We included in our study 138 pituitary gland specimens, prelevated during
surgery for pituitary macro-adenomas, previously diagnosed by clinical, hormonal
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Reticular fibres within pituitary adenomas

Figure 1. (a) Normal hyphophysis in haematoxylin-eosin stain. HE, x 100. (b) Silver stain, normal
hyphophysis, x 100. (c) Pituitary hyperplasia; haematoxylin-eosin stain, x 400. (d) Pituitary hyperplasia
- the reticular fibers network thinner in comparison with normal cases. Silver stain, x 400.

and imagistic methods in the department of neuroendocrine and pituitary diseases

of the National Institute of Endocrinology, Bucharest. The specimens were fixed in
10% buffered formalin for 48 hours and parrafin embedded using the routine
histological procedure in the Histology department, Victor Babes University of
Medicine and Pharmacy, Timisoara. Five micrometers thick serial sections were
performed from each paraffin block and sections were mounted on silanized slides.
Sections from each case were stained with routine haematoxylin and eosin method
for pathological evaluation and Gordon-Sweet silver stain (Sigma Reagents).
Microscopic observation was performed by three independent observers using
Nikon Eclipse E600 (Nikon Corporation, Tokio, Japan). Images were captured and
processed with Lucia G software system.
We evaluated only the specimens which showed positive control of the staining
in the connective tissue surrounding small blood vessels. In the normal or pathological
pituitary we described the presence and patterns of reticular fibers network.
The study was performed according with agreement no. 16605/2009 obtained
from Ethical Commitee of Carol Davila University of Medicine and Pharmacy
Bucharest, Romania.

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RESULTS
Microscopic examination of 138 cases revealed: 135 cases of abnormal pituitary
lesions (pituitary adenoma, n=133, pituitary apoplexy, n=1; pituitary hyperplasia, n=1)
and 3 specimens of normal hypophysis. Among the cases with pituitary adenoma, 12
specimens showed both areas of pituitary adenoma and normal hypophysis.
Normal hypophysis was recognized by its specific arhitectural pattern composed
of nested cells surrounded by a rich capillary network (Fig. 1a). Gordon-Sweet silver
staining method highlighted numerous thin and branched reticular fibers disposed as

Figure 2. (a) Pituitary adenoma; haematoxylin eosin stain, x100. (b) Pituitary adenoma- numerous reticulin
fibres wich delineate networks and form pseudo- networks; silver stain, x 100. (c) Pituitary adenoma;
haematoxilin eosin stain, x100.(d) Pituitary adenoma- reticulin fibres disposed to a network with disorderly
aspect, without completely surround nests cell; silver stain, x 100.(e) Pituitary adenoma; haematoxylin eosin
stain, x 100. (f) Pituitary adenoma- reticular network which mimics the normal distribution by delimitation
of pseudonests of hypophyseal cells, x400.
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Reticular fibres within pituitary adenomas

Figure 3. (a) Pituitary adenoma; haematoxylin eosin stain, x 100. (b) Pituitary adenoma- reticular
fibres
with variable thickness, located in the adenoma areas and without defined nests, silver
stain, x 400. (c) Pituitary adenoma - long and thin reticular fibres
without network distribution,
disposed isolately between adenoma cells, silver stain, x100.

a continous layer around the nests of pituitary hormone-producing cells (acinar

disposition) (Fig. 1b). Pituitary hyperplasia (1 case) was characterized by the
preservation of the nested (acinar) architecture but the nests were larger than normal,
irregular, and contained considerably more larger cells than usual. The reticular fibers
network is thinner as compared with the normal hypophysis (Fig. 1 c,d).
According to the growth pattern, pathological evaluation of pituitary adenomas
(n= 132) reclassified them as follows: solid (86 cases), trabecular (22 cases), papillary
(7 cases), diffuse (6 cases), alveolar (2 cases), mixed (solid and papillary 5 cases, solid
and trabecular 2 cases, difuse and trabecular 1 case, follicular and trabecular 1case,
trabecular and papillary 1 case).
Many of Gordon-Sweet silver stained pituitary adenomas showed positive
staining for reticulin fibers restricted around small blood vessels and negative inside
the adenoma. Eighteen from 133 cases of pituitary adenomas (13.53%) were
positive for silver staining inside the adenoma tissue. Taking into account the
growth pattern, we classified the eighteen cases of pituitary adenomas as follow:
solid pattern (15 cases), difuse (1 case), nodular (1 case). We have noticed five
distribution patterns of reticular fibers in pituitary adenomas.
The first pattern was represented by numerous reticular fibres which delineate
networks and form pseudo-networks. These mimic network of reticular fibres which
surround cell nests in normal hypophysis (Fig. 2 a,b). The second pattern was
characterised by short, sinuous, disorderly reticular fibres. We found a higher density
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of reticular fibres with sinuous aspect and tendency to form networks. We noticed
reticular fibres disposed into a network with disorderly aspect, without completely
surrounding nests cell; meshs network contained 2 to 4 hypophyseal cells (Fig. 2 c,d).
The third pattern had normal-like aspect. We noticed reticular network which mimics
the normal distribution by delimitation of pseudonests of hyphophyisis cells; this
delimitation is incomplete with inconstant pattern (Fig. 2 e,f). In fourth pattern we
notice short, non-anastomosated reticular fibres with variable thickness, disposed
inside of adenoma areas and without defined nests (Fig. 3 a,b). The fifth pattern of
reticular fibres which we noticed was characterised by long, thin, reticular fibres,
without forming networks, disposed isolately between adenoma cells (Fig. 3c).

DISCUSSION
The term reticular was coined in 1892 by M. Siegfried (13). The cells of the
normal adenohypophysis are organized in nests, each nest consisting of an admixture
of different cell types surrounded by well defined reticular rich network (14). All of
our 3 specimens with normal pituitary gland showed a regular reticular fibers network
both around blood vessels and endocrine nests. For pituitary hyperplasia, the GordonSweet silver stain demonstrated the preservation of the nest architecture. However, the
nests were larger than normal, irregular, and contained considerably more cells than
usual (15). In the only one case of pituitary hyperplasia included in this study we
noticed this aspects and a thin reticular network in comparison with normal cases.
In 1977, Velasco et al (16) described the use of a rapid silver stain to diagnose
adenomas on frozen section. The stain highlights reticular fibers, which are sparser in
adenomas compared with normal pituitary gland (17). Asa et al (18) demonstrated on
a transgenic mouse that pituitary reveals hyperplasia and an area of disrupted
architecture composed of sheets of large cells without acini. We found these aspects in
the case of hyperplasia and in some cases of adenomas. Beside these aspects in 18
cases of pituitary adenomas we observed the persistence of reticular fibers inside the
adenomas with disorganized arrangement. We described five particular distribution
patterns for the remanent reticular fibers in pituitary adenomas.
The presence of scanty reticular fibers network is highlighted and accepted in the
diagnosis of pituitary apoplexy (19) and hyperplasia but not for pituitary adenomas.
Despite the fact that most of the pituitary adenomas included in the study had no
reticular fibers inside the tumor mass, a small percent of the cases were positive with
silver stain methods. For eighteen cases of pituitary adenomas, tumor masses included
positive reticular fibers other than those disposed around blood vessels.
Our findings are in contradiction with widely accepted opinion that the lack of the
reticular fibers network in pituitary adenomas compared with normal pituitary gland
represents a compulsory diagnostic criteria to differentiate between these two conditions
(2). However limits and controversies around the diagnosis of pituitary adenomas were
reported also using more sophisticated techniques. Discrepancies between
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immunohistochemistry and electronmicroscopy have also been described (20).

In conclusion, we consider that the use of silver stain as an exclusive
diagnostic tool to differentiate between pituitary adenomas and normal or
hyperplastic pituitary is not enough to characterize hypophyseal lesions. Persistence
of reticular fibers in a small number of pituitary adenomas could be a possible pitfall
in the discrimination between the normal hypophysis and pituitary adenoma.
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