Headache Medicine, v.10, n.2, p.60-62, 2019

ABSTRACT

RESUMO

Descritores: Mastócito; Dura-máter; Humano; Artéria meningeal; Enxaqueca.

SHORT COMMUNICATION

Histomorphometric analysis of mast cells in different

regions of human intracranial dura mater

Análise histomorfométrica de mastócitos em diferentes

regiões da dura-máter intracraniana humana

Emanuela Paz Rosas

Silvania Tavares Paz

Ana Clara de Souza Neta

Raisa Ferreira Costa

Ana Paula Fernandes da Silva

Manuela Figueiroa Lyra de Freitas

Marcelo Moraes Valença

1,4

Programa de Pós-graduação em Biologia

Aplicada à Saúde-LIKA/UFPE;

Programa de Pós-graduação em Saúde

Translacional – CCS/UFPE;

Departamento de Nutrição – CCS/UFPE;

Programa de Pós-graduação em Ciências

Biológicas – CB/UFPE.

Departamento de Anatomia – CB/UFPE,

Recife, Pernambuco, Brazil.

*Correspondence

Emanuela Paz Rosas

E-mail: manu_pathy@hotmail.com

Received: April 2, 2019.

Accepted: May 4, 2019.

Objective: To analyze mast cell histomorphometry in three different regions

of the human intracranial dura mater. Method: Three specimens of dura

mater were collected after approval by the Ethics Committee (CAAE No.

57692216.5.0000.5208). Each dura mater was obtained from human cadavers

between 7 and 24 hours after death. After collection, the samples were xed,

cut into two fragments and longitudinally placed in the following way: external

(periosteum) and internal (meningeal) sides. The fragments (1.5 cm

) were taken

from three different regions: proximity of the right middle meningeal artery, the

proximity of the left middle meningeal artery and superior sagittal sinus. These

fragments were submitted to microtomy (10 µm), stained with 0.1% toluidine

blue and analyzed by optical microscopy. The histomorphometric parameters

adopted were: the distance from the mast cells to the vessels, the number and

if the mast cells were degranulated. Five elds from each case were analyzed.

For this analysis, the Image J 1.52a 2019 software was used. Results: A higher

number of mast cells was observed in the periosteal layer when compared with

the meningeal layer (p=0.026). When the distribution of the mast cells was

evaluated, we observed that the cells were localized in the proximity of the

middle meningeal artery (p<0.05). Conclusion: In human dura mater, the mast

cells are localized in the proximity of dural arteries.

Keywords: Mast cell; Dura mater; Human; Meningeal artery; Migraine.

Objetivo: Analisar a histomorfometria dos mastócitos em três regiões

diferentes da dura-máter intracraniana humana. Método: Três amostras de

dura-máter foram coletadas após aprovação pelo Comitê de Ética (CAAE nº

57692216.5.0000.5208). Cada dura-máter foi obtida de cadáveres humanos

entre 7 e 24 horas após a morte. Após a coleta, as amostras foram xadas,

cortadas em dois fragmentos e dispostas longitudinalmente da seguinte

maneira: face externa (periósteo) e interna (meníngeo). Os fragmentos

(1,5 cm

) foram retirados de três regiões diferentes: proximidade da artéria

meníngea média direita, proximidade da artéria meníngea média esquerda e

seio sagital superior. Esses fragmentos foram submetidos à microtomia (10 µm),

corados com azul de toluidina a 0,1% e analisados por microscopia óptica. Os

parâmetros histomorfométricos adotados foram: distância dos mastócitos aos

vasos, número e se os mastócitos estavam desgranulados. Foram analisados

cinco campos de cada espécime. Para esta análise, foi utilizado o software

Image J 1.52a 2019. Resultados: Observou-se maior número de mastócitos

na camada periosteal quando comparada à camada meníngea (p = 0,026).

Quando avaliada a distribuição dos mastócitos, observamos que as células

estavam localizadas nas proximidades da artéria meníngea média (p <0,05).

Conclusão: Na dura-máter humana, os mastócitos estão localizados nas

proximidades das artérias durais.

10(2).indb 60 21/10/2019 18:25:26

Histomorphometric analysis of mast cells in different regions

Rosas EP, et al.

Headache Medicine, v.10, n.2, p.60-62, 2019

INTRODUCTION

Recent evidence strongly suggests a vital role of

dura mater mast cell in the genesis of migraine.

1-5

Gupta and Harvima described the mast cells as

a “powerhouse” since they release “algogenic and

pruritogenic mediators, which initiate a reciprocal

communication with specic nociceptors on sensory

nerve bers.”

Mast cells are cells found in abundance in the

dura mater and by local mechanism regulates vascular

and neural functions, releasing substances such as

histamine. Scientic evidence suggests that mast cells

participate in the pathophysiology of triggering a

migraine attack by inducing local sterile inammation

near the dura mater nociceptors.

Curiously, the mast cells are located in the proximity

of the arteries in the dura mater, in close association

with neurons. These cells appear to be activated

through the trigeminal nerve. It is postulated that many

neuropeptides, namely calcitonin gene-related peptide

(CGRP), hemokinin A, neurotensin (NT), pituitary

adenylate cyclase-activating peptide (PACAP), and

substance P may activate mast cells, resulting in the

release of vasoactive and pro-inammatory mediators,

involved in the pathophysiology of migraine.

Mast cells

can also release substances with pro-inammatory

and vasoactive actions (e.g., interleukin-6 and vascular

endothelial growth factor (VEGF).

The objective of the present study was to analyze

mast cell histomorphometry in three different regions of

the human intracranial dura mater.

METHOD

Three specimens of dura mater were collected

after approval by the Ethics Committee (CAAE No.

57692216.5.0000.5208).

Each dura mater was obtained from human cadavers

between 7 and 24 hours after death.

After collection, the samples were xed, cut into two

fragments and longitudinally placed in the following way:

external (periosteum) and internal (meningeal) sides.

The fragments (1.5 cm

) were taken from three

different regions: proximity of the right middle meningeal

artery, the proximity of the left middle meningeal artery

and superior sagittal sinus.

These fragments were submitted to microtomy

(10 µm), stained with 0.1% toluidine blue and analyzed

by optical microscopy. The histomorphometric

parameters adopted were: the distance from the mast

cells to the vessels, the number and if the mast cells

were degranulated. Five elds from each case were

analyzed. For this analysis, the Image J 1.52a 2019

software was used.

RESULTS AND DISCUSSION

A higher number of mast cells was observed in the

periosteal layer when compared with the meningeal

layer (p=0.026).

When the distribution of the mast cells was evaluated,

we observed that the cells were localized in the proximity

of the middle meningeal artery (Figure 1), suggesting

that there is a signicant role played by the mast cells in

dura mater to regulate vascular function. Probably the

relationship between mast cells and meningeal arteries

is an essential component in the migraine pathogenesis.

Figure 1. Mast cell density per mm2 in relation to the

distance to the vessel. P versus 0-100 µm group, Kruskal-

Wallis test and Dunn´s multiple compatisons test.

The distance between the artery and the mast cell

was measured in 153 cells (57 65 m, min 0 - max 247;

median 33, 95%IC 46-67). No differences were observed

in the concentration of mast cells in convexity of the dura

mater versus the superior sagittal sinus.

In his study, with human dura mater postmortem,

60-70% of the mast cells were degranulated. Migraine is a

disorder with signicant autonomic dysfunction. Clinically,

long-lasting ushing suggests degranulation of mast cells.

In conclusion, in human dura mater, the mast cells

are localized in the proximity of dural arteries.

REFERENCES

1. Okragly AJ, Morin SM, DeRosa D, Martin AP, Johnson KW,

Johnson MP, Benschop RJ. Human mast cells release the

10(2).indb 61 21/10/2019 18:25:27

Histomorphometric analysis of mast cells in different regions

Rosas EP, et al.

Headache Medicine, v.10, n.2, p.60-62, 2019

migraine-inducing factor pituitary adenylate cyclase-activating

polypeptide (PACAP). Cephalalgia. 2018 Aug;38(9):1564-1574.

doi: 10.1177/0333102417740563. Epub 2017 Nov 5.

2. Kilinc E, Dagistan Y, Kukner A, Yilmaz B, Agus S, Soyler G,

Tore F. Salmon calcitonin ameliorates migraine pain through

modulation of CGRP release and dural mast cell degranulation

in rats. Clin Exp Pharmacol Physiol. 2018 Jun;45(6):536-546.

doi: 10.1111/1440-1681.12915. Epub 2018 Feb 13.

3. Baun M, Pedersen MH, Olesen J, Jansen-Olesen I. Dural mast

cell degranulation is a putative mechanism for headache

induced by PACAP-38. Cephalalgia. 2012 Mar;32(4):337-45.

doi: 10.1177/0333102412439354.

4. Theoharides TC, Donelan J, Kandere-Grzybowska K,

Konstantinidou A. The role of mast cells in migraine

pathophysiology. Brain Res Brain Res Rev. 2005

Jul;49(1):65-76.

5. Jansen-Olesen I, Hougaard Pedersen S. PACAP and its

receptors in cranial arteries and mast cells. J Headache

Pain. 2018 Feb 20;19(1):16. doi: 10.1186/s10194-017-0822-2.

6. Kalpna Gupta, Ilkka T. Harvima. Mast cell-neural

interactions contribute to pain and itch Immunol Rev.

Author manuscript; available in PMC 2019 Mar 1. Published

in nal edited form as: Immunol Rev. 2018 Mar; 282(1): 168–

187. doi: 10.1111/imr.12622

10(2).indb 62 21/10/2019 18:25:27