Home About us Editorial board Ahead of print Current issue Archives Submit article Instructions Subscribe Contacts Login 
  • Users Online: 388
  • Home
  • Print this page
  • Email this page


 
 Table of Contents  
SHORT COMMUNICATION
Year : 2016  |  Volume : 23  |  Issue : 1  |  Page : 44-46

The utility of azan trichrome staining in Ameloblastoma


1 Department of Oral Pathology, College of Medicine, University of Ibadan, Ibadan, Nigeria
2 Department of Oral and Maxillofacial Surgery, College of Dentistry, King Khalid University, Abha, Saudi Arabia
3 Department of Oral, Cranio Maxillofacial and Facial Plastic Surgery, Medical Center of the Goethe University Frankfurt, Frankfurt am Main; REPAIR Lab, Institute of Pathology, University Medical Center, Johannes Gutenberg University, Mainz, Germany

Date of Web Publication13-Apr-2016

Correspondence Address:
Shahram Ghanaati
Department of Oral, Cranio Maxillofacial and Facial Plastic Surgery, Medical Center of the Goethe University Frankfurt, Frankfurt am Main; REPAIR Lab, Institute of Pathology, University Medical Center, Johannes Gutenberg University, Mainz
Germany
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1117-1936.180187

Rights and Permissions
  Abstract 

Background: It is occasionally difficult to distinguish the stellate reticulum-like region of ameloblastoma from the fibrous connective tissue stroma. This difficulty is further pronounced in the plexiform variant of ameloblastoma that has very sparse fibrous connective tissue.
Aim: To test the utility of Azan trichrome stain in marking tumour regions and the peri-tumour environment of ameloblastoma.
Materials and Methods: Sections were prepared for 18 formalin fixed paraffin-embedded blocks of ameloblastoma cases and stained with Azan trichrome stain according to the manufacturer's specification.
Results and Conclusions: The tumour areas were stained mostly brown, with the ameloblasts mainly marked as deep brown while the stellate reticulum-like region was light brown. The structures in the peri-tumour region were marked with different shades of blue. Azan trichrome staining was able to distinguish between the fibrous connective tissue and the stellate reticulum-like areas in 100% of the cases.

Keywords: Ameloblastoma, Azan trichrome stain, connective tissue, stellate reticulum


How to cite this article:
Adisa AO, Udeabor SE, Kubesch A, Barbeck M, Ghanaati S. The utility of azan trichrome staining in Ameloblastoma. Niger Postgrad Med J 2016;23:44-6

How to cite this URL:
Adisa AO, Udeabor SE, Kubesch A, Barbeck M, Ghanaati S. The utility of azan trichrome staining in Ameloblastoma. Niger Postgrad Med J [serial online] 2016 [cited 2022 Jan 27];23:44-6. Available from: https://www.npmj.org/text.asp?2016/23/1/44/180187


  Introduction Top


It is occasionally difficult to distinguish the stellate reticulum-like region of ameloblastoma from the fibrous connective tissue stroma area. This difficulty is further pronounced in the plexiform variant of ameloblastoma that has intricate interlacing islands, strands and cords of the tumour and connective tissue, such that in some cases it is virtually impossible to distinguish the fibrous connective tissue from the stellate reticulum-like region. It has been shown that the estimated monthly growth rate of desmoplastic ameloblastoma, which has abundant dense fibrous connective tissue stroma, is significantly lower than other conventional variants of ameloblastoma. [1] Hence, it can be projected that the less the fibrous connective tissue stroma, the faster the rate of monthly growth of ameloblastoma. Compared to other conventional variants, the fibrous connective tissue stroma of plexiform ameloblastoma is very sparse indeed. It may thus be useful to be able to estimate the area covered by connective tissue in this variant as a possible predictor of growth rate. Plexiform ameloblastoma is reported to be the second most recurrent variant after the follicular variant. [2]

Trichrome staining is a method in which three anionic dyes are used to mark a varied number of structures histologically. [3] Heidenhain Azan trichrome staining is an improvement over the traditional Mallory's method. [3] It is commonly used to distinguish cells from extracellular components and stains muscle fibres red, cartilage and bone matrix blue. [3]

Our study tested the utility of Azan trichrome stain in marking tumour regions and the peri-tumour environment of ameloblastoma, especially to distinguish the stellate reticulum-like region from the fibrous connective tissue stroma area.


  Materials and Methods Top


Eighteen formalin fixed paraffin-embedded blocks of ameloblastoma cases from the Oral Pathology Department of University College Hospital, Ibadan, Nigeria, were sectioned and stained with haematoxylin and eosin for re-evaluation and inclusion in the study. At the REPAIR Laboratory, Institute of Pathology, School of Medicine, University of Mainz Germany, sections were prepared for Azan trichrome staining using the following procedure: The sections were de-paraffinised using xylene and hydrated with descending alcohol series (100%, 90%, 70% and 60%) each for 15 min. Thereafter, they were rinsed in distilled water for 3 min and then immersed in the nuclear fast red solution for 30 min, rinsed two times under running water and once in distilled water. The sections were differentiated in 0.1% aniline alcohol solution for 3 min; this was followed by one change of acetic alcohol solution to terminate the differentiation. They were immersed in 5% aqueous phosphotungstic acid for 10 min and rinsed in distilled water, after which they were then immersed for 5 min in 1 part aniline blue-orange mixture to three parts distilled water and briefly rinsed again in distilled water. The section was then taken through an ascending series of alcohol solution (60%, 70%, 80%, 90% and 100%) to dehydrate the sections through two changes of each solution for 2 min each. The sections were finally cleared in xylene three times for 3 min each and then mounted with distyrene, plasticiser and xylene, then cover slips were placed. The staining pattern was varied but consisted of brown, blue and reddish colours. Two of the authors examined the slides together and resolved the controversy by a third review.


  Results Top


Eighteen cases of ameloblastoma comprising seven plexiform, three follicular, six cystic and two haemangiomatous variants were selected for inclusion in the study. There was no relationship between histological variant and azan staining colouration/pattern. The tumour areas were stained mostly brown [Figure 1] and [Figure 2], with the ameloblasts mainly marked as deep brown (61.1%) while the stellate reticulum-like region was light brown (72.2%) [Table 1]. The fibrous connective tissue, blood vessels and bone in the peri-tumour region were marked with different shades of blue [Table 1] and [Figure 1],[Figure 2],[Figure 3] and [Figure 4].
Figure 1: Plexiform ameloblastoma showing light blue fibrous connective tissue, light brown stellate reticulum-like region and palisaded deep brown ameloblasts (×100). Note the relative absence of blood vessel in the stellate reticulum compared to the fibrous connective tissue area. LBl: Light blue, DBr: Deep brown, LBr: Light brown

Click here to view
Figure 2: Plexiform ameloblastoma with cystic degeneration showing light blue fibrous connective tissue, light brown stellate reticulum-like region and palisaded deep brown ameloblasts (×100). LBl: Light blue, DBr: Deep brown, LBr: Light brown

Click here to view
Figure 3: Follicular ameloblastoma with the ameloblasts and stellate reticulum appearing deep purple and light purple, respectively (×200). The fibrous connective tissue is deep blue and regions of haemorrhage appear light brown to orange. LP: Light purple, DP: Deep purple, DB: Deep blue

Click here to view
Figure 4: Plexiform ameloblastoma showing the reversed nuclei of ameloblasts taking up a bright orange colour (x400). The thin light blue line marks the fibrous connective tissue stroma, and the light brown region is the stellate reticulum

Click here to view
Table 1: Azan colouration of tumour and peri - tumour areas of ameloblastoma

Click here to view



  Discussion Top


In this report, we have demonstrated the staining characteristics of Azan in ameloblastoma. We did not find any other such study in English literature either describing this pattern in ameloblastoma or linking any deductions with biologic behaviour or outcome after management. This was a retrospective study and since all the cases were treated by surgical resection with an apparently normal margin of at least 1.5 cm, follow-up and outcome were not factored into the analysis. The clinical-pathological relevance, or lack thereof, of our description may be clarified by future longitudinal studies.


  Conclusions Top


Azan trichrome staining was able to distinguish between the fibrous connective tissue and the stellate reticulum-like areas in all of the cases in the present study.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Effiom OA, Odukoya O. Desmoplastic ameloblastoma: Analysis of 17 Nigerian cases. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2011;111:e27-31.  Back to cited text no. 1
    
2.
Reichart PA, Philipsen HP, Sonner S. Ameloblastoma: Biological profile of 3677 cases. Eur J Cancer B Oral Oncol 1995;31B: 86-99.  Back to cited text no. 2
    
3.
Kiernan JA. Histological and Histochemical Methods. Theory and Practice. 4 th ed. Bloxham, UK: Scion; 2008.  Back to cited text no. 3
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1]


This article has been cited by
1 Augmented BMP4 signal impairs tongue myogenesis
Jian Zhang,Chensheng Lin,Yingnan Song,Yanding Zhang,Jiang Chen
Journal of Molecular Histology. 2021;
[Pubmed] | [DOI]
2 Inhibition of autophagy in theca cells induces CYP17A1 and PAI-1 expression via ROS/p38 and JNK signalling during the development of polycystic ovary syndrome
Mutsumi Kobayashi,Osamu Yoshino,Akitoshi Nakashima,Masami Ito,Kazuyuki Nishio,Yosuke Ono,Tae Kusabiraki,Chisato Kunitomi,Nozomi Takahashi,Miyuki Harada,Katsushige Hattori,Makoto Orisaka,Yutaka Osuga,Shigeru Saito
Molecular and Cellular Endocrinology. 2020; : 110792
[Pubmed] | [DOI]
3 Dilated cardiomyopathy-linked heat shock protein family D member 1 mutations cause up-regulation of reactive oxygen species and autophagy through mitochondrial dysfunction
Hirokazu Enomoto,Nishant Mittal,Takayuki Inomata,Takuro Arimura,Tohru Izumi,Akinori Kimura,Keiichi Fukuda,Shinji Makino
Cardiovascular Research. 2020;
[Pubmed] | [DOI]
4 Osteocalcin, Azan and Toluidine blue staining in fibrous dysplasia and ossifying fibroma of the jaws
Samuel Ebele Udeabor,Akinyele Olumuyiwa Adisa,Anna Orlowska,Poju Chia,Robert A. Sader,Shahram Ghanaati
Alexandria Journal of Medicine. 2018;
[Pubmed] | [DOI]



 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Materials and Me...
Results
Discussion
Conclusions
References
Article Figures
Article Tables

 Article Access Statistics
    Viewed3667    
    Printed69    
    Emailed0    
    PDF Downloaded397    
    Comments [Add]    
    Cited by others 4    

Recommend this journal


[TAG2]
[TAG3]
[TAG4]