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INVESTIGATION OF THE MICROSCOPIC STRUCTURE OF ..., Study notes of Zoology

Zoology

SCRIPTA MEDICA (BRNO) – 76 (4): 215–220, September 2003. INVESTIGATION OF THE MICROSCOPIC STRUCTURE. OF RABBIT COMPACT BONE TISSUE.

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SCRIPTA MEDICA(BRNO) – 76 (4): 215–220, September 2003
INVESTIGATION OF THE MICROSCOPIC STRUCTURE
OF RABBIT COMPACT BONE TISSUE
MARTINIAKOVÁ M.1, VONDRÁKOVÁ M.1, FABI· M.2
1Department of Zoology and Anthropology, Constantine the Philosopher University, Nitra,
Slovak Republic
2Department of Animal Physiology, Slovak Agricultural University, Nitra, Slovak Republic
Abstract
The detailed microscopic structure (qualitative and quantitative characteristics) of 10 rabbit thigh
bones was investigated. Femur diaphysis from each individual was sectioned at its smallest breadth.
The final thickness of the sections was approximately 100 microns. The average areas, perimeters,
minimal and maximal diameters of 200 vascular canals of primary osteons, 40 haversian canals of
secondary osteons, and 40 secondary osteons were measured on digital images. According to our
study the investigated bone tissue is in general composed of primary longitudinal bone tissue. Some
areas of dense haversian remodelling occur mainly in the posteromedial and posterolateral sides.
Haversian canals of the secondary osteons (like the vascular canals of the primary osteons) are short.
Key words
Microstructure, Femur diaphysis, Rabbit, Histomorphometry
INTRODUCTION
The long bone diaphysis of juvenile and adult mammals is composed of
compact bone tissue which builds the wall of the shaft. The important elements of
its structural organisation are primary and secondary (haversian) osteons.
Histological research of the compact bone tissue microstructure can be carried out
in two ways: qualitatively and quantitatively. Qualitative characteristics describe
the type of the bone tissue from the medullary cavity towards the periosteal
surface. The qualitative approach counts and measures (e.g., area, perimeter,
minimal and maximal diameter of osteons or haversian canals).
The aim of our work was to analyse the microstructure of rabbit femur
diaphysis. Microscopic structure of the compact bone was evaluated from the
point of view of qualitative and quantitative characteristics.
MATERIALAND METHODS
Our research focused on 10 femurs of five 5–7 months old female rabbits of New Zealand White
albino breed. Each of the bones was sectioned at the smallest breadth (SB) of its diaphysis where
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SCRIPTA MEDICA (BRNO) – 76 (4): 215–220, September 2003

INVESTIGATION OF THE MICROSCOPIC STRUCTURE

OF RABBIT COMPACT BONE TISSUE

MARTINIAKOVÁ M. 1 , VONDRÁKOVÁ M. 1 , FABI· M. 2

(^1) Department of Zoology and Anthropology, Constantine the Philosopher University, Nitra, Slovak Republic (^2) Department of Animal Physiology, Slovak Agricultural University, Nitra, Slovak Republic

A b s t r a c t

The detailed microscopic structure (qualitative and quantitative characteristics) of 10 rabbit thigh bones was investigated. Femur diaphysis from each individual was sectioned at its smallest breadth. The final thickness of the sections was approximately 100 microns. The average areas, perimeters, minimal and maximal diameters of 200 vascular canals of primary osteons, 40 haversian canals of secondary osteons, and 40 secondary osteons were measured on digital images. According to our study the investigated bone tissue is in general composed of primary longitudinal bone tissue. Some areas of dense haversian remodelling occur mainly in the posteromedial and posterolateral sides. Haversian canals of the secondary osteons (like the vascular canals of the primary osteons) are short.

K e y w o r d s

Microstructure, Femur diaphysis, Rabbit, Histomorphometry

INTRODUCTION

The long bone diaphysis of juvenile and adult mammals is composed of

compact bone tissue which builds the wall of the shaft. The important elements of

its structural organisation are primary and secondary (haversian) osteons.

Histological research of the compact bone tissue microstructure can be carried out

in two ways: qualitatively and quantitatively. Qualitative characteristics describe

the type of the bone tissue from the medullary cavity towards the periosteal

surface. The qualitative approach counts and measures (e.g., area, perimeter,

minimal and maximal diameter of osteons or haversian canals).

The aim of our work was to analyse the microstructure of rabbit femur

diaphysis. Microscopic structure of the compact bone was evaluated from the

point of view of qualitative and quantitative characteristics.

MATERIAL AND METHODS

Our research focused on 10 femurs of five 5–7 months old female rabbits of New Zealand White albino breed. Each of the bones was sectioned at the smallest breadth (SB) of its diaphysis where

the compact bone is thick and provides a large area for the study of the bone tissue microstructure. In total, 10 transversal sections of the femur diaphysis were cut. The obtained segments were macerated and degreased. Later the samples were glued (Eukitt) onto a matted slide and cut using a diamond disk. Afterwards the bone fragments were ground by a laboratory grinder (Montasupal). The final thickness of the sections was approximately 100 microns. For the examination, an optical microscope Jenaval (Carl Zeiss Jena) with a digital CCD camera (Mintrow) at a magnification of 200x were used. Photographic documentation of the slides was made using computer programs Ati Player 5.2 (Ati Technol. Inc.) and Adobe Photoshop 5.0. Altogether 40 digital images were obtained. The qualitative characteristics were determined according to the generally known and internationally accepted classifications by Enlow and Brown (1956 ), Rämsch and Zerndt (1963), and Gladuhsew (1964). The quantitative characteristics were found out using the computer software Scion Image (Scion Corporation). The following parameters were measured: area, perimeter, minimal and maximal diameters of 200 primary vascular canals of primary osteons, of 40 haversian canals of secondary osteons, and of 40 secondary osteons. The basic statistical characteristics of the position and variability for each of the particular parameters were counted using the Excel 2000 software package.

RESULTS

The femurs of all the analysed animals had the following microstructure in

common. The arrangement and distribution of different bone tissue types is given

from the medullary cavity towards the periosteal surface:

  • the inner layer surrounding the medullary cavity is formed by a zone of

lamellar bone tissue which contains longitudinally arranged primary vascular

canals. The areas between the neighbouring primary osteons are often very large

and thus give an avascular appearance to the lamellar tissue mainly in the

anteromedial sides.

  • then there follows the layer of irregular haversian bone tissue. This is

characterised by scattered, isolated and relatively scarce haversian systems. Some

areas of dense haversian remodelling occur mainly in the posteromedial and

posterolateral sides ( Fig. 1 ).

  • further towards the periosteal surface, the haversian tissue is gradually

replaced by primary vascular radial ( Fig. 2 ), but predominantly longitudinal bone

tissue.

  • finally, primary vascular longitudinal bone tissue appears on the dorsal parts

of all the investigated bones. The vascular canals run in a direction essentially

parallel to the long axis of the bone ( Fig. 3 ).

All in all, 200 primary vascular canals of primary osteons, 40 haversian canals

of secondary osteons, and 40 secondary osteons were measured. The results are

shown in Table 1.

DISCUSSION

The results given in Table 1 showed that the mean diameter of primary vascular

canal of primary osteon (counted as arithmetic mean of minimal and maximal

diameter of 200 primary vascular canals of 200 primary osteons) is 12.49 μm.

Fig. 1 Dense Haversian bone tissue (magnification 200x)

Fig. 2 Primary vascular radial bone tissue (magnification 200 x)

Fig. 3 Primary vascular longitudinal bone tissue (magnification 200x)

rabbit femur diaphysis of New Zealand White albino breed. The results could be

applied in archaeozoology for the identification of species from bone fragments.

CONCLUSION

We investigated qualitative and quantitative characteristics of the rabbit femur

diaphysis microstructure. Our results revealed that the investigated bone tissue is

in general composed of primary longitudinal bone tissue. In posteromedial and

posterolateral sides there are areas of dense haversian bone tissue. The secondary

osteons like the primary osteons contain short vascular canals as documented in

Table 1.

A c k n o w l e d g e m e n t

We thank Ing. Peter Chrenek, PhD. (Research Institute of Animal Production, Nitra) for providing the rabbit skeletal material, and RNDr. Ján Spi‰iak, CSc. (Institute of Geology, Slovak Acad. Sci., Banská Bystrica) for assistance with thin section grinding. We acknowledge the kind assistance of RNDr. Roman Kuna, PhD. and RNDr. Radoslav Omelka (Department of Botany and Genetics, Constantine the Philosopher University, Nitra) with photographic documentation. This study was supported by the UKF grant: GAM 21/2002/FPV