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. 2015 Mar-Apr;23(2):135-44.
doi: 10.1590/1678-775720140319.

Evaluation of protein undernourishment on the condylar process of the Wistar rat mandible correlation with insulin receptor expression

Marcelo Arthur Cavalli  1 , Aline Gonçalves  1 , Joice Naiara Bertaglia Pereira  1 , Jodonai Barbosa da Silva  1 , Silvia de Campos Boldrini  2 , Edson Aparecido Liberti  2
Affiliations

Evaluation of protein undernourishment on the condylar process of the Wistar rat mandible correlation with insulin receptor expression

Marcelo Arthur Cavalli et al. J Appl Oral Sci. 2015 Mar-Apr.

Abstract

The mandible condylar process cartilage (CP) of Wistar rats is a secondary cartilage and acts as a mandibular growth site. This phenomenon depends on adequate proteins intake and hormone actions, including insulin.

Objectives: The present study evaluated the morphological aspects and the expression of the insulin receptor (IR) in the cartilage of the condylar process (CP) of rats subjected to protein undernourishment.

Material and methods: The nourished group received a 20% casein diet, while the undernourished group (U) received a 5% casein diet. The re-nourished groups, R and RR, were used to assess the effects of re-nutrition during puberty and adulthood, respectively. CPs were processed and stained with picro-sirius red, safranin-O and azocarmine. Scanning electron microscopy and immunohistochemistry were also performed.

Results: The area of the CP cartilage and the number of cells in the chondroblastic layer decreased in the U group, as did the thickness of the CP layer in the joint and hypertrophic layer. Renourishment during the pubertal stage, but not during the adult phase, restored these parameters. The cell number was restored when re-nutrition occurred in the pubertal stage, but not in the adult phase. The extracellular matrix also decreased in the U group, but was restored by re-nutrition during the pubertal stage and further increased in the adult phase. IR expression was observed in all CPs, being higher in the chondroblastic and hypertrophic cartilage layers. The lowest expression was found in the U and RR groups.

Conclusions: Protein malnutrition altered the cellularity, the area, and the fibrous cartilage complex, as well as the expression of the IRs.

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Figures

Figure 1
Figure 1. Schematic representation of experimental group formation
Figure 2
Figure 2. Sagittal mandible condylar process cartilage (CP) of Wistar rats. Picro-sirius red under light polarized coloring method. Collagen type I (yellow, orange and red) and III (green) detection. A: Articular layers (a), pre-chondroblastic (p), chondroblastic (c) and hypertrophic (h) of the CP; B - F (40X): Distribution of collagen types I and III in the matrix of fibrous cartilage of CP in groups N, nourished from day 21 to day 60; U, undernourished from day 21 to day 60; R, renourished from day 21 to day 60; NN, nourished until day 100; and RR, renourished from day 21 to day 100
Figure 3
Figure 3. Scanning electron microscopy (SEM) of the mandible condylar process cartilage (CP) of Wistar rats of groups N, nourished from day 21 to day 60; U, undernourished from day 21 to day 60; R, renourished from day 21 to day 60; and NN, nourished until day 100. A - D: 500X; E - H: 700X; I - L: 1200X
Figure 4
Figure 4. Analysis of mandible condylar process cartilage layers thickness (TCPC)
Figure 5
Figure 5. Light microscopy of the mandible condylar process cartilage (CP) of Wistar rats. Azo-carmine staining. A: group N, nourished from day 21 to day 60, with CP layers: articular (a), pre-chondroblastic (p), chondroblastic (c) and hypertrophic (h); B: group U, undernourished from day 21 to day 60; C: group R, renourished from day 21 to day 60; D: group NN, nourished until day 100; and E: group RR, renourished from day 21 to day 100
Figure 6
Figure 6. Insulin Receptors (IR) of the mandible condylar process cartilage (CP) of Wistar rats. A - C: IR immunohistochemistry; D - F: IR immunofluorescence. The arrows show the sensitized cells to IR (immunostained) and arrowheads point to cells not sensitized
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