Altered Cellular Kinetics in the Growth Plate of the Femoral Head of Spontaneously Hypertensive Rats

PURPOSE: Pathologic changes in the growth plate remain unknown in Legg-Calve-Perthes (LCP) disease. Spontaneously hypertensive rats have proven to be a good model for studying LCP disease. This study investigated the histopathologic changes and the expression of vascular endothelial growth factor in the growth plate of spontaneously hypertensive rats (SHR). MATERIALS AND METHODS: Sixty SHR rats were divided into two groups: those showing osteonecrosis (SHR+n group: 32), and those showing normal ossification (SHR-n group: 28). Thirty Wister Kyoto rats served as a control. For histomorphological measurement, the length of each zone of the growth plate was measured. Cell kinetics was measured by 5-bromo-2'-deoxyuridin (BrdU) immunohistochemistry and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assays. Vascular endothelial growth factor (VEGF) immunohistochemistry was used to identify of expression of VEGF. RESULTS: The lengths of growth plates of the SHR+n group were significantly shorter in the initial growth period than those of the other groups. The lowest proliferative rate and the highest apoptosis rate were observed in the SHR+n group at the initial growth period. The expression of VEGF in the growth plate of the SHR group was lower than the control group, and it was lower in the SHR+n group than in the SHR-n group. CONCLUSION: The growth plate of the SHR+n group was found to be affected by disease process of ischemic necrosis of the femoral head, and this might explain the relative overgrowth of the greater trochanter in the later stages of LCP disease.

Altered Cellular Kinetics in Growth Plate according to Alterations in Weight Bearing

PURPOSE: To examine the effects of change in weight bearing on the growth plate metabolism, a simulated animal model of weightlessness was introduced and the chondrocytes' cellular kinetics was evaluated. MATERIALS AND METHODS: Unloading condition on the hind-limb of Sprague-Dawley rats was created by fixing a tail and lifting the hind-limb. Six rats aged 6 weeks old were assigned to each group of unloading, reloading, and control groups of unloading or reloading. Unloading was maintained for three weeks, and then reloading was applied for another one week thereafter. Histomorphometry for the assessment of vertical length of the growth plate, 5-bromo-2'-deoxyuridin immunohistochemistry for cellular kinetics, and biotin nick end labeling transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay for chondrocytes apoptosis in the growth plate were performed. RESULTS: The vertical length of the growth plate and the proliferative potential of chondrocytes were decreased in the unloading group compared to those of control groups. Inter-group differences were more significant in the proliferative and hypertrophic zones. Reloading increased the length of growth plate and proliferative potential of chondrocytes. However, apoptotic changes in the growth plate were not affected by the alterations of weight bearing. CONCLUSION: Alterations in the weight bearing induced changes in the chondrocytic proliferative potential of the growth plate, however, had no effects on the apoptosis. This may explain why non-weight bearing in various clinical situations hampers normal longitudinal bone growth. Further studies on the factors for reversibility of chondrocytic proliferation upon variable mechanical stresses are needed.

Caracterizaçäo da cultura primária de condrócitos a partir da cartilagem de costela de ratos / Characterization of a chondrocyte primary culture from rat rib resting cartilage

Neste trabalho, procurou-se padronizar e caracterizar a cultura primária de condrócitos obtidos a partir da cartilagem de costelas de ratos. As células foram cultivadas em meio de Eagle modificado por Dulbecco (DME), suplementado com 10% de soro Fetal Bovino e mantidas, durante os período de incunbaçäo, em atmosfera de ar contendo 5% de CO2. Pode-se obter alto rendimento (0,99 ñ 0,18 . 10**6 células/rato) e viabilidade celular (91.7%) pela digestäo enzimática da cartilagem pela colagenase. Após 2 a 3 dias do plaqueamento, as células aderem ao substrato e começam a se dividir formando monocamada. O tempo de adesäo mostrou-se independente da densidade de plaqueamento. O tempo de multiplicaçäo da cultura foi de 23,19 horas. A cultura é de fácil manutençäo, demonstrando ser, a cartilagem costal de rato, boa fonte de condrócitos