ABSTRACT
Children with chronic renal failure in general present growth retardation that is aggravated by corticosteroids. We describe here the effects of methylprednisolone (MP) and recombinant human growth hormone (rhGH) on the growth plate (GP) of uremic rats. Uremia was induced by subtotal nephrectomy in 30-day-old rats, followed by 20 IU kg-1 day-1 rhGH (N = 7) or 3 mg kg-1 day-1 MP (N = 7) or 20 IU kg-1 day-1 rhGH + 3 mg kg-1 day-1 MP (N = 7) treatment for 10 days. Control rats with intact renal function were sham-operated and treated with 3 mg kg-1 day-1 MP (N = 7) or vehicle (N = 7). Uremic rats (N = 7) were used as untreated control animals. Structural alterations in the GP and the expression of anti-proliferating cell nuclear antigen (PCNA) and anti-insulin-like growth factor I (IGF-I) by epiphyseal chondrocytes were evaluated. Uremic MP rats displayed a reduction in the proliferative zone height (59.08 ± 4.54 vs 68.07 ± 7.5 æm, P < 0.05) and modifications in the microarchitecture of the GP. MP and uremia had an additive inhibitory effect on the proliferative activity of GP chondrocytes, lowering the expression of PCNA (19.48 ± 11.13 vs 68.64 ± 7.9 percent in control, P < 0.0005) and IGF-I (58.53 ± 0.96 vs 84.78 ± 2.93 percent in control, P < 0.0001), that was counteracted by rhGH. These findings suggest that in uremic rats rhGH therapy improves longitudinal growth by increasing IGF-I synthesis in the GP and by stimulating chondrocyte proliferation.
Subject(s)
Animals , Female , Humans , Rats , Glucocorticoids/pharmacology , Growth Plate/drug effects , Human Growth Hormone/pharmacology , Methylprednisolone/pharmacology , Uremia/metabolism , Autoantibodies/metabolism , Cell Proliferation , Chondrocytes/drug effects , Growth Plate/metabolism , Growth Plate/pathology , Insulin-Like Growth Factor I/metabolism , Proliferating Cell Nuclear Antigen/metabolism , Rats, Wistar , Tibia/drug effects , Tibia/pathology , Uremia/pathologyABSTRACT
The precise nature of hormones and growth factors directly responsible for cartilage maturation is still largely unclear. Since longitudinal bone growth occurs through endochondral bone formation, excess or deficiency of most hormones and growth factors strongly influences final adult height. The structure and composition of the cartilaginous extracellular matrix have a critical role in regulating the behavior of growth plate chondrocytes. Therefore, the maintenance of the three-dimensional cell-matrix interaction is necessary to study the influence of individual signaling molecules on chondrogenesis, cartilage maturation and calcification. To investigate the effects of insulin on both proliferation and induction of hypertrophy in chondrocytes in vitro we used high-density micromass cultures of chick embryonic limb mesenchymal cells. Culture medium was supplemented with 1 percent FCS + 60 ng/ml (0.01 æM) insulin and cultures were harvested at regular time points for later analysis. Proliferating cell nuclear antigen immunoreactivity was widely detected in insulin-treated cultures and persisted until day 21 and [ H]-thymidine uptake was highest on day 14. While apoptosis increased in control cultures as a function of culture time, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-labeled cells were markedly reduced in the presence of insulin. Type II collagen production, alkaline phosphatase activity and cell size were also lower in insulin-treated cultures. Our results indicate that under the influence of 60 ng/ml insulin, chick chondrocytes maintain their proliferative potential but do not become hypertrophic, suggesting that insulin can affect the regulation of chondrocyte maturation and hypertrophy, possibly through an antiapoptotic effect
Subject(s)
Animals , Chick Embryo , Cell Differentiation , Chondrocytes , Insulin , Mesoderm , Apoptosis , Cell Culture Techniques , Cell Division , Extracellular Matrix , Extremities , MesodermABSTRACT
Dow's syndrome (DS) is associated with cellular and humoral immunological abnormalities, and altered histology of the lymphoid and epithelial compartments of the thymus has been reported. In the present study, we investigated the extracellular matrix (ECM) of six DS thymuses. A novel fibroblast-containing network was demonstrated in the cortical region with anti-fibronectin, anti-type I collagen and anti-type IV collagen antibodies. Rupture sites of the baement membrane were also observed. Since the ECM is essential to normal thymic ontogeny, it is likely that the severe intrathymic ECM structure changes occuring in DS may be linked to abnormal cortical thymocyte depletion