Effect of X-ray Irradiation on Extracellular Matrix Components of Rat Osteoblasts and Femurs

The effect of X-irradiation on extracellular matrix components was examined in cultured rat fetal osteoblasts and rat femurs. The osteoblasts were X-irradiated at 2 Gy or 5 Gy, and gene expression of matrix metalloproteinase (MMP)-2, MMP-13, type-I collagen and decorin were analyzed by real-time polymerase chain reaction. Hyaluronan in the medium was also determined after 48 h of culture. Expression of the MMP-13 gene was found to be increased, whereas those of the type-I collagen and decorin genes tended to be decreased. Synthesis of hyaluronan and expression of the MMP-2 gene were not significantly affected. We then subjected SpragueDawley rats to X-irradiation at 2 Gy or 5Gy. After 1-3 months, the rats were euthanatized and the glycosaminoglycan fractions were purified from their femurs. These fractions were analyzed for their uronic acid content, sensitivity to mucopolysaccharidases, glycosaminoglycan chain length, and unsaturated disaccharide units. It was found that the uronic acid content was decreased by X-irradiation. The main glycosaminoglycan in rat femur was identified as dermatan sulfate, and its chain length was not affected by X-irradiation. The unsaturated disaccharide unit of the glycosaminoglycan was found to be ∆Di-4S, and no apparent structural alteration of the glycosaminoglycan was observed after X-irradiation. The core protein and dermatan sulfate chain of decorin have been reported to bind with type-I collagen and to type-IX collagen, which binds to type-I collagen, respectively. The present results thus suggest that the decrease in both decorin and type I-collagen levels in rat bone after X-irradiation may contribute to a loss of mechanical strength.

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