Effects of bone CS-proteoglycans, DS-decorin, and DS-biglycan on hydroxyapatite formation in a gelatin gel
Effects of bone CS-proteoglycans, DS-decorin, and DS-biglycan on hydroxyapatite formation in a gelatin gel
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DOI：
10.1007/s002239900339
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发表时间：
1997-10-01
影响因子：
4.2
通讯作者：
Rosenberg, L
中科院分区：
文献类型：
--
作者：
Boskey, AL;Spevak, L;Rosenberg, L
富含亮氨酸的小骨蛋白多糖、双糖链蛋白聚糖和核心蛋白聚糖可以通过羟基磷灰石柱色谱进行纯化，证明它们对骨磷灰石具有潜在的亲和力。为了确定它们对体外磷灰石形成和生长的影响，在明胶凝胶扩散系统中分析了分别从皮肤和关节软骨中获得的硫酸软骨素（CS）骨蛋白聚糖的混合物或含有蛋白聚糖、DS-核心蛋白和DS-双聚糖的硫酸皮肤素（DS）的纯化级分，其中 在没有蛋白质的情况下，3.5 天内会形成磷灰石。 3.5天时，低浓度的骨CS-蛋白聚糖混合物和低浓度的DS-双聚糖(5-25μg/ml)相对于无蛋白聚糖的对照增加了磷灰石的形成。 CS-蛋白多糖混合物在50μg/ml时的效果低于在10μg/ml时的效果。同样，DS-双糖链蛋白在 5-25 μg/ml 时最有效。第5天：当评估磷灰石生长和增殖时，10和50μg/ml的CS-骨蛋白聚糖和DS-双糖链蛋白聚糖均增加了矿物质产量。相比之下，DS-核心蛋白聚糖在任何这些浓度下对矿物质积累都没有显着影响。在种子生长实验中，1和10μg/ml CS-蛋白聚糖和10和50μg/ml DS-双糖链蛋白是矿物质沉积的显着有效抑制剂，而DS-核心蛋白聚糖没有表现出抑制种子生长的倾向。使用摩尔消光系数确定浓度，并使用 Langmuir 吸附等温线模型确定 DS-双糖链蛋白聚糖和 DS-核心蛋白聚糖与磷灰石（比表面积 54 m(2)/g）的结合。 DS-双糖链蛋白聚糖对磷灰石的亲和力比 DS-核心蛋白聚糖更高（0.285 ml/mu mol 与 0.0098 ml/mu mol）。 DS-双糖链蛋白聚糖结合具有更高的特异性，结合位点较少（3.5 μ mol/m(2)，而 DS-核心蛋白聚糖为 18.2 μ mol/m(2)）。数据表明，在小蛋白聚糖中，双聚糖在矿化调节中可能比核心蛋白聚糖发挥更重要的作用。
The small leucine-rich bone proteoglycans, biglycan and decorin, can be purified by chromatography on hydroxyapatite columns, demonstrating their potential affinities for bone apatite. To determine their effects on in vitro apatite formation and growth, a mixture of the chondroitin-sulfate (CS) bone proteoglycans, or purified fractions of the dermatan sulfate (DS) containing proteoglycans, DS-decorin and DS-biglycan obtained from skin and articular cartilage, respectively, were analyzed in a gelatin gel diffusion system in which apatite formation occurs in the absence of proteins in a 3.5 day period. Low concentrations of the bone CS-proteoglycan mixture and low DS-biglycan concentrations (5-25 mu g/ml) increased apatite formation relative to proteoglycan-free controls at 3.5 days. The CS-proteoglycan mixture was less effective at 50 mu g/ml than at 10 mu g/ml. DS-biglycan was similarly most effective at 5-25 mu g/ml. At 5 days: when apatite growth and proliferation were assessed, 10 and 50 mu g/ml of both CS-bone proteoglycan and DS-biglycan increased mineral yields. DS-decorin, in contrast, had no significant effect on mineral accumulation at any of these concentrations. In seeded growth experiments, 1 and 10 mu g/ml CS-proteoglycan and 10 and 50 mu g/ml DS-biglycan were significant effective inhibitors of mineral accretion, whereas DS-decorin showed no tendency to inhibit seeded growth. Using molar extinction coefficients to determine concentrations, the binding of DS-biglycan and DS-decorin to apatite (specific surface 54 m(2)/g) was determined using a Langmuir adsorption iso-therm model. DS-biglycan had a greater affinity for apatite than DS-decorin (0.285 ml/mu mol versus 0.0098 ml/mu mol). DS-biglycan binding was more specific with fewer binding sites (3.5 mu mol/m(2) compared with 18.2 mu mol/m(2) for DS-decorin). Data suggest that of the small proteoglycans, biglycan may play a more significant role than decorin in the regulation of mineralization.