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Collagen metabolism of human osteoarthritic articular cartilage as modulated by bovine collagen hydrolysates

机译:牛胶原蛋白水解物调节人骨关节炎关节软骨的胶原蛋白代谢

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摘要

Destruction of articular cartilage is a characteristic feature of osteoarthritis (OA). Collagen hydrolysates are mixtures of collagen peptides and have gained huge public attention as nutriceuticals used for prophylaxis of OA. Here, we evaluated for the first time whether different bovine collagen hydrolysate preparations indeed modulate the metabolism of collagen and proteoglycans from human OA cartilage explants and determined the chemical composition of oligopeptides representing collagen fragments. Using biophysical techniques, like MALDI-TOF-MS, AFM, and NMR, the molecular weight distribution and aggregation behavior of collagen hydrolysates from bovine origin (CH-Alpha®, Peptan™ B 5000, Peptan™ B 2000) were determined. To investigate the metabolism of human femoral OA cartilage, explants were obtained during knee replacement surgery. Collagen synthesis of explants as modulated by 0–10 mg/ml collagen hydrolysates was determined using a novel dual radiolabeling procedure. Proteoglycans, NO, PGE2, MMP-1, -3, -13, TIMP-1, collagen type II, and cell viability were determined in explant cultures. Groups of data were analyzed using ANOVA and the Friedman test (n = 5–12). The significance was set to p=0.05. We found that collagen hydrolysates obtained from different sources varied with respect to the width of molecular weight distribution, average molecular weight, and aggregation behavior. None of the collagen hydrolysates tested stimulated the biosynthesis of collagen. Peptan™ B 5000 elevated NO and PGE2 levels significantly but had no effect on collagen or proteoglycan loss. All collagen hydrolysates tested proved not to be cytotoxic. Together, our data demonstrate for the first time that various collagen hydrolysates differ with respect to their chemical composition of collagen fragments as well as by their pharmacological efficacy on human chondrocytes. Our study underscores the importance that each collagen hydrolysate preparation should first demonstrate its pharmacological potential both in vitro and in vivo before being used for both regenerative medicine and prophylaxis of OA.
机译:关节软骨的破坏是骨关节炎(OA)的特征。胶原蛋白水解物是胶原蛋白肽的混合物,作为预防骨关节炎的营养剂已引起了公众的广泛关注。在这里,我们首次评估了不同的牛胶原蛋白水解产物制剂是否确实调节了人OA软骨外植体的胶原蛋白和蛋白聚糖的代谢,并确定了代表胶原蛋白片段的寡肽的化学组成。使用生物物理技术(如MALDI-TOF-MS,AFM和NMR),测定了源自牛的胶原蛋白水解物的分子量分布和聚集行为(CH-Alpha®,Peptan™B 5000,Peptan™B 2000)。为了研究人股骨OA软骨的代谢,在膝关节置换手术中获得了外植体。使用新型的双重放射标记方法确定了受0-10 mg / ml胶原水解物调节的外植体胶原合成。在外植体培养物中确定蛋白聚糖,NO,PGE2,MMP-1,-3,-13,TIMP-1,II型胶原和细胞活力。使用方差分析和弗里德曼检验分析数据组(n = 5–12)。显着性设定为p = 0.05。我们发现,从不同来源获得的胶原蛋白水解物的分子量分布宽度,平均分子量和聚集行为各不相同。测试的胶原蛋白水解产物均未刺激胶原蛋白的生物合成。 Peptan™B 5000显着提高了NO和PGE2的水平,但对胶原蛋白或蛋白聚糖的损失没有影响。测试的所有胶原蛋白水解物均证明没有细胞毒性。总之,我们的数据首次证明了各种胶原蛋白水解产物在胶原蛋白片段的化学组成以及它们对人软骨细胞的药理作用方面均不同。我们的研究强调了每种胶原蛋白水解物制剂在用于再生医学和预防OA之前首先应在体内和体外证明其药理潜力的重要性。

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