Malemud C J, Papay R S, Hering T M, Holderbaum D, Goldberg V M, Haqqi T M
Department of Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4946, USA.
Osteoarthritis Cartilage. 1995 Dec;3(4):227-38. doi: 10.1016/s1063-4584(05)80014-7.
The proteoglycans synthesized by human osteoarthritic femoral head cartilage and nonarthritic articular cartilage age-matched to the osteoarthritic cartilage specimens was studied in explant cultures and in chondrocytes generated by explant outgrowth from the cartilages. Twenty-four hours after explanation, both nonarthritic articular cartilage and osteoarthritic cartilage synthesized principally one large proteoglycan core protein that migrated on 3-5% acrylamide gels with an apparent molecular mass (M(r)) of approximately 520 kDa after enzymatic digestion with chondroitinase ABC and keratanase. The proteoglycan was found in both the explant itself and in the medium compartment of the culture as well. This proteoglycan contained chondroitin-6-sulfate, keratan sulfate and the hyaluronan binding region as evidenced by immunoblotting with murine anti-proteoglycan monoclonal antibodies indicating that the proteoglycan was aggrecan. To a much lesser extent two additional proteoglycan core proteins were also found in the explant but were not seen in the culture medium compartment. These proteoglycans possessed apparent M(r)'s of approximately 480 kDa and approximately 390 kDa on 3-5% acrylamide gels after chondroitinase ABC and keratanase digestion. The medium compartment contained principally the approximately 520 kDa proteoglycan core protein. In osteoarthritic cartilage explants, the pattern of newly synthesized proteoglycans recovered from the tissue as assessed on 3-16% polyacrylamide gradient gels remained relatively the same from day 1 after explantation up to 36 days of culture. By contrast, the proteoglycans recovered from the culture medium contained chondroitin sulfate and keratan sulfate after 1, 7, and 21 days in culture but by 36 days appeared to contain only chondroitin sulfate. Chondrocytes generated from osteoarthritic cartilage and age-matched nonarthritic articular cartilage synthesized different patterns of large (greater than 200 kDa) proteoglycan. Whereas chondrocytes derived from osteoarthritic cartilage continued to synthesize principally the approximately 520 kDa proteoglycan core protein, the chondrocytes derived from nonarthritic cartilage synthesized in addition to this proteoglycan, abundant amounts of the other two proteoglycan core proteins as well.
对人骨关节炎股骨头软骨以及与骨关节炎软骨标本年龄匹配的非关节炎关节软骨所合成的蛋白聚糖，在组织外植体培养物以及由软骨外植体生长产生的软骨细胞中进行了研究。在取材24小时后，非关节炎关节软骨和骨关节炎软骨主要合成了一种大的蛋白聚糖核心蛋白，在用软骨素酶ABC和角蛋白酶进行酶消化后，该蛋白聚糖在3 - 5%丙烯酰胺凝胶上迁移，其表观分子量（M(r)）约为520 kDa。这种蛋白聚糖在外植体本身以及培养物的培养基部分均有发现。通过用鼠抗蛋白聚糖单克隆抗体进行免疫印迹证明，这种蛋白聚糖含有硫酸软骨素-6-硫酸酯、硫酸角质素和透明质酸结合区域，表明该蛋白聚糖是聚集蛋白聚糖。在程度上要小得多的情况下，在外植体中还发现了另外两种蛋白聚糖核心蛋白，但在培养基部分未观察到。在用软骨素酶ABC和角蛋白酶消化后，这些蛋白聚糖在3 - 5%丙烯酰胺凝胶上的表观M(r)约为480 kDa和约390 kDa。培养基部分主要含有约520 kDa的蛋白聚糖核心蛋白。在骨关节炎软骨外植体中，从组织中回收的新合成蛋白聚糖的模式，在取材后第1天直至培养36天，在3 - 16%聚丙烯酰胺梯度凝胶上评估时相对保持不变。相比之下，从培养基中回收的蛋白聚糖在培养1、7和21天后含有硫酸软骨素和硫酸角质素，但到36天时似乎只含有硫酸软骨素。由骨关节炎软骨和年龄匹配的非关节炎关节软骨产生的软骨细胞合成了不同模式的大（大于200 kDa）蛋白聚糖。源自骨关节炎软骨的软骨细胞继续主要合成约520 kDa的蛋白聚糖核心蛋白，而源自非关节炎软骨的软骨细胞除了这种蛋白聚糖外，还合成了大量的另外两种蛋白聚糖核心蛋白。
