HYDROXYAPATITE FORMATION IN A DYNAMIC COLLAGEN GEL SYSTEM - EFFECTS OF TYPE-I COLLAGEN, LIPIDS, AND PROTEOGLYCANS
HYDROXYAPATITE FORMATION IN A DYNAMIC COLLAGEN GEL SYSTEM - EFFECTS OF TYPE-I COLLAGEN, LIPIDS, AND PROTEOGLYCANS
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DOI：
10.1021/j100341a086
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发表时间：
1989-02-23
影响因子：
--
通讯作者：
BOSKEY, AL
中科院分区：
文献类型：
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作者：
BOSKEY, AL
在变性胶原凝胶系统中监测羟基磷灰石的形成，通过该系统，钙和磷酸盐溶液以恒定速率从“无限储液器”循环。当使用2-6 mL体积的10%明胶凝胶时，钙和磷酸盐的扩散系数分别为6.0 × 10 ~（-6）和3.9 × 10 ~（-6）cm ~ 2/s。在不存在任何其他大分子的情况下，在5.5±0.5天后，在距离钙溶液循环通过的末端1.54±0.02 mL（3.08 cm）的点处，在3 mL凝胶中可检测到羟基磷灰石形成。在该时间点，观察到的中央沉淀剂带附近的钙和磷酸盐含量为37 mM 2。通过X射线衍射、电子显微镜和化学分析证实了羟基磷灰石的存在。在对照凝胶中发生矿化的部位加入0.1 mL的乳酸I型胶原纤维（1 mg/mL）或合成复合酸性磷脂（0.3-1.2 mg/mL），可减少形成第一个可观察到的矿物存款所需的时间。第5天，脂质增加了相对于对照凝胶的矿物质形成量。包含0.1 mL的4-10 mg/mL关节软骨蛋白聚糖聚集体或单体制剂可在5天期间防止矿物质沉积。0.1 mL中央带中包含的羟基磷灰石种子（0.5-5 mg/mL）增殖，在2-6天内显示出高度可重复、可检测的矿物质含量增加。这种独特的动态凝胶系统的优点是在其他大分子存在下研究羟基磷灰石的形成和/或增殖，包括重现性和只需要少量的大分子。
Hydroxyapatite formation was monitored in a denatured collagen gel system through which calcium and phosphate solutions circulated at a constant rate from an “infinite reservoir”. With the use of 10% gelatin gels, 2-6 mL in volume, the diffusion coefficients for calcium and phosphate were 6.0 X10-6 and 3.9 X 10-6 cm2/s, respectively. In the absence of any other macromolecules, hydroxyapatite formation was detectable in the 3-mL gels at a point 1.54±0.02 mL (3.08 cm) from the end through which the calcium solution was being circulated after 5.5±0.5 days. At this time point the observed calcium and phosphate content adjacent to the centralprecipitant band was 37 mM2. The presence of hydroxyapatite was verified by X-ray diffraction, electron microscopy, and chemical analyses. Inclusionof 0.1 mL of lathyritic type I collagen fibers (1 mg/mL) or synthetic complexed acidic phospholipids (0.3-1.2 mg/mL) at the site where mineralization occurred in control gels decreased the time required for the formation of the first observable mineral deposit. The lipids increased the amount of mineralformed relative to the control gels atday 5. Inclusion of 0.1 mL of 4-10 mg/mL articular cartilage proteoglycan aggregate or monomer preparations prevented mineral deposition during the 5-day period. Hydroxyapatite seeds (0.5-5 mg/mL) included in the 0.1-mL central band proliferated, showing highly reproducible, detectable increases in mineral content at 2-6 days. The advantages of this unique dynamic gel system for the study of hydroxyapatite formation and/or proliferation in the presence of other macromolecules include reproducibility and the need for only smallamounts of macromolecules.