DEVELOPMENT OF AN IN-VITRO CULTURE SYSTEM FOR THE STUDY OF OSTEOCLAST ACTIVITY AND FUNCTION
DEVELOPMENT OF AN IN-VITRO CULTURE SYSTEM FOR THE STUDY OF OSTEOCLAST ACTIVITY AND FUNCTION
复制标题
DOI：
10.1016/s0099-2399(06)80094-4
复制
发表时间：
1994-07-01
影响因子：
4.2
通讯作者：
WINDELER, AS
中科院分区：
文献类型：
--
作者：
GILLES, JA;CARNES, DL;WINDELER, AS
由于矿化组织的复杂结构，吸收是一个很难在体内研究的过程。本研究的目的是建立一种研究破骨细胞附着和活性的体外培养系统。使用射频溅射技术在聚碳酸酯过滤器上涂覆一层薄薄的羟基磷灰石，以制备透明、微孔的基片，先前涂覆了I型胶原。从炎性吸收的大鼠磨牙根表面分离出破骨细胞和从大鼠长骨内膜表面分离的破骨细胞分别培养在实验基质上。该培养系统被证明是成功的，支持破骨细胞的活性和活性，扫描电子显微镜下观察到细胞附着和铺展。通过使用荧光探针丫啶橙监测产酸，进一步证明了活性。当分离的破骨细胞或牙本质破骨细胞与合成的底物结合时，观察到的反应机制与体内发生的机制相似。因此，开发的系统允许在体外系统中对破骨细胞进行基本观察。
Resorption is a difficult process to study in vivo because of the complex organization of mineralized tissue. The objective of this study was to develop an in vitro culture system for studying osteoclast attachment and activity. Transparent, microporous substrata were prepared using radio frequency-sputtering techniques to apply a thin layer of hydroxyapatite on polycarbonate filters previously coated with Type I collagen. Dentinoclasts isolated from the root surface of rat molar teeth undergoing inflammatory resorption and osteoclasts isolated from the endosteal surfaces of rat long bones were incubated on the experimental substrata. The culture system proved successful, supporting osteoclast viability and activity as demonstrated by cellular attachment and spreading seen by scanning electron microscopy. Activity was further demonstrated by monitoring acid production using a fluorescent probe, acridine orange. Isolated osteoclasts or dentinoclasts, when combined with synthetically prepared substrata, were observed to respond by mechanisms comparable to those that occur in vivo. Therefore, the system developed permits basic observations of osteoclasts in an in vitro system.