Cam Y, Meyer J M, Staubli A, Ruch J V
Arch Anat Microsc Morphol Exp. 1986;75(2):75-89.
The functional differentiation of odontoblasts requires specific interactions between these cells and the extracellular matrix. To further analyze these phenomena we studied the effects of a "dental papillae biomatrix" on isolated dental papillae cultured in vitro. The dental papillae biomatrix was extracted from EDTA-dissociated day-18 mouse dental papillae by homogenization, NaCl and enzymatic treatments, and deposited on Millipore filters. This biomatrix was studied by means of transmission electron microscopy and indirect immunofluorescence: it contained collagen fibrils, type IV collagen, fibronectin and laminin; cellular residues were also observed. The dental papillae were isolated by trypsin treatment of homologous tooth germs and cultured on uncoated (control) and coated filters. As shown by histological and cytological data, odontoblast-like cells never differentiated in control cultures. In presence of biomatrix and serum, polarized functional cells were observed. The functional state of these cells was enhanced by the addition of ascorbic acid to the culture media. Study of the incorporation of 3H-proline in cultured dental papillae and in macromolecules secreted into the culture media corroborated the morphological findings.
成牙本质细胞的功能分化需要这些细胞与细胞外基质之间的特定相互作用。为了进一步分析这些现象,我们研究了“牙乳头生物基质”对体外培养的分离牙乳头的影响。牙乳头生物基质是通过匀浆、氯化钠和酶处理从第18天的EDTA解离的小鼠牙乳头中提取的,并沉积在微孔滤膜上。通过透射电子显微镜和间接免疫荧光对这种生物基质进行了研究:它含有胶原纤维、IV型胶原、纤连蛋白和层粘连蛋白;还观察到了细胞残留物。通过用胰蛋白酶处理同源牙胚分离出牙乳头,并在未包被(对照)和包被的滤膜上进行培养。组织学和细胞学数据显示,在对照培养物中从未分化出成牙本质样细胞。在存在生物基质和血清的情况下,观察到了极化的功能细胞。向培养基中添加抗坏血酸可增强这些细胞的功能状态。对培养的牙乳头和分泌到培养基中的大分子中3H-脯氨酸掺入情况的研究证实了形态学发现。