The cloning and application of growth factors in restorative dentine

这项研究首先试图找到未知的基因，这些基因专门表达成牙本质母细胞产生的修复牙本质。接下来，为了证实外源基因可以转移到大鼠牙髓中，并阐明腺病毒载体在体内的转移有效性。当用钻石点法制备牙齿时，我们通过消减杂交从牙髓中检测到14个特异性克隆，并将携带LacZ基因的重组腺病毒注射到6周龄Wistar大鼠的牙洞中。注射后1周，解剖牙体，X-Gal染色检测LacZ的表达。提取小鼠肝、肾、心、脑组织总RNA，RT-PCR检测LacZ基因在其他组织中的表达。洞内可见少量成牙本质细胞表达LacZ。在其他器官中未观察到外源基因的表达。基于这些发现，用腺病毒载体将基因直接导入牙洞是一种有效的体内方法。

This study first sought to find unknown genes that expressed specifically to be made the restorative dentine by odontblast. Next, to confirm that foreign genes can be transferred to dental pulp of rats and to clarify the in vivo transfer availability of the adenovirus vectors.When tooth was prepared the cavity by diamond points, we detected the specific 14 clones from dental pulp by subtractive hybridization.Recombinant adenovirus harboring LacZ gene was injected into the prepared tooth cavity of 6-week-old Wistar rats. At 1week after injection, the tooth was dissected and X-gal staining examined the expression of delivered LacZ. To investigate the expression of transferred gene in other organs, total RNA was extracted from liver, kidney, heart, and brain and expression of LacZ mRNA were analysed by RT-PCR. Expression of LacZ was observed a few odontblasts in the cavity. In the other organs, expression of the delivered transgene was not observed. Based on these findings, direct gene delivery into the tooth cavity using adenovirus vector is feasible as an effective in vivo method.