Boabaid Fernanda, Berry Janice E, Koh Amy J, Somerman Martha J, McCcauley Laurie K
Department of Periodontics/Prevention/Geriatrics, School of Dentistry, University of Michigan, Ann Arbor, MI 48109-1078, USA.
J Periodontol. 2004 Sep;75(9):1247-54. doi: 10.1902/jop.2004.75.9.1247.
Parathyroid hormone-related protein (PTHrP) promotes osteoclastogenesis by inhibiting expression of osteoprotegerin (OPG), a decoy receptor for the receptor activator of nuclear factor kappa B (RANK), and by enhancing production of RANK ligand (RANKL) by osteoblasts. However, little is known regarding the role of PTHrP in regulating cementoblast-mediated osteoclastogenesis.
This study determined the impact of PTHrP on osteoclastogenesis using: 1) OCCM-30 (immortalized murine cementoblasts), 2) RAW 264.7 cells (murine myeloid cells), or 3) OCCM-30 plus RAW 264.7 cells. Cells were treated with PTHrP (1-34), RANKL, or PTHrP and RANKL combined. Enzyme-linked immunosorbent assays (ELISAs) for OPG and RANKL were performed on media and cell lysates, and tartrate-resistant acid phosphatase (TRAP) and mRNA detection for the osteoclast associated receptor (OSCAR) were performed.
The highest numbers of TRAP-positive cells and cells expressing OSCAR were found in the RAW cell group treated with either RANKL alone or RANKL and PTHrP. TRAP-positive cells were fewer when OCCM cells were co-cultured with RAW, but the greatest numbers were still with both PTHrP and RANKL. OPG levels were highest from OCCM cells and PTHrP decreased these levels. In contrast, RANKL levels were low in OCCM cell lysates and PTHrP increased RANKL. In vivo studies also revealed high osteoclastic activity surrounding developing teeth in mice administered PTH.
These results demonstrate that PTHrP influences the balance of OPG and RANKL production by cementoblasts, and further indicate that this effect, in the context of surrounding cells, might have a significant impact on osteoclastogenesis, root resorption, and tooth eruption.
甲状旁腺激素相关蛋白(PTHrP)通过抑制骨保护素(OPG)的表达促进破骨细胞生成,OPG是核因子κB受体激活剂(RANK)的诱饵受体,并且通过增强成骨细胞产生RANK配体(RANKL)来促进破骨细胞生成。然而,关于PTHrP在调节成牙骨质细胞介导的破骨细胞生成中的作用知之甚少。
本研究使用以下方法确定PTHrP对破骨细胞生成的影响:1)OCCM-30(永生化小鼠成牙骨质细胞),2)RAW 264.7细胞(小鼠骨髓细胞),或3)OCCM-30加RAW 264.7细胞。细胞用PTHrP(1-34)、RANKL或PTHrP与RANKL联合处理。对培养基和细胞裂解物进行OPG和RANKL的酶联免疫吸附测定(ELISA),并进行抗酒石酸酸性磷酸酶(TRAP)和破骨细胞相关受体(OSCAR)的mRNA检测。
在用单独的RANKL或RANKL与PTHrP处理的RAW细胞组中发现TRAP阳性细胞和表达OSCAR的细胞数量最多。当OCCM细胞与RAW细胞共培养时,TRAP阳性细胞较少,但数量最多的仍是同时使用PTHrP和RANKL的情况。OPG水平在OCCM细胞中最高,PTHrP降低了这些水平。相反,OCCM细胞裂解物中RANKL水平较低,PTHrP增加了RANKL。体内研究还显示,给予PTH的小鼠发育牙齿周围的破骨细胞活性较高。
这些结果表明PTHrP影响成牙骨质细胞产生OPG和RANKL的平衡,并进一步表明在周围细胞的背景下,这种作用可能对破骨细胞生成、牙根吸收和牙齿萌出有重大影响。