Department of Morphology, Laboratory of Histology and Embryology, Dental School, UNESP - Univ. Estadual Paulista, Araraquara, São Paulo, Brazil.
J Anat. 2010 Aug;217(2):116-25. doi: 10.1111/j.1469-7580.2010.01249.x. Epub 2010 Jun 16.
During the active tooth eruption process, structural changes in the lamina propria are necessary to provide extracellular matrix remodelling and for the establishment of the eruptive pathway. A large number of resident cells, recruited cells and proteases have been demonstrated in the eruptive process, but the participation of MMP-9 and mast cells has not yet been demonstrated. In this study, we set out to evaluate the intensity of MMP-9 immunoexpression, the frequency of mast cells and the correlation between the incidence of mast cells and bone resorption in different phases of tooth eruption. Fragments of maxilla containing first molars, obtained from 9-, 11-, 13- and 16-day-old rats, were fixed in 4% formaldehyde, decalcified and embedded in paraffin. Sagittal sections were stained with Masson's trichrome or submitted to the tartrate-resistant acid phosphatase method for quantification of osteoclasts. Sections stained by 1% toluidine blue were used for quantification of metachromatic mast cells mm(-2) of lamina propria. The expression of MMP-9 in the lamina propria was evaluated by immunohistochemistry. In the 9-day-old rats, the lamina propria contained few mast cells and occasional osteoclasts were found in the bone surface overlying the occlusal portion of the tooth germs. Otherwise, a significant increase in the number of mast cells was observed in the intra-osseous phase of tooth eruption (11-day-old rats), period in which numerous TRAP-positive osteoclasts were found in the bone surface. MMP-9 immunolabelling was detected in fibroblasts, mast cells and macrophage-like cells of the lamina propria in all ages studied. However, an enhanced immunolabelling was evident in the advanced phase of tooth eruption (16-day-old rats). During the intra-osseous phase, the parallel between the high frequency of both mast cells and osteoclasts suggests that mast cells could exert a paracrine function on the osteoclasts and then stimulate bone resorption. The immunoexpression of MMP-9 in different cells of lamina propria, including mast cells, indicates that this enzyme participates in the degradation of ECM, mainly during late phase of mucosal penetration. Thus mast cells and MMP-9 are involved in the complex process of degradation of the eruptive pathway extracellular matrix.
在牙齿的主动萌出过程中,固有层的结构变化对于提供细胞外基质重塑和建立萌出途径是必要的。在萌出过程中已经证明了大量的固有层常驻细胞、募集细胞和蛋白酶的存在,但 MMP-9 和肥大细胞的参与尚未得到证实。在这项研究中,我们旨在评估 MMP-9 免疫表达的强度、肥大细胞的频率以及不同萌出阶段肥大细胞的发生率与骨吸收之间的相关性。从 9、11、13 和 16 天大的大鼠获得含有第一磨牙的上颌骨片段,用 4%甲醛固定,脱钙并包埋在石蜡中。矢状切片用 Masson 三色染色或用抗酒石酸酸性磷酸酶法染色以定量破骨细胞。用 1%甲苯胺蓝染色的切片用于测量固有层的变色肥大细胞 mm(-2)。通过免疫组织化学评估 MMP-9 在固有层中的表达。在 9 天大的大鼠中,固有层含有少量肥大细胞,在覆盖牙胚咬合部分的骨表面偶尔可以发现破骨细胞。否则,在牙齿萌出的骨内阶段(11 天大的大鼠)观察到肥大细胞数量显著增加,在此期间在骨表面发现了大量的 TRAP 阳性破骨细胞。在所有研究的年龄中,MMP-9 免疫标记都在固有层的成纤维细胞、肥大细胞和巨噬细胞样细胞中检测到。然而,在牙齿萌出的晚期(16 天大的大鼠),免疫标记明显增强。在骨内阶段,肥大细胞和破骨细胞高频出现的平行关系表明,肥大细胞可能对破骨细胞发挥旁分泌作用,然后刺激骨吸收。固有层中不同细胞的 MMP-9 免疫表达,包括肥大细胞,表明该酶参与 ECM 的降解,主要在黏膜穿透的晚期阶段。因此,肥大细胞和 MMP-9 参与了萌出途径细胞外基质的降解这一复杂过程。