Accorsi-Mendonça Thais, Zambuzzi Willian Fernando, da Silva Paiva Katiúcia Batista, Pereira Lauris José Roberto, Cestari Tânia Mary, Taga Rumio, Granjeiro José Mauro
Department of Biological Sciences, Bauru Dental School, University of Sao Paulo, Brazil.
J Mol Histol. 2005 May;36(4):311-6. doi: 10.1007/s10735-005-7018-9.
The purpose of the study was to analyze the involvement of metalloproteinase 2 (MMP-2) and macrophages in the tissue and cell response to the organic graft material produced from bovine cancellous bone. Thirty adult male white Wistar rats (Rattus norvegicus) received implants of blocks of demineralized bovine bone matrix between the fasciae of the quadriceps muscle. The specimens collected at 3, 7, 14, 21 and 28 days after implantation (n = 6/period). Sections of 6 microm thick were stained with hematoxylin and eosin and immunolabeled with anti-MMP-2 and anti-CD68 using standard avidin-biotin-peroxidase method. The tissue response to the material was initially mediated by polymorphonuclear neutrophils, evolving to a mononuclear inflammatory infiltrate with macrophages and few lymphocytes and plasma cells and presence of inflammatory multinucleated giant cells (GC) in contact with the material that exhibited signs of resorption. The number of cells immunolabeled to MMP-2 was highest at day 7 (103.2 +/- 39.1), but significantly decreased (F = 3.67; p = 0.044) until day 28 (45.9 +/- 13.1). CD68 immunostaining also significantly decreased (F = 6.75; p = 0.007) from day 7 (49.5 +/- 10.4) to day 28 (19.5 +/- 8.9). A positive and statistically significant correlation was observed between the evolutions of these two variables. The material had been almost completely resorbed at day 28. Among cells present at the granuloma, anti-MMP-2 immunostaining was predominant and more intense in macrophages, yet lightly immunolabeled multinucleated giant cells were found in close contact with the material. Thus, considering the experimental limitations of this study, we concluded that MMP-2 produced by macrophages participates in the resorption of demineralized bovine bone.
本研究的目的是分析金属蛋白酶2(MMP-2)和巨噬细胞在对牛松质骨产生的有机移植材料的组织和细胞反应中的作用。30只成年雄性白色Wistar大鼠(褐家鼠)在股四头肌筋膜间植入脱矿牛骨基质块。在植入后3、7、14、21和28天收集标本(每个时间段n = 6)。6微米厚的切片用苏木精和伊红染色,并使用标准抗生物素蛋白-生物素-过氧化物酶方法用抗MMP-2和抗CD68进行免疫标记。对该材料的组织反应最初由多形核中性粒细胞介导,逐渐演变为单核炎性浸润,伴有巨噬细胞、少量淋巴细胞和浆细胞,并且存在与表现出吸收迹象的材料接触的炎性多核巨细胞(GC)。免疫标记MMP-2的细胞数量在第7天最高(103.2±39.1),但到第28天显著减少(F = 3.67;p = 0.044)(45.9±13.1)。CD68免疫染色也从第7天(49.5±10.4)到第28天(19.5±8.9)显著减少(F = 6.75;p = 0.007)。观察到这两个变量的变化之间存在正相关且具有统计学意义。该材料在第28天时几乎已完全吸收。在肉芽肿中存在的细胞中,抗MMP-2免疫染色在巨噬细胞中占主导且更强,但发现与材料紧密接触的轻度免疫标记的多核巨细胞。因此,考虑到本研究的实验局限性,我们得出结论,巨噬细胞产生的MMP-2参与脱矿牛骨的吸收。