Oliveira Danila de, Hassumi Jaqueline Suemi, Gomes-Ferreira Pedro Henrique da Silva, Polo Tárik Ocon Braga, Ferreira Gabriel Ramalho, Faverani Leonardo Perez, Okamoto Roberta
Universidade Estadual Paulista, Faculdade de Odontologia de Araçatuba, Departamento de Ciências Básicas, Araçatuba, SP, Brasil.
Universidade Estadual Paulista, Faculdade de Odontologia de Araçatuba, Departamento de Cirurgia e Clínica Integrada, Araçatuba, SP, Brasil.
J Appl Oral Sci. 2017 Jan-Feb;25(1):42-52. doi: 10.1590/1678-77572016-0165.
The aim of this study was to evaluate the bone repair process at the bone/implant interface of osteoporotic rats treated with sodium alendronate through the analysis of microtomography, real time polymerase chain reactions and immunohistochemistry (RUNX2 protein, bone sialoprotein (BSP), alkaline phosphatase, osteopontin and osteocalcin).
A total of 42 rats were used and divided in to the following experimental groups: CTL: control group (rats submitted to fictitious surgery and fed with a balanced diet), OST: osteoporosis group (rats submitted to a bilateral ovariectomy and fed with a low calcium diet) and ALE: alendronate group (rats submitted to a bilateral ovariectomy, fed with a low calcium diet and treated with sodium alendronate). A surface treated implant was installed in both tibial metaphyses of each rat. Euthanasia of the animals was conducted at 14 (immunhostochemistry) and 42 days (immunohistochemistry, micro CT and PCR). Data were subjected to statistical analysis with a 5% significance level.
Bone volume (BV) and total pore volume were higher for ALE group (P<0.05). Molecular data for RUNX2 and BSP proteins were significantly expressed in the ALE group (P<0.05), in comparison with the other groups. ALP expression was higher in the CTL group (P<0.05). The immunostaining for RUNX2 and osteopontin was positive in the osteoblastic lineage cells of neoformed bone for the CTL and ALE groups in both periods (14 and 42 days). Alkaline phosphatase presented a lower staining area in the OST group compared to the CTL in both periods and the ALE at 42 days.
There was a decrease of osteocalcin precipitation at 42 days for the ALE and OST groups. Therefore, treatment with short-term sodium alendronate improved bone repair around the implants installed in the tibia of osteoporotic rats.
本研究旨在通过显微断层扫描、实时聚合酶链反应和免疫组织化学(RUNX2蛋白、骨唾液蛋白(BSP)、碱性磷酸酶、骨桥蛋白和骨钙素)分析,评估阿仑膦酸钠治疗的骨质疏松大鼠骨/植入物界面的骨修复过程。
共使用42只大鼠,分为以下实验组:CTL:对照组(接受假手术并喂食均衡饮食的大鼠),OST:骨质疏松组(接受双侧卵巢切除术并喂食低钙饮食的大鼠)和ALE:阿仑膦酸钠组(接受双侧卵巢切除术、喂食低钙饮食并接受阿仑膦酸钠治疗的大鼠)。在每只大鼠的双侧胫骨干骺端植入一个表面处理过的植入物。在14天(免疫组织化学)和42天(免疫组织化学、显微CT和PCR)对动物实施安乐死。数据进行统计学分析,显著性水平为5%。
ALE组的骨体积(BV)和总孔隙体积更高(P<0.05)。与其他组相比,ALE组中RUNX2和BSP蛋白的分子数据显著表达(P<0.05)。CTL组中碱性磷酸酶(ALP)表达更高(P<0.05)。在两个时期(14天和42天),CTL组和ALE组新形成骨的成骨细胞谱系细胞中RUNX2和骨桥蛋白的免疫染色均为阳性。在两个时期,OST组碱性磷酸酶的染色面积均低于CTL组,在42天时低于ALE组。
ALE组和OST组在42天时骨钙素沉淀减少。因此,短期阿仑膦酸钠治疗改善了骨质疏松大鼠胫骨植入物周围的骨修复。