葛根素联合Ag-HA涂层植入对种植体周围炎大鼠炎症水平和骨矿物质密度的影响

Effects of puerarin combined with Ag-HA coating implantation on inflammation and bone mineral density in rats with peri-implantitis

摘要:

目的 探讨葛根素联合银-羟基磷灰石（Ag-HA）涂层植入对种植体周围炎大鼠的炎症水平、骨矿物质密度及骨生物学参数的影响。通过比较不同治疗组的临床表现、炎症因子水平、基因和蛋白表达及骨密度指标，为种植体周围炎的治疗提供新的思路和实验依据。方法 选取雄性大鼠50只为研究对象复制牙周炎模型。模型复制成功后将牙周炎模型大鼠随机分为牙周炎组10只、Ag-HA组20只、联合组20只。牙周炎组大鼠不接受干预，Ag-HA组大鼠接受Ag-HA涂层种植体治疗，联合组大鼠接受葛根素联合Ag-HA涂层种植体治疗。检测并比较术前及术后1、2、4、8周大鼠龈沟出血指数、牙龈指数、探诊深度、血清炎症因子水平、骨结合能力。收集3组大鼠术后8周牙周组织，采用实时荧光定量聚合酶链反应检测相关mRNA的表达，Western blotting检测相关蛋白的表达。对大鼠模型进行micro CT三维重建后统计并比较骨生物学参数。结果 牙周炎组、Ag-HA组、联合组术前及术后1、2、4、8周的龈沟出血指、牙龈指数、探诊深度、人肿瘤坏死因子-α（TNF-α）、人白细胞介素-6（IL-6）、人白细胞介素-1β（IL-1β）、碱性磷酸酶（ALP）、骨结合率比较，结果 ①不同时间点龈沟出血指、牙龈指数、探诊深度、TNF-α、IL-6、IL-1β、ALP、骨结合率比较，差异均有统计学意义（P <0.05）；②牙周炎组、Ag-HA组、联合组龈沟出血指、牙龈指数、探诊深度、TNF-α、IL-6、IL-1β、ALP、骨结合率比较，差异均有统计学意义（P <0.05），联合组龈沟出血指、牙龈指数、探诊深度、TNF-α、IL-6、IL-1β水平较低，ALP、骨结合能力较高，相对治疗效果较好；③3组龈沟出血指、牙龈指数、探诊深度、TNF-α、IL-6、IL-1β、骨结合率变化趋势比较，差异均有统计学意义（P <0.05）；3组骨ALP变化趋势比较，差异无统计学意义（P >0.05）。联合组大鼠牙周组织IL-1β、CXCL12、CXCR4 mRNA和蛋白相对表达量均低于Ag-HA组（P <0.05），OPG mRNA和蛋白相对表达量均高于Ag-HA组（P <0.05）。联合组大鼠组织矿物质密度、骨体积分数、骨小梁厚度均高于Ag-HA组（P <0.05），联合组大鼠骨矿物质密度、上颌第一磨牙釉牙骨质界至牙槽嵴顶的距离均低于Ag-HA组（P <0.05）。结论 葛根素联合Ag-HA涂层植入能显著改善种植体周围炎大鼠的炎症水平、牙龈指数、探诊深度及骨结合能力。联合治疗较单独治疗在抑制炎症反应、提高骨密度及改善牙周组织修复方面效果更显著。特别是在OPG蛋白表达和骨生物学参数方面，联合治疗表现出较高的优势，提示其在种植体周围炎治疗中的潜在应用价值。

Abstract:

Objective To investigate the effects of puerarin combined with silver-hydroxyapatite (Ag-HA) coating implantation on inflammations, bone mineral density (BMD), and bone biological parameters in rats with peri-implantitis, and to provide new insights and experimental evidence for the treatment of peri-implantitis by comparing the clinical manifestations, gene and protein expression of inflammatory cytokines, and BMD across different treatment groups.Methods Fifty male rats were selected for this study and induced with peri-implantitis. After successful modeling, the rats were randomly divided into three groups: the peri-implantitis group (10 rats), the Ag-HA group (20 rats), and the combination group (20 rats). The peri-implantitis group received no intervention, the Ag-HA group was treated with Ag-HA coating implantation, and the combination group received both puerarin and Ag-HA coating implantation. The rats were evaluated preoperatively and at 1, 2, 4, and 8 weeks postoperatively for sulcus bleeding index (SBI), gingival index (GI), probing depth (PD), serum levels of inflammatory factors, and osseointegration capacity. Periodontal tissues were collected from all three groups of rats at 8 weeks postoperatively, and mRNA levels of relevant genes were detected using real-time quantitative PCR. Western blotting was performed to assess the expression of associated proteins. Micro-CT 3D reconstruction was carried out on the rat models, and bone biological parameters were statistically analyzed and compared.Results Comparisons were made on the SBI, GI, PD, levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β, and alkaline phosphatase (ALP), and the bone-implant contact rate among the three groups preoperatively and at 1, 2, 4, and 8 weeks postoperatively. The results exhibited that these indicators were statistically significant different across the time points (P < 0.05) and among the groups (P < 0.05), where the SBI, GI, PD, and levels of TNF-α, IL-6, and IL-1β were lower and the ALP level and the bone-implant contact rate were higher in the combination group, indicating better therapeutic outcomes. The change trends of all these indicators (P < 0.05) but ALP were significantly different among the groups (P > 0.05). The relative mRNA and protein expression levels of IL-1β, CXCL12, and CXCR4 in the periodontal tissues of rats in the combination group were lower than those in the Ag-HA group (P < 0.05), while the relative mRNA and protein expression levels of osteoprotegerin (OPG) in the combination group were higher than those in the Ag-HA group (P < 0.05). The tissue mineral density, bone volume-to-total volume ratio (BV/VT), and trabecular thickness (Tb.Th) of rats in the combination group were higher than those in the Ag-HA group (P < 0.05), whereas the BMD and distance from cementoenamel junction to alveolar bone crest (CEJ-ABC) were lower in the combination group than those in the Ag-HA group (P < 0.05).Conclusion Puerarin combined with Ag-HA coating implantation significantly improves inflammation levels, GI, PD, and osseointegration capacity in peri-implantitis rats. The combination treatment demonstrates more pronounced effects in suppressing inflammation, enhancing bone density, and promoting periodontal tissue repair compared to Ag-HA coating implantation alone. Notably, the combination therapy exhibits superior effects on OPG protein expression and bone biological parameters, suggesting their potential applicability in the treatment of peri-implantitis.