Department of Endodontics, School of Dentistry, São Paulo State University (UNESP), Araçatuba, SP, Brazil.
Department of Restorative Dentistry, School of Dentistry, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG, Brazil.
Int Endod J. 2019 Dec;52(12):1723-1737. doi: 10.1111/iej.13190. Epub 2019 Aug 13.
To investigate hydrogen peroxide (H O )-induced responsiveness in pulp cells using heme oxygenase-1 (HO-1) immunolabelling, Jun-D immunolabelling to study the effects of H O on odontoblastic differentiation and CD90+/CD73+/CD105+/CD45- cell counting for in vivo identification of mesenchymal stem cells in the pulp.
The maxillary molars of 50 rats were treated with a bleaching gel (35% H O , 1 × 30 min) or placebo gel (control groups). At 2, 3, 7, 15 and 30 days after the treatment (n = 10), inflammation in pulp tissue was analysed by haematoxylin-eosin staining, HO-1- and Jun-D-immunolabelled cells were counted in each third of the pulp chamber, and the number of CD90+/CD73+/CD105+/CD45- cells was quantified by immunofluorescence. The results were assessed using the Paired t-test or Wilcoxon signed-rank test (P < 0.05).
Significant H O -induced inflammation was noted at 2 and 3 days (P < 0.05), with tertiary dentine formation occurring from 7 days. The bleached specimens had greater HO-1 immunolabelling in the middle and cervical thirds of the coronal pulp at 2 and 3 days, in all thirds at 7 days, and in the occlusal third at 15 days (P < 0.05), and significant nuclear Jun-D immunolabelling in the cervical third at 2 and 3 days and in the occlusal and middle thirds at 7 days (P < 0.05). Bleached and control groups had low numbers of CD90+/CD73+/CD105+/CD45- cells in the pulp at all periods (P > 0.05).
Pulp cells responded to oxidative stress by expressing HO-1 during the post-bleaching inflammation phase until the beginning of the repair phase. Jun-D expression occurred during the reduction of inflammation and the beginning of tertiary dentine production. The presence of oxidative stress did not influence the number of CD90+/CD73+/CD105+/CD45- cells identified in vivo in the dental pulp.
通过血红素加氧酶-1(HO-1)免疫标记研究牙髓细胞中过氧化氢(H 2 O 2 )诱导的反应性,用 Jun-D 免疫标记研究 H 2 O 2 对成牙本质细胞分化的影响,并通过 CD90+/CD73+/CD105+/CD45-细胞计数鉴定牙髓中的间充质干细胞。
50 只大鼠的上颌磨牙用漂白凝胶(35%H 2 O 2 ,1×30 分钟)或安慰剂凝胶(对照组)处理。治疗后 2、3、7、15 和 30 天(n=10),通过苏木精-伊红染色分析牙髓组织炎症,在牙髓腔的每三分之一中计数 HO-1 和 Jun-D 免疫标记细胞,并通过免疫荧光定量 CD90+/CD73+/CD105+/CD45-细胞的数量。采用配对 t 检验或 Wilcoxon 符号秩检验(P<0.05)评估结果。
治疗后 2 天和 3 天观察到明显的 H 2 O 2 诱导炎症(P<0.05),7 天开始形成第三期牙本质。漂白标本在治疗后 2 天和 3 天的冠髓中、颈髓三分之一以及治疗后 7 天的所有三分之一以及治疗后 15 天的牙合三分之一的 HO-1 免疫标记增加(P<0.05),在治疗后 2 天和 3 天的颈髓三分之一以及治疗后 7 天的牙合三分之一和中三分之一的核 Jun-D 免疫标记增加(P<0.05)。在所有时期,漂白组和对照组牙髓中的 CD90+/CD73+/CD105+/CD45-细胞数量均较低(P>0.05)。
牙髓细胞在漂白后炎症期到修复期开始时通过表达 HO-1 对氧化应激作出反应。Jun-D 的表达发生在炎症减轻和第三期牙本质产生开始时。氧化应激的存在并不影响体内鉴定的牙髓中 CD90+/CD73+/CD105+/CD45-细胞的数量。