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新型根修复材料对转染 BMP-2 人成骨细胞的细胞毒性和生物活性潜力。

Cytotoxicity and bioactive potential of new root repair materials for use with BMP-2 transfected human osteoblast cells.

机构信息

Universidade Estadual Paulista - Unesp, Schoool of Dentistry, Department of Restorative Dentistry, Araraquara, SP, Brazil.

Universidade Estadual Paulista - Unesp, Schoool of Dentistry, Department of Diagnosis and Surgery, Araraquara, SP, Brazil.

出版信息

Braz Oral Res. 2022 May 2;36:e063. doi: 10.1590/1807-3107bor-2022.vol36.0063. eCollection 2022.

Abstract

Modified formulations of calcium silicate repair materials with additives have been developed to enhance handling, consistency, biocompatibility and bioactivity. Considering the relevance of osteoblastic cell response to mineralized tissue repair, human osteoblastic cells (Saos-2 cells overexpressing BMP-2) were exposed to mineral trioxide aggregate (MTA) (with calcium tungstate - CaWO4), MTA HP Repair, Bio-C Repair and Bio-C Pulpo. Cell viability was assessed by 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) and neutral red (NR), and cell death, by flow cytometry. Gene expression of bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX-2), and alkaline phosphatase (ALP) osteogenic markers were evaluated by real-time polymerase chain reaction (RT-qPCR). ALP activity and alizarin red staining (ARS) were used to detect mineralization nodule deposition. Bioactive cements presented no cytotoxic effect, and did not induce apoptosis at the higher dilution (1:12). MTA, Bio-C Repair and Bio-C Pulpo exhibited higher ALP activity than the control group (P < 0.05) after 7 days. MTA, MTA HP and Bio-C Pulpo affected the formation of mineralized nodules (p < 0.05). Exposure to all cement extracts for 1 day increased BMP-2 gene expression. RUNX-2 mRNA was greater in MTA, MTA HP and Bio-C Repair. MTA, MTA HP and Bio-C Pulpo increased the ALP mRNA expression, compared with BMP-2 unexposed cells (P < 0.05). Calcium silicate cements showed osteogenic potential and biocompatibility in Saos-2 cells transfected BMP-2, and increased the mRNA expression of BMP-2, RUNX-2, and ALP osteogenic markers in the BMP-2 transfected system, thereby promoting a cellular response to undertake the mineralized tissue repair.

摘要

已开发出具有添加剂的硅酸钙修复材料的改良配方,以增强其操作性、一致性、生物相容性和生物活性。鉴于成骨细胞反应与矿化组织修复的相关性,将人成骨细胞(过表达 BMP-2 的 Saos-2 细胞)暴露于矿化三氧化物聚合体(MTA)(含钨酸钙 - CaWO4)、MTA HP 修复剂、Bio-C 修复剂和 Bio-C 糊剂中。通过 3-(4,5-二甲基噻唑-2,5-二苯基四氮唑溴盐(MTT)和中性红(NR)评估细胞活力,并通过流式细胞术评估细胞死亡。通过实时聚合酶链反应(RT-qPCR)评估骨形态发生蛋白 2(BMP-2)、 runt 相关转录因子 2(RUNX-2)和碱性磷酸酶(ALP)成骨标志物的基因表达。ALP 活性和茜素红染色(ARS)用于检测矿化结节的沉积。生物活性水泥在较高稀释度(1:12)下没有细胞毒性作用,也不会诱导细胞凋亡。与对照组相比,MTA、Bio-C 修复剂和 Bio-C 糊剂在第 7 天表现出更高的 ALP 活性(P < 0.05)。MTA、MTA HP 和 Bio-C 糊剂影响矿化结节的形成(p < 0.05)。暴露于所有水泥提取物 1 天可增加 BMP-2 基因表达。在 MTA、MTA HP 和 Bio-C 修复剂中,RUNX-2 mRNA 更高。与未暴露于 BMP-2 的细胞相比,MTA、MTA HP 和 Bio-C 糊剂增加了 ALP mRNA 的表达(P < 0.05)。在转染 BMP-2 的 Saos-2 细胞中,硅酸钙水泥显示出成骨潜力和生物相容性,并增加了转染 BMP-2 系统中成骨标志物 BMP-2、RUNX-2 和 ALP 的 mRNA 表达,从而促进细胞对矿化组织修复的反应。

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