Chuang Shu-Fen, Chen Yu-Hsuan, Ma Peter X, Ritchie Helena H
School of Dentistry and Institute of Oral Medicine, National Cheng Kung University, No. 1 Universal Road, Tainan 70101, Taiwan.
Department of Stomatology, National Cheng Kung University Hospital, 138 ShengLi Road, Tainan 70403, Taiwan.
Polymers (Basel). 2022 Sep 3;14(17):3656. doi: 10.3390/polym14173656.
Conventional direct pulp capping, such as calcium hydroxide (Ca(OH)) or silicate products, usually induces an inflammatory reaction to provoke pulp regeneration. Phosphophoryn (PP) and dentin sialoprotein (DSP), the two most abundant non-collagenous proteins in the dentin matrix, are responsible for dentin mineralization, pulp cell migration, and differentiation. Here we examined the PP and combined DSP/PP as bio-inductive pulp capping materials by in vitro and in vivo tests. Firstly, the effects of the PP dose on pulp cell migration and matrix protein expression were examined by an agarose bead test. Secondly, the role of recombinant DSP (recDSP) and recDSP/PP on stimulating DSP-PP transcript expression was examined by RT-PCR. DSPP mRNA was also knocked down by RNA interference (RNAi) to examine their functions on dentin matrix mineralization. Finally, we used ferret animal models to test PP and recDSP/PP acting as capping agents on in vivo pulp responses and reparative dentin formation. The result showed that intermediate-dose PP was the most effective to enhance cell migration and differentiation. RecDSP/PP strongly enhanced the DSP-PP transcript expression, while inhibition of DSPP mRNA expression by siRNAs partially or completely affected dental pulp cell mineralization. The in vivo results showed that intermediate-dose PP and recDSP/PP proteins induced less pulp inflammation and promoted reparative dentin formation. Contrarily, conventional calcium hydroxide induced severe pulp inflammation. With these findings, DSP and PP could serve as capping agents for pulp capping therapy.
传统的直接盖髓术,如使用氢氧化钙(Ca(OH))或硅酸盐产品,通常会引发炎症反应以促进牙髓再生。磷蛋白(PP)和牙本质涎蛋白(DSP)是牙本质基质中两种含量最丰富的非胶原蛋白,它们负责牙本质矿化、牙髓细胞迁移和分化。在此,我们通过体外和体内试验研究了PP以及联合使用的DSP/PP作为生物诱导性盖髓材料的效果。首先,通过琼脂糖珠试验研究了PP剂量对牙髓细胞迁移和基质蛋白表达的影响。其次,通过逆转录聚合酶链反应(RT-PCR)研究了重组DSP(recDSP)和recDSP/PP对刺激DSP-PP转录表达的作用。还通过RNA干扰(RNAi)敲低DSPP mRNA以研究它们在牙本质基质矿化中的功能。最后,我们使用雪貂动物模型来测试PP和recDSP/PP作为盖髓剂对体内牙髓反应和修复性牙本质形成的作用。结果表明,中等剂量的PP在增强细胞迁移和分化方面最有效。RecDSP/PP强烈增强了DSP-PP转录表达,而小干扰RNA(siRNAs)抑制DSPP mRNA表达部分或完全影响了牙髓细胞矿化。体内结果表明,中等剂量的PP和recDSP/PP蛋白诱导的牙髓炎症较少,并促进了修复性牙本质的形成。相反,传统的氢氧化钙会引发严重的牙髓炎症。基于这些发现,DSP和PP可作为盖髓治疗的盖髓剂。