Association of Dental Pulp Stem Cells and Ricinus Bone Compound in a Model of Bone Defect

Alan Araújo de Jesus, A. Jesus, Flávia Oliveira de Lima, Luíz A. R. Freitas, Cassio S Meira, R. Ribeiro-dos-Santos, M. Soares
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Abstract

Autogenous bone grafting is needed in some bone tissue defects; however, it causes secondary surgical wounds and morbidity. Tissue bioengineering may be an alternative approach for bone regeneration. Here we investigated the osteogenic potential of dental pulp stem cells from deciduous teeth (DPSC) in association with a Ricinus bone compound (RBC) in a model of bone defect. The influence of the biomaterial RBC on the proliferation and osteogenic differentiation of DPSC was assessed in vitro by MTT metabolism and alizarin red staining, respectively. The morphologic analysis was performed using the optic and scanning electron (SEM) microscopies. For the in vivo study, 54 Wistar rats submitted to calvarial defects were filled with RBC or RBC+DPSC. A control group had the defects filled only with blood clots. Analyses were performed 15, 30 and 60 days after treatment using digital radiography, optical microscopy, SEM and chemical analysis by electron dispersive spectroscopy. The Ricinus bone compound (RBC) did not inhibit the osteogenic differentiation in vitro. No spontaneous regeneration was observed in the control group. The area of the calvarial defect of the RBC+DPSC group showed greater radiopacity on day 15. The RBC presented no reabsorption, was biocompatible and showed osteointegration, working as a mechanical filling. Only sparse ossification areas were found and those were larger and more developed on the RBC+DPSC group when compared to animals treated only with RBC. RBC in association with DPSC is a promising combination for applications in bone regeneration.  
牙髓干细胞与蓖麻骨化合物在骨缺损模型中的关联
部分骨组织缺损需要自体骨移植;然而,它会引起继发性手术伤口和发病率。组织生物工程可能是骨再生的一种替代方法。在这里,我们研究了乳牙牙髓干细胞(DPSC)与蓖麻骨化合物(RBC)在骨缺损模型中的成骨潜能。体外MTT代谢法和茜素红染色法观察生物材料红细胞对DPSC增殖和成骨分化的影响。形态学分析使用光学和扫描电子显微镜(SEM)进行。在体内研究中,54只Wistar大鼠颅骨缺损被灌注RBC或RBC+DPSC。对照组的缺陷只填满了血凝块。治疗后15、30和60天分别采用数字x线摄影、光学显微镜、扫描电镜和电子色散光谱化学分析进行分析。蓖麻骨化合物(RBC)对体外成骨分化无抑制作用。对照组未见自发再生。RBC+DPSC组颅骨缺损面积在第15天显示出更大的放射不透明。RBC无重吸收,具有生物相容性和骨整合性,可作为机械填充物。与仅接受RBC治疗的动物相比,RBC+DPSC组只发现了稀疏的骨化区域,并且这些区域更大、更发达。RBC与DPSC联合应用是骨再生的一个很有前途的组合。
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