Conditioned Media from Human Pulp Stem Cell Cultures Improve Bone Regeneration in Rat Calvarial Critical-Size Defects

来自人牙髓干细胞培养物的条件培养基可改善大鼠颅骨临界尺寸缺损的骨再生

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作者:Leonardo Fernandes Buss, Gustavo Sigrist de Martin, Elizabeth Ferreira Martinez, Isabela Amanda de Abreu Araújo Porcaro Filgueiras, José Luiz Magnabosco, Bruno Frenhan Alves, Bruno de Macedo Almeida, Tatiana Kotaka, Marcelo Lucchesi Teixeira, José Ricardo Muniz Ferreira, Daniel Navarro da Rocha, Rau

Abstract

The aim of this study was to test whether lyophilized conditioned media from human dental pulp mesenchymal stem cell cultures promote the healing of critical-size defects created in the calvaria of rats. Prior to the surgical procedure, the medium in which dental pulp stem cells were cultured was frozen and lyophilized. After general anesthesia, an 8 mm diameter bone defect was created in the calvaria of twenty-four rats. The defects were filled with the following materials: xenograft alone (G1) or xenograft associated with lyophilized conditioned medium (G2). After 14 or 42 days, the animals were euthanized, and the specimens processed for histologic and immunohistochemical analysis. Bone formation at the center of the defect was observed only in the G2 at 42 days. At both timepoints, increased staining for VEGF, a marker for angiogenesis, was observed in G2. Consistent with this, at 14 days, G2 also had a higher number of blood vessels detected by immunostaining with an anti-CD34 antibody. In conclusion, conditioned media from human dental pulp mesenchymal stem cell cultures had a positive effect on the regenerative process in rat critical-size bone defects. Both the formation of bone and enhancement of vascularization were stimulated by the conditioned media.

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