Archives of oral biology · 2026
Murine undifferentiated or differentiated dental pulp cells secretomes have opposite effects on cell viability and clastic differentiation in mouse marrow cultures.
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Sarra Giovanna, Kokado Matheus Yuki Ferreira, Bolsonello Carolina Gomes, Chinen Elisângela, Bombonato-Prado Karina Fittipaldi, Arana-Chavez Victor Elias
Аннотация
OBJECTIVE (S): This study aimed to evaluate the role of undifferentiated or differentiated dental pulp cells in the cell viability, differentiation and function of clastic precursors from mouse marrow cultures.
DESIGN: Undifferentiated mesenchymal cells isolated from the dental pulp of mice incisors (undifferentiated dental pulp cells, UDPCs) and odontoblastic-like cells from the MDPC-23 murine lineage (differentiated dental pulp cells, DDPCs) were cultured and the media containing their secretomes were collected. Undifferentiated bone marrow cells containing clastic precursors were treated with clonogenic culture medium (controls), UDPCs or DDPCs secretomes with or without the supplementation with vitamin D (VD) as a clastic stimulus. Cell viability was assessed through MTT assay; clastic differentiation was evaluated through TRAP histochemical staining, immunofluorescence for RANKL, and gene expression analysis for OPG and RANKL; and scanning electron microscopy was performed to evaluate resorption lacunae on dentin substrates. Statistical analysis was performed using ANOVA followed by Tukey's test, with a significance level set at 5% (p < 0.05).
RESULTS: The UDPCs secretome increased cell viability, reduced the number of TRAP-positive cells even in the presence of VD, decreased RANKL gene expression in the presence of VD, and reduced the presence of RANKL-positive cells. The DDPCs secretome decreased cell viability, increased the number of TRAP-positive cells in the absence of VD, increased the presence of RANKL-positive cells and led to larger resorption lacunae in the presence of VD.
CONCLUSIONS: The UDPCs secretome promoted cell proliferation while attenuating clastic differentiation, whereas the DDPCs secretome reduced cell proliferation while improving clastic differentiation.
DESIGN: Undifferentiated mesenchymal cells isolated from the dental pulp of mice incisors (undifferentiated dental pulp cells, UDPCs) and odontoblastic-like cells from the MDPC-23 murine lineage (differentiated dental pulp cells, DDPCs) were cultured and the media containing their secretomes were collected. Undifferentiated bone marrow cells containing clastic precursors were treated with clonogenic culture medium (controls), UDPCs or DDPCs secretomes with or without the supplementation with vitamin D (VD) as a clastic stimulus. Cell viability was assessed through MTT assay; clastic differentiation was evaluated through TRAP histochemical staining, immunofluorescence for RANKL, and gene expression analysis for OPG and RANKL; and scanning electron microscopy was performed to evaluate resorption lacunae on dentin substrates. Statistical analysis was performed using ANOVA followed by Tukey's test, with a significance level set at 5% (p < 0.05).
RESULTS: The UDPCs secretome increased cell viability, reduced the number of TRAP-positive cells even in the presence of VD, decreased RANKL gene expression in the presence of VD, and reduced the presence of RANKL-positive cells. The DDPCs secretome decreased cell viability, increased the number of TRAP-positive cells in the absence of VD, increased the presence of RANKL-positive cells and led to larger resorption lacunae in the presence of VD.
CONCLUSIONS: The UDPCs secretome promoted cell proliferation while attenuating clastic differentiation, whereas the DDPCs secretome reduced cell proliferation while improving clastic differentiation.