PRF preserved DPSC viability and stemness markers under MTA, while CEM alone was cytotoxic in vitro

What was tested

Pourmohammadi-Nejad and colleagues isolated dental pulp stem cells (DPSCs) from impacted third molars of three healthy donors aged 18 to 25, confirmed the typical mesenchymal immunophenotype (CD90 and CD105 positive, CD34 and CD45 negative), and verified adipogenic and osteogenic differentiation. They then exposed passage 3 to 6 cells for 72 h to extracts of mineral trioxide aggregate (MTA), calcium-enriched mixture cement (CEM), or each cement combined with 50 percent platelet-rich fibrin (PRF) conditioned medium. Endpoints were MTT viability, colony formation after 10 to 14 days, and real-time PCR for OCT4, SOX2, and NANOG, with three biological replicates per group. The study is entirely in vitro.

Viability and colony formation

After 72 h, MTA extract and MTA plus PRF did not differ significantly from the untreated control. CEM extract alone, however, significantly reduced DPSC viability compared with control (p < 0.05). Adding PRF to CEM partially restored viability, though the rescue did not reach statistical significance versus control. In the colony formation assay, CEM alone also reduced colony number, while CEM plus PRF produced more colonies than CEM alone. MTA and MTA plus PRF formed colonies comparable to or greater than control. These findings are reported in Figures 4 and 5 of the primary paper.

Stemness gene expression

The authors measured mRNA for OCT4, SOX2, and NANOG and normalized expression to a housekeeping gene using the delta-delta Ct method. OCT4 did not change significantly across any condition. NANOG was significantly higher in the MTA group than in control (p < 0.01) and highest in the MTA plus PRF group, which exceeded control and all other groups (p < 0.001). SOX2 was significantly elevated only in the MTA plus PRF group (p < 0.05). The paper frames these three genes as “pluripotency markers,” although DPSCs are multipotent mesenchymal cells; the practical point is that MTA plus PRF shifted the expression profile toward a less differentiated state.

What it does not show

This is a cell-culture experiment with no pulp injury model, no animal, and no patient. The authors explicitly note that they did not include a PRF-only group, so the independent effect of PRF cannot be separated from the cement extracts. They also did not measure odontogenic differentiation, mineralization, inflammatory cytokines, or long-term outcomes. The reduced viability with CEM alone is an early in vitro observation and does not contradict clinical reports that CEM can perform well in direct pulp capping; the paper itself cites a clinical study in which CEM and MTA success rates were similar over 18 months.

Where this sits

For the pulp and dentin repair program, the result adds a cell-biology rationale for combining MTA with autologous PRF: the combination kept cells alive and raised NANOG and SOX2 expression in a culture dish. It does not change the program’s tier, because the strongest human evidence in this route remains the randomized 26-patient autologous pulp stem cell graft trial, and this new study offers no human or animal efficacy data. The current field assessment is at /field/.

Provenance: every claim above was checked against the full open-access report at PubMed Central (PMC13469813), including the abstract, methods, results, figure captions, and limitations, per our method at /method/.