Evaluation of the combined assessment of two digital enhancement filters in periapical radiographs obtained with different projection angles in the detection of simulated dental root fractures.

OBJECTIVE: To evaluate the diagnostic efficacy of the combined assessment of the original radiographic image with the Invert or Emboss digital enhancement filters in periapical radiographs obtained with different horizontal projection angles in the detection of simulated dental root fracture. MATERIALS AND METHODS: Thirty-four single-rooted teeth were selected, out of which 17 teeth were subjected to root fracture. Each tooth was individually placed in an empty socket of a dry human maxilla or mandible and X-rayed following the paralleling technique at three horizontal projections: mesial, right angle, distal. Then, the Invert and Emboss enhancement filters were applied. Five examiners independently evaluated all the images and rated the fractures using a 5-point scale. Weighted kappa test assessed the intra- and interexaminer agreements. Diagnostic values were calculated and the areas under the receiver operating characteristic curve (AUC) were compared using two-way ANOVA with Tukey test as post hoc (α = 0.05). RESULTS: The inter- and intraexaminer agreement ranged from moderate to almost perfect and from substantial to almost perfect, respectively. Diagnostic values were considerably high for all conditions with no significant difference between the AUC values (p > 0.05). CONCLUSIONS: The combined use of the original radiographic image with the Invert or Emboss digital enhancement filters in periapical radiographs obtained with different projection angles did not influence the detection of simulated dental root fracture.

Influence of tumor cell-derived TGF-ß on macrophage phenotype and macrophage-mediated tumor cell invasion.

In oral squamous cell carcinoma (OSCC), macrophages are the most abundant immune cell type in the tumor microenvironment (TME). Macrophage infiltration is inversely proportional to prognosis and disease survival, particularly when these tumor-associated macrophages (TAM) assume an M2-like phenotype. This phenotype is determined by cues from the microenvironment, especially tumor cell-secreted molecules, and is associated with increased production of extracellular-matrix-degrading enzymes, angiogenic molecules and immunosuppressing cytokines. This study investigates, in vitro and in vivo, the relative contribution of OSCC cell-secreted transforming growth factor beta (TGF-ß) on the phenotype of macrophages and on macrophage-facilitated tumor invasion. TCGA database shows a positive correlation between high expression of TGFB1 and macrophage infiltrate in Head and neck squamous cell carcinoma (HNSCC). THP-1 derived-macrophages were exposed to the secretome of two OSCC cell lines using two strategies to block the effects of neoplastic cell-secreted TGF-ß: pre-treatment with a TGF-ß receptor type I kinase inhibitor (LY364947) and antibody-mediated depletion. RT-qPCR, ELISA and flow cytometry determined macrophage phenotype after exposure to conditioned medium (CM) from H-314 (TGF-ßhigh) or SCC-9 (TGF-ßlow) cell lines. The influence of TGF-ß on macrophage-mediated tumor cell invasion (myogel and CAM assays) and chemotaxis (Boyden chamber) was assessed using co-cultures of macrophages and OSCC cells in which macrophages were pre-conditioned with the secretome of OSCC cells in the presence and absence of LY364947. Blocking the effects of TGF-ß skewed macrophages to the M1 end of the phenotype by differential effects depending on the strategy for inhibiting the influence of TGF-ß and on the neoplastic cell secretome. In vitro and in vivo invasion of H-314 cell line was reduced by inhibiting TGFBR1 signaling in macrophages, whereas SCC-9 cell invasion was not affected. SCC-9/macrophage reciprocal chemotaxis were enhanced by inhibiting TGFBR1 signaling in macrophages, whereas only macrophage chemotaxis to H314 products was inhibited by inhibiting TGFBR1. In summary, blocking the effects of OSCC cell-secreted TGF-ß in macrophages attenuates M2-like phenotypical traits of macrophages and can impact invasion and chemotaxis of tumor cells differentially.