Cleft palate represents one of the most common congenital birth defects. Transforming growth factor ␤ (TGF␤) signaling plays crucial functions in regulating craniofacial development, and loss of TGF␤ receptor type II in cranial neural crest cells leads to craniofacial malformations, including cleft palate in mice (Tgfbr2 fl/fl ;Wnt1-Cre mice). Here we have identified candidate target genes of TGF␤ signaling during palatal formation. These target genes were selected based on combining results from gene expression profiles of embryonic day 14.5 palates from Tgfbr2 fl/fl ;Wnt1-Cre mice and previously identified cleft palate phenotypes in genetically engineered mouse models. We found that fibroblast growth factor 9 (Fgf9) and transcription factor pituitary homeobox 2 (Pitx2) expressions are significantly down-regulated in the palate of Tgfbr2 fl/fl ;Wnt1-Cre mice, and Fgf9 and Pitx2 loss of function mutations result in cleft palate in mice. Pitx2 expression is down-regulated by siRNA knockdown of Fgf9, suggesting that Fgf9 is upstream of Pitx2. We detected decreased expression of both cyclins D1 and D3 in the palates of Tgfbr2 fl/fl ;Wnt1-Cre mice, consistent with the defect in cell proliferation. Significantly, exogenous FGF9 restores expression of cyclins D1 and D3 in a Pitx2-dependent manner and rescues the cell proliferation defect in the palatal mesenchyme of Tgfbr2 fl/fl ; Wnt1-Cre mice. Our study indicates that a TGF␤-FGF9-PITX2 signaling cascade regulates cranial neural crest cell proliferation during palate formation.Cleft palate represents one of the most frequent congenital birth defects in the human population (1). The causes of cleft palate remain largely unknown, but they appear to be complex, including genetic and environmental factors (2, 3). Cleft palate causes many clinical symptoms and complications, such as difficulties with suckling and eating, dysfunction of tongue and oral muscles, dental abnormalities, and speech and language delay (4). Therefore, prevention of cleft palate is the ultimate objective, and a prerequisite of this aim is to elucidate the mechanisms of healthy palate development and the causes of cleft palate.The majority of cells in the craniofacial region are derived from cranial neural crest cells (CNCC), 2 which produce the facial skeleton, smooth muscle, and sensory nerve. Transforming growth factor ␤ (TGF␤) signaling plays crucial functions in craniofacial development, including palate formation by mediating cell proliferation, differentiation, and extracellular matrix formation via regulation of downstream target genes (5-7). TGF␤ ligands activate the membrane receptor serine/threonine kinase complex composed of TGF␤ receptor type II (T␤RII) and TGF␤ receptor type I (T␤RI/ALK5). The ligandreceptor complex phosphorylates SMAD2 and SMAD3, which form a transcriptional complex with SMAD4, and then the transcriptional complex translocates into the nucleus to control the expression of downstream target genes (8 -10). T␤RII is expressed in the CNCC-derived palatal mesenchyme (11,1...

Improving soft tissue attachment to implant abutments is a crucial factor for enduring health and maintenance of soft peri-implant tissue health. In this in vitro study we aimed to compare the biocompatibility of three different abutment surfaces: titanium, zirconia and modified polyetheretherketone (PEEK). Surface topography, roughness and wettability were investigated with scanning electron microscopy, profilometer and contact angle meter, respectively. Human gingival epithelial keratinocytes were examined for viability, morphology, proliferation and migration by using tetrazolium salt colorimetric assay, scanning electron microscopy imaging, immunofluorescence bromodeoxyuridine analysis and scratch wound healing assays. Roughness measurements revealed differences between the investigated surfaces. Keratinocytes cultured on all examined surfaces indicated adhesion and attachment by means of scanning electron microscopy imaging. Cell viability assays showed no significant differences between the groups (p > 0.05). The modified PEEK surface similarly improved surface roughness in comparison to titanium and zirconia, which resulted in greater and equivalent cell proliferation and migration. The study methodology showed here may emphasize the importance of cell interactions with different abutment materials, which in part increases the changes of implant success. PEEK, titanium and zirconia surface types used in this study showed mostly similar epithelial biological responses.

The use of automated biometrics-based personal identification systems is a ubiquitous procedure in present times. Biometrics has certain limitations, such as in cases when bodies are decomposed, burned, or only small fragments of calcified tissues remain. Dental enamel is the most mineralized tissue of organisms and resists post-mortem degradation. It is characterized by layers of prisms of regularly alternating directions, known as Hunter-Schreger bands (HSB). In this article, we show that the pattern variation of the HSB, referred here as toothprint, can be used as a biometric-based parameter for personal identification in automated systems.

Periodontal ligament stem cells (PDLSC) play an important role in periodontal tissue homeostasis/turnover and could be applied in cell-based periodontal regenerative therapy. Bacterial supernatants secreted from diverse periodontal bacteria induce the production of cytokines that contribute to local periodontal tissue destruction. However, little is known about the impact of whole bacterial toxins on the biological behavior of PDLSC. Therefore this study investigated whether proliferation, migration, inflammatory cytokines expression and transcriptional profile would be affected by exposure to endotoxins from bacterial species found in the subgingival plaque. PDLSC were cultured with the following bacterial supernatants: S . mutans , S . anginosus , P . intermedia , F . nucleatum , P . gingivalis and T . denticola . These supernatants were prepared in dilutions of 1:1000, 1:500, 1:300 and 1:50. Using quantitative RT-PCR, gene expression of selected inflammatory cytokines ( IL-6 , IL-8 and IL-1β ) and cell-surface receptors ( TLR2 , TLR4 ) showed upregulation of ≈2.0- to 3.0-fold, when exposed to P . intermedia , F . nucleatum , P . gingivalis and T . denticola . However, supernatants did not affect proliferation (MTT) and migration (wound scratch assays) of PDLSC. Next generation RNA sequencing confirmed modified lineage commitment of PDLSC by stimulating chondrogenesis, adipogenesis and inhibition of osteogenesis under P . gingivalis supernatant treatment compared to control. Taken together, this study shows stem cell immunomodulatory response to different periodontal bacteria supernatant and suggests that stem cell transcriptional capacity, migration/proliferation and osteogenesis may differ in the presence of those pathogens. These results bring into question stem cell contribution to periodontal tissue regeneration and onset of inflammation.

Periodontitis is a common immune-inflammatory oral disease. Early detection plays an important role in its prevention and progression. Saliva is a reliable medium that mirrors periodontal health and is easily obtainable for identifying periodontal biomarkers in point-of-care diagnostics. The aim of this study is to evaluate the effectiveness of diagnostic salivary tests to determine periodontal status. Whole saliva (stimulated/unstimulated) from twenty healthy and twenty stage III grade B generalized periodontitis patients was tested for lactoferrin, alkaline phosphatase, calcium, density, osmolarity, pH, phosphate, buffer capacity, salivary flow rate and dynamic viscosity. A semi-quantitative urinary strip test was used to evaluate markers of inflammation in saliva (erythrocytes, leukocytes, urobilinogen, nitrite, glucose, bilirubin, and ketones), clinical periodontal parameters and pathogenic bacteria. Concentrations of lactoferrin, hemoglobin, and leukocytes were found to be significantly higher in the stimulated and unstimulated saliva in periodontitis patients compared to healthy patients, whereas alkaline phosphatase levels were higher in unstimulated saliva of periodontitis patients (p < 0.05). Periodontal biomarker analysis using test strips may be considered rapid and easy tool for distinguishing between periodontitis and healthy patients. The increase in lactoferrin, hemoglobin, and leucocytes—determined by strip tests—may provide a non-invasive method of periodontal diagnosis.