Human dental pulp derived from exfoliated deciduous teeth has been described as a promising alternative source of multipotent stem cells. While these cells share certain similarities with mesenchymal stem-like cells (MSC) isolated from other tissues, basically they are still poorly characterized. In this study, for the first time, a proteomic map of abundantly expressed proteins in stromal cells derived from the dental pulp of human exfoliated deciduous teeth (SHED) was established. We also analyzed proteomic signatures of 2 clonal strains derived from SHEDs by single-cell cloning. The SHEDs were established from enzyme-disaggregated deciduous dental pulp from 6-year-old children. They had typical fibroblastoid morphology and high colony-forming efficiency index (16.4%). Cloning was performed at the second passage using limiting dilution in a 96-well plate (0.3 cell/well). Differentiation assessment revealed strong osteogenic but no adipogenic potential of the SHEDs in either clonal strain. The cells expressed characteristic antigens of MSC-like cells, including CD73, CD90, CD105, CD146, and did not express hematopoietic markers CD14, CD34, and CD45, as assessed with FACS analysis. For proteomic studies, cytosolic and nuclear proteins were analyzed with 2-dimensional gel electrophoresis (2-DE) and identified using matrix-assisted laser desorption/ionization (MALDI)-time of fl ight (TOF)-mass spectrometry (MS). All proteins were identified with high level of confidence (the lowest sequence coverage was 27%). Identification of highly expressed proteins in SHEDs revealed proteomic profiles very similar to that of MSC-like cells derived from other tissues. We also found a high degree of similarity between proteomic signatures of primary SHEDs and clonal cell strains. Thus, our data confirm a close resemblance between SHEDs and MSC-like cells from other tissues and may serve as starting point for creating-comprehensive proteomic maps.

Modern dental treatment with standard screw-type implants leave some cases unaddressed in patients with extreme jaw bone resorption. Custom-made subperiosteal dental implant could be an alternative treatment modality to sinus lift, nerve lateralization or zygomatic implant techniques. Subperiosteal dental implants were utilized for many years to treat such patients. A combination of traditional subperiosteal implant designs with current advancements in 3D imaging, design and printing allow to reduces treatment time and provides abutments for prostheses in cases where other techniques do not provide satisfactory results. The data manipulation and design software are important aspects in the manufacturing of custom implants. Programs that are specialized for industrial or medical design typically cost tens of thousands of US dollars. In this work I establish and test steps for design and production of a custom medical device (subperiosteal implant) from patient computed tomography (CT) data. Work stages to be defined are: selection of necessary software, CT data processing, 3D virtual model creation, modeling technique for custom implant and data file preparation for printing. Patient CT data was successfully converted into a watertight STL (Standard Tessellation Language) model of the maxilla. Error corrections and design were completed using freely available programs from Autodesk Inc.. The implant was produced in Ti64 (a type 5 titanium alloy) using three-dimensional (3D) printing DMLS (direct metal laser sintering) process. The avoidance of high cost software makes this treatment modality more accessible to smaller clinics or mid-size production facilities and subsequently more available to patients.

AAT at physiologic and inflammatory concentrations positively modulates the proliferation and motility of SHEDs in vitro. These results suggest the importance of AAT in the maintenance and regulation of tissue progenitor cells in vivo.

The acetylation of histones and transcription factors plays a crucial role in cell functional activity. Balance between histone acetyltransferases (HATs) and histone deacetylases (HDACs) is closely related to diverse cellular processes, including differentiation, proliferation, and apoptosis. Expression of different classes of HDACs is strictly dependent on the cell type, and differentiated cells possess different members of the HDAC family. In our study, we employed mesenchymal stem-like cells (MSC) derived from human exfoliated deciduous teeth. Expression of HDACs at mRNA level was analysed using the RT-PCR method. Next, we have assessed the protein levels of different classes of HDACs and their localization within the cell. The cytoplasm and nuclear fractions of cell lysates were subjected to western blot analysis. Since the functional activity of class II HDACs is determined by their shuttling between the nucleus and the cytoplasm, we have analysed the subcellular localization of HDAC4 in response to the wellknown activators of differentiation, i. e. bone morphogenetic protein 2 (BMP-2) and phorbol 12-myristate-13-acetate (PMA), using fluorescent confocal microscopy. Our results suggest that class I and II HDAC family members are expressed and produced in MSC-like cells. The predicted HDAC4 translocation from the cell nucleus was not induced by PMA or BMP-2 in our model. However, our findings open new insights into a possible targeting for HDACs in future transcription activations / derepression studies.