Petros Koidis

Comparative analysis of in vitro osteo/odontogenic differentiation potential of human dental pulp stem cells (DPSCs) and stem cells from the apical papilla (SCAP).

OBJECTIVE The aim of this study was to compare the in vitro osteo/odontogenic differentiation potential of mesenchymal stem cells (MSCs) derived from the dental pulp (dental pulp stem cells - DPSCs) or the apical papilla (stem cells from the apical papilla - SCAP) of permanent developing teeth. DESIGN DPSCs and SCAP cultures were established from impacted third molars of young healthy donors at the stage of root development. Cultures were analysed for stem cell markers, including STRO-1, CD146, CD34 and CD45 using flow cytometry. Cells were then induced for osteo/odontogenic differentiation by media containing dexamethasone, KH(2)PO(4) and β-glycerophosphate. Cultures were analysed for morphology, growth characteristics, mineralization potential (Alizarin Red method) and differentiation markers (dentine sialophosphoprotein-DSPP, bone sialoprotein-BSP, osteocalcin-OCN, alkaline phosphatase-ALP), using immunocytochemistry and reverse transcriptase-polymerase chain reaction. RESULTS All DPSCs and SCAP cultures were positive for STRO-1, CD146 and CD34, in percentages varying according to cell type and donor, but negative for CD45. Both types of MSCs displayed an active potential for cellular migration, organization and mineralization, producing 3D mineralized structures. These structures progressively expressed differentiation markers, including DSPP, BSP, OCN, ALP, having the characteristics of osteodentin. SCAP, however, showed a significantly higher proliferation rate and mineralization potential, which might be of significance for their use in bone/dental tissue engineering. CONCLUSIONS This study provides evidence that different types of dental MSCs can be used in tissue engineering/regeneration protocols as an approachable stem cell source for osteo/odontogenic differentiation and biomineralization that could be further applied for stem cell-based clinical therapies.

Hydroxy Carbonate Apatite Formation on Particulate Bioglass In Vitro as a Function of Time

The mechanism of bonding of bioactive glasses with living tissues has been reported to be associated with the development of a layer consisting of carbonate-containing hydroxyapatite similar to that of bone on the surface of the materials. This layer is also formed in vitro, in solutions with ion concentrations similar to those of human blood plasma, like SBF (Simulated Body Fluid). The development of HCAp (Hydroxy carbonate apatite) layer on the surface of a commercially available Bioglass® Synthetic Bone Graft Particulate (Perioglas®) after immersion in SBF solution using Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM) with associated Energy Dispersive Spectroscopy (EDS). PerioGlas® powder with particle size 20-63μm, pressed in a vacuum press in order to produce pellets. The pellets were soaked in SBF for 12, 18, 24 and 48 hours at 37°C. Results revealed the formation of an amorphous CaO-P 2 O 5 - rich layer on the surface of the specimens after 12 hours in the solution and a well crystalline HCAp layer after 24 hours immersion time.

Effects of HEMA and TEDGMA on the in vitro odontogenic differentiation potential of human pulp stem/progenitor cells derived from deciduous teeth

OBJECTIVES The aim of this study was to investigate the effects of HEMA and TEGDMA on the odontogenic differentiation potential of dental pulp stem/progenitor cells. METHODS Dental stem/progenitor cell cultures were established from pulp biopsies of human deciduous teeth of 1-3 year-old children (Deciduous Teeth Stem Cells-DTSCs). Cultures were characterized for stem cell markers, including STRO-1, CD146, CD34, CD45 using flow cytometry. Cytotoxicity was evaluated with the MTT assay. DTSCs were then induced for osteo/odontogenic differentiation by media containing dexamethasone, KH(2)PO(4),β-glycerophosphate and L-ascorbic acid phosphate in the presence of nontoxic concentrations of HEMA (0.05-0.5mM) and TEGDMA (0.05-0.25mM) for 3 weeks. Additionally, the effects of a single exposure (72 h) to higher concentrations of HEMA (2mM) and TEGDMA (1mM) were also evaluated. RESULTS DTSCs cultures were positive for STRO-1 (7.53±2.5%), CD146 (91.79±5.41%), CD34 (11.87±3.02%) and negative for CD45. In the absence of monomers cell migration, differentiation and production of mineralized dentin-like structures could be observed. Cells also progressively expressed differentiation markers, including dentin sialophosphoprotein-DSPP, bone sialoprotein-BSP, osteocalcin-OCN and alkaline phosphatase-ALP. On the contrary, long-term exposure to nontoxic concentrations of HEMA and TEGDMA significantly delayed the differentiation and mineralization processes of DTSCs, whereas, one time exposure to higher concentrations of these monomers almost completed inhibited mineral nodule formation. BSP, OCN, ALP and DSPP expression were also significantly down-regulated. SIGNIFICANCE These findings suggest that HEMA and TEGDMA can severely disturb the odontogenic differentiation potential of pulp stem/progenitor cells, which might have significant consequences for pulp tissue homeostasis and repair.

Effect of Lactobacillus-protective cultures with bacteriocin-like inhibitory substances¿ producing ability on microbiological, chemical and sensory changes during storage of refrigerated vacuum-packaged sliced beef

Aims:  To investigate the effect of applying two different Lactobacillus‐protective cultures, with bacteriocin‐like inhibitory substances’ (BLIS) producing ability, individually or in combination, on microbiological, chemical and sensory changes during storage of refrigerated vacuum‐packaged sliced beef meat.

Effect of age and sex on craniomandibular disorders

The purpose of this study was to investigate the effect of age and sex on craniomandibular disorders. A total of 195 subjects were examined, 148 women and 47 men. The range of age was 16 to 70 years. Clinical and anamnestic examination data established the ratio of women to men as 4:1. The prevalence of clicking, headache, teeth-clenching, hypomobility, difficulty in mastication, and neuromuscular symptoms was greater among young women than in other age and sex groups. There was a statistically significant correlation between severity of symptoms and age among women, and a relative decline by age in the prevalence of symptoms for both sexes. Finally, younger patients showed a greater prevalence of neuromuscular symptoms, whereas older patients had more peripheral symptoms.

Computer methods for automating preoperative dental implant planning: Implant positioning and size assignment

The paper presents computer-aided methods that allocate a dental implant and suggest its size, during the pre-operative planning stage, in conformance with introduced optimization criteria and established clinical requirements. Based on computed tomography data of the jaw and prosthesis anatomy, single tooth cases are planned for the best-suited implant insertion at a user-defined region. An optimum implantation axis line is produced and cylindrical implants of various candidate sizes are then automatically positioned, while their occlusal end is leveled to bone ridge, and evaluated. Radial safety margins are used for the assessment of the implant safety distance from neighboring anatomical structures and bone quantity and quality are estimated and taken into consideration. A case study demonstrates the concept and allows for its discussion.

Is there a potential for durable adhesion to zirconia restorations? A systematic review

STATEMENT OF PROBLEM With a number of zirconia ceramic materials currently available for clinical use, an overview of the scientific literature on the adhesion methods and their potential influence is indicated. PURPOSE The purpose of this systematic review was to classify and analyze the existing methods and materials proposed to improve adhesion to zirconia surfaces. MATERIAL AND METHODS The current literature of in vitro studies examining the bond strength on zirconia ceramics, including clinical studies from 1998 until 2014, was analyzed. A search of the English language literature was undertaken using MEDLINE and PubMed, and a hand search was made for any relevant research paper from the library of a dental school. Papers evaluating only alumina restoration bond or ceramic-zirconia bond were excluded. RESULTS A total of 134 publications were identified for analysis. Different adhesive techniques with different testing methods were reviewed. Results were difficult to compare in that the parameters varied in each research protocol. CONCLUSIONS Airborne-particle abrasion and tribochemical silica coating are reference pretreatment methods. Adhesive monomers are necessary for chemical bonding. Surface contamination and aging have negative effects on adhesion to zirconia. Many factors influence each combination of zirconia material, such as surface treatment, adhesive medium, and aging conditions. Laboratory studies should be confirmed by clinical trials.

Effect of in vitro aging on the flexural strength and probability to fracture of Y-TZP zirconia ceramics for all-ceramic restorations.

OBJECTIVES Dental zirconia restorations should present long-term clinical survival and be in service within the oral environment for many years. However, low temperature degradation could affect their mechanical properties and survival. The aim of this study was to investigate the effect of in vitro aging on the flexural strength of yttrium-stabilized (Y-TZP) zirconia ceramics for ceramic restorations. METHODS One hundred twenty bar-shaped specimens were prepared from two ceramics (ZENO Zr (WI) and IPS e.max(®) ZirCAD (IV)), and loaded until fracture according to ISO 6872. The specimens from each ceramic (nx=60) were divided in three groups (control, aged for 5h, aged for 10h). One-way ANOVA was used to assess statistically significant differences among flexural strength values (P<0.05). The variability of the flexural strength values was analyzed using the two-parameter Weibull distribution function, which was applied for the estimation of Weibull modulus (m) and characteristic strength (σ0). The crystalline phase polymorphs of the materials (tetragonal, t, and monoclinic, m, zirconia) were investigated by X-ray diffraction (XRD) analysis, Raman spectroscopy and Fourier transform infrared (FTIR) spectroscopy. RESULTS A slight increase of the flexural strength after 5h, and a decrease after 10h of aging, was recorded for both ceramics, however statistically significant was for the WI group (P<0.05). Both ceramics presented a t→m phase transformation, with the m-phase increasing from 4 to 5% at 5h to around 15% after 10h. SIGNIFICANCE The significant reduction of the flexural strength after 10h of in vitro aging, suggests high fracture probability for one of the zirconia ceramics tested.

Comparative characterization of STRO-1neg/CD146pos and STRO-1pos/CD146pos apical papilla stem cells enriched with flow cytometry

OBJECTIVE Stem Cells residing in the Apical Papilla (SCAP) of human permanent teeth represent a promising cell source for dental tissue regeneration. Therefore, the functional and molecular properties of specific subpopulations existing within heterogeneous cultures should be further investigated to give insight whether their selection could be beneficial for targeted therapeutic applications. DESIGN In this study we extensively characterized SCAP cultures established from 10 healthy subjects, as well as their STRO-1(pos/)CD146(pos) and STRO-1(neg/)CD146(pos) subpopulations isolated with fluorescence-activated cell sorting. SCAP were analyzed for embryonic (Nanog, Oct3/4, SSEA-3, TRA-1-60), mesenchymal (STRO-1, CD146/MUC18, CD105/endoglin, CD24, CD90/Thy-1, CD81-TAPA, CD34, CD49f/a6-integrin), neural (CD271/NGFR, nestin) and hematopoietic (CD117/c-kit, CD45) stem cell (SC) markers using flow cytometry. Multipotentiality was evaluated with culture specific staining (Alizarin-Red-S, Oil- Red-O) and RT-PCR analysis for osteo/odontogenic (DSPP, BSP, ALP, osteocalcin, osteonectin, BMP-2, Runx2), adipogenic (lipoprotein-lipase-LPL) and neurogenic (Neurofilament/NFL-L, nestin, β-tubulin-III, NCAM) markers. RESULTS Our results showed that the STRO-1(pos)/CD146(pos) subpopulation demonstrated higher CFU efficiency and much higher expression of several embryonic and mesenchymal SC markers compared to the non-sorted SCAP. They also showed enhanced odontogenic differentiation potential, as evidenced by higher mineralization capacity and expression of osteo/odontogenic markers. By contrast, absence of STRO-1 in the STRO-1(neg)/CD146(pos) subpopulation yielded the opposite results and was associated with significant downgrading of the above-mentioned properties. CONCLUSIONS These results suggest that STRO-1(pos)/CD146(pos) SCAP cells represent a very promising adult MSCs source with enhanced multipotent SC properties that could be easily isolated with simple flow cytometric methods to be used for tissue engineering applications.

Comparison of cell proliferation on modified dental ceramics

The aim of this study was to investigate the influence of substrate characteristics such as chemical composition and surface morphology of dental ceramics to support cell attachment and proliferation. Thus, body (B) and shoulder (S) porcelain differing on their surface morphology and composition were treated with oxides CaO or CaO and P(2)O(5) and four modified ceramics BCa, BCaP, SCa, SCaP were constructed, respectively. The modified ceramics differ from their controls concerning their surface morphology as evaluated by Scanning Electron Microscope (SEM), and their surface chemical composition (Na, KP and Ca) as evaluated by Energy Dispersing Spectroscopy (EDS). All modified ceramics support better than the control ceramics the cell proliferation over 72 h incubation period. Furthermore, higher rates of cell proliferation was detected in shoulder modified ceramics (SCa and SCaP) than in all other cases.