Hermann Götz

Effect of diode laser irradiation on root surfaces in vitro.

OBJECTIVE The objective of this study was to evaluate possible morphological alterations of root surfaces after GaAlAs-diode laser (809 nm) irradiation under standardized in vitro conditions. MATERIALS AND METHODS Root specimens obtained from extracted periodontally diseased teeth were scaled and root planed with curettes followed by air-powder abrasive treatment prior to lasing. The variable parameters were power output (0.5-2.5 W) and exposure time (10-30 sec per specimen). Additionally, the effect of a saline solution and a human blood film on the root surface was investigated. The root segments were analyzed by means of a reflected light microscope. Photographs before and after irradiation were taken and evaluated. The scale of carbonization was quantified using a grid laid over the photographs. Specimens with distinct morphological changes were analyzed with a scanning electron microscope. RESULTS Lasing dry specimens and specimens moistened with saline resulted in no detectable alterations, irrespective of irradiation time and power output applied. Depending on different settings, irradiation caused severe damages to the root surface when segments were covered by a thin blood film. Irradiation at a power output of 1 Watt and below had barely any negative effect on the root surface, whereas lasing at 1.5, 2.0, and 2.5 Watt resulted in partial or total carbonizations of the root samples. The angle of irradiation had a significant effect on the scale of the root surface damage (Mann-Whitney U test,p < 0.05). CONCLUSION The diode laser may cause damage to periodontal hard tissues if irradiation parameters are not adequate.

In vitro evaluation of various bioabsorbable and nonresorbable barrier membranes for guided tissue regeneration

BackgroundDifferent types of bioabsorbable and nonresorbable membranes have been widely used for guided tissue regeneration (GTR) with its ultimate goal of regenerating lost periodontal structures. The purpose of the present study was to evaluate the biological effects of various bioabsorbable and nonresorbable membranes in cultures of primary human gingival fibroblasts (HGF), periodontal ligament fibroblasts (PDLF) and human osteoblast-like (HOB) cells in vitro.MethodsThree commercially available collagen membranes [TutoDent® (TD), Resodont® (RD) and BioGide® (BG)] as well as three nonresorbable polytetrafluoroethylene (PTFE) membranes [ACE (AC), Cytoplast® (CT) and TefGen-FD® (TG)] were tested. Cells plated on culture dishes (CD) served as positive controls. The effect of the barrier membranes on HGF, PDLF as well as HOB cells was assessed by the Alamar Blue fluorometric proliferation assay after 1, 2.5, 4, 24 and 48 h time periods. The structural and morphological properties of the membranes were evaluated by scanning electron microscopy (SEM).ResultsThe results showed that of the six barriers tested, TD and RD demonstrated the highest rate of HGF proliferation at both earlier (1 h) and later (48 h) time periods (P < 0.001) compared to all other tested barriers and CD. Similarly, TD, RD and BG had significantly higher numbers of cells at all time periods when compared with the positive control in PDLF culture (P ≤ 0.001). In HOB cell culture, the highest rate of cell proliferation was also calculated for TD at all time periods (P < 0.001). SEM observations demonstrated a microporous structure of all collagen membranes, with a compact top surface and a porous bottom surface, whereas the nonresorbable PTFE membranes demonstrated a homogenous structure with a symmetric dense skin layer.ConclusionResults from the present study suggested that GTR membrane materials, per se, may influence cell proliferation in the process of periodontal tissue/bone regeneration. Among the six membranes examined, the bioabsorbable membranes demonstrated to be more suitable to stimulate cellular proliferation compared to nonresorbable PTFE membranes.

Long‐term response of osteogenic cells on micron and submicron‐scale‐structured hydrophilic titanium surfaces: sequence of cell proliferation and cell differentiation

OBJECTIVE Modifications of surface topography and surface chemistry are key factors for guiding target cells during dental implant healing. Recent in vitro studies confirmed promotion of early osteogenic cell differentiation on submicron scaled surfaces in particular when hydrophilized. However, no long-term observations on both osteogenic cell proliferation as well as on cell maturation have been reported for respectively modified surfaces. Aim of this study was to monitor osteogenic cell proliferation and expression of specific osteogenic cell differentiation markers on a protein level over an extended period of 3 weeks with respect to surface modifications. MATERIAL AND METHODS Modified titanium (Ti) disks were obtained from Institute Straumann, representing the following surfaces: smooth pretreatment (PT), sandblasted/acid etched (SLA), and hydrophilized (modSLA). Surface topography was analyzed by scanning electron microscopy, surface elemental composition was assessed by X-Ray Photoelectronic Spectroscopy (XPS). Tissue culture polystyrene (TCPS) served as a control substrate. Human osteogenic cells (HOB-c) were cultivated on the respective substrates. After 24 hrs, 48 hrs, 72 hrs, 7 d, 14 d and 21 d, cell count was assessed as well as osteogenic cell differentiation utilizing cellular Quantitative Immuno-Cytochemistry (QIC) assay for collagen type I (COL), alkaline phosphatase (AP), osteopontin (OPN) and osteocalcin (OC). Data were normalized with respect to internal controls. RESULTS In contrast to the other modified Ti disks, modSLA stands out due to low surface carbon contamination. TCPS and PT surfaces preserved a rather immature, mitotic active osteogenic phenotype (high proliferation rates, no increase of OC production), SLA and especially modSLA surfaces promoted the maturation of osteogenic precursors into post-mitotic osteoblasts. In detail, modSLA resulted in lowest cell proliferation rates, but exhibited highest expression rates of OC. DISCUSSION Our results, which confirm previous studies, reveal long-term promotion of osteogenic cell maturation by topography (micron and submicron scale roughness) and surface hydrophilicity.

Bestimmung der Größe der spezifischen Oberfläche von Knochenersatzmaterialien mittels Gasadsorption

Fragestellung: Die Oberfläche sowie die Mikro- und Mesoporosität von Knochenersatzmaterialien beeinflussen deren chemische und biologische Eigenschaften. Daher wurden in dieser Untersuchung die Größe der spezifischen Oberfläche sowie die Verteilung der Porendurchmesser (Poren < 1 μm) in Knochenersatzmaterialien bestimmt. Material und Methode: Die untersuchten Hydroxylapatite waren synthetischen, bovinen und phytotrophen Ursprungs. Die Trikalziumphosphate und die Biogläser umfassten nur rein synthetische Materialien. ¶Die Gasadsorptionsuntersuchung je einer Probe erfolgte mit Hilfe eines volumetrischen N 2 -Kr-Systems (ASAP 2010, Micromeritics). Zusätzlich wurde für Materialien mit spezifischen Oberflächen ¶> 2 m 2 /g die Porengrößenverteilung nach der BJH-Methode ermittelt. Ergebnisse: 1. Spezifische Oberfläche: 2 der Materialien zeigten eine auffallend große spezifische Oberfläche (BioOss 79,7 m 2 /g, Algipore neu 14,6 m 2 /g). Eine mittlere Oberfläche zeigten Algipore alt (4,9 m 2 /g) und Interpore 200 (2,64 m 2 /g). Die übrigen Materialien zeigten nur kleine Oberflächen (Ceros 80 1,8 m 2 /g, Ceros 82 ¶1,31 m 2 /g, Cerasorb 1,2 m 2 /g, Biobase 0,7 m 2 /g, Endobone 0,7 m 2 /g, Perioglas 0,6 m 2 /g, Allotropat 50 0,2 m 2 /g, Biogran 0,2 m 2 /g). 2. Häufigkeitsverteilung des Porendurchmessers: Die Materialien mit großen und mittleren spezifischen Oberflächen zeigten folgende Porendurchmesser: BioOss 2–50 nm, Algipore neu 2–¶100 nm, Algipore alt 5–50 nm, Interpore 200 2–100 nm. Porengrößen < 2 nm fanden sich kaum. Schlussfolgerungen: Die Materialien BioOss, Algipore alt und neu und Interpore 200 haben ein großes interkonnektierendes Mesoporensystem (Durchmesser < 1 μm). Für die Materialien Biobase, Endobone, Perioglas, Allotropat 50 und Biogran ist dies nicht anzunehmen. Die Materialien Ceros 80, Ceros 82 und Cerasorb zeigen eine dazwischen liegende spezifische Oberflächengröße und weisen einen mäßigen Anteil von solchen interkonnektierenden Poren auf. Ein Einfluss der interkonnektierenden Porosität und der deutlich unterschiedlichen spezifischen Oberflächen auf das Verhalten der Knochenersatzmaterialien in vivo ist nahe liegend. The surface area and the microporosity of bone regeneration materials influence their chemical and ¶biological properties. Therefore, the size of the specific surface area and the distribution of the pore diameters (pores < 1 μm) of bone regeneration materials were analyzed within this study. The analyzed hydroxyapatites were of synthetic, bovine, and phytotroph origin. The tricalcium phosphates and the bioglasses included only synthetic materials. The gas adsorption of each specimen was analyzed using a volumetric N2/Kr system (ASAP 2010, Micromeritics). Additionally, for materials with a specific surface area (> 2 m2/g) the pore size distribution was evaluated by the BJH-method. Two of the ¶materials evaluated astonishingly large dimensions of the specific surface area (BioOss 79.7 m2/g, Algipore new 14.6 m2/g). A medium ¶surface area was found for Algipore old (4.9 m2/g) and Interpore200 ¶(2.64 m2/g). All other included materials showed only small sizes of the specific surface area (Ceros80 ¶1.8 m2/g, Ceros82 1.31 m2/g, Cerasorb 1.2 m2/g, Biobase 0.7 m2/g, Endobone 0.7 m2/g, Perioglas 0.6 m2/g, Allotropat50 0.23 m2/g, Biogran ¶0.2 m2/g). The materials with large and medium sizes of the specific surface area evaluated the following pore diameters: BioOss 2–50 nm, Algipore new 2–100 nm, Algipore old 5–50 nm, Interpore200 2–100 nm. Pore sizes less than 2 nm were not found in relevant numbers. The materials BioOss, old and new Algipore, and Interpore200 contain a large interconnecting mesopore system (diameter < 1 μm). For the materials Biobase, Endobone, Perioglas, Allotropat 50, and Biogran this cannot be assumed. The materials Ceros80, Ceros82, and Cerasorb evaluated a specific surface area between those and might include only a small part of these interconnecting pores. An influence of the interconnecting porosity and the different sizes of the specific surface areas on the biological behavior of the bone regeneration materials can be suggested.

Axial growth of hexactinellid spicules: Formation of cone-like structural units in the giant basal spicules of the hexactinellid Monorhaphis

The glass sponge Monorhaphis chuni (Porifera: Hexactinellida) forms the largest bio-silica structures on Earth; their giant basal spicules reach sizes of up to 3m and diameters of 8.5mm. Previously, it had been shown that the thickness growth proceeds by appositional layering of individual lamellae; however, the mechanism for the longitudinal growth remained unstudied. Now we show, that the surface of the spicules have towards the tip serrated relief structures that are consistent in size and form with the protrusions on the surface of the spicules. These protrusions fit into the collagen net that surrounds the spicules. The widths of the individual lamellae do not show a pronounced size tendency. The apical elongation of the spicule proceeds by piling up cone-like structural units formed from silica. As a support of the assumption that in the extracellular space silicatein(-like) molecules exist that associate with the external surface of the respective spicule immunogold electron microscopic analyses were performed. With the primmorph system from Suberites domuncula we show that silicatein(-like) molecules assemble as string- and net-like arrangements around the spicules. At their tips the silicatein(-like) molecules are initially stacked and at a later stay also organized into net-like structures. Silicatein(-like) molecules have been extracted from the giant basal spicule of Monorhaphis. Applying the SDS-PAGE technique it could be shown that silicatein molecules associate to dimers and trimers. Higher complexes (filaments) are formed from silicatein(-like) molecules, as can be visualized by electron microscopy (SEM). In the presence of ortho-silicate these filaments become covered with 30-60nm long small rod-like/cuboid particles of silica. From these data we conclude that the apical elongation of the spicules of Monorhaphis proceeds by piling up cone-like silica structural units, whose synthesis is mediated by silicatein(-like) molecules.

Comparative analysis of numerical and experimental data of orthodontic mini-implants

The purpose of this study was to compare numerical simulation data derived from finite element analysis (FEA) to experimental data on mini-implant loading. Nine finite element (FE) models of mini-implants and surrounding bone were derived from corresponding experimental specimens. The animal bone in the experiment consisted of bovine rib. The experimental groups were based on implant type, length, diameter, and angle of insertion. One experimental specimen was randomly selected from each group and was digitized in a microCT scanner. The FE models consisted of bone pieces containing Aarhus mini-implants with dimensions 1.5 × 7 mm and 1.5 × 9 mm or LOMAS mini-implants (dimensions 1.5 × 7 mm, 1.5 × 9 mm, and 2 × 7 mm). Mini-implants were inserted in two different ways, perpendicular to the bone surface or at 45 degrees to the direction of the applied load. Loading and boundary conditions in the FE models were adjusted to match the experimental situation, with the force applied on the neck of the mini-implants, along the mesio-distal direction up to a maximum of 0.5 N. Displacement and rotation of mini-implants after force application calculated by FEA were compared to previously recorded experimental deflections of the same mini-implants. Analysis of data with the Altman-Bland test and the Youden plot demonstrated good agreement between numerical and experimental findings (P = not significant) for the models selected. This study provides further evidence of the appropriateness of the FEA as an investigational tool in relevant research.

Histotomography of the odontoblast processes at the dentine–enamel junction of permanent healthy human teeth in the confocal laser scanning microscope

Abstract The translucency of teeth allows the non-destructive subsurface visualisation of their microstructure by confocal laser scanning microscopy (CLSM) at a level of about 150 μm below the surface. The dentine–enamel junction (DEJ) is accessible only directly adjacent to the cervix of the tooth. Therefore teeth have to be sectioned for studying marginal areas of the dental hard tissue. The potential of the technique for (pseudo) three-dimensional visualisation allows the study of an array of individual confocal images, the interpretation of which is similar to that of macroscopic tomographs (CT-scan, MRI). Additionally, the extended focus mode yields the overlay of individual confocal images in the form of a two-dimensional projection. This mode of operation proved to be particularly suited for the visualisation of odontoblast processes in their whole extension. The three-dimensional junction between enamel and dentine, the branches of the odontoblast processes and their interactions with the DEJ is demonstrable by CLSM without staining or other procedures of sample preparation. The direct microscopic comparison between samples, either fresh or kept in a humid chamber, and Technovit-embedded sample blocks gives evidence that the risk of artefacts by sample storage or by the embedding procedure is minimal. The tomographs limited to subsurface areas of the tissue also exclude mechanical surface artefacts due to grinding or cutting.

Haloperoxidase Mimicry by CeO2−x Nanorods Combats Biofouling

CeO2-x nanorods are functional mimics of natural haloperoxidases. They catalyze the oxidative bromination of phenol red to bromophenol blue and of natural signaling molecules involved in bacterial quorum sensing. Laboratory and field tests with paint formulations containing 2 wt% of CeO2-x nanorods show a reduction in biofouling comparable to Cu2 O, the most typical biocidal pigment.

Circumferential spicule growth by pericellular silica deposition in the hexactinellid sponge Monorhaphis chuni

SUMMARY The giant basal spicule of the hexactinellid sponge Monorhaphis chuni represents the longest natural siliceous structure on Earth. This spicule is composed of concentrically arranged lamellae that are approximately 10 μm thick. In the present study, we investigated the formation of outer lamellae on a cellular level using microscopic and spectroscopic techniques. It is shown that the formation of an outermost lamella begins with the association of cell clusters with the surface of the thickening and/or growing spicule. The cells release silica for controlled formation of a lamella. The pericellular (silica) material fuses to a delimited and textured layer of silica with depressions approximately 20–30 μm in diameter. The newly formed layer initially displays 40 μm wide, well-structured banded ribbons and only attains its plain surface in a final step. The chemical composition in the depressions was studied using energy dispersive X-ray spectroscopy and by staining with Texas Red. The data suggest that those depressions are the nests for the silica-forming cells and that silica formation starts with a direct association of silica-forming cells with the outer surface of the spicule, where they remain and initiate the development of the next lamellae.

Surface Functionalization of Orthopedic Titanium Implants with Bone Sialoprotein

Orthopedic implant failure due to aseptic loosening and mechanical instability remains a major problem in total joint replacement. Improving osseointegration at the bone-implant interface may reduce micromotion and loosening. Bone sialoprotein (BSP) has been shown to enhance bone formation when coated onto titanium femoral implants and in rat calvarial defect models. However, the most appropriate method of BSP coating, the necessary level of BSP coating, and the effect of BSP coating on cell behavior remain largely unknown. In this study, BSP was covalently coupled to titanium surfaces via an aminosilane linker (APTES), and its properties were compared to BSP applied to titanium via physisorption and untreated titanium. Cell functions were examined using primary human osteoblasts (hOBs) and L929 mouse fibroblasts. Gene expression of specific bone turnover markers at the RNA level was detected at different intervals. Cell adhesion to titanium surfaces treated with BSP via physisorption was not significantly different from that of untreated titanium at any time point, whereas BSP application via covalent coupling caused reduced cell adhesion during the first few hours in culture. Cell migration was increased on titanium disks that were treated with higher concentrations of BSP solution, independent of the coating method. During the early phases of hOB proliferation, a suppressive effect of BSP was observed independent of its concentration, particularly when BSP was applied to the titanium surface via physisorption. Although alkaline phosphatase activity was reduced in the BSP-coated titanium groups after 4 days in culture, increased calcium deposition was observed after 21 days. In particular, the gene expression level of RUNX2 was upregulated by BSP. The increase in calcium deposition and the stimulation of cell differentiation induced by BSP highlight its potential as a surface modifier that could enhance the osseointegration of orthopedic implants. Both physisorption and covalent coupling of BSP are similarly effective, feasible methods, although a higher BSP concentration is recommended.