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Biomedical subjects

Gottfried Schmalz

Publications and source records attributed to Gottfried Schmalz.

At least 19 recordsLinked to original sources

Chemiluminescence-based detection and comparison of protein amounts adsorbed on differently modified silica surfaces.

The biological consequences of protein adsorption on biomaterial surfaces are considered to be of utmost importance for their biocompatibility. A new method based on amino group-labeling coupled to a chemiluminescence reaction for direct determination of proteins adsorbed on material surfaces was employed. This method was used to explore the effects of surface chemistry and surface roughness on protein adsorption in a silicon oxide model system. Corundum sandblasting was applied to silicon wafers to create roughened surfaces while immobilization of fluorocarbon-, hydrocarbon-, and poly(ethylene glycol)-containing silanes produced surfaces of varying wettability. The adsorption behavior of two complex body fluids, human serum and saliva, and of two purified components, human serum albumin and fibronectin, was strongly influenced by the surface parameters. A general tendency to higher amounts of adsorbed protein was found on roughened surfaces and modification with poly(ethylene glycol) or with fluorocarbon moieties reduced protein adsorption. The values obtained with the new method could be confirmed by a colorimetric determination of protein amounts adsorbed on identically modified silica beads and were in accordance with those previously reported utilizing established methods for protein quantification. The presented method, which was methodically simple to perform and allowed the simultaneous measurement of a large number of samples, may be of future value for high-throughput surveying of the protein adsorption characteristics of biomaterials.

Adsorption↗

Two-year clinical performance of cast gold vs ceramic partial crowns.

Cast gold partial crowns (CGPC) are an accepted means of restoring posterior teeth with extended lesions. However, for esthetic reasons, CGPC are being increasingly substituted with partial ceramic crowns (PCC). The aim of the present prospective split-mouth study was to compare the clinical performance of PCC and CGPC. There were 29 patients (male 12, female 17) who participated in the investigation for a total of 58 restorations. In each patient, one CGPC (Degulor C) and one PCC (Vita MarkII/Cerec III) were placed. CGPC were inserted using conventional zinc-phosphate cement (Harvard); PCC were adhesively luted to the cavities (Variolink II/Excite). The restorations were clinically rated using modified United States Public Health Service (USPHS) criteria at baseline and 1 and 2 years after placement. The median patient age was 38 years (range 25-54). There were 29 of the CGPC and 14 PCC placed in molars, while 15 PCC were placed in premolars. All patients were available for the 1- and the 2-year recall. One PCC (1.7%) failed and had to be replaced after 2 years in situ. The rest of the restorations were functional without need of replacement. The evaluation using USPHS criteria revealed no statistically significant differences between CGPC and PCC with the exception of anatomic form: PCC showed occlusal chipping in two cases without need of replacement. From these data, it can be concluded that PCC may provide an esthetic and tissue-conservative alternative to CGPC. However, long-term studies comparing the clinical performance and longevity of cast gold and ceramic partial crowns for posterior teeth are desirable.

Adult↗

Proteome analysis of glandular parotid and submandibular-sublingual saliva in comparison to whole human saliva by two-dimensional gel electrophoresis.

The secretions of the salivary parotid and submandibular-sublingual (SMSL) glands constitute the main part of whole human saliva (WS) in which proline-rich proteins (PRPs) and mucins represent dominant groups. Although proteome analysis had been performed on WS, no identification of PRPs or mucins by 2-DE and MS was achieved in WS and no comprehensive analysis of both glandular secretions is available so far. The aim of this study was to compare the protein map of WS to parotid and SMSL secretions for the display of PRPs and mucins. WS and glandular secretions were subjected to 2-DE and spots were analyzed by MALDI-MS. New components identified in WS were cyclophilin-B and prolyl-4-hydroxylase. Also acidic and basic PRPs as well as the proline-rich glycoprotein (PRG) could now be mapped in WS. Acidic PRPs were found equally in parotid and SMSL secretions, whereas basic PRPs and PRG were found primarily in parotid secretion. Salivary mucin MUC7 was identified in SMSL secretion. Thus, the more abundant proteins of WS can be explained mainly by mixed contributions of parotid and SMSL secretions with only few components remaining that may be derived from local sources in the oral cavity.

Amino Acid Sequence↗

Human pulp-derived cells immortalized with Simian Virus 40 T-antigen.

Primary cells in culture have a limited capacity to divide and soon reach a non-proliferative state. This cellular senescence limits the investigation of cells derived from human pulp concerning cellular pathways, gene regulation, mechanisms of dentin formation, or responses to material exposure. To overcome this problem, primary human pulp-derived cells were established and transfected with a plasmid containing coding sequences of Simian Virus 40 (SV40) large T-antigen. This resulted in the establishment of several cell clones showing an extension of life span. Expression of T-antigen transcripts and protein was verified by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. Primary human pulp cells were cultured until senescence (i.e. up to passage 7) and transfected cells could be cultured to passage 18 after transfection, when a cellular crisis with massive cell death occurred. One clone escaped from crisis and has been maintained in culture for 55 wk. Experiments were performed to characterize transfected cells in comparison to primary cells. Cell morphology and proliferation were analyzed, and expression of cell-specific gene transcripts and proteins (including collagen types I and III, alkaline phosphatase, bone sialoprotein, osteocalcin, and dentin sialophosphoprotein and dentin matrix protein I) was detected by RT-PCR and immunohistochemistry. Transfection of human pulp-derived cells resulted in an immortalized cell line retaining many of the phenotypic characteristics observed in primary cells.

Adolescent↗

Effect of N-acetyl-L-cysteine on ROS production and cell death caused by HEMA in human primary gingival fibroblasts.

Previous investigations have shown that 2-hydroxyethyl methacrylate (HEMA) causes reactive oxygen species (ROS) production, which in turn affects cell survival and cell death. The purpose of this study was to evaluate the effects of the antioxidant N-acetyl-L-cysteine (NAC) on HEMA-induced toxicity in human primary gingival fibroblasts (HGF). HGF were treated with various concentrations of HEMA (0-12 mm) in the absence and presence of NAC (1, 5, and 10 mm). The 3-(4,5 dimethyiazol-2-1)-2-5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the mitochondrial dehydrogenase activity after HEMA exposure. Viability and cell death were determined by flow cytometry using Annexin V and PI staining. ROS production was detected by the increasing fluorescence of the oxidation-sensitive dye 2',7'-dichlorofluorescein diacetate (DCFH-DA) after HEMA treatment. After a 24h incubation period, HEMA concentrations higher then 10mm caused a decrease of cell viability, mitochondrial activity, and an increase of cell death. HEMA concentrations of 4-12 mm markedly increased ROS levels in a dose-dependent manner. High NAC concentrations (5 and 10 mm) significantly reduced cell death, and restored the mitochondrial activity after a 24 h co-treatment, but 1 mm NAC increased HEMA toxicity (p<0.05). All NAC concentrations significantly reduced ROS levels induced by HEMA after a 2 h exposure (p<0.05), but no such reduction was observed after a 4 h treatment. Furthermore, treatment with 10 mm HEMA and 1 mm NAC for 6h caused an increase in ROS levels compared to 10 mm HEMA alone (p<0.05). In conclusion, our results suggest that high NAC concentrations protect HGF against HEMA cytotoxicity by reducing the induced ROS levels.

Acetylcysteine↗

Bond strength of a new universal self-adhesive resin luting cement to dentin and enamel.

The objective of this study was to assess the bonding performance of a new universal self-adhesive cement RelyX Unicem (RXU) to dentin and enamel compared to four currently used luting systems, using a shear bond strength test with and without thermocycling. Median bond strengths were determined after 24 h storage, and after thermocycling (6,000 cycles, 5-55 degrees C) for RXU and compared to Syntac/Variolink II (SynC/V) as a standard for luting conventional ceramics, ED-Primer II/Panavia F2.0 (EDII/PF2), Prime and Bond NT/Dyract Cem Plus (PBNT/DyCP), and a glass ionomer cement, Ketac Cem (KetC), as a standard for luting high-strength ceramic and metal-based restorations. Data (n=10 per group) were statistically analyzed using the Mann-Whitney-Wilcoxon test at the 0.05 level of significance. The bond strength (MPa) of RXU to dentin (10.8) was not statistically different from those of SynC/V (15.1), EDII/PF2 (10.5) or PBNT/DyCP (10.1), and statistically higher than KetC (4.1). The bond strength of RXU to enamel (14.5) was significantly lower than those of SynC/V (32.8), EDII/PF2 (23.6), and PBNT/DyCP (17.8), but higher than KetC (6.1). After thermocycling, the bond strength of RXU to enamel significantly decreased, but was still significantly higher than that of KetC. RelyX Unicem may be considered an alternative to Ketac Cem for high-strength ceramic or metal-based restorations, and may be used for luting conventional ceramic crowns with little or no enamel left.

Compomers↗

Cytotoxic and mutagenic effects of dental composite materials.

Mutagenicity of single compounds of dental resinous materials has been investigated on many occasions before, but the induction of mutagenic effects by extracts of clinically used composites is still unknown. Here, cytotoxic effects and the formation of micronuclei were determined in V79 fibroblasts after exposure to extracts of modern composite filling materials (Solitaire, Solitaire 2, Tetric Ceram, Dyract AP, Definite). For cytotoxicity testing, test specimens were aged for various time periods (0, 24, and 168 h), and V79 cells were then exposed to dilutions of the original extracts for 24, 48, and 72 h. The ranking of the cytotoxic effects of the composites according to EC50 values after a 24-h exposure period was as follows: Solitaire (most toxic)=Solitaire 2<Tetric Ceram<Dyract AP<Definite (least toxic). Cytotoxicity was independent of the period of aging for each composite, but varied with exposure periods. The cytotoxic effect of Solitaire increased about two-fold between exposure periods of 24, 48, and 72 h, no changes were observed with Solitaire 2, and cytotoxicity of Tetric Ceram, Dyract AP, and Definite was reduced. Even eight-fold diluted original extracts of freshly mixed Solitaire test specimens increased the numbers of micronuclei about 10-fold, and Solitaire 2 was slightly less effective. The mutagenic effects of these materials were reduced in the presence of a rat liver homogenate (S9). Weak increases of the numbers of micronuclei were detected only with undiluted extracts of Tetric Ceram and Dyract AP, but Definite was not effective. Our findings suggest that mutagenic components of biologically active composite resins should be replaced by more biocompatible substances to avoid risk factors for the health of patients and dental personnel.

Animals↗

The effect of triethylene glycol dimethacrylate on the cell cycle of mammalian cells.

The induction of DNA damage by a genotoxic agent is a signal leading to cell cycle delay, and thereby enables and induces DNA repair prior to cell cycle progression. Triethylene glycol dimethacrylate (TEGDMA), a monomer of dental resinous materials, caused mutagenic effects in mammalian cells probably as a consequence of DNA damage. Therefore, we hypothesized that TEGDMA will induce a cell cycle delay in mammalian cells. Here, cell lines deficient and proficient of a functional p53 tumor suppressor protein were used to study the effects of TEGDMA on the various phases of the cell cycle. V79 Chinese hamster lung fibroblasts (p53 deficient), N1 human skin fibroblasts (p53 proficient), and primary human pulp fibroblasts (p53 proficient) were exposed to increasing TEGDMA concentrations (0-3 mmol/l). Cell survival and vitality were determined after a 24-h exposure period and a 24-h recovery period, and the distribution of cells between the phases of the cell cycle in untreated and TEGDMA-treated cultures was analyzed by flow cytometry. The majority of the TEGDMA-treated V79 cells accumulated in G2 phase. In contrast, about 30% of human N1 fibroblasts were reversibly blocked in G1 phase by 0.5-3.0 mmol/l TEGDMA. The fraction of G2-phase cells was increased only by high TEGDMA concentrations. The percentage of human pulp cells in G1 phase increased very slightly with 1 mmol/l TEGDMA, but cell numbers in G1 phase were reduced by 10-20% by 1.5-3 mmol/l TEGDMA. The percentage of pulp cells in G2 phase increased about 2-fold without any obvious effect of a 24-h recovery period. Therefore, TEGDMA caused cell cycle delays through p53-dependent and independent pathways in the various cell lines. From these results, we conclude that TEGDMA may influence physiological processes like cell growth and differentiation of human pulp cells in vivo.

Animals↗

Effect of temporary cements on the bond strength of ceramic luted to dentin.

OBJECTIVES: The purpose of this study was to evaluate the effect of either eugenol-containing or eugenol-free temporary cement removal by excavator or sandblasting on the shear bond strength of ceramic luted to dentin. METHODS: A self-etching primer system, Panavia F 2.0, Kuraray Medical (PF2), and a total-etch bonding system, Excite/Variolink II, Vivadent (EXV), were used. One hundred and forty human molars, ground to expose dentin surfaces, were divided into 14 groups (seven groups for each adhesive system). For each adhesive system, either a eugenol-containing (Temp bond) or a eugenol-free (Temp bond NE) temporary cement was applied to the dentin surface for 7 days, then removed by an excavator or sandblasting (four groups). Three control groups were studied where fresh dentin was either scratched by excavator or sandblasted, or underwent no surface treatment. After application of the adhesives, ceramic cones (Cerafil inserts) were adhesively luted to standardized dentin areas. After 24 h storage in distilled water, the shear bond strengths were determined at a cross-head speed of 0.75 mm/min. RESULTS: For each adhesive system, neither the method of temporary cement removal nor the type of temporary cement affected the bond strength significantly (P<or=0.05). EXV showed statistically higher bond strengths (26.6-31.6 MPa) than PF2 (8.6-12.9 MPa) within all groups. SIGNIFICANCE: The use of temporary cements, either containing eugenol or not, does not alter the retentive strength of ceramic restorations luted to dentin using the tested adhesive systems, whether the temporary cements are removed by excavator or sandblasting.

Air Abrasion, Dental↗

Influence of dentin on the effectiveness of antibacterial agents.

The influence of dentin on the effectiveness of three antibacterial agents (triclosan, glutaraldehyde, NaOCl) on Streptococcus mutans, S. sobrinus, and Lactobacillus acidophilus was tested using the agar diffusion method with and without bovine dentin discs (200 mum and 500 mum thickness) placed between bacteria and test substances. The effect of 0.3% triclosan on all tester strains (100%) was reduced after passage through 500 mum dentin discs to 0% (L. acidophilus) and to 22% and 28% (S. mutans and S. sobrinus). Seal&Protect (Dentsply, Konstanz, Germany), a triclosan containing dental bonding agent, produced inhibition zones only against S. mutans, but no zone when applied on 200 mum dentin discs. The inhibition zones for 1% NaOCl and 5% glutaraldehyde against all tester strains were significantly increased up to 230% (glutaraldehyde) and 236% (NaOCl) when applied on dentin discs, compared to direct application (100%). Dentin may either decrease or increase the inhibitory effect of antibacterial agents.

Animals↗

Selective influence of dentin thickness upon cytotoxicity of dentin contacting materials.

The effect of three dentin contacting materials on three-dimensional cultures of pulp-derived cells was evaluated in a dentin barrier test device. The test materials (Syntac Classic, Prompt L-Pop, Vitrebond) were applied on dentin disks of different thicknesses ranging from 100 to 500 microm. After 24 h of exposure with and without perfusion of the test chamber, cell survival was evaluated using the MTT assay and related to a nontoxic control material. Syntac Classic decreased cell activity significantly (p </= 0.0003), independently of the dentin thickness. For Prompt L-Pop and Vitrebond a significant influence of dentin thickness was found on the cell reaction. After exposure of the control material, photometric readings showed no dependency of the cellular reaction on dentin thickness (p > 0.05). It could be demonstrated that dentin acts as a barrier, decreasing the elicited cytotoxicity with increasing thickness. This effect is material related, showing little influence for the nontoxic or the glutaraldehyde containing material.

Animals↗

Morphological characterization of the resin-dentin interface in primary teeth.

The effects of self-etching adhesives on primary teeth were evaluated. Reports in the literature suggest differences between the first and second dentition regarding the composition and the morphology and, therefore, a possible difference in the performance of dental adhesives. The purpose of this study was to evaluate the effects of self-etching adhesives on primary dentin. Eight dentin disks were obtained from the occlusal surface of primary molars. The disks were divided between four dental adhesives [Prompt L-PoP (LP), Clearfil SE BOND (SE), Etch&Prime 3.0 (EP) and Prime&Bond NT (PB) + H(3)PO(4) (Control) and "restored" with a composite (Pertac II)]. After sectioning, fixation and HMDS drying, specimens were polished and Field Emission SEM examinations were carried out. Clearly visible hybrid layer formation was found for PB, LP and EP. An undoubtedly detectable interdiffusion zone was not evident after the use of SE. A clearly visible adhesive layer was recognizable for PB, EP and SE, but not continuously detectable for LP. Debonded regions were observed for all systems evaluated, but distinct differences in the failure mode were detected. The evaluated dental adhesives did not generate completely sealed interfaces between the composite resin and the dentin of primary teeth in vitro.

Acid Etching, Dental↗

Mutagenic activity of structurally related oxiranes and siloranes in Salmonella typhimurium.

Ring-opening molecules like oxiranes (epoxides) maybe suitable for the development of non-shrinking dental composite materials. Since oxiranes are reactive molecules, they can cause adverse biological effects in living organisms. The introduction of siloranes, a merger of silane and oxirane, may solve this problem. Here, new oxiranes and siloranes were analyzed for the induction of mutations in Salmonella typhimurium (TA97a, TA98, TA100, and TA102), and a reactive oxirane molecule served as a reference. This chemical, epoxy cyclohexyl methyl-epoxy cyclohexane carboxylate (Est-Ep) tested positive in S. typhimurium TA100. The numbers of mutants were about 3-10-fold higher than controls in the presence of a metabolically active S9 fraction isolated from rat liver. Only a weak mutagenic effect was observed after direct testing (without S9). Di(cyclohexene-epoxidemethyl)ether (Eth-Ep) also caused a slight increase of mutant numbers in TA100 both in the presence and absence of S9. In contrast, no effects were detected with the large oxirane molecules, 2,2-bis(4,1-phenylenoxy-3,1-propanediyl-3-oxatricyclo [3.2.1.0(2,4)]octylcarboxy) propylidene (Nor-BP-Ep) and 2,2-bis(4,1-phenylenoxy-3,1-propanediyl-3,4-epoxycyclo-hexylcarboxylic-acid) propylidene (Est-BP-Ep). As to the siloranes, 1,4-bis(2,3-epoxypropyloxypropyl-dimethylsilyl)-benzene (Phen-Glyc) was a direct mutagen in S. typhimurium TA100 and TA102. This weak but dose-related increase of revertants was even enhanced by S9. Other siloranes, like di-3,4-epoxy cyclohexylmethyl-dimethyl-silane (DiMe-Sil), methyl-bis[2-(7-oxabicyclo[4.1.0]hept-3-yl)phenyl silane (Ph-Sil), and 1,3,5,7-tetrakis(ethyl cyclohexane epoxy)-1,3,5,7-tetramethyl-cyclotetrasiloxane (TET-Sil) tested negative in all S. typhimurium strains. All compounds will be further analyzed for the formation of chromosomal aberrations in mammalian cell cultures.

Animals↗

Bond strength of self-etching adhesives to dental hard tissues.

Since most cavity preparations show not only areas of exposed enamel and superficial dentin but also of deep dentinal areas, it would be desirable to use a dental adhesive which can obtain equally good bond strength to these hard tissues. The objective of this study was to measure the bond strength of self-etching adhesives to enamel and superficial and deep dentin. Human molars were ground to expose a surface of enamel or superficial (2.0-2.5 mm distance to pulp) or deep dentin (0.5-1.0 mm), embedded in acrylic resin and assigned to three groups (total n=90). The adhesives tested were: Clearfil SE Bond (SE), Prompt L-Pop (LP), and Etch&Prime 3.0 (EP). After application of the adhesives, an inverted, truncated cone of composite (Pertac II, ESPE) was cured. After storage, specimens were debonded in tension at a cross-head speed of 0.5 mm/min. The median tensile bond strength values to enamel were SE 29.1 MPa, LP 22.4 MPa, and EP 21.5 MPa. The median bond strengths for superficial dentin were SE 27.3 MPa, LP 8.4 MPa, and EP 3.2 MPa. The median bond strengths for deep dentin were SE 20.2 MPa, LP 5.7 MPa, and EP 1.3 MPa. The adhesives tested bonded effectively to enamel, but only SE achieved promising bond strengths to superficial and deep dentin.

Acid Etching, Dental↗

Microleakage in class II composite resin restorations: application of a clearing protocol.

Microleakage at the cervical tooth/restoration interface of class II restorations was evaluated, either on multiple sections or on teeth rendered transparent, in order to determine whether a modified clearing protocol could be established as a routine method for microleakage evaluation in class II restorations. Forty-eight class II cavities were restored with a composite/bonding agent (Tetric Universal/Heliobond, Vivadent, Schaan, Liechtenstein) in bulk technique (BT) (n=24) and with a composite/dentin adhesive system (Tetric Universal/Heliobond/Syntac Classic) in increments (IT) (n=24). The 24 specimens were assigned to two groups. Group I: dye penetration (India ink), multiple sectioning (conventional method). Group II: dye penetration (India ink) and modified clearing protocol. Linear microleakage (% dye/section) along the cervical tooth/restoration interface on multiple sections, and microleakage patterns (% dye/surface) at the cervical surface on transparent teeth were evaluated with an image analyzing system. The data were statistically analyzed with the Mann Whitney U-test, Wilcoxon's rank sum test, and the error rates method. The application of the modified clearing protocol allowed for distinct evaluation of microleakage patterns. Either protocol revealed lower penetration values for IT than for BT. With the conventional protocol, the median (25-75% percentiles) percentage of dye penetration was 2% (1-46%) for IT and 40% (23-85%) for BT. The clearing protocol revealed dye penetration of 5% (1-22%) for IT and 16% (4-24%) for BT. The clearing protocol together with the use of India ink as tracer allows for evaluation and visualization of the continuous distribution of microleakage at the cervical tooth/restoration interface without loss of information due to sectioning, although this method seems to be less discriminative than the conventional sectioning technique.

Acrylates↗

Biological interactions of dental cast alloys with oral tissues.

OBJECTIVE: All dental cast alloys release metal ions into the oral environment which have the potential to interact with the oral tissues. Amount and type of metal elements released are varying and not directly related to the composition of the alloy. The aim of the present literature survey was to describe the interactions of dental cast alloys with living tissues and to relate them to clinically adverse local reactions of the oral tissues. RESULTS: Interactions of dental cast alloys with the oral tissues take place by different mechanisms; e.g. bacterial adherence promotion, toxic and subtoxic effects and allergy. Whereas bacterial adhesion promotion may be counteracted by adequate oral hygiene measures, the other mechanisms may lead to clinically adverse local reactions due to the metal present. However, the number of cases, where such a relation can be safely diagnosed, is small. Safe ground is felt with proven allergies. The comparatively high allergy rate towards nickel should be the impetus to replace those alloys whenever there is a suitable alternative. Medical and dental impairments as well as general medication may lead to symptoms allegedly attributed to metal restorations. CONCLUSIONS AND SIGNIFICANCE: Patients relating oral symptoms to metal restorations should be subjected to a thorough dental and general medical examination in order to exclude non-material related diseases being the cause for their complaints/symptoms. These cases are a challenge for the collaboration between the medical and dental professions.

Animals↗

Analysis of odontoblast gene expression using a novel approach, laser capture microdissection.

Studying the mechanisms of molecular interactions in developing tissues demands sensitive molecular biological in vivo and in vitro techniques. Laser capture microdissection (LCM) allows for the isolation of mRNA in histological sections even from single cells, thus enabling the identification of in vivo gene expression products in closely circumscribed tissue areas. The aims of this study were to assess the optimal fixation, processing, and staining conditions to retrieve RNA from microdissected odontoblasts. Fluorometric assays and RT-PCR analysis of alpha 1(I) collagen, dentin sialophosphoprotein (Dspp), and osteocalcin (OC) confirmed that the total RNA isolated from day 0 and day 3 captured odontoblasts was sufficient in quantity and quality. Our results indicate that individual odontoblasts obtained by LCM are morphologically intact and chemically unaltered, allowing accurate molecular and biochemical analyses.

Animals↗