Cobalt chromium alloys in fixed prosthodontics: Investigations of mechanical properties and microstructure.

STATEMENT OF PROBLEM: Cobalt chromium (Co-Cr) alloys possess beneficial mechanical properties because alloys, even in thin sections, can resist high mastication forces and exhibit an acceptable bond to the surface porcelain layer. Traditional manufacturing techniques of Co-Cr alloys such as casting have been replaced with newer fabrication techniques, such as milling, laser melting, and presintered milling. Despite scarce documentation, these new manufacturing techniques are being used to fabricate dental and implant constructions. PURPOSE: This in vitro study investigates the hardness, yield strength, elastic modulus, and microstructure of the most commonly used Co-Cr alloys for fixed prosthodontics based on manufacturing technique. In addition, this study investigates the effect of heat treatment on the mechanical properties and microstructure of these materials. MATERIAL AND METHODS: Five Co-Cr alloys were included (dumbbell and rectangular shaped) based on four manufacturing techniques: cast, milled, laser melted, and presintered milled. Commercially pure titanium grade 4 and titanium-6 aluminum-4 vanadium ELI (extra low interstitial) were included for comparison, and yield strength and elongation after fracture were evaluated. The specimens were tested for hardness using the Vickers test and for elastic modulus using a nondestructive impulse excitation technique. The microstructure of selected specimens was analyzed using focused ion beam-scanning electron microscopy (FIB-SEM) and energy dispersive X-ray spectroscopy (EDS). RESULTS: The mechanical properties depend on the manufacturing technique used; the laser-melted and presintered Co-Cr specimens demonstrated the highest mechanical properties, followed by the milled and cast groups. Both the laser-melted and the presintered milled Co-Cr specimens showed smaller grain size compared with the cast and milled Co-Cr specimens. The titanium-6 aluminum-4 vanadium ELI demonstrated higher hardness and yield strength compared to commercially pure titanium grade 4. No major differences were observed for the selected materials regarding the mechanical properties and microstructural appearance after heat treatment. CONCLUSIONS: The laser melting and presintered milling techniques produced higher mechanical properties compared with the cast and milled Co-Cr. These findings were confirmed through microstructural analysis with respect to the grain size, precipitation, and number of pores.

Early Cup Loosening After Metal-on-Metal Total Joint Replacement of the Trapeziometacarpal Joint: A Retrieval Study.

PURPOSE: To investigate the possible mechanisms behind early cup loosening in a metal-on-metal trapeziometacarpal joint replacement. METHODS: The trapezia from 5 female patients were removed as part of a salvage procedure after a median of 22 months (range, 7-43 months) after implantation. Three osteoarthritic patients with symptomatic cup loosening and 2 with instability had a median age of 62 years (range, 59-65 years) at primary surgery. The trapezia with cups in situ were preserved and processed for histomorphometry. Studies with laser ablation inductively coupled plasma mass spectrometry and scanning electron microscopy with energy dispersive x-ray spectroscopy were also performed on 2 of the specimens. RESULTS: In all 5 specimens, osteolytic lesions undermined the cups and were also seen at the cup edges, completely surrounding the loose cups. Large amounts of dark particular material were seen in the periprosthetic tissues, mostly internalized by macrophages. The presence of chrome and cobalt in these regions was confirmed. Four of the 5 cups showed marked or complete loss of hydroxyapatite. CONCLUSIONS: We have found several possible explanations for the poor performance of this cup, including its cannulated design and metal-on-metal bearing. The changes seen are early and advanced, raising serious concerns about the implant and particularly the articulation. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic V.

Bone tissue response following local drug delivery of bisphosphonate through titanium oxide nanotube implants in a rabbit model.

OBJECTIVES: The objective of this study was to evaluate whether surface chemistry-controlled TiO2 nanotube structures may serve as a local drug delivery system for zoledronic acid improving implant-bone support. METHODS: Twenty-four screw-shaped Ti implants with surface chemistry-controlled TiO2 nanotube structures were prepared and divided into a zoledronic acid-formatted test and a native control group. The implants were inserted into contra-lateral femoral condyles in 12 New Zealand White rabbits. Bone support was evaluated using resonance frequency analysis (RFA) and removal torque (RTQ), as well as histometric analysis following a 3-weeks healing interval. RESULTS: Zoledronic acid-formatted TiO2 nanotube test implants showed significantly improved implant stability and osseointegration measured using RFA and RTQ compared with control (p < 0.05), and showed significantly enhanced new bone formation within the root of the threads compared with control (p < 0.05). CONCLUSIONS: TiO2 nanotube implants may prove to be a significant delivery system for drugs or biologic agents aimed at supporting local bone formation. Additional study of candidate drugs/agents, optimized dosage and release kinetics is needed prior to evaluation in clinical settings.

Physical exercise increases adult hippocampal neurogenesis in male rats provided it is aerobic and sustained.

KEY POINTS: Aerobic exercise, such as running, enhances adult hippocampal neurogenesis (AHN) in rodents. Little is known about the effects of high-intensity interval training (HIT) or of purely anaerobic resistance training on AHN. Here, compared with a sedentary lifestyle, we report a very modest effect of HIT and no effect of resistance training on AHN in adult male rats. We found the most AHN in rats that were selectively bred for an innately high response to aerobic exercise that also run voluntarily and increase maximal running capacity. Our results confirm that sustained aerobic exercise is key in improving AHN. ABSTRACT: Aerobic exercise, such as running, has positive effects on brain structure and function, such as adult hippocampal neurogenesis (AHN) and learning. Whether high-intensity interval training (HIT), referring to alternating short bouts of very intense anaerobic exercise with recovery periods, or anaerobic resistance training (RT) has similar effects on AHN is unclear. In addition, individual genetic variation in the overall response to physical exercise is likely to play a part in the effects of exercise on AHN but is less well studied. Recently, we developed polygenic rat models that gain differentially for running capacity in response to aerobic treadmill training. Here, we subjected these low-response trainer (LRT) and high-response trainer (HRT) adult male rats to various forms of physical exercise for 6-8 weeks and examined the effects on AHN. Compared with sedentary animals, the highest number of doublecortin-positive hippocampal cells was observed in HRT rats that ran voluntarily on a running wheel, whereas HIT on the treadmill had a smaller, statistically non-significant effect on AHN. Adult hippocampal neurogenesis was elevated in both LRT and HRT rats that underwent endurance training on a treadmill compared with those that performed RT by climbing a vertical ladder with weights, despite their significant gain in strength. Furthermore, RT had no effect on proliferation (Ki67), maturation (doublecortin) or survival (bromodeoxyuridine) of new adult-born hippocampal neurons in adult male Sprague-Dawley rats. Our results suggest that physical exercise promotes AHN most effectively if the exercise is aerobic and sustained, especially when accompanied by a heightened genetic predisposition for response to physical exercise.

Inflammatory Response to Wear Particles: Comparisons Between Zirconia and Titanium In Vitro.

PURPOSE: To compare the inflammatory responses of peripheral blood mononuclear cells (PBMCs) subjected to titanium (Ti) and/or zirconia (Zr) particles while growing on Ti or Zr discs. MATERIALS AND METHODS: In total, 240 discs were fabricated at a size of 2 mm in height and 5 mm in diameter. Of the 240 discs, 120 discs were made of Ti (commercially pure [CP] grade 4), and 120 discs were made of Zr (3 mol% yttria-stabilized zirconia polycrystals [3Y-TZP]). The PBMCs were cultured on the two-disc materials, and particles with a size up to 20 mm Ti (99.5% Ti) and 0.1 to 0.2 mm Zr (3Y-TZP) were added to the cultures. The concentration levels of inflammatory cytokines in culture supernatants were measured through Bio-Plex assay (Bio-Rad Laboratories), and light microscopic analysis was performed to detect cell attachment and characterize particle shape and cell-particle interaction. RESULTS: The inflammatory responses of PBMCs were generally higher when cells were cultured on a Ti surface compared to a Zr surface. In addition, higher cytokine levels were seen when cells were cultured in the presence of Ti particles compared to Zr particles when no discs were used. However, there were only significantly increased levels for three cytokines (MCP-1, IFN-γ, and TNF-α) when particles were added to Ti discs. Higher release of neutrophil extracellular traps (NETs) from neutrophils were seen in presence of Zr particles compared to Ti particles. A reduction in cell death was observed in the presence of Zr particles compared to Ti particles and unstimulated control samples. CONCLUSIONS: The type of growth material and presence of particle affects PBMCs in vitro. Cells seeded on Ti discs and together with Ti particles generated higher levels of inflammatory cytokines compared to the Zr counterparts.

Inflammatory Gene Profile and Particle Presence in Peri-Implant Mucosa: a Pilot Study on 9 Patients.

Objectives: The purpose of this pilot study is to compare gene expression in mucosa around dental implants with zirconia abutment to titanium and investigate presence of particles in mucosa samples and on implant heads. Material and Methods: Ten patients with a single implant supported prosthesis connected to zirconia or titanium abutments were invited at the five-year control. A clinical examination and a survey on experience of function and appearance were conducted. A mucosa biopsy taken in close vicinity to the implant were analysed by real-time polymerase chain reaction (qPCR) and presence of particles in a scanning electron microscope/energy-dispersive X-ray spectroscope (SEM/EDX). Cytological smear samples were collected and analysed through inductively coupled plasma mass spectrometry (ICP-MS) to investigate presence of particles on implant heads. Results: In total, 9 patients participated in the study, five with titanium abutments and four with zirconia abutments. All patients were satisfied with function and aesthetics. Titanium and iron particles were detected in mucosa biopsies. The ICP - MS analysis demonstrated presence of zirconia and titanium. Several proinflammatory genes were upregulated in the zirconia abutment group. Conclusions: Around zirconia abutments a slight increase in proinflammatory response and amount of wear particles was seen as compared to titanium. Wear particles of titanium were present in all soft tissue samples, however zirconia particles only in the samples from implants heads/mucosa with zirconia abutments.

Comparison of porcelain veneer fracture in implant-supported fixed full-arch prostheses with a framework of either titanium, cobalt-chromium, or zirconia: An in vitro study.

OBJECTIVES: The aim of this study was to compare porcelain veneer strength on screw-retained implant-supported fixed full-arch prostheses with a framework of either milled titanium (Ti), cobalt-chromium (CoCr), and yttria-stabilized zirconia (Y-TZP) in an in vitro loading model. MATERIALS AND METHODS: Fifteen screw-retained maxillary implant-supported full-arch prostheses (FDP), five each of Ti, CoCr, and Y-TZP frameworks with porcelain veneers were included. All FDPs were subjected to thermocycling before loading until fracture of the veneer. The load was applied at the distal fossa of the occlusal area of the pontic replacing 24. Fracture loads were analyzed, and the fracture quality was assessed. Statistical analysis on the fracture load was performed using Kruskal-Wallis test. The statistical significance was set at p < .05. RESULTS: There was no statistical significance found between the groups regarding fracture load. The highest and lowest load was seen within the CoCr FDP, varying between 340 and 1484 N. Different types of fracture appearances were seen. The Y-TZP FDPs had a higher number of fractures locally in the loaded area while CoCr and Ti more often showed cracks in the anterior region, at a distance from the loaded area. CONCLUSIONS: Within the limitations of this study, the conclusion was that framework material may affect the fracture behavior of maxillary full-arch bridges; however, there were no differences in veneer fracture strength when frameworks of Ti, CoCr, or Y-TZP were compared.

Inflammatory Response to Cobalt-Chromium Alloys Fabricated With Different Techniques.

OBJECTIVES: To explore the in vitro cytokine expression of human peripheral blood mononuclear cells exposed to cobalt-chromium alloys, manufactured with different techniques, in comparison with commercially pure titanium grade 4 and titanium alloy grade 23. MATERIAL AND METHODS: Peripheral blood mononuclear cells (PBMC) were collected from 10 healthy blood donors and exposed to machine-ground coin-shaped: (a) cobalt-chromium (Co-Cr) specimens (n = 5) manufactured by four techniques, i.e. cast, milled, laser melted and presintered milled; (b) commercially pure titanium grade 4; and (c) titanium alloy grade 23. The cells were cultured for 4, 24 and 72 hours followed by investigations of pro- and anti-inflammatory cytokine release using Bio-Plex Pro™. RESULTS: In general, the PBMC produced significantly more cytokines when exposed to the cast and presintered milled Co-Cr materials compared to laser melted, milled Co-Cr and titanium materials. CONCLUSIONS: Within the limitation of the present study, it may be suggested that cast and presintered milled cobalt-chromium alloys provoke a stronger inflammatory response compared to milled and laser melted cobalt-chromium alloys and titanium materials.

Cobalt-chromium alloys fabricated with four different techniques: Ion release, toxicity of released elements and surface roughness.

OBJECTIVE: To investigate the metal ion release, surface roughness and cytoxicity for Co-Cr alloys produced by different manufacturing techniques before and after heat treatment. In addition, to evaluate if the combination of materials affects the ion release. METHODS: Five Co-Cr alloys were included, based on four manufacturing techniques. Commercially pure titanium, CpTi grade 4 and a titanium alloy were included for comparison. The ion release tests involved both Inductive Coupled Plasma Optical Emission Spectrometry and Inductive Coupled Plasma Mass Spectrometry analyses. The surface analysis was conducted with optical interferometry. Cells were indirectly exposed to the materials and cell viability was evaluated with the MTT (3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide) method. RESULTS: All alloys showed a decrease of the total ion release when CpTi grade 4 was present. The total ion release decreased over time for all specimens and the highest ion release was observed from the cast and milled Co-Cr alloy in acidic conditions. The cast and laser-melted Co-Cr alloy and the titanium alloy became rougher after heat treatment. All materials were within the limits of cell viability according to standards. SIGNIFICANCE: The ion release from Co-Cr alloys is influenced by the combination of materials, pH and time. Surface roughness is influenced by heat treatment. Furthermore, both ion release and surface roughness are influenced by the manufacturing technique and the alloy type. The clinical implication needs to be further investigated.

The effect of housing environment on bone healing in a critical radius defect in New Zealand White rabbits.

In animal studies on bone healing, the effect of housing space and physical activity are seldom taken into account. Bone formation was evaluated in New Zealand White rabbits (mean ± SEM BW: 3.9 ± 0.11 kg) with a critical bone defect after 12 weeks of rehabilitation in pair-housing in 3 m2 large floor pens (Floor, n = 10) or standard single housing in 0.43 m2 cages (Cage, n = 10). In the randomised full-factorial study, a bone replica of calcium phosphate cement (CPC, n = 10) or autologous bone (AB, n = 10) was implanted in the unilateral 20 mm radius defect. Post-mortem, the oxidative capacity was measured by citrate synthase (CS) activity in M. quadriceps and the defect filling volume and density evaluated by microcomputer tomography (µ-CT). Histology sections were evaluated by subjective scoring and histomorphometry. Fourteen rabbits remained until the end of the study. Group Floor (n = 7; 3 CPC + 4 AB) had a higher CS activity and a larger bone defect filling volume and lower density by µ-CT measurements than group Cage (n = 7; 3 CPC + 4 AB). Three out of four rabbits in AB-Floor presented fusion of the defect with reorganisation of trabecular bone, whereas three of four in AB-Cage showed areas of incomplete healing. Floor rabbits had a higher score of bony fusion between the radius and ulna than Cage rabbits. There were no differences between groups in histomorphometry. The study found that a larger housing space increased physical activity and promoted bone formation.

Distinguishing vegan-, vegetarian-, and omnivorous diets by hair isotopic analysis.

BACKGROUND: Dietary risks contribute heavily to disability-adjusted life years (DALYs), being more important than hypertension, obesity, or smoking. To measure dietary exposure remains a challenge in nutrition research. AIMS: The aim of the present study was to test the hypothesis that isotope ratios of 15N and 13C in human hair could distinguish between subjects adhering to different habitual diets. METHODS: 20 male and 29 female subjects average 31 years old (range 19-53), with stable dietary habits volunteered. Diets were vegan, vegetarian and omnivorous. Hair samples were processed on an elemental analyser coupled to isotope-ratio mass spectrometry. RESULTS: δ15N differed between vegan, vegetarian and omnivorous diets, p < 0.05 for all. δ13C differed between vegan and omnivorous diets, p < 0.05, but neither of these diets were separated from the vegetarian diet. CONCLUSION: Elemental Analysis of δ13C and especially δ15N with isotope ratio mass spectrometry seems to be a promising, non-invasive and objective way to distinguish groups of subjects on different habitual diets, at least if n=>10.

Aseptic Ligatures Induce Marginal Peri-Implant Bone Loss-An 8-Week Trial in Rabbits.

The clinical value of ligature-induced experimental peri-implantitis studies has been questioned due to the artificial nature of the model. Despite repeated claims that ligatures of silk, cotton and other materials may not induce bone resorption by themselves; a recent review showed that the tissue reaction toward them has not been investigated. Hence, the current study aimed to explore the hard and soft tissue reactions toward commonly used ligature materials. A total of 60 dental implants were inserted into the femur (n = 20) and tibia (n = 40) of 10 rabbits. The femoral implants were ligated with sterile 3-0 braided silk in one leg and sterile cotton retraction chord in the other leg. The tibial implants were ligated with silk or left as non-ligated controls. All wounds were closed in layers. After a healing time of 8 weeks, femoral (silk versus cotton) and proximal tibial (silk versus non-ligated control) implants were investigated histologically. Distal tibial (silk versus non-ligated control) implants were investigated with real time polymerase chain reaction (qPCR). The distance from the implant-top to first bone contact point was longer for silk ligated implants compared to non-ligated controls (p = 0.007), but did not vary between cotton and silk. The ligatures triggered an immunological reaction with cell infiltrates in close contact with the ligature materials, adjacent soft tissue encapsulation and bone resorption. qPCR further demonstrated an upregulated immune response toward the silk ligatures compared to non-ligated controls. Silk and cotton ligatures provoke foreign body reactions of soft tissue encapsulation type and bone resorption around implants in the absence of plaque.

Inflammatory cytokine release from human peripheral blood mononuclear cells exposed to polyetheretherketone and titanium-6 aluminum-4 vanadium in vitro.

Objective To investigate the cytokine expression profiles of blood cells exposed to polyetheretherketone and titanium-6 aluminum-4 vanadium materials in vitro. Materials and methods Coin-shaped samples composed of titanium-6 aluminum-4 vanadium, polyetheretherketone, and blasted polyetheretherketone were manufactured. The surfaces of the coins were characterized using optical interferometry, scanning electron microscopy, and contact angle measurements. Peripheral blood mononuclear cells collected from 10 blood donors were cultured for one, three, and six days in the presence or absence of the coins, and then assayed for cytokine production. Quantification of the peripheral blood mononuclear cells attached to the coins was performed using confocal microscopy after immunofluorescence staining. Results The machined titanium-6 aluminum-4 vanadium coins had a smoother surface topography compared to the machined polyetheretherketone and blasted polyetheretherketone. The highest mean contact angle was noted for the blasted polyetheretherketone, followed by the machined polyetheretherketone and titanium-6 aluminum-4 vanadium. The peripheral blood mononuclear cells produced significantly more proinflammatory cytokines when exposed to the polyetheretherketone surface compared to the titanium-6 aluminum-4 vanadium surface, while the blasted polyetheretherketone induced the highest level of proinflammatory cytokine release from the peripheral blood mononuclear cells. Significantly more cells attached to both polyetheretherketone surfaces, as compared to the titanium-6 aluminum-4 vanadium surface. Conclusion Polyetheretherketone induces a stronger inflammatory response from peripheral blood mononuclear cells than does titanium-6 aluminum-4 vanadium. Surface topography has an impact on cytokine release from peripheral blood mononuclear cells.

Retrieved implants from irradiated sites in humans: a histologic/histomorphometric investigation of oral and craniofacial implants.

PURPOSE: The aim of this report was to quantitatively and qualitatively evaluate the tissue response to bone-anchored implants retrieved from irradiated sites in patients. MATERIALS AND METHODS: The material consists of 23 consecutively received Brånemark implants (Nobel Biocare AB, Göteborg, Sweden) placed in pre- or postoperatively irradiated sites. Twenty-two of the 23 implants were suitable for histologic evaluation of undecalcified sections in the light microscope. RESULTS: The oral implants with shorter time in situ demonstrated sparse bone to implant contact with mainly dense connective tissue in the interface. However, for implants with longer time in situ, high amounts of bone-implant contact and bone fill of threads were noted. The mean values of bone-implant contact and bone area within the thread were calculated to 40% (16-94) and 70% (13-96), respectively. The craniofacial implants, with the exception of two implants lined with a capsular formation, demonstrated mature and newly formed bone at the bone-implant interface. The mean value for bone-metal contact was calculated to 45 and 53% for two specimens. The mean value for bone area within the thread ranged from 65 to 88% for three specimens. CONCLUSION; The possibility to achieve bone anchorage of implants in irradiated tissue was supported by the findings in this study. However, due to limited material, conclusions with regard to radiation dose and bone tissue response to implants cannot be stated.

Histologic evaluation of Brånemark clinic oral implants retrieved from grafted sites.

PURPOSE: The aim of this report is to quantitatively and qualitatively describe the bone tissue response to Brånemark implants retrieved from grafted sites in patients. MATERIALS AND METHOD: The material consists of consecutively received Brånemark implants retrieved from grafted sites. Thirty-five of these implants, retrieved from 16 patients, were suitable for the histologic evaluation of undecalcified sections in the light microscope. RESULTS: The unloaded implants were mainly lined with soft tissue, and sparse bone-implant contact was observed only in some sections. The loaded implants, with the exception of one implant removed due to mobility, had mature and new bone-implant contact. Resorption of graft through cutting cone structures was detected. Cement lines were found separating bone-like tissue albeit no cellular content and bone tissue with detectable osteocytes. CONCLUSION: In this heterogeneous group of implants from grafted sites, the unloaded implants showed limited bone-implant contact. The autografts showed seemingly mixed viability as judged by the cell content in the osteocyte lacunae and cement lines separating areas with filled and empty lacunae.

Nanoporous TiO(2) thin film on titanium oral implants for enhanced human soft tissue adhesion: a light and electron microscopy study.

BACKGROUND: Previous experimental studies have demonstrated direct soft tissue attachment for nanoporous titanium dioxide (TiO(2) ) thin film on implants, while implants without TiO(2) thin film have not shown this capability. PURPOSE: The aims were to evaluate and compare TiO(2) surface-modified experimental microimplants with unmodified microimplants with respect to tissue interaction of the human oral mucosa evaluated by light microscopy on ground sections and semithin sections and transmission electron microscopy on ultrathin sections, and to characterize the inflammatory response and the level of the marginal bone resorption. MATERIALS AND METHODS: The study was a single-center, randomized, comparative, clinical investigation with intrasubject comparison of implants with and without TiO(2) thin film in 15 patients. RESULTS: Two comparator microimplants showed mild erythema and expulsion of fluids. The surrounding tissues around all test implants were clinically healthy. The oral mucosa in contact with the abutment part of the microimplant was 72% for the test implants and 48% for the comparator implants, a statistically significant difference (p =.0268). No statistically significant difference was found in other histological variables. The marginal bone loss in 14 weeks was 0.5 mm for the stable test (n = 11) and 1.7 mm for the stable comparator implants (n = 9; p = .0248). CONCLUSIONS: The nanoporous TiO(2) surface modification has potential clinical benefits because of increased adherence of soft tissue and possible reduced bone resorption.

Interaction of Helicobacter pylori with sialylated carbohydrates: the dependence on different parts of the binding trisaccharide Neu5Ac{alpha}3Gal{beta}4GlcNAc.

We have recently shown that binding of Helicobacter pylori to sialylated carbohydrates is dependent on the presence of the carboxyl group and the glycerol chain of Neu5Ac. In this work we investigated the importance of GlcNAc in the binding trisaccharide Neu5Acalpha3Galbeta4GlcNAc and the role of the N-acetamido groups of both Neu5Ac and GlcNAc. An important part of the project was epitope dissection, that is chemical derivatizations of the active carbohydrate followed by binding studies. In addition we used a panel of various unmodified carbohydrate structures in the form of free oligosaccharides or glycolipids. These were tested for binding by hemagglutination inhibition assay, TLC overlay tests, and a new quantitative approach using radiolabeled neoglycoproteins. The studies showed that the N-acetamido group of Neu5Ac is important for binding by H. pylori, whereas the same group of GlcNAc is not. In addition, Fuc attached to GlcNAc, as tested with sialyl-Lewis x, did not affect the binding. Free Neu5Ac was inactive as inhibitor, and Neu5Acalpha3Gal turned out to be active. The binding preference for neolacto structures was confirmed, although one strain also was inhibited by lacto chains. The combined results revealed that an intact Neu5Ac is crucial for the interactions with H. pylori. Parts of Gal also seem to be necessary, whereas the role of the GlcNAc is secondary. GlcNAc does influence binding, however, primarily serving as a guiding carrier for the binding epitope rather than being a part of the binding structure.

The temperature dependence and involvement of mitochondria permeability transition and caspase activation in damage to organotypic hippocampal slices following in vitro ischemia.

The aggravating effect of hyperglycemia on ischemic brain injury can be mimicked in a model of in vitro ischemia (IVI) using murine hippocampal slice cultures. Using this model, we found that the damage in the CA1 region following IVI in the absence or presence of 40 mm glucose (hyperglycemia) is highly temperature dependent. Decreasing the temperature from 35 to 31 degrees C during IVI prevented cell death, whereas increasing the temperature by 2 degrees C markedly aggravated damage. As blockade of the mitochondrial permeability transition (MPT) is equally effective as hypothermia in preventing ischemic cell death in vivo, we investigated whether inhibition of MPT or of caspases was protective following IVI. In the absence of glucose, the MPT blockers cyclosporin A and MeIle4-CsA but not the immunosuppressive compound FK506 diminished cell death. In contrast, following hyperglycemic IVI, MPT blockade was ineffective. Also, the pan-caspase inhibitor Boc-Asp(OMe)fluoromethyl ketone did not decrease cell death in the CA1 region following IVI or hyperglycemic IVI. We conclude that cell death in the CA1 region of organotypic murine hippocampal slices following IVI is highly temperature dependent and involves MPT. In contrast, cell death following hyperglycemic IVI, although completely prevented by hypothermia, is not mediated by mechanisms that involve MPT or caspase activation.