Fibrous cartilage of human menisci is less shock-absorbing and energy-dissipating than hyaline cartilage.

PURPOSE: To test meniscal mechanical properties such as the dynamic modulus of elasticity E* and the loss angle δ at two loading frequencies ω at different locations of the menisci and compare it to E* and δ of hyaline cartilage in indentation mode with spherical indenters. METHODS: On nine pairs of human menisci, the dynamic E*-modulus and loss angle δ (as a measure of the energy dissipation) were determined. The measurements were performed at two different strain rates (slow sinusoidal and fast single impact) to show the strain rate dependence of the material. The measurements were compared to previous similar measurements with the same equipment on human hyaline cartilage. RESULTS: The resultant E* at fast indentation (median 1.16 MPa) was significantly higher, and the loss angle was significantly lower (median 10.2°) compared to slow-loading mode's E* and δ (median 0.18 MPa and 16.9°, respectively). Further, significant differences for different locations are shown. On the medial meniscus, the anterior horn shows the highest resultant dynamic modulus. CONCLUSION: In dynamic measurements with a spherical indenter, the menisci are much softer and less energy-dissipating than hyaline cartilage. Further, the menisci are stiffer and less energy-dissipating in the middle, intermediate part compared to the meniscal base. In compression, the energy dissipation of meniscus cartilage plays a minor role compared to hyaline cartilage. At high impacts, energy dissipation is less than on low impacts, similar to cartilage.

Seven hours to adequate antimicrobial therapy in urosepsis using isothermal microcalorimetry.

Urosepsis can progress toward severe sepsis, septic shock, and, ultimately, death. Rapid antimicrobial susceptibility testing is crucial to decrease mortality and morbidity. This report shows that isothermal microcalorimetry can provide an antibiogram within 7 h with a sensitivity of 95% and specificity of 91% using Vitek-2 system as a reference.

The subtalar and talonavicular joints: a way to access the long-term load intake using conventional CT-data.

PURPOSE: The aim of this study was to investigate the distribution of density of the subchondral bone plate within the articular surfaces of the subtalar and talonavicular joint regarding to its mineralisation and to verify whether a correlation to the mechanical bone strength exists. METHODS: A total of 21 cadaverous lower leg specimens were investigated. Computed tomography osteo-absorptiometry (CT-OAM) was used to display the mineralisation of the subchondral bone plate analysing its density. The mechanical strength was measured by means of indentation testing. The distribution pattern was analysed regarding their dissemination with the main focus on number and location of their maxima. The correlation of both parameters was evaluated by linear regression. RESULTS: The mineralisation and the mechanical strength were not distributed homogenously throughout the articular surfaces but showed unique and reproducible patterns. The range of absolute values for density and strength varied in between the samples and joint surfaces, but the number and location of the maxima evaluated by both methods showed to be concurring. The coefficient of correlation of both datasets ranged from 0.76 to 0.95 (median 0.88) and showed a linear dependency. CONCLUSIONS: Density distribution and mechanical strength of the subchondral bone plate are significantly associated and can be seen as a mirror of the long-term load intake of a joint. It can be concluded that CT-OAM as a tool to visualize subchondral bone plate density distribution regarding to its mineralisation can be used to indirectly gain information about joint biomechanics in vivo by the use of conventional CT-data.

Again and Again-Survival of Candida albicans in Urine Containing Antifungals.

BACKGROUND: Relapse of Candida albicans urinary tract infection (UTI) is frequent despite appropriate treatment, as commonly used antifungals such fluconazole and flucytosine are only fungistatics. To improve treatment of Candida UTI and decrease relapses, understanding the long-term metabolic activity and survival of C. albicans in urine containing antifungals at minimal inhibitory concentration (MIC) is needed. METHODS: we monitored the survival, metabolic activity and consumption of glucose and proteins by C. albicans using conventional methods and isothermal microcalorimetry (IMC). We also investigated the influence of dead Candida cells on the growth of their living counterparts. RESULTS: For 33 days, weak activity was observed in samples containing antifungals in which C. albicans growth rate was reduced by 48%, 60% and 88%, and the lag increased to 172 h, 168 h and 6 h for amphotericin, flucytosine and fluconazole, respectively. The metabolic activity peaks corresponded to the plate counts but were delayed compared to the exhaustion of resources. The presence of dead cells promoted growth in artificial urine, increasing growth rate and reducing lag in similar proportions. CONCLUSIONS: Even with antifungal treatment, C. albicans relapses are possible. The low metabolic activity of surviving cells leading to regrowth and chlamydospore formation possibly supported by autophagy are likely important factors in relapses.

Rapid detection of urinary tract pathogens using microcalorimetry: principle, technique and first results.

UNLABELLED: What's known on the subject? and What does the study add? Microcalorimetry has been applied in several microbiological studies, but never in a clinical urological context. In addition, basic knowledge on the growth of urinary pathogens in urine is still scarce and data regarding the growth rate of many urinary pathogens in urine are still not available. The study demonstrates that this innovative application of microcalorimetry is useful in (i) investigating the growth of urinary pathogens in sterilized urine and (ii) as a rapid tool for diagnosis of urinary infection as well as for further identification of the causative infectious agent. OBJECTIVE: To investigate the value of isothermal microcalorimetry (IMC) in the detection and differentiation of common urinary tract pathogens in urine. IMC is a non-specific analytical tool for the measurement of heat in the microwatt range. PATIENTS AND METHODS: A microcalorimeter equipped with 48 channels was used. Detection was accomplished, and growth was monitored for four bacterial strains in sterilized urine at 37 °C by measuring metabolic heat flow (µW = µJ/s) as a function of time. The strains were Escherichia coli, Proteus mirabilis, Enterococcus faecalis and Staphylococcus aureus. RESULTS: Bacterial growth was detected after 3.1 to 17.1 h with decreasing inocula. The detection limit was 1 colony-forming unit (CFU)/mL for E. coli, 10 CFU/mL for P. mirabilis and E. faecalis and 10(3) CFU/mL for S. aureus. The total heat was highest in P. mirabilis ranging from 10 to 12 J, followed by E. coli (3-4 J), S. aureus (2-3 J) and E. faecalis (1.3-1.5 J). The shape of the heat flow curves was characteristic for each species independent of its initial concentration. CONCLUSIONS: IMC allows rapid detection of bacteriuria, much faster than conventional culture. Urinary tract pathogen detection after only 3.1 h is realistic. Clearly different heat flow patterns enable accurate pathogen differentiation. Due to expeditious identification of urine samples that contain only low colony counts (i.e. less than 10(3) CFU/mL), IMC may become a valuable screening tool for detecting the presence of significant bacteriuria.

Correlation between mineralization and mechanical strength of the subchondral bone plate of the humeral head.

BACKGROUND: One of the main problems in shoulder arthroplasty is the fixation of the prosthesis, where the subchondral bone plate plays an important role. Subchondral mineralization patterns represent the loading history of a joint and give information about the individual biomechanical situation. The objective of this study was to determine if a correlation between subchondral mineralization and mechanical strength in the humeral head exists. MATERIALS AND METHODS: Subchondral mineralization of 32 shoulder specimens was investigated by use of computed tomography (CT) osteoabsorptiometry. The previously dissected specimens were scanned axially in a CT scanner, and the obtained data sets were transferred into an image analyzing system. Maximum intensity projection was used to evaluate the density distribution of the subchondral bone plate. To get information about mechanical strength of the subchondral bone, each specimen was investigated at 29 predefined points by means of an indentation-testing machine. RESULTS: The maximum strength was mostly detected in the center (monocentric pattern) or in anterior and posterior areas of the articular surface (bicentric pattern). The distribution of mineralization showed the same 2 reproducible patterns. The coefficient of correlation between mechanical strength and mineralization shown on CT was between 0.59 and 0.96. The obtained information was statistically significant (P < .01). CONCLUSION: Mechanical strength and subchondral mineralization in the humeral head are significantly associated (P < .01). As a consequence of these findings, CT osteoabsorptiometry can be indirectly used to give information about bone quality in vivo. Our findings could be useful for the development of new fixation methods in shoulder surgery (eg, humeral resurfacing arthroplasty).

Density and strength distribution in the human subchondral bone plate of the patella.

PURPOSE: The aim of this study was to map the strength distribution of the human patella and correlate it to the subchondral bone plate density obtained by means of computed tomographyosteoabsorptiometry (CT-OAM). METHODS: Measurements were performed at 34 standardized points on each patella. The mineralization patterns of the subchondral bone plate of 20 patellae were displayed with the help of CT-OAM. False-coloured distribution patterns for our measurements were generated. The mechanical strength was determined at the same points by indentation testing. RESULTS: We showed that neither the density nor the mechanical strength is distributed homogeneously but exhibited regular, reproducible distribution patterns which mirror long-term stress distribution in articular surfaces. A direct correlation was found between both parameters in the subchondral bone plate. CONCLUSION: The correlation of density and mechanical strength makes CT-OAM a valuable tool to assess and monitor changes in the strength of the subchondral bone plate in vivo.

Quantification of vital adherent Streptococcus sanguinis cells on protein-coated titanium after disinfectant treatment.

The quantification of vital adherent bacteria is challenging, especially when efficacy of antimicrobial agents is to be evaluated. In this study three different methods were compared in order to quantify vital adherent Streptococcus sanguinis cells after exposure to disinfectants. An anaerobic flow chamber model accomplished initial adhesion of S. sanguinis on protein-coated titanium. Effects of chlorhexidine, Betadine®, Octenidol®, and ProntOral® were assessed by quantifying vital cells using Live/Dead BacLight™, conventional culturing and isothermal microcalorimetry (IMC). Results were analysed by Kruskal-Wallis one-way analysis of variance. Live/dead staining revealed highest vital cell counts (P < 0.05) and demonstrated dose-dependent effect for all disinfectants. Microcalorimetry showed time-delayed heat flow peaks that were proportioned to the remaining number of viable cells. Over 48 h there was no difference in total heat between treated and untreated samples (P > 0.05), indicating equivalent numbers of bacteria were created and disinfectants delayed growth but did not eliminate it. In conclusion, contrary to culturing, live/dead staining enables detection of cells that may be viable but non-cultivable. Microcalorimetry allows unique evaluation of relative disinfectant effects by quantifying differences in time delay of regrowth of remaining vital cells.

Intra-operative femoral condylar stress during arthroscopy: an in vivo biomechanical assessment.

PURPOSE: Excessive varus and valgus stress forces during arthroscopy might exceed minimal compressive strength of cancellous bone. In extreme cases, this could lead to post-arthroscopic osteonecrosis. It was our purpose to measure the valgus and varus stress forces during arthroscopy and draw conclusions on the development of osteonecrosis. METHODS: On 24 consecutive patients undergoing arthroscopy, the maximum varus and valgus stress forces (N) were measured in vivo using a strain gauge mounted to a leg holder. The forces (N) and contact stresses (kPa) on the femoral condyles were calculated based on the measured acting lateral force at the femur fixation based on the lever principle. RESULTS: The maximum contact stress during varus on the medial condyle was significantly lower in patients with intact meniscus (mean ± standard error of the mean: 243 ± 29 kPa) than in patients with meniscus-deficient knees (520 ± 61 kPa; P < 0.01). A similar finding was obtained for the maximum contact stress during valgus on the lateral condyle: 630 ± 72 kPa in patients with intact meniscus compared to 2,173 ± 159 kPa in patients with meniscus-deficient knees (P < 0.01). In 19 patients (79%), the maximum contact stress was higher during valgus than during varus. The maximum contact stress on the lateral condyle during valgus was significantly higher for more experienced surgeons (P = 0.01). CONCLUSION: The maximum contact stresses in knees with intact menisci did not exceed the critical threshold of the compressive strength in cancellous bone. However, the maximum contact stresses in meniscus-deficient knees were frequently higher than the threshold. However, these stresses were much lower than those during daily activities and therefore unlikely to lead to post-arthroscopic osteonecrosis. LEVEL OF EVIDENCE: Diagnostic study, Level II.

Mineralisation and mechanical strength of the glenoid cavity subchondral bone plate.

PURPOSE: Failures in total shoulder replacements are often due to aseptic loosening of the glenoid component; the subchondral bone plate is an important factor governing primary fixation of implant materials. Therefore, we investigated characteristic mineralisation patterns of the subchondral bone plate, which demonstrate long-term stress on articular surfaces, age-related changes, postsurgical biomechanical situations and regions of fixation. Using computed tomography osteo-absorptiometry (CT-OAM), these distribution patterns can be demonstrated in vivo. The aim of this study was to investigate the relationship between subchondral bone-plate mineralisation measured with CT-OAM and the mechanical strength measured by indentation. METHODS: A total of 32 cadaverous glenoid cavities were evaluated by CT-OAM and indentation testing. Linear regression was used to compare mineralisation and strength of the subchondral bone plate. RESULTS: Results showed two patterns of mineralisation distribution. Twenty-eight cavities were related to bicentric distribution pattern and four showed a single maximum. The correlation coefficient between CT-OAM density and subchondral bone-plate strength was determined to be between 0.62 and 0.96 (P < 0.02). CONCLUSIONS: Long-term stress affects not only the subchondral but also the underlying cancellous bone. It therefore can be assumed that mineralisation patterns of the subchondral bone plate continue in cancellous bone. Areas of high density could serve as anchoring locations for orthopaedic implants in resurfacing the glenoid cavity.

Micro- and nanomechanical analysis of articular cartilage by indentation-type atomic force microscopy: validation with a gel-microfiber composite.

As documented previously, articular cartilage exhibits a scale-dependent dynamic stiffness when probed by indentation-type atomic force microscopy (IT-AFM). In this study, a micrometer-size spherical tip revealed an unimodal stiffness distribution (which we refer to as microstiffness), whereas probing articular cartilage with a nanometer-size pyramidal tip resulted in a bimodal nanostiffness distribution. We concluded that indentation of the cartilage's soft proteoglycan (PG) gel gave rise to the lower nanostiffness peak, whereas deformation of its collagen fibrils yielded the higher nanostiffness peak. To test our hypothesis, we produced a gel-microfiber composite consisting of a chondroitin sulfate-containing agarose gel and a fibrillar poly(ethylene glycol)-terephthalate/poly(butylene)-terephthalate block copolymer. In striking analogy to articular cartilage, the microstiffness distribution of the synthetic composite was unimodal, whereas its nanostiffness exhibited a bimodal distribution. Also, similar to the case with cartilage, addition of the negatively charged chondroitin sulfate rendered the gel-microfiber composite's water content responsive to salt. When the ionic strength of the surrounding buffer solution increased from 0.15 to 2 M NaCl, the cartilage's microstiffness increased by 21%, whereas that of the synthetic biomaterial went up by 31%. When the nanostiffness was measured after the ionic strength was raised by the same amount, the cartilage's lower peak increased by 28%, whereas that of the synthetic biomaterial went up by 34%. Of interest, the higher peak values remained unchanged for both materials. Taken together, these results demonstrate that the nanoscale lower peak is a measure of the soft PG gel, and the nanoscale higher peak measures collagen fibril stiffness. In contrast, the micrometer-scale measurements fail to resolve separate stiffness values for the PG and collagen fibril moieties. Therefore, we propose to use nanostiffness as a new biomarker to analyze structure-function relationships in normal, diseased, and engineered cartilage.

Biomedical use of isothermal microcalorimeters.

Isothermal microcalorimetry is becoming widely used for monitoring biological activities in vitro. Microcalorimeters are now able to measure heat production rates of less than a microwatt. As a result, metabolism and growth of relatively small numbers of cultured bacteria, protozoans, human cells and even small animals can be monitored continuously and extremely accurately at any chosen temperature. Dynamic effects on these organisms of changes in the culture environment--or of additions to it--are easily assessed over periods from hours to days. In addition microcalorimetry is a non-destructive method that does not require much sample preparation. It is also completely passive and thus allows subsequent evaluations of any kind on the undisturbed sample. In this review, we present a basic description of current microcalorimetry instruments and an overview of their use for various biomedical applications. These include detecting infections, evaluating effects of pharmaceutical or antimicrobial agents on cells, monitoring growth of cells harvested for tissue eingineering, and assessing medical and surgical device material physico-chemical stability and cellular biocompatibility.

Isothermal micro calorimetry--a new method for MIC determinations: results for 12 antibiotics and reference strains of E. coli and S. aureus.

BACKGROUND: Antimicrobial susceptibility testing of microorganisms is performed by either disc diffusion or broth dilution tests. In clinical use, the tests are often still performed manually although automated systems exist. Most systems, however, are based on turbidometric methods which have well-known drawbacks. RESULTS: In this study we evaluated isothermal micro calorimetry (IMC) for the determination of minimal inhibitory concentrations (MICs) of 12 antibiotics for five micro-organisms. Here we present the data for the 12 antibiotics and two representative microorganisms E. coli (a Gram-) and S. aureus (a Gram+). IMC was able to determine the MICs correctly according to CLSI values. Since MICs require 24 hours, time was not reduced. However, IMC provided new additional data - a continuous record of heat-producing bacterial activity (e.g. growth) in calorimetry ampoules at subinhibitory antibiotic concentrations. Key features of the heatflow (P) and aggregate heat (Q) vs. time curves were identified (t delay and Delta Q/Delta t respectively). Antibiotics with similar modes of action proved to have similar effects on t delay and/or Delta Q/Delta t. CONCLUSION: IMC can be a powerful tool for determining the effects of antibiotics on microorganisms in vitro. It easily provides accurate MICs - plus a potential means for analyzing and comparing the modes of action of antibiotics at subinhibitory concentrations. Also IMC is completely passive, so after evaluation, ampoule contents (media, bacteria, etc.) can be analyzed by any other method desired.

Rapid differentiation of methicillin-susceptible Staphylococcus aureus from methicillin-resistant S. aureus and MIC determinations by isothermal microcalorimetry.

In this study, the use of isothermal microcalorimetry (IMC) for differentiation between methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) and MIC determination was evaluated. It was possible to differentiate between MRSA and MSSA within 4 h, whereas the standard method required 24 h. The MICs of cefoxitin were successfully determined for MRSA and MSSA by using IMC.

Use of hydrodynamic forces to engineer cartilaginous tissues resembling the non-uniform structure and function of meniscus.

The aim of this study was to demonstrate that differences in the local composition of bi-zonal fibrocartilaginous tissues result in different local biomechanical properties in compression and tension. Bovine articular chondrocytes were loaded into hyaluronan-based meshes (HYAFF-11) and cultured for 4 weeks in mixed flask, a rotary Cell Culture System (RCCS), or statically. Resulting tissues were assessed histologically, immunohistochemically, by scanning electron microscopy and mechanically in different regions. Local mechanical analyses in compression and tension were performed by indentation-type scanning force microscopy and by tensile tests on punched out concentric rings, respectively. Tissues cultured in mixed flask or RCCS displayed an outer region positively stained for versican and type I collagen, and an inner region positively stained for glycosaminoglycans and types I and II collagen. The outer fibrocartilaginous capsule included bundles (up to 2 microm diameter) of collagen fibers and was stiffer in tension (up to 3.6-fold higher elastic modulus), whereas the inner region was stiffer in compression (up to 3.8-fold higher elastic modulus). Instead, molecule distribution and mechanical properties were similar in the outer and inner regions of statically grown tissues. In conclusion, exposure of articular chondrocyte-based constructs to hydrodynamic flow generated tissues with locally different composition and mechanical properties, resembling some aspects of the complex structure and function of the outer and inner zones of native meniscus.

Clinical development and current status: Europe.

This article reviews the development and current status of cemented fixation in total hip replacement in Europe. Key points include the wide country-to-country variation in use of cemented vs. non-cemented fixation and the largely overlooked importance of the choice of bone cement as a factor highly correlated with clinical outcome. Laboratory studies by the authors are also reviewed. Results suggest that the type of acrylic bone cement used affects wear phenomena at the implant/cement interface. Further studies by microcalorimetry suggest that certain aspects of acrylic starting materials (low molecular weight and use of radiation sterilization) affect long-term physico-chemical stability and may thus influence clinical outcomes.

[Measurement of the isometric dorsiflexion and plantar flexion force in the ankle joint]. / Messung der isometrischen Dorsalextensions- und Plantarflexions- Kraft in den Sprunggelenken.

This article describes an easy to use test equipment for measuring the isometric force in the ankle joints in dorsiflexion and plantar flexion. The combination of the test equipment for measuring the voluntary maximal isometric muscle force in the ankle joint, the surface electromyograms and the motion analysis of the measured leg allow an objective comparison of the strength of the muscular force between the left and right leg. It might be also used as a control setup during rehabilitation after surgical treatment or injuries.

Isothermal microcalorimetry accurately detects bacteria, tumorous microtissues, and parasitic worms in a label-free well-plate assay.

Isothermal microcalorimetry is a label-free assay that allows monitoring of enzymatic and metabolic activities. The technique has strengths, but most instruments have a low throughput, which has limited their use for bioassays. Here, an isothermal microcalorimeter, equipped with a vessel holder similar to a 48-well plate, was used. The increased throughput of this microcalorimeter makes it valuable for biomedical and pharmaceutical applications. Our results show that the sensitivity of the instrument allows the detection of 3 × 10(4) bacteria per vial. Growth of P. mirabilis in Luria Broth medium was detected between 2 and 9 h with decreasing inoculum. The culture released 2.1J with a maximum thermal power of 76 µW. The growth rate calculated using calorimetric and spectrophotometric data were 0.60 and 0.57 h(-1) , respectively. Additional insight on protease activities of P. mirabilis matching the last peak in heat production could be gathered as well. Growth of tumor microtissues releasing a maximum thermal power of 2.1 µW was also monitored and corresponds to a diameter increase of the microtissues from ca. 100 to 428 µm. This opens new research avenues in cancer research, diagnostics, and development of new antitumor drugs. For parasitic worms, the technique allows assessment of parasite survival using motor and metabolic activities even with a single worm.

Novel microcalorimetric assay for antibacterial activity of implant coatings: The cases of silver-doped hydroxyapatite and calcium hydroxide.

Biomaterials with antimicrobial properties are now commonly used in different clinical specialties including orthopedics, endodontic, and traumatology. As a result, assessing the antimicrobial effect of coatings applied on implants is of critical importance. In this study, we demonstrate that isothermal microcalorimetry (IMC) can be used for monitoring bacterial growth and biofilm formation at the surface of such coatings and for determining their antimicrobial effects. The antibacterial effects of silver doped hydroxyapatite (HA) and calcium hydroxide coatings on Staphylococcus epidermidis were determined with a minimal workload. Using the Gompertz growth model we determined biofilm growth rates close to those values reported in the literature. Furthermore, we were able to estimate the reduction in the bacterial inocula originally applied at the surface of the coatings. Therefore, in addition to monitoring the antimicrobial effect of silver doped HA and calcium hydroxide coatings, we also demonstrate that IMC might be a valuable tool for assessing such antimicrobial properties of implant coatings at a minimal workload.

Generation and characterization of osteochondral grafts with human nasal chondrocytes.

We investigated whether nasal chondrocytes (NC) can be used to generate composite constructs with properties necessary for the repair of osteochondral (OC) lesions, namely maturation, integration and capacity to recover from inflammatory burst. OC grafts were fabricated by combining engineered cartilage tissues (generated by culturing NC or articular chondrocytes - AC - onto Chondro-Gide® matrices) with devitalized spongiosa cylinders (Tutobone®). OC tissues were then exposed to IL-1ß for three days and cultured for additional 2 weeks in the absence of IL-1ß. Cartilage maturation extent was assessed (immune) histologically, biochemically and by delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) while cartilage/bone integration was assessed using a peel-off mechanical test. The use of NC as compared to AC allowed for more efficient cartilage matrix accumulation and superior integration of the cartilage/bone layers. dGEMRIC and biochemical analyzes of the OC constructs showed a reduced glycosaminoglycan (GAG) contents upon IL-1ß administration. Cartilaginous matrix contents and integration forces returned to baseline up on withdrawal of IL-1ß. By having a cartilage layer well developed and strongly integrated to the subchondral layer, OC tissues generated with NC may successfully engraft in an inflammatory post-surgery joint environment.