Femoral subchondral bone properties of patients with cam-type femoroacetabular impingement.

OBJECTIVE: Morphological deformities of the hip, such as femoroacetabular impingement (FAI) may be responsible for up to 80% of hip osteoarthritis. In cam type FAI, the pathomechanism has been attributed to repeated abnormal contact between the femur and the antero-superior acetabular rim, resulting in cartilage and labrum degeneration. Subchondral bone stiffness likely plays a major role in the process, but little is known of the mechanical properties of the cam deformity. The purpose of this study was to determine tissue modulus and the trabecular micro-architecture of the subchondral bone of the cam deformity of patients undergoing resection surgery as well as comparing these parameters to healthy aged matched controls. DESIGN: Twelve osteochondral bone biopsies were obtained from symptomatic FAI patients and ten osteochondral control specimens were harvested from cadaveric femurs. A combination of mechanical testing, micro-CT and finite element (FE) analysis were used to determine tissue modulus, bone volume fraction, trabecular thickness, trabecular and spacing, and trabecular number. RESULTS: The mean tissue modulus of the cam-type FAI deformities (E = 5.4 GPa) was significantly higher than normal controls (E = 2.75 GPa, P = 0.038), but no statistically significant differences were found in bone micro-architectural parameters. CONCLUSIONS: The data suggests that subchondral bone of the cam deformity consists of older secondary mineralized bone. This supports the notion that the cam deformity is a primary malformation with intrinsic biomechanical abnormalities rather than a secondary deformity as part of the degenerative process of the covering cartilage or remodeling due to repeated impingement.

Towards characterization of photo-excited electron transfer and catalysis in natural and artificial systems using XFELs.

The ultra-bright femtosecond X-ray pulses provided by X-ray Free Electron Lasers (XFELs) open capabilities for studying the structure and dynamics of a wide variety of biological and inorganic systems beyond what is possible at synchrotron sources. Although the structure and chemistry at the catalytic sites have been studied intensively in both biological and inorganic systems, a full understanding of the atomic-scale chemistry requires new approaches beyond the steady state X-ray crystallography and X-ray spectroscopy at cryogenic temperatures. Following the dynamic changes in the geometric and electronic structure at ambient conditions, while overcoming X-ray damage to the redox active catalytic center, is key for deriving reaction mechanisms. Such studies become possible by using the intense and ultra-short femtosecond X-ray pulses from an XFEL, where sample is probed before it is damaged. We have developed methodology for simultaneously collecting X-ray diffraction data and X-ray emission spectra, using an energy dispersive spectrometer, at ambient conditions, and used this approach to study the room temperature structure and intermediate states of the photosynthetic water oxidizing metallo-protein, photosystem II. Moreover, we have also used this setup to simultaneously collect the X-ray emission spectra from multiple metals to follow the ultrafast dynamics of light-induced charge transfer between multiple metal sites. A Mn-Ti containing system was studied at an XFEL to demonstrate the efficacy and potential of this method.

Increased acetabular subchondral bone density is associated with cam-type femoroacetabular impingement.

OBJECTIVE: Femoroacetabular impingement (FAI) has been associated with significant acetabular cartilage damage and subsequent degenerative arthritis. Subchondral bone, often neglected in osteoarthritis studies, may play an important role in the degenerative cascade. Hence the goal of this study was to assess acetabular subchondral bone mineral density (BMD) in subjects with asymptomatic or symptomatic cam deformities compared to normal control subjects. The relationship between BMD and the alpha angle, a quantitative measure of the deformity, was also analyzed. METHODS: Patients diagnosed with symptomatic cam FAI were recruited ('Surgical') as well as subjects from the general asymptomatic population, classified from CT imaging as normal ('Control') or having a cam deformity ('Bump') based on their alpha angle measurement. There were 12 subjects in each group. All subjects underwent a CT scan with a calibration phantom. BMD was calculated in regions of interest around the acetabulum from CT image intensity and the phantom calibration. BMD was compared between groups using spine BMD as a covariate. The relationship between BMD and alpha angle was assessed by linear regression. RESULTS: In the antero-superior regions bone density was 15-34% higher in the Bump group (P < 0.05) and 14-38% higher in the Surgical group (P < 0.05) compared to Controls. BMD correlated positively with the alpha angle measurements (R(2) = 0.44, P < 0.001). CONCLUSION: BMD was elevated in subjects with cam-type deformities, with the severity of the deformity more correlative than symptom status. Similarities to the symptomatic group suggest that hips with an asymptomatic deformity may already be in early stages of joint degeneration.

Bone density is higher in cam-type femoroacetabular impingement deformities compared to normal subchondral bone.

OBJECTIVE: Cam-type femoroacetabular impingement (FAI) deformities have been associated with early osteoarthritic degeneration of the hip. Degeneration depends on many factors such as joint morphology and dynamics of motion. Bone mineral density (BMD) appears to be a manifestation of the above, and may be a potentiator. Thus the goal of this study was to assess subchondral BMD of cam deformities in symptomatic and asymptomatic FAI subjects, and to compare to normal controls. METHODS: Subjects undergoing surgical correction of a symptomatic cam-type deformity were recruited ("Surgical"). Asymptomatic volunteers were also recruited and classified as normal ("Control") or having a deformity ("Bump") based on their alpha angle measurement. All subjects (n = 12 per group) underwent computed tomography (CT) with a calibration phantom. BMD was determined in volumes of interest around the femoral head and neck to a depth of 5 mm. BMD was compared between groups in each section using spine BMD as a covariate. RESULTS: No differences were seen between groups in the peripheral bearing surface. The Bump group exhibited higher BMD than Controls within the head/neck junction (P < 0.05). When compared to normal subchondral bone in the peripheral level of Controls, BMD in the deformity was up to 78% higher in Bump subjects and up to 47% higher in Surgical subjects (P < 0.05). CONCLUSION: Subchondral BMD of cam deformities is higher than that of normal subchondral bone in the peripheral region of the femoral head, regardless of symptom status. The expected increased subchondral stiffness may increase contact stresses in the joint tissues leading to accelerated degeneration.

Opposite repertory-rubrics in Bayesian perspective.

INTRODUCTION: Hitherto entries have been added to a rubric in the repertory when patients responding well to a specific medicine showed the corresponding symptom. Continuing like this, theoretically every medicine will eventually appear in every rubric. METHOD: This becomes clear if we compare opposite symptom-rubrics. Polarity Analysis (PA) subtracts opposite rubrics and has been shown to improve clinical results. CONCLUSION: The source of this problem and the reason for the success of PA are clear from Bayesian perspective. A reliable repertory should be based on Bayesian principles.

Cement penetration and primary stability of the femoral component after impaction allografting. A biomechanical study in the cadaveric femur.

This study explored the relationship between the initial stability of the femoral component and penetration of cement into the graft bed following impaction allografting. Impaction allografting was carried out in human cadaveric femurs. In one group the cement was pressurised conventionally but in the other it was not pressurised. Migration and micromotion of the implant were measured under simulated walking loads. The specimens were then cross-sectioned and penetration of the cement measured. Around the distal half of the implant we found approximately 70% and 40% of contact of the cement with the endosteum in the pressure and no-pressure groups, respectively. The distal migration/micromotion, and valgus/varus migration were significantly higher in the no-pressure group than in that subjected to pressure. These motion components correlated negatively with the mean area of cement and its contact with the endosteum. The presence of cement at the endosteum appears to play an important role in the initial stability of the implant following impaction allografting.

Detecting structural variances of Co3O4 catalysts by controlling beam-induced sample alterations in the vacuum of a transmission electron microscope.

This article summarizes core aspects of beam-sample interactions in research that aims at exploiting the ability to detect single atoms at atomic resolution by mid-voltage transmission electron microscopy. Investigating the atomic structure of catalytic Co3O4 nanocrystals underscores how indispensable it is to rigorously control electron dose rates and total doses to understand native material properties on this scale. We apply in-line holography with variable dose rates to achieve this goal. Genuine object structures can be maintained if dose rates below ~100 e/Å2s are used and the contrast required for detection of single atoms is generated by capturing large image series. Threshold doses for the detection of single atoms are estimated. An increase of electron dose rates and total doses to common values for high resolution imaging of solids stimulates object excitations that restructure surfaces, interfaces, and defects and cause grain reorientation or growth. We observe a variety of previously unknown atom configurations in surface proximity of the Co3O4 spinel structure. These are hidden behind broadened diffraction patterns in reciprocal space but become visible in real space by solving the phase problem. An exposure of the Co3O4 spinel structure to water vapor or other gases induces drastic structure alterations that can be captured in this manner.

The acetabular labrum: a review of its function.

UNLABELLED: The acetabular labrum is a soft-tissue structure which lines the acetabular rim of the hip joint. Its role in hip joint biomechanics and joint health has been of particular interest over the past decade. In normal hip joint biomechanics, the labrum is crucial in retaining a layer of pressurised intra-articular fluid for joint lubrication and load support/distribution. Its seal around the femoral head is further regarded as a contributing to hip stability through its suction effect. The labrum itself is also important in increasing contact area thereby reducing contact stress. Given the labrum's role in normal hip joint biomechanics, surgical techniques for managing labral damage are continuously evolving as our understanding of its anatomy and function continue to progress. The current paper aims to review the anatomy and biomechanical function of the labrum and how they are affected by differing surgical techniques. TAKE HOME MESSAGE: The acetabular labrum plays a critical role in hip function and maintaining and restoring its function during surgical intervention remain an essential goal. Cite this article: Bone Joint J 2016;98-B:730-5.

Proliferation of lymphoid precursor cells in the bone marrow of patients with various disorders of the immune system.

In the human bone marrow the nuclear enzyme terminal deoxynucleotidyl transferase (TdT) is expressed by cells during early stages of lymphocyte differentiation. In order to investigate a possible regulation of lymphopoiesis at this level of differentiation, the relative frequency and the in vitro 3H-thymidine labeling index (3HdT-LI) of TdT-positive bone marrow cells were assessed in patients with different functional activities of the immune system. TdT-positive lymphoid precursor cells could be detected in the bone marrow of all children investigated, including six patients with various forms of immunodeficiency. Neither a transient hyperfunction of the immune system during the immunological rebound after cessation of long-term chemotherapy for acute lymphoblastic leukemia, nor a congenital or acquired hypofunction of the immune system had any detectable influence on the invariably high in vitro 3HdT-LI of TdT-positive bone marrow cells, a phenomenon possibly related to an autonomous and high turnover of this precursor cell compartment in the human bone marrow.

Observing gas-catalyst dynamics at atomic resolution and single-atom sensitivity.

Transmission electron microscopy (TEM) has become an indispensable technique for studying heterogeneous catalysts. In particular, advancements of aberration-corrected electron optics and data acquisition schemes have made TEM capable of delivering images of catalysts with sub-Ångström resolution and single-atom sensitivity. Parallel developments of differentially pumped electron microscopes and of gas cells enable in situ observations of catalysts during the exposure to reactive gas environments at pressures of up to atmospheric levels and temperatures of up to several hundred centigrade. Here, we outline how to take advantage of the emerging state-of-the-art instrumentation and methodologies to study surface structures and dynamics to improve the understanding of structure-sensitive catalytic functionality. The concept of using low electron dose-rates in TEM in conjunction with in-line holography and aberration-correction at low voltage (80 kV) is introduced to allow maintaining atomic resolution and sensitivity during in situ observations of catalysts. Benefits are illustrated by exit wave reconstructions of TEM images of a nanocrystalline Co3O4 catalyst material acquired in situ during their exposure to either a reducing or oxidizing gas environment.

Structure/activity relationships of the genotoxic potencies of sixteen pyrrolizidine alkaloids assayed for the induction of somatic mutation and recombination in wing cells of Drosophila melanogaster.

Sixteen pyrrolizidine alkaloids (PAs) were examined for their genotoxic potency in the wing spot test of Drosophila melanogaster following oral application. This in vivo assay tests for the induction of somatic mutation and mitotic recombination in cells of the developing wing primordia. All PAs tested except the C9-monoester supinine were clearly genotoxic. Depending on their chemical structure, however, genotoxicity of the PAs varied widely in a range encompassing about three orders of magnitude. In general, macrocyclic diester-type PAs were the most and 7-hydroxy C9-monoester types the least genotoxic representatives studied, while open diesters were intermediate in this respect. Stereoisomeric PAs mostly showed similar, but sometimes also clearly unequal genotoxicity. An increasing number of hydroxy groups in the PA molecule seemed to reduce its genotoxic potency. With respect to the structure/activity relationships, there appears to be a good correlation between hepatotoxicity of PAs in experimental rodents and genotoxicity in the wing spot test of Drosophila. This suggests that PAs are bioactivated along similar pathways in the mammalian liver and in the somatic cells of Drosophila. The genotoxic potential of PAs in the Drosophila wing spot test and their carcinogenic potential in mammals also seem correlated, although the information in the literature on carcinogenicity of the non-macrocyclic PAs with moderate to low genotoxic potency is concededly limited. Comparisons with other genotoxicity tests suggest that the wing spot test is particularly suitable for genotoxins like PAs, on the one hand because of the versatile metabolic bioactivation system of Drosophila and on the other hand also because of its excellent sensitivity to the crosslinking agents among the genotoxins.

Optimal experimental design and sample size for the statistical evaluation of data from somatic mutation and recombination tests (SMART) in Drosophila.

In genetic toxicology it is important to know whether chemicals should be regarded as clearly hazardous or whether they can be considered sufficiently safe, which latter would be the case from the genotoxicologist's view if their genotoxic effects are nil or at least significantly below a predefined minimal effect level. A previously presented statistical decision procedure which allows one to make precisely this distinction is now extended to the question of how optimal experimental sample size can be determined in advance for genotoxicity experiments using the somatic mutation and recombination tests (SMART) of Drosophila. Optimally, the statistical tests should have high power to minimise the chance for statistically inconclusive results. Based on the normal test, the statistical principles are explained, and in an application to the wing spot assay, it is shown how the practitioner can proceed to optimise sample size to achieve numerically satisfactory conditions for statistical testing. The somatic genotoxicity assays of Drosophila are in principle based on somatic spots (mutant clones) that are recovered in variable numbers on individual flies. The underlying frequency distributions are expected to be of the Poisson type. However, some care seems indicated with respect to this latter assumption, because pooling of data over individuals, sexes, and experiments, for sample, can (but need not) lead to data which are overdispersed, i.e., the data may show more variability than theoretically expected. It is an undesired effect of overdispersion that in comparisons of pooled totals it can lead to statistical testing which is too liberal, because overall it yields too many seemingly significant results. If individual variability considered alone is not in contradiction with Poisson expectation, however, experimental planning can help to minimise the undesired effects of overdispersion on statistical testing of pooled totals. The rule for the practice is to avoid disproportionate sampling. It is recalled that for optimal power in statistical testing, it is preferable to use equal total numbers of flies in the control and treated series. Statistical tests which are based on Poisson expectations are too liberal if there is overdispersion in the data due to excess individual variability. In this case we propose to use the U test as a non-parametric two-sample test and to adjust the estimated optimal sample size according to (i) the overdispersion observed in a large historical control and (ii) the relative efficiency of the U test in comparison to the t test and related parametric tests.

Statistical methods to decide whether mutagenicity test data from Drosophila assays indicate a positive, negative, or inconclusive result.

Two alternative hypotheses are used to distinguish among the possibilities of a positive, inconclusive, or negative result in Drosophila mutagenicity tests. In the null hypothesis one assumes that there is no difference in the mutation frequency between control and treated series. The alternative hypothesis postulates a priori that the treatment results in an increased mutation frequency that is m times the spontaneous frequency. To test against the hypotheses, the conditional binomial test according to Kastenbaum and Bowman or the chi 2 test for proportions may be applied. These 2 methods are in principle equivalent. An alternative method which is based on determining confidence limits of observed mutation frequencies also leads to the same conclusions. The practical calculations are formulated and an application is shown with a test example demonstrating the genotoxicity of the pyrrolizidine alkaloid 7-acetylintermedine in the somatic wing mosaic test. In the Appendix, the calculus for the 3 testing methods is explained with a numerical example.

The vicinal chloroalcohols 1,3-dichloro-2-propanol (DC2P), 3-chloro-1,2-propanediol (3CPD) and 2-chloro-1,3-propanediol (2CPD) are not genotoxic in vivo in the wing spot test of Drosophila melanogaster.

In this study, the vicinal chloroalcohols 1,3-dichloro-2-propanol (DC2P), 3-chloro-1,2-propanediol (3CPD) and 2-chloro-1,3-propanediol (2CPD) were investigated for genotoxicity in the wing spot test of Drosophila. DC2P is an important starting material in many processes of synthesis in chemical industry. 3CPD as well as some related glycerol chlorohydrins were identified in protein hydrolysates industrially used for the production of food items such as seasonings, sauces and soups. The wing spot test is a somatic mutation and recombination test (SMART) and is a sensitive in vivo assay for the detection of mutagens and promutagens. The test was applied here in its standard version with normal bioactivation and in a variant with increased cytochrome P450-dependent bioactivation capacity. All three compounds were clearly non-genotoxic in these in vivo assays. The results are in agreement with recent findings which strongly suggest that positive genotoxicity results in in vitro testing of vicinal chloroalcohols such as DC2P are due to directly acting genotoxic intermediates arising from a chemical reaction with the culture medium rather than from enzymatic biotransformation.

Genetic toxicity of six carcinogens and six non-carcinogens in the Drosophila wing spot test.

Six rodent carcinogens, 5 of which are also human carcinogens, and 6 compounds recognized as non-carcinogens were tested for their genotoxic activity in the Drosophila melanogaster wing spot test. 72-h-old larvae trans-heterozygous for the recessive wing cell markers 'multiple wing hairs' (mwh) and 'flare' (flr3) were fed various concentrations of the test compounds for a period of 48 h. With amitrole and 4-aminobiphenyl, larvae of the same age were also given an acute treatment of 6 h with higher concentrations, and, in addition, 48-h-old larvae were fed for a longer period of 72 h. Repeats of all experiments document the good reproducibility of the results in the wing spot test. Amitrole and 4-aminobiphenyl were genotoxic after both 48-h and 72-h treatments, but their activity could not be detected following acute exposure of only 6 h. Chlorambucil and melphalan were clearly genotoxic. The carcinogens sodium arsenite and sodium arsenate, however, which are highly toxic to Drosophila, could only be tested at low exposure levels and were negative under these treatment conditions. The 6 non-carcinogens (ascorbic acid, 2-aminobiphenyl, mannitol, piperonyl butoxide, stannous chloride and titanium dioxide) were all definitely non-genotoxic in the Drosophila wing spot test. The data for the non-carcinogens demonstrate that non-genotoxic compounds can be identified in the wing spot test with a reasonable experimental effort.

The genotoxicity of the anti-cancer drug mitoxantrone in somatic and germ cells of Drosophila melanogaster.

The novel antineoplastic drug mitoxantrone was studied for its genotoxic effects in Drosophila melanogaster. In male germ cells, the clinical preparation Novantrone, the dihydrochloride salt of mitoxantrone, did not induce sex-linked recessive lethal mutations in feeding and injection experiments with adult flies, although statistically the results were inconclusive rather than truly negative. However, the free base mitoxantrone was weakly, but significantly genotoxic in this test (0.14% lethals/mM exposure concentration); this is most probably the result of prolonged exposure. On the other hand, both forms of mitoxantrone assayed were clearly genotoxic in the somatic mutation and recombination test of the wing. This test assays the cells of the proliferating imaginal wing discs of larvae. Depending on the feeding method used, the overall clone induction frequency was in the range of about 2-6 x 10(-5) per cell and cell generation and per mM exposure dose. Correction of these frequencies according to mean clone size led to slightly higher estimates (by about 5-25% higher). Although the majority of the clone induction events are due to mitotic recombination, a significant proportion can be attributed to mutational events (gene and chromosome mutations). The genotoxicity of mitoxantrone seems to depend mainly on impaired DNA synthesis in cycling cells owing to the compound's ability to inhibit topoisomerase II by intercalation into DNA.

Thirty compounds tested in the Drosophila wing spot test.

The Drosophila wing somatic mutation and recombination test (SMART) was evaluated for its suitability in genotoxicity screening by testing 30 chemicals. Of the 2 crosses used, the mwh-flr3 cross turned out to be more convenient than the previously used mwh-flr cross. Based on the experience gained with both acute exposures and chronic exposures of different duration, we suggest that the optimal strategy in genotoxicity screening is to start with chronic exposure of 3-day-old larvae for 48 h (that is, until pupation). Only for unstable compounds and very volatile compounds and gases are acute treatments, including inhalation, recommended. In general, a qualitative evaluation of the genotoxicity of a compound in the wing assay is possible with as few as 1-2 different exposure concentrations. A more quantitative evaluation of genotoxicity, based upon dose-response data, can often be achieved with as few as 3-4 concentrations. The results reported here were obtained in 2 different laboratories, demonstrating that the wing spot test is easily transferable to other laboratories. The experience gained indicates that the assay has now been developed to an extent that a coordinated international comparative validation study is desirable.

The effect of nucleotomy on lumbar spine mechanics in compression and shear loading.

STUDY DESIGN: An in vitro biomechanical investigation on human cadaveric specimens was conducted before and after nucleotomy. Endplate and vertebral body deformation patterns were measured under compression and shear loading, in addition to kinematics and disc pressure. OBJECTIVE: The working hypotheses of this study were that in compression, nucleotomy results in an altered deformation pattern of the endplate and that in shear, nucleotomy does not result in an altered endplate deformation pattern or disc pressure. SUMMARY OF BACKGROUND DATA: The pressure distributions within the intervertebral disc have been studied in compression loading but not in shear loading. Severe degeneration and surgical nucleotomy result in small nuclear pressure and altered loading distribution in compression. The effect of these changes on the vertebral endplate and the response under shear loads are not well understood. METHODS: Five L3-L4 and two L4-L5 functional spinal units were tested under compression and shear loading, intact and after nucleotomy. Vertebral body deformations, intradiscal pressure, and intervertebral kinematics were measured. A series of compression-type (maximum 1000 N) and shear-type (maximum 500 N) loads were applied. RESULTS: With nucleotomy, the disc pressure and the endplate strains decreased under compression, but the vertebral rim strains did not change. In shear, the vertebral rim and endplate strains did not change with nucleotomy. Disc pressure was lower in shear than in compression. CONCLUSION: Nucleotomy resulted in decreased disc pressure, decreased endplate deformation, and modified loading patterns onto the inferior vertebra in compression loading. However, nucleotomy did not appreciably affect the behavior of the disc in shear loading.

Allograft impaction and cement penetration after revision hip replacement. A histomorphometric analysis in the cadaver femur.

We studied various aspects of graft impaction and penetration of cement in an experimental model. Cancellous bone was removed proximally and local diaphyseal lytic defects were simulated in six human cadaver femora. After impaction grafting the specimens were sectioned and prepared for histomorphometric analysis. The porosity of the graft was lowest in Gruen zone 4 (52%) and highest in Gruen zone 1 (76%). At the levels of Gruen zones 6 and 2 the entire cross-section was almost filled with cement. Cement sometimes reached the endosteal surface in other Gruen zones. The mean peak impaction forces exerted with the impactors were negatively correlated with the porosity of the graft.

[Metal ions: important co-players in aseptic loosening]. / Metallionen: Wichtige Mitspieler in der aseptischen Lockerung.

AIM: The aims of this review were to discuss the different mechanisms of biocorrosion of orthopaedic metal implants in the human body, as well as the effects of the released metal ions on bone metabolism and the immune system in regard to their involvement in the pathophysiological mechanisms of aseptic loosening and metal hypersensitivity. Implant failure due to aseptic loosening is thought to occur in about 10-15% of cases. METHODS: A review of the literature (using PubMed with the search terms: biocorrosion, metal ions and bone metabolism) was performed. Additionally, we discuss our research results in the field of aseptic loosening. RESULTS: Despite a great effort in developing new implants, metal devices used in orthopaedic and trauma surgery remain prone to biocorrosion by several mechanisms including the direct corrosion by osteoclasts, leading to the production of significant amounts of wear particles and metal ions. In addition to the well documented increased osteolytic activity caused by large (in the nanometer range) wear particles, increasing evidence strongly suggests that the released metal ions contribute to the pathophysiological mechanism of aseptic loosening. Metal ions stimulate both the immune system and bone metabolism through a series of direct and indirect pathways leading to an increased osteolytic activity at the bone-implant interface. CONCLUSION: To date, revision surgery remains the only option for the treatment of a failed orthopaedic implant caused by aseptic loosening. A better understanding of the complex pathophysiological mechanisms (including the effects caused by the released metal ions) of aseptic loosening may have a significant potential in developing novel implants and therapies in order to reduce the incidence of this complication.