Ultrasonographic Assessment of the Normal Femoral Articular Cartilage of the Knee Joint: Comparison with 3D MRI.

OBJECTIVE: Ultrasonography (US) has a promising role in evaluating the knee joint, but capability to visualize the femoral articular cartilage needs systematic evaluation. We measured the extent of this acoustic window by comparing standardized US images with the corresponding MRI views of the femoral cartilage. DESIGN: Ten healthy volunteers without knee pathology underwent systematic US and MRI evaluation of both knees. The femoral cartilage was assessed on the oblique transverse axial plane with US and with 3D MRI. The acoustic window on US was compared to the corresponding views of the femoral sulcus and both condyles on MRI. The mean imaging coverage of the femoral cartilage and the cartilage thickness measurements on US and MRI were compared. RESULTS: Mean imaging coverage of the cartilage of the medial femoral condyle was 66% (range 54%-80%) and on the lateral femoral condyle 37% (range 25%-51%) compared with MRI. Mean cartilage thickness measurement in the femoral sulcus was 3.17 mm with US and 3.61 mm with MRI (14.0% difference). The corresponding measurements in the medial femoral condyle were 1.95 mm with US and 2.35 mm with MRI (21.0% difference), and in the lateral femoral condyle, they were 2.17 mm and 2.73 mm (25.6% difference), respectively. CONCLUSION: Two-thirds of the articular cartilage of the medial femoral condyle, and one-third in the lateral femoral condyle, can be assessed with US. The cartilage thickness measurements seem to be underestimated by US. These results show promise for the evaluation of the weight-bearing cartilage of the medial femoral condyle with US.

Comparison of ultrasonographic, radiographic and intra-operative findings in severe hip osteoarthritis.

Aim of this study was to assess the US findings of patients with late-stage hip OA undergoing total hip arthroplasty (THA), and to associate the US findings with conventional radiography (CR) and intraoperative findings. Moreover, the inter-rater reliability of hip US, and association between the US and Oxford Hip Score (OHS) were evaluated. Sixty-eight hips were included, and intraoperative findings were available on 48 hips. Mean patient age was 67.6 years and 38% were males. OA findings-osteophytes at femoral collum and anterosuperior acetabulum, femoral head deformity and effusion-were assessed on US, CR and THA. The diagnostic performance of US and CR was compared by applying the THA findings as the gold standard. Osteoarthritic US findings were very common, but no association between the US findings and OHS was observed. The pooled inter-rater reliability (n = 65) varied from moderate to excellent (k = 0.538-0.815). When THA findings were used as the gold standard, US detected femoral collum osteophytes with 95% sensitivity, 0% specificity, 81% accuracy, and 85% positive predictive value. Concerning acetabular osteophytes, the respective values were 96%, 0%, 88% and 91%. For the femoral head deformity, they were 92%, 36%, 38% and 83%, and for the effusion 49%, 85%, 58% and 90%, respectively. US provides similar detection of osteophytes as does CR. On femoral head deformity, performance of the US is superior to CR. The inter-rater reliability of the US evaluation varies from moderate to excellent, and no association between US and OHS was observed in this patient cohort.

Association between grayscale sonographic and clinical findings in severe knee osteoarthritis.

PURPOSE: To assess whether ultrasonographic (US) findings associate with clinical findings in severe knee osteoarthritis (OA). Association of US findings with side-of-knee pain and inter-reader agreement of knee US were also evaluated. METHODS: One-hundred-two patients (in total 123 knees) with severe knee OA were recruited for this cross-sectional study. US was performed by a single observer, and on 53 knees by two independent observers to assess inter-reader reliability. Preoperative clinical data was available for 69 knees. Cutoff values were applied to dichotomize US and clinical findings. The Chi-square test, Mann-Whitney test, and prevalence- and bias-adjusted kappa (PABAK) were applied for statistical analyses. RESULTS: Seven of 99 associations tested were statistically significant. Associations were observed between range of flexion and lateral femoral (P = .009) and tibial (P = .001) osteophytes, mediolateral instability and damage to the lateral femoral cartilage (P = .014) and damage to the lateral meniscus (P = .031), and alignment and damage to the lateral femoral cartilage (P < .001), lateral tibial osteophytes (P = .037), and damage to the lateral meniscus (P < .001). A strong association was observed between medial-sided pain and same-sided cartilage damage and osteophytes (P < .001). That inter-reader agreement was excellent on the medial side of the knee joint (PABAK = 0.811-0.887). CONCLUSIONS: US findings show a rather poor association with clinical OA findings. Inter-reader agreement of knee US is excellent on the medial side.

Osteoclasts secrete osteopontin into resorption lacunae during bone resorption.

Osteopontin (OPN) is a non-collagenous extracellular sialylated glycoprotein located in bone. It is believed to be one of the key components in osteoclast attachment to bone during resorption. In this study, we characterized OPN and other glycoproteins found in the resorption lacunae to confirm the role of osteoclasts in OPN secretion using electron microscopy and mass spectrometry. Additionally, we examined the glycan epitopes of resorption pits and the effects of different glycan epitopes on the differentiation and function of osteoclasts. Osteoarthritic femoral heads were examined by immunohistochemistry to reveal the presence of OPN in areas of increased bone metabolism in vivo. Our results demonstrate that human osteoclasts secrete OPN into resorption lacunae on native human bone and on carbonated hydroxyapatite devoid of natural OPN. OPN is associated with an elevated bone turnover in osteoarthritic bone under experimental conditions. Our data further confirm that osteoclasts secrete OPN into the resorption pit where it may function as a chemokine for subsequent bone formation. We show that α2,3- and α2,6-linked sialic acids have a role in the process of osteoclast differentiation. OPN is one of the proteins that has both of the above sialic residues, hence we propose that de-sialylation can effect osteoclast differentiation in bone.

Ultrasonography of the late-stage knee osteoarthritis prior to total knee arthroplasty: comparison of the ultrasonographic, radiographic and intra-operative findings.

The purpose of this study was to assess the effectiveness of the ultrasonography (US) on detecting osteoarthritis of the knee, and compare US and radiographic findings to intraoperative total knee arthroplasty (TKA) findings. Fifty-seven late-stage osteoarthritic knees undergoing TKA were evaluated with US and radiography. Standard knee US assessing femoral cartilage damage, osteophytes, effusion, synovitis, and meniscal extrusion was performed. On radiographs, osteophytes, joint space narrowing, and Kellgren-Lawrence grade were evaluated. Corresponding intra-operative findings were assessed during TKA as the gold standard. On the damage of the medial femoral condyle cartilage, the sensitivity of US was high (92%), whereas on the lateral condyle and sulcus area, sensitivities were 58% and 46%, respectively. On osteophytes, the detection rate of the US was remarkable especially on the medial side yielding sensitivities of 90-95%. The sensitivities for detecting effusion and synovitis were also excellent (97%). US detection rate of femoral cartilage damage was in concordance with the radiographic joint space narrowing. For the detection of osteophytes, US provided superior results to radiography particularly on the medial side. In conclusion, US can reliably assess the late-stage OA changes of the knee especially on the medial side of the knee joint.

Osteoclasts in the interface with electrospun hydroxyapatite.

Electrospinning is a method to produce lightweight, resorbable and bioinspired scaffolds for tissue engineering. Here we investigated the influence of electrospun hydroxyapatite fibers (HA) on macrophages and osteoclasts. A mouse macrophage cell line (RAW 264.7) and human bone marrow derived primary osteoclasts (hOC) were cultured with electrospun HA fibers embedded in Matrigel. Cell morphology and the secretion of pro-inflammatory cytokines (IL-6 and TNF-α) were analyzed using macrophages. Both fluorescent microscopy and scanning electron microscopy indicated that the cell morphology differed on the various materials (HA fibers on Matrigel, pure Matrigel and a glass control). Control macrophages were activated with bacterial lipopolysaccharide (LPS) but electrospun HA did not provoke an inflammatory response. Cytokine secretion detected with enzyme-linked immunosorbent assay (ELISA) also supported this observation. LPS, but not HA fibers, stimulated TNF-α and IL-6 secretion by macrophages at the 2 day time point. After 4 days in culture there was an increasing trend in cytokine secretion in the HA fiber samples. Human bone marrow myeloid precursor cells were able to fuse and differentiate on the fibrous mineral scaffold to form functional multinuclear osteoclasts that were able to resorb the HA nanofibers. This indicates that osteoclasts do not necessarily need a continuous bone surface but osteoclast ruffled border membranes can form a resorption interface with a fibrous mineral scaffold.

Preparation and bioactive properties of nanocrystalline hydroxyapatite thin films obtained by conversion of atomic layer deposited calcium carbonate.

Nanocrystalline hydroxyapatite thin films were fabricated on silicon and titanium by atomic layer deposition (ALD) of CaCO3 and its subsequent conversion to hydroxyapatite by diammonium hydrogen phosphate (DAP) solution. The effects of conversion process parameters to crystallinity and morphology of the films were examined. DAP concentration was found to be critical in controlling the crystal size and homogeneity of the films. The hydroxyapatite phase was identified by XRD. ToF-elastic recoil detection analysis studies revealed that the films are calcium deficient in relation to hydroxyapatite with a Ca/P ratio of 1.39 for films converted with 0.2 M DAP at 95 °C. The coatings prepared on titanium conformally follow the rough surface topography of the substrate, verifying that the good step coverage of the ALD method was maintained in the conversion process. The dissolution tests revealed that the coating was nondissolvable in the cell culture medium. Annealing the coated sample at 700 °C for 1 h seemed to enhance its bonding properties to the substrate. Also, the biocompatibility of the coatings was confirmed by human bone marrow derived cells in vitro. The developed method provides a new possibility to produce thin film coatings on titanium implants with bone-type hydroxyapatite that is biocompatible with human osteoblasts and osteoclasts.

An in vitro study into the effect of zinc substituted hydroxyapatite on osteoclast number and activity.

Zinc ions have been shown to inhibit osteoclast development and proliferation both in vitro and in vivo. The same inhibiting effect has been observed in vitro when zinc was substituted into tri-calcium phosphate (TCP). Because of the solubility of TCP it is not an ideal candidate for a material to inhibit osteoclast activity in the long term. Hydroxyapatite (HA) is less soluble and so potentially offers a more long-term, sustainable effect. Previous work has shown that zinc can successfully be substituted into HA and still retain phase purity after heat treatment. The study reported here presents the effects of zinc substituted HA on the development and activity of osteoclast-like cells. It was found that increasing zinc substitution levels led to a decrease in the number of these cells present after 21 days. When resorption activity was investigated it was found that an increase in the amount of zinc present in the discs led to a significant decrease in the amount of resorption taking place on the discs. These results provide evidence for the potential of zinc substituted HA as a material to reduce resorptive activity to provide long-term bonding of implant to bone.

Osteoclastogenesis is influenced by modulation of gap junctional communication with antiarrhythmic peptides.

Osteoclasts are formed by the fusion of mononuclear precursor cells of the monocyte-macrophage lineage. Among several putative mechanisms, gap-junctional intercellular communication (GJC) has been proposed to have a role in osteoclast fusion and bone resorption. We examined the role of GJC in osteoclastogenesis and in vitro bone resorption with mouse bone marrow hematopoietic stem cells and RAW 264.7 cells. Blocking of gap junctions with 18-α-glycyrrhetinic acid (18GA) led to inhibition of osteoclastogenesis and in vitro bone resorption. Similarly, the GJC inhibitor GAP27 inhibited osteoclast formation. GJC modulation with the antiarrhythmic peptides (AAPs) led to increased amounts of multinuclear RAW 264.7 osteoclasts as well as increased number of nuclei per multinuclear cell. In the culture of bone marrow hematopoietic stem cells in the presence of bone marrow stromal cells AAP reduced the number of osteoclasts, and coculture of MC3T3-E1 preosteoblasts with RAW 264.7 macrophages prevented the action of AAPs to promote osteoclastogenesis. The present data indicate that AAPs modulate the fusion of the pure culture of cells of the monocyte-macrophage lineage. However, the fusion is influenced by GJC in cells of the osteoblast lineage.

Affecting osteoblastic responses with in vivo engineered potato pectin fragments.

Pectins, complex plant-derived polysaccharides, are novel candidates for biomaterial nanocoatings. Pectic rhamnogalacturonan-I regions (RG-I) can be enzymatically treated to so-called modified hairy regions (MHR). We surveyed the growth and differentiation of murine preosteoblastic MC3T3-E1 cells on Petri dishes coated with RG-Is from native or genetically engineered potato tubers. Uncoated tissue culture polystyrene (TCPS) and aminated (AMI) dishes served as controls. MHRPTR_GAL sample was depleted of galactose (9 mol % galactose; 23 mol % arabinose) and MHRPTR_ARA of arabinose (61 mol % galactose; 6 mol % arabinose). Wild-type (modified hairy region from potato pectin (MHRP)_WT) fragment contained default amounts (58 mol % galactose; 13 mol % arabinose) of both sugars. Focal adhesions (FAs) indicating cellular attachment were quantified. Reverse transcriptase polymerase chain reaction (RT-PCR) of alkaline phosphatase and osteocalcin genes indicating osteoblastic differentiation was performed along with staining the produced calcium with tetracycline as an indicator of osteoblastic differentiation. Osteoblasts proliferated on all the samples to some extent. The control surfaces performed better than any of the pectin samples, of which the MHRP_WT seemed to function best. FA length was greater on MHRPTR_GAL than on other pectin samples, otherwise the mutants did not significantly deviate. RT-PCR results indicate that differences between the samples at the gene expression level might be even subtler. However, tetracycline-stained calcium-containing mineral was detected merely only on uncoated TCPS. These results indicate the possibility to affect bone cell growth with in vivo-modified pectin fragments, consecutively providing information on the significance of certain monosaccharides on the biocompatibility of these polysaccharides.