Biomedical subjects

Harrie Weinans

Publications and source records attributed to Harrie Weinans.

16 recordsLinked to original sources

PubMed2007

Histogram-based selective deblurring to improve computed tomography imaging of calcifications.

OBJECTIVES: Computed tomography (CT) imaging of small high-density structures, eg, calcifications, is hampered by image blur. This study aims to deconvolve calcifications in the transverse and longitudinal directions while avoiding noise amplification and edge-ringing artifacts in the surrounding low-density structures. MATERIALS AND METHODS: A method referred to as histogram-based selective deblurring (HiSD) has been developed to generate a restored image by combining the low-intensity (ie, Hounsfield Units) information of the original image with the high-intensity information of the deconvolved image. HiSD is evaluated on phantom and in vitro atherosclerotic plaque CT images by comparing the original and restored images with their corresponding reference micro-CT images both qualitatively and quantitatively. RESULTS: HiSD reduces calcification blur in the transverse and longitudinal directions without introducing noise and ringing-artifacts in the surrounding tissues. Calcification area and volume measurements are significantly improved in the restored images (reducing on average overestimation by 32% and 83%, respectively). CONCLUSIONS: HiSD significantly improves CT visualization and quantification of small high-density structures imaged in vitro.

Algorithms↗

PubMed2006

Long-term serotonin administration leads to higher bone mineral density, affects bone architecture, and leads to higher femoral bone stiffness in rats.

New evidence suggests a control of bone mass by the central nervous system. We have previously shown that functional serotonin receptors are present in bone cells and that serotonin stimulates proliferation of osteoblast precursor cells in vitro. In the present study we investigated the effects of serotonin on bone tissue in vivo. Ten, 2-month-old female Sprague-Dawley rats were injected with serotonin subcutaneously (s.c.) (5 mg/kg) once daily for 3 months, controls received saline. Using microdialysis and HPLC, free circulating serotonin levels were measured. DXA scans were made after 3 months of serotonin administration. Bone architecture and mechanical properties were investigated by micro-computed tomography (microCT), histomorphometry, and mechanical testing. A long-lasting hyperserotoninemia with a >10-fold increase in serotonin appeared. Total body BMD was significantly higher (0.1976+/-0.0015 vs. 0.1913+/-0.0012 g/cm2) in rats receiving serotonin. Cortical thickness (Ct.Th) measured by microCT analysis was also higher, whereas trabecular bone volume (BV) was lower. Interestingly, the perimeter and cross-sectional moment of inertia (MOI), a proxy for geometrical bone strength, were the same in both groups. These data suggest that serotonin reduces resorption or/and increases apposition of endosteal bone. Mechanical testing showed that femoral stiffness was higher in serotonin-dosed animals. The energy absorption also seemed slightly, but not significantly higher. In conclusion, hyperserotoninemia led to a higher BMD, altered bone architecture and higher femural bone stiffness in growing rats, demonstrating that serotonin may have important effects on bone in vivo.

Animals↗

PubMed2006

Ultrasonic characterization of human trabecular bone microstructure.

New quantitative ultrasound (QUS) techniques involving ultrasound backscattering have been introduced for the assessment of bone quality. QUS parameters are affected by the transducer characteristics, e.g. frequency range, wave and pulse length. Although frequency-dependent backscattering has been studied extensively, understanding of the ultrasound scattering phenomenon in trabecular bone is still limited. In the present study, the relationships between QUS parameters and the microstructure of human trabecular bone were investigated experimentally and by using numerical simulations. Speed of sound (SOS), normalized broadband ultrasound attenuation (nBUA), average attenuation, integrated reflection coefficient (IRC) and broadband ultrasound backscatter (BUB) were measured for 26 human trabecular bone cylinders. Subsequently, a high-resolution microCT system was used to determine the microstructural parameters. Moreover, based on the sample-specific microCT data, a numerical model for ultrasound propagation was developed for the simulation of experimental measurements. Experimentally, significant relationships between the QUS parameters and microstructural parameters were demonstrated. The relationships were dependent on the frequency, and the strongest association (r = 0.88) between SOS and structural parameters was observed at a centre frequency of 5 MHz. nBUA, average attenuation, IRC and BUB showed somewhat lower linear correlations with the structural properties at a centre frequency of 5 MHz, as compared to those determined at lower frequencies. Multiple regression analyses revealed that the variation of acoustic parameters could best be explained by parameters reflecting the amount of mineralized tissue. A principal component analysis demonstrated that the strongest determinants of BUB and IRC were related to the trabecular structure. However, other structural characteristics contributed significantly to the prediction of the acoustic parameters as well. The two-dimensional numerical model introduced in the present study demonstrated good agreement with the experimental measurements. However, further studies with the simulation model are warranted to systematically investigate the relation between the structural parameters and ultrasound scattering.

Absorptiometry, Photon↗

PubMed2006

Bone loss dynamics result in trabecular alignment in aging and ovariectomized rats.

Because of the destructive nature of techniques used to measure bone morphometry, studies of architectural changes and bone loss have utilized cross-sectional study designs, with all its inherent limitations in nuances. Here, the results of a longitudinal study using in vivo micro-CT are presented elucidating the dynamics of bone loss and architectural adaptation in rat models of aging and postmenopausal bone loss. Using 3-D methodology, we observed the changes in bone architecture in the proximal tibia of normally aging and ovariectomized rats for 54 weeks. Spatial patterns in bone resorption were observed that were similar for both groups. Remaining trabeculae increased in thickness or were remodeled into new trabecular structures, especially in the ovariectomized animals. The combination of bone loss and bone formation resulted in alignment of trabeculae across the growth plate. Cortical modeling that was associated with growth continued after cessation of longitudinal growth in the ovariectomized animals, resulting in shape changes of the proximal tibia. The organized nature of the changes in bone architecture that occurred after ovariectomy and the high similarity with the changes observed in the normally aging animals, suggest that estrogen depletion resulted in an acceleration of a normal bone adaptation process. The observed aligning of trabeculae suggests regulation through mechanical loading.

Aging↗

PubMed2006

Stretch-induced modulation of matrix metalloproteinases in mineralizing osteoblasts via extracellular signal-regulated kinase-1/2.

Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) produced by osteoblasts play an essential role in bone remodeling. Hence, these proteins could provide an interesting means by which mechanical loading leads to adaptation of bone. Here, we examined the effect of stretch on MMP-1, -2, -3, -8, -9, -13, and -14, as well as TIMP-1 and -2 gene expression in differentiating, mineralizing, and nonmineralizing human SV-40 immortalized preosteoblast cells. In the mineralizing osteoblast culture, but not in the nonmineralizing cultures, cyclic stretch for only 15 min resulted in an increase of MMP-1 (fourfold) and -3 (depending on differentiation stage up to 25-fold) transcript abundance. No clear effect was observed for other MMPs, TIMP-1 or -2. The increase of MMP-1 and -3 was confirmed on the protein level. Stretching experiments performed in the presence of a specific inhibitor of extracellular signal-regulated kinase (ERK) showed a strong suppression of the stretch-induced increase in MMP-1 and -3. In conclusion, we show that MMP-1 and MMP-3 are mechanosensitive genes in mineralizing the human osteoblast, and that the mechano-induction of these genes is mediated via the ERK pathway. Our findings implicate that these MMPs are important factors in the mechanoregulation of bone turnover. With the ability to generate MMPs at highly stretched sites, osteoblasts can potantially direct osteoclasts to specific bone surface areas prepared for resorption.

Butadienes↗

PubMed2006

Prediction of torsional failure in 22 cadaver femora with and without simulated subtrochanteric metastatic defects: a CT scan-based finite element analysis.

BACKGROUND: In metastatic bone disease, prophylactic fixation of impending long bone fracture is preferred over surgical treatment of a manifest fracture. There are no reliable guidelines for prediction of pathological fracture risk, however. We aimed to determine whether finite element (FE) models constructed from quantitative CT scans could be used for predicting pathological fracture load and location in a cadaver model of metastatic bone disease. MATERIAL AND METHODS: Subject-specific FE models were constructed from quantitative CT scans of 11 pairs of human femora. To simulate a metastatic defect, a transcortical hole was made in the subtrochanteric region in one femur of each pair. All femora were experimentally loaded in torsion until fracture. FE simulations of the experimental set-up were performed and torsional stiffness and strain energy density (SED) distribution were determined. RESULTS: In 15 of the 22 cases, locations of maximal SED fitted with the actual fracture locations. The calculated torsional stiffness of the entire femur combined with a criterion based on the local SED distribution in the FE model predicted 82% of the variance of the experimental torsional failure load. INTERPRETATION: In the future, CT scan-based FE analysis may provide a useful tool for identification of impending pathological fractures requiring prophylactic stabilization.

Aged↗

PubMed2006

The novel vitamin D analog ZK191784 as an intestine-specific vitamin D antagonist.

Vitamin D [1,25(OH)2D3] plays a crucial role in Ca2+ homeostasis by stimulating Ca2+ (re)absorption and bone turnover. The 1,25(OH)2D3 analog ZK191784 was recently developed to dissociate the therapeutic immunomodulatory activity from the hypercalcemic side effects of 1,25(OH)2D3 and contains a structurally modified side chain characterized by a 22,23-double bond, 24R-hydroxy group, 25-cyclopropyl ring, and 5-butyloxazole unit. We investigated the effect of ZK191784 on Ca2+ homeostasis and the regulation of Ca2+ transport proteins in wild-type (WT) mice and mice lacking the renal epithelial Ca2+ channel TRPV5 (TRPV5-/-). The latter display hypercalciuria, hypervitaminosis D, increased intestinal expression of the epithelial Ca2+ channel TRPV6, the Ca2+-binding protein calbindin-D(9K), and intestinal Ca2+ hyperabsorption. ZK191784 normalized the Ca2+ hyperabsorption and the expression of intestinal Ca2+ transport proteins in TRPV5-/- mice. Furthermore, the compound decreased intestinal Ca2+ absorption in WT mice and reduced 1,25(OH)2D3-dependent 45Ca2+ uptake by Caco-2 cells, substantiating a 1,25(OH)2D3-antagonistic action of ZK191784 in the intestine. ZK191784 increased renal TRPV5 and calbindin-D(28K) expression and decreased urine Ca2+ excretion in WT mice. Both 1,25(OH)2D3 and ZK191784 enhanced transcellular Ca2+ transport in primary cultures of rabbit connecting tubules and cortical collecting ducts, indicating a 1,25(OH)2D3-agonistic effect in kidney. ZK191784 enhanced bone TRPV6 mRNA levels and 1,25(OH)2D3 as well as ZK191784 stimulated secretion of the bone formation marker osteocalcin in rat osteosarcoma cells, albeit to a different extent. In conclusion, ZK191784 is a synthetic 1,25(OH)2D3 ligand displaying a unique tissue-specific profile when administered in vivo. Because ZK191784 acts as an intestine-specific 1,25(OH)2D3 antagonist, this compound will be associated with less hypercalcemic side effects compared with the 1,25(OH)2D3 analogs currently used in clinical practice.

Animals↗

PubMed2005

Identification of acid-sensing ion channels in bone.

Bone balances serum pH variations and both osteoclasts and osteoblasts are regulated by subtle changes in pH. The aim of the current study was to identify molecules in bone that can sense pH. Interesting candidates are the acid-sensing ion channels (ASICs). In bone, ASIC2 and ASIC3 were most abundant, while in chondrocytes it was ASIC1. Isolated human monocytes expressed ASIC1, -2, and -3, which persisted after induction to osteoclast differentiation, albeit to a lower level. In human osteoblasts ASIC1, ASIC2, and ASIC3 mRNAs were shown. Western blot and immunostaining confirmed this at protein level. ASIC4 expression was always very low abundant. For the first time, we demonstrated ASICs in human skeleton, providing a means to sense and respond to differences in extracellular pH.

Acid Sensing Ion Channels↗

PubMed2005

Hypervitaminosis D mediates compensatory Ca2+ hyperabsorption in TRPV5 knockout mice.

Vitamin D plays an important role in Ca(2+) homeostasis by controlling Ca(2+) (re)absorption in intestine, kidney, and bone. The epithelial Ca(2+) channel TRPV5 mediates the Ca(2+) entry step in active Ca(2+) reabsorption. TRPV5 knockout (TRPV5(-/-)) mice show impaired Ca(2+) reabsorption, hypercalciuria, hypervitaminosis D, and intestinal hyperabsorption of Ca(2+). Moreover, these mice demonstrate upregulation of intestinal TRPV6 and calbindin-D(9K) expression compared with wild-type mice. For addressing the role of the observed hypervitaminosis D in the maintenance of Ca(2+) homeostasis and the regulation of expression levels of the Ca(2+) transport proteins in kidney and intestine, TRPV5/25-hydroxyvitamin-D(3)-1alpha-hydroxylase double knockout (TRPV5(-/-)/1alpha-OHase(-/-)) mice, which show undetectable serum 1,25(OH)(2)D(3) levels, were generated. TRPV5(-/-)/1alpha-OHase(-/-) mice displayed a significant hypocalcemia compared with wild-type mice (1.10 +/- 0.02 and 2.54 +/- 0.01 mM, respectively; P < 0.05). mRNA levels of renal calbindin-D(28K) (7 +/- 2%), calbindin-D(9K) (32 +/- 4%), Na(+)/Ca(2+) exchanger (12 +/- 2%), and intestinal TRPV6 (40 +/- 8%) and calbindin-D(9K) (26 +/- 4%) expression levels were decreased compared with wild-type mice. Hyperparathyroidism and rickets were present in TRPV5(-/-)/1alpha-OHase(-/-) mice, more pronounced than observed in single TRPV5 or 1alpha-OHase knockout mice. It is interesting that a renal Ca(2+) leak, as demonstrated in TRPV5(-/-) mice, persisted in TRPV5(-/-)/1alpha-OHase(-/-) mice, but a compensatory upregulation of intestinal Ca(2+) transporters was abolished. In conclusion, the elevation of serum 1,25(OH)(2)D(3) levels in TRPV5(-/-) mice is responsible for the upregulation of intestinal Ca(2+) transporters and Ca(2+) hyperabsorption. Hypervitaminosis D, therefore, is of crucial importance to maintain normocalcemia in impaired Ca(2+) reabsorption in TRPV5(-/-) mice.

Animals↗

PubMed2005

The effect of glucosamine sulphate on osteoarthritis: design of a long-term randomised clinical trial [ISRCTN54513166].

BACKGROUND: Pharmacological treatment for osteoarthritis (OA) can be divided into two groups: symptom-modifying drugs and disease-modifying drugs. Symptom-modifying drugs are currently the prescription of choice for patients with OA, as disease-modifying drugs are not yet available in usual care. However, there has recently been a lot of debate about glucosamine sulphate (GS), a biological agent that is thought to have both symptom-modifying and disease-modifying properties. This assumption has yet to be proved. The objective of this article is to present the design of a blind randomised clinical trial that examines the long-term symptom-modifying and disease-modifying effectiveness of GS in patients with hip OA. This trial is ongoing and will finish in March 2006. METHODS/DESIGN: Patients with hip OA meeting the ACR-criteria are randomly allocated to either 1500 mg of oral GS or placebo for the duration of two years. The primary outcome measures, which are joint space narrowing (JSN), and change in the pain and function score of the Western Ontario McMaster Universities Osteoarthritis index (WOMAC), are determined at baseline and after two years of follow-up during the final assessment. Intermediate measures at three-month intervals throughout the trial are used to study secondary outcome measures. Secondary outcome measures are changes in WOMAC stiffness score, quality of life, medical consumption, side effects and differences in biomarker CTX-II.

Antirheumatic Agents↗

PubMed2005

Prediction of density and mechanical properties of human trabecular bone in vitro by using ultrasound transmission and backscattering measurements at 0.2-6.7 MHz frequency range.

The ultrasound (US) backscattering method has been introduced as an alternative for the through-transmission measurement of sound attenuation and speed in diagnosis of osteoporosis. Both attenuation and backscattering depend strongly on the US frequency. In this study, 20 human trabecular bone samples were measured in transmission and pulse-echo geometry in vitro. The aim of the study was to find the most sensitive frequency range for the quantitative ultrasound (QUS) analyses. Normalized broadband US attenuation (nBUA), speed of sound (SOS), broadband US backscatter (BUB) and integrated reflection coefficient (IRC) were determined for each sample. The samples were spatially scanned with five pairs of US transducers covering a frequency range of 0.2-6.7 MHz. Furthermore, mechanical properties and density of the same samples were determined. At all frequencies, SOS, BUB and IRC showed statistically significant linear correlations with the mechanical properties or density of human trabecular bone (0.51 < r < 0.82, 0.54 < r < 0.81 and 0.70 < r < 0.85, respectively). In contrast to SOS, IRC and BUB, nBUA showed statistically significant correlations with mechanical parameters or density at the centre frequency of 1 MHz only. Our results suggest that frequencies up to 5 MHz can be useful in QUS analyses for the prediction of bone mechanical properties and density. Since the use of higher frequencies provides better axial and spatial resolution, improved structural analyses may be possible. While extensive attenuation of high frequencies in trabecular bone limits the clinically feasible frequency range, selection of optimal frequency range for in vivo QUS application should be carefully considered.

Algorithms↗

PubMed2005

Lordotic vertebrae in sea bass (Dicentrarchus labrax L.) are adapted to increased loads.

Lordosis in fish is an abnormal ventral curvature of the vertebral column, accompanied by abnormal calcification of the afflicted vertebrae. Incidences of lordosis are a major problem in aquaculture and often correlate with increased swimming activity. To understand the biomechanical causes and consequences of lordosis, we mapped the morphological changes that occur in the vertebrae of European sea bass during their development from larva to juvenile. Our micro-CT analysis of lordotic and non-lordotic vertebrae revealed significant differences in their micro-architecture. Lordotic vertebrae have a larger bone volume, flattened dorsal zygapophyses and extra lateral ridges. They also have a larger second moment of area (both lateral and dorso-ventral) than non-lordotic vertebrae. This morphology suggests lordotic vertebrae to be adapted to an increased bending moment, caused by the axial musculature during increased swimming activity. We hypothesize the increase in swimming activity to have a two-fold effect in animals that become lordotic. The first effect is buckling failure of the axial skeleton due to an increased compressive load. The second effect is extra bone deposition as an adaptive response of the vertebrae at the cellular level, caused by an increased strain and strain rate in these vertebrae. Lordosis thus comprises both a buckling failure of the vertebral column and a molecular response that adapts the lordotic vertebrae to a new loading regime.

Adaptation, Physiological↗

PubMed2004

Improved cartilage integration and interfacial strength after enzymatic treatment in a cartilage transplantation model.

The objective of the present study was to investigate whether treatment of articular cartilage with hyaluronidase and collagenase enhances histological and mechanical integration of a cartilage graft into a defect. Discs of 3 mm diameter were taken from 8-mm diameter bovine cartilage explants. Both discs and annulus were either treated for 24 hours with 0.1% hyaluronidase followed by 24 hours with 10 U/ml collagenase or left untreated (controls). Discs and annulus were reassembled and implanted subcutaneously in nude mice for 5 weeks. Integration of disc with surrounding cartilage was assessed histologically and tested biomechanically by performing a push-out test. After 5 weeks a significant increase in viable cell counts was seen in wound edges of the enzyme-treated group as compared with controls. Furthermore, matrix integration (expressed as a percentage of the total interface length that was connected; mean +/- standard error) was 83 +/- 15% in the treated samples versus 44 +/- 40% in the untreated controls. In the enzyme-treated group only, picro-Sirius Red staining revealed collagen crossing the interface perpendicular to the wound surface. Immunohistochemical analyses demonstrated that the interface tissue contained cartilage-specific collagen type II. Collagen type I was found only in a small region of fibrous tissue at the level of the superficial layer, and collagen type III was completely absent in both groups. A significant difference in interfacial strength was found using the push-out test: 1.32 +/- 0.15 MPa in the enzyme-treated group versus 0.84 +/- 0.14 MPa in the untreated controls. The study shows that enzyme treatment of cartilage wounds increases histological integration and improves biomechanical bonding strength. Enzymatic treatment may represent a promising addition to current techniques for articular cartilage repair.

Animals↗

PubMed2004

An improved segmentation method for in vivo microCT imaging.

UNLABELLED: Image segmentation methods for microCT can influence the accuracy of bone morphometry calculations. A new automated segmentation method is introduced, and its performance is compared with standard segmentation methods. The new method can improve the results of in vivo microCT, where the need to keep radiation dose low limits scan quality. INTRODUCTION: An important topic for microCT analysis of bone samples is the segmentation of the original reconstructed grayscale data sets to separate bone from non-bone. Problems like noise, resolution limitations, and beam-hardening make this a nontrivial issue. Inappropriate segmentation methods will reduce the potential power of microCT and may introduce bias in the architectural measurements, in particular, when new in vivo microCT with its inherent limitations in scan quality is used. Here we introduce a new segmentation method using local thresholds and compare its performance to standard global segmentation methods. MATERIAL AND METHODS: The local threshold method was validated by comparing the result of the segmentation with histology. Furthermore, the effect of choosing this new method versus standard segmentation methods using global threshold values was investigated by studying the sensitivity of these methods to signal to noise ratio and resolution. RESULTS: Using the new method on high-quality scans yielded accurate results and virtually no differences between histology and the segmented data sets could be observed. When prior knowledge about the volume fraction of the bone was available the global threshold also resulted in appropriate results. Degrading the scan quality had only minor effects on the performance of the new segmentation method. Although global segmentation methods were not sensitive to noise, it was not possible to segment both lower mineralized thin trabeculae and the higher mineralized cortex correctly with the same threshold value. CONCLUSION: At high resolutions, both the new local and conventional global segmentation methods gave near exact representations of the bone structure. When scanned samples are not homogenous (e.g., thick cortices and thin trabeculae) and when resolution is relatively low, the local segmentation method outperforms global methods. It maximizes the potential of in vivo microCT by giving good structural representation without the need to use longer scanning times that would increase absorption of harmful X-ray radiation by the living tissue.

Algorithms↗

PubMed2003

The dependence of the elastic properties of osteoporotic cancellous bone on volume fraction and fabric.

Osteoporosis is a progressive systemic skeletal condition characterized by low bone mass and microarchitectural deterioration, with a consequent increase in susceptibility to fracture. Hence, osteoporosis would be best diagnosed by in vivo measurements of bone strength. As this is not clinically feasible, our goal is to estimate bone strength through the assessment of elastic properties, which are highly correlated to strength. Previously established relations between morphological parameters (volume fraction and fabric) and elastic constants could be applied to estimate cancellous bone stiffness in vivo. However, these relations were determined for normal cancellous bone. Cancellous bone from osteoporotic patients may require different relations. In this study we set out to answer two questions. First, can the elastic properties of osteoporotic cancellous bone be estimated from morphological parameters? Second, do the relations between morphological parameters and elastic constants, determined for normal bone, apply to osteoporotic bone as well? To answer these questions we used cancellous bone cubes from femoral heads of patients with (n=26) and without (n=32) hip fractures. The elastic properties of the cubes were determined using micro-finite element analysis, assuming equal tissue moduli for all specimens. The morphological parameters were determined using microcomputed tomography. Our results showed that, for equal tissue properties, the elastic properties of cancellous bone from fracture patients could indeed be estimated from morphological parameters. The morphology-based relations used to estimate the elastic properties of cancellous bone are not different for women with or without fractures.

Bone and Bones↗

PubMed2003

Living cells in 1 of 2 frozen femoral heads.

Allogeneic, frozen bone is now the most commonly grafted tissue (Norman-Taylor and Villar 1997). Tissue banks collect bone material according to protocols developed with the aim of maintaining osseoinductive properties of grafts as well as preventing transmission of viral or bacterial diseases (Standards from the American Association of Tissue Banks (AATB) or from the European Association for Musculo-skeletal Transplanting (EAMST)). Standard procedures include cryopreservation of tissue at -80 degrees C, which is generally considered to devitalize the bone by killing all cells present, resulting in reduced immunogenicity of the graft. The osseoinductive properties of frozen, allogeneic bone grafts have therefore mainly been attributed to the dead bone matrix, that may provide osteoblast-stimulating growth factors and other essential proteins, and/or an osteoclast substrate to direct bone remodeling (Aspenberg et al. 1996, Kingsmill et al. 1999). Recently however, it was suggested that some cells in bone biopsies may survive standard bone bank freezing procotols. It is unclear whether vital cells are present in other bone banks and whether these cells can contribute to the clinical outcome of frozen allogeneic bone grafting. In this report, we show that frozen bone biopsies, obtained from the Erasmus Medical Center bone bank may contain living cells that can be cultured in vitro. These cultured cells were found to originate from the donor by genotyping.

Bone Banks↗