Biomedical subjects

S A Goldstein

Publications and source records attributed to S A Goldstein.

At least 19 recordsLinked to original sources

PubMed2000

Proton block and voltage gating are potassium-dependent in the cardiac leak channel Kcnk3.

Potassium leak conductances were recently revealed to exist as independent molecular entities. Here, the genomic structure, cardiac localization, and biophysical properties of a murine example are considered. Kcnk3 subunits have two pore-forming P domains and unique functional attributes. At steady state, Kcnk3 channels behave like open, potassium-selective, transmembrane holes that are inhibited by physiological levels of proton. With voltage steps, Kcnk3 channels open and close in two phases, one appears to be immediate and one is time-dependent (tau = approximately 5 ms). Both proton block and gating are potassium-sensitive; this produces an anomalous increase in outward flux as external potassium levels rise because of decreased proton block. Single Kcnk3 channels open across the physiological voltage range; hence they are "leak" conductances; however, they open only briefly and rarely even after exposure to agents that activate other potassium channels.

Amino Acid Sequence↗

PubMed2000

Effect of compressive loading on chondrocyte differentiation in agarose cultures of chick limb-bud cells.

It is well established that mechanical loading is important to homeostasis of cartilage tissue, and growing evidence suggests that it influences cartilage differentiation as well. Whereas the effect of mechanical forces on chondrocyte biosynthesis and gene expression has been vigorously investigated, the effect of the mechanical environment on chondrocyte differentiation has received little attention. The long-term objective of this research is to investigate the regulatory role of mechanical loading in cell differentiation. The goal of this study was to determine if mechanical compression could modulate chondrocyte differentiation in vitro. Stage 23/24 chick limb-bud cells, embedded in agarose gel, were subjected to either static (constant 4.5-kPa stress) or cyclic (9.0-kPa peak stress at 0.33 Hz) loading in unconfined compression during the initial phase of commitment to a phenotypic lineage. Compared with nonloaded controls, cyclic compressive loading roughly doubled the number of cartilage nodules and the amount of sulfate incorporation on day 8, whereas static compression had little effect on these two measures. Neither compression protocol significantly affected overall cell viability or the proliferation of cells within nodules. Since limb-bud mesenchymal cells were seeded directly into agarose, an assessment of cartilage nodules in the agarose reflects the proportion of the original cells that had given rise to chondrocytes. Thus, the results indicate that about twice as many mesenchymal cells were induced to enter the chondrogenic pathway by cyclic mechanical compression. The coincidence of the increase in sulfate incorporation and nodule density indicates that the primary effect of mechanical compression on mesenchymal cells was on cellular differentiation and not on their subsequent metabolism. Further studies are needed to identify the primary chondrogenic signal associated with cyclic compressive loading and to determine the mechanism by which it influences commitment to or progression through the chondrogenic lineage, or both.

Animals↗

PubMed2000

Vertebral trabecular bone microscopic tissue elastic modulus and hardness do not change in ovariectomized rats.

Ovariectomized rats have been used extensively and have received substantial acceptance as animal models for postmenopausal osteoporosis. However, little is known about the microscopic tissue properties of rat vertebral bone, especially during osteoporosis caused by estrogen depletion. This study applied a new nanoindentation technique to quantify the microscopic mechanical properties of vertebral trabecular bone tissue in ovariectomized rats. Seventeen-week-old Sprague-Dawley rats underwent an ovariectomy. After death at 37 weeks, the fraction of the trabecular bone area of the lumbar vertebrae (L4) was measured with scanning electron microscopy and the elastic modulus and hardness were determined with the nanoindentation technique. The bone area fraction was reduced significantly after the ovariectomy. However, the elastic modulus and hardness did not change significantly at the microscopic level. The results indicate that estrogen-dependent osteoporosis in rats manifests in a loss of bone mass whereas the elastic and hardness properties of the surviving bone tissue remain relatively unchanged.

Animals↗

PubMed2000

Heterogeneity of bone lamellar-level elastic moduli.

Advances in our ability to assess fracture risk, predict implant success, and evaluate new therapies for bone metabolic and remodeling disorders depend on our understanding of anatomically specific measures of local tissue mechanical properties near and surrounding bone cells. Using nanoindentation, we have quantified elastic modulus and hardness of human lamellar bone tissue as a function of tissue microstructures and anatomic location. Cortical and trabecular bone specimens were obtained from the femoral neck and diaphysis, distal radius, and fifth lumbar vertebra of ten male subjects (aged 40-85 years). Tissue was tested under moist conditions at room temperature to a maximum depth of 500 nm with a loading rate of 10 nm/sec. Diaphyseal tissue was found to have greater elastic modulus and hardness than metaphyseal tissues for all microstructures, whereas interstitial elastic modulus and hardness did not differ significantly between metaphyses. Trabecular bone varied across locations, with the femoral neck having greater lamellar-level elastic modulus and hardness than the distal radius, which had greater properties than the fifth lumbar vertebra. Osteonal, interstitial, and primary lamellar tissues of compact bone had greater elastic moduli and hardnesses than trabecular bone when comparing within an anatomic location. Only femoral neck interstitial tissue had a greater elastic modulus than its osteonal counterpart, which suggests that microstructural distinctions can vary with anatomical location and may reflect differences in the average tissue age of cortical bone or mineral and collagen organization.

Adult↗

PubMed2000

Immediate effect of Biosense guided percutaneous direct myocardial revascularization with holmium:yttrium aluminium garnet laser on myocardial contractility assessed by transesophageal echocardiography.

OBJECTIVE: To evaluate immediate changes in left ventricular wall motion in patients treated using Biosense direct myocardial revascularization laser system. METHODS: Regional wall motion in 10 patients undergoing catheter-based direct myocardial revascularization using a holmium:yttrium aluminium garnet laser was assessed by transesophageal echocardiography before and immediately after the procedure. RESULTS: Mild deterioration in wall-motion score occurred rarely for only three of 160 (1.9%) segments and did not induce clinical heart failure. CONCLUSION: With the current catheter-based laser myocardial revascularization strategy, mild deterioration in wall motion of treated segments was rarely observed and did not effect overall left ventricular function or induce clinical congestive heart failure.

Aged↗

PubMed2000

Variations in three-dimensional cancellous bone architecture of the proximal femur in female hip fractures and in controls.

Cubes of cancellous bone were obtained from proximal femora of women with hip fractures (n = 26) and from female cadaveric controls (n = 32) to compare architecture and mechanics between groups. Specimens were scanned on a microcomputed tomography system. Stereologic algorithms and model-based estimates were applied to the data to characterize the three-dimensional cancellous microstructure. Cubes were mechanically tested to failure to obtain mechanical properties. Specimens from control subjects had significantly higher bone volume fraction, trabecular number, and connectivity than specimens from patients with hip fractures; no difference in trabecular thickness was observed between groups. Both maximum modulus and ultimate stress were significantly higher in the control than in the fracture group, consistent with the higher bone volume found in the control group. No statistical differences in any of these architectural or mechanical variables were found when groups were matched for bone volume. Specimens from both patients with hip fractures and controls demonstrated strong relationships between trabecular number and bone volume fraction that were statistically equivalent, suggesting that for a given bone mass, both groups have the same overall number of trabeculae. However, there was an architectural difference between fracture and control groups in terms of the three-dimensional spatial arrangement of trabeculae. Fracture specimens had a significantly more anisotropic (oriented) structure than control specimens, with proportionately fewer trabecular elements transverse to the primary load axis, even when matched for bone volume. Relationships between mechanical and architectural parameters were significantly different between groups, suggesting that fracture and control groups have different structure-mechanics relationships, which we hypothesize may be a consequence of the altered three-dimensional structure between groups.

Algorithms↗

PubMed2000

Increased marrow-derived osteoprogenitor cells and endosteal bone formation in mice lacking thrombospondin 2.

The phenotype of thrombospondin 2 (TSP2)-null mice includes abnormalities in collagen fibrils and increases in ligamentous laxity, vascular density, and bleeding time. In this study, analyses by computerized tomography (CT) revealed that cortical density was increased in long bones of TSP2-null mice. Histomorphometric analysis showed that the mid-diaphyseal endosteal bone formation rate (BFR) of TSP2-null mice was increased in comparison with that of wild-type (WT) animals. Although microgeometric analysis showed that periosteal and endosteal radii were reduced, the mechanical properties of femurs from TSP2-null mice were not significantly different from those of controls, presumably because of the concomitant increase in endosteal bone mass. Bone loss in ovariectomized mice was equivalent for WT and mutant mice, a finding that indicates that TSP2-null animals are capable of normal bone resorption. To further explore the cellular basis for the increased endosteal BFR in TSP2-null mice, marrow stromal cells (MSCs) were isolated and examined in vitro. These cells were found to be present in increased numbers in a colony forming unit (CFU) assay and showed an increased rate of proliferation in vitro. We conclude that TSP2 regulates the proliferation of osteoblast progenitors, directly or indirectly, and that in its absence endosteal bone formation is increased.

Animals↗

PubMed2000

Increased distraction rates influence precursor tissue composition without affecting bone regeneration.

The effect of increased distraction rate on bony tissue differentiation was studied using a paired bilateral model of rat femur lengthening. After a 6-day latency period, one randomly selected femur for each rat was distracted at 0.5 mm/day (normal rate) for 12 days, and the contralateral femur was distracted at 1.5 mm/day (increased rate) for 4 days. Femoral lengthening for each side was 6.0 mm, leaving the increased rate leg with an extra 8 days of consolidation compared with the normal rate limb. Group I rats (n = 9) were killed at day 18 postsurgery and analyzed for cartilage tissue composition and distribution. Group II rats (n = 7) were killed on day 36 postsurgery and analyzed by three-dimensional microcomputed tomography (MCT) for changes in new bone volume. Digital color analysis of slides stained with type II collagen antibody showed increases in cartilaginous tissue formation on the increased rate side (1.51 mm2 vs. 0.83 mm2; p = 0.10). No differences in new bone volume were detected between increased rate limbs and their contralateral controls (46.13 mm3 vs. 42.69 mm3; p = 0.63). These findings suggest that intermediate distraction rates may influence precursor tissue composition without affecting the final amount of new bone formed. Because damage to the tissue was not detected at either time point, these changes in chondrogenesis may reflect sensitivity of the pluripotential gap tissue to tension accumulation during lengthening. Future work with this in vivo model is focused on improving our understanding of the mechanisms behind this strain sensitivity.

Animals↗

PubMed2000

An in vivo model for investigations of mechanical signal transduction in trabecular bone.

The premise that bone cells are able to perceive and respond to mechanical forces is well accepted. This article describes the use of an in vivo hydraulic bone chamber for investigations of mechanical signal transduction. The servohydraulic loading mechanism was activated to apply a controlled compressive load to the woven trabecular bone that formed in one chamber, while the contralateral chamber served as an unloaded control. Specimens were harvested at a series of postload time points, and the cellular response to loading was evaluated by cytochemical, histomorphometric, and Northern blot analysis. A repetitive daily load stimulus elicited osteoblast biosynthetic activity characterized by an initial increase in type I procollagen by day 3 and a subsequent rise in alkaline phosphatase (ALP) activity after the sixth daily load episode. Application of a single load episode induced a biphasic pattern of c-fos and zif-268 gene expression with up-regulation at 30 minutes, down-regulation at 12 h, and up-regulation 24 h after the mechanical stimulus. The results show that a synchronized pattern of bone cell activity and gene expression occurs in response to controlled mechanical stimulation and that candidate load-responsive molecular mediators can be evaluated easily by this model.

Alkaline Phosphatase↗

PubMed2000

A new technique for the quantitative analysis of cranial suture biology.

OBJECTIVE: Our objective was to assess the ability of the microcomputed tomography scanner to correctly image normal and synostosed cranial sutures at the ultrastructural level. DESIGN AND METHODS: Two specimens of coronal sutures were collected from operative specimens. After appropriate preparation, histological sections were obtained and stained with toluene blue for evaluation. Representative histological sections were compared to microcomputed tomography slices. RESULTS AND CONCLUSIONS: With microcomputed tomography, we successfully imaged one normal and one synostosed human coronal suture and performed a quantitative analysis of these specimens. Microcomputed tomography scanning was found to be a highly accurate imaging device for the evaluation of cranial suture development. Microcomputed tomography offers three-dimensional imaging at the microscopic level and allows for rapid quantitative analysis of bone architecture, including several measurements unavailable through histologic analysis. We believe that microcomputed tomography can play an important role in imaging and in the quantitative analysis of the stereology of bone microarchitecture. Among its advantages, microcomputed tomography is able to image many more slices than are obtainable through histology, and the method is not prone to human error. Microcomputed tomography slices are generated without destruction of the specimen and without loss or corruption of reproducible data. Structure-oriented slices from microcomputed tomography together with cellular-oriented sections from histology are complementary in the overall quantitative analysis of cranial sutures.

Child, Preschool↗

PubMed2000

Effects of long-term administration of vitamin D3 analogs to mice.

This study explores the effects of chronic administration of vitamin D(3) compounds on several biological functions in mice. Knowledge of long-term tolerability of vitamin D(3) analogs may be of interest in view of their potential clinical utility in the management of various pathologies such as malignancies, immunological disorders and bone diseases. Four unique vitamin D(3) analogs (code names, compounds V, EO, LH and LA) and 1,25-dihydroxyvitamin D(3) (1, 25(OH)(2)D(3)) were administered i.p. for 55 weeks to Balb/c mice. Each analog had previously been shown to have potent in vitro activities. After 55 weeks of administration, the mice had a profound decrease in their serum levels of interleukin-2 (IL-2). Likewise, several analogs depressed serum immunoglobulin G concentrations (compounds LH and LA), but levels of blood lymphocytes and splenic lymphocyte subsets (CD4, CD8 and CD19) were not remarkably depressed. The percent of committed myeloid hematopoietic stem cells was 4- to 5-fold elevated in the bone marrow of the mice that received analogs LH and V; nevertheless, their peripheral blood white and red cell counts and platelets were not significantly different in any of the groups. The mice that received 1,25(OH)(2)D(3) had a decrease in bone quantity and quality with a decrease in cross-sectional area and cortical thickness, and a 50% reduction in both stiffness and failure load compared with the control group. In contrast, the cohort that received a fluorinated analog (compound EO) developed bones with significantly larger cross-sectional area and cortical thickness as well as stronger mechanical properties compared with the control group. At the conclusion of the study, body weights were significantly decreased in all experimental mice. Their blood chemistries were normal. Extensive gross and microscopic autopsy analyses of the mice at the conclusion of the study were normal, including those of their kidneys. In conclusion, the vitamin D(3) analogs were fairly well tolerated. They did suppress immunity as measured by serum IL-2 and may provide a means to depress the immune response after organ transplantation and for autoimmune diseases. Use of these analogs prevented the detrimental effects of vitamin D(3) administration on mechanical and geometric properties of bone, while one analog (compound EO) actually enhanced bone properties. These results suggest that long-term clinical trials with the analogs are feasible.

Animals↗

PubMed1999

A molecular target for viral killer toxin: TOK1 potassium channels.

Killer strains of S. cerevisiae harbor double-stranded RNA viruses and secrete protein toxins that kill virus-free cells. The K1 killer toxin acts on sensitive yeast cells to perturb potassium homeostasis and cause cell death. Here, the toxin is shown to activate the plasma membrane potassium channel of S. cerevisiae, TOK1. Genetic deletion of TOK1 confers toxin resistance; overexpression increases susceptibility. Cells expressing TOK1 exhibit toxin-induced potassium flux; those without the gene do not. K1 toxin acts in the absence of other viral or yeast products: toxin synthesized from a cDNA increases open probability of single TOK1 channels (via reversible destabilization of closed states) whether channels are studied in yeast cells or X. laevis oocytes.

Animals↗

PubMed1999

MiRP1 forms IKr potassium channels with HERG and is associated with cardiac arrhythmia.

A novel potassium channel gene has been cloned, characterized, and associated with cardiac arrhythmia. The gene encodes MinK-related peptide 1 (MiRP1), a small integral membrane subunit that assembles with HERG, a pore-forming protein, to alter its function. Unlike channels formed only with HERG, mixed complexes resemble native cardiac IKr channels in their gating, unitary conductance, regulation by potassium, and distinctive biphasic inhibition by the class III antiarrhythmic E-4031. Three missense mutations associated with long QT syndrome and ventricular fibrillation are identified in the gene for MiRP1. Mutants form channels that open slowly and close rapidly, thereby diminishing potassium currents. One variant, associated with clarithromycin-induced arrhythmia, increases channel blockade by the antibiotic. A mechanism for acquired arrhythmia is revealed: genetically based reduction in potassium currents that remains clinically silent until combined with additional stressors.

Amino Acid Sequence↗

PubMed1999

Marrow-derived progenitor cell injections enhance new bone formation during distraction.

Bilateral femoral distraction was performed in rats to investigate whether injections of marrow-derived mesenchymal progenitor cells could be used to facilitate new bone formation. The cells were isolated from whole marrow of 2-6-month-old Sprague-Dawley rats. One-year-old recipient Sprague-Dawley rats were divided into five experimental groups. Rats in groups I, II, and III received injections of mesenchymal progenitor cells on days 6 (beginning), 12 (middle), and 18 (end of distraction) after surgery, respectively. Those in group IV received injections of serum and carrier gel alone, and those in group V received no injections. Distraction zones were harvested at 36 days and analyzed for new bone volume within the distraction gap by three-dimensional microcomputed tomography. Significant increases in new bone volume were observed for femora injected with marrow-derived progenitor cells compared with contralateral femora and controls (no injection). The timing of the cell injections appeared to have no effect on the experimental outcome. Histologic analyses demonstrated active formation of new trabecular bone with marked osteoblastic activity and osteoid production. No qualitative differences in histologic appearances of new bone among rats in any of the five groups were seen. The results of in vitro lysis assays indicated that donor and recipient rats were not completely syngenic, leaving some doubt as to the reasons for observed increases in new bone formation. Future work will focus on attempting to repeat these experiments in a fully syngenic rat model. This rat distraction model can be used to explore the molecular and cellular behavior of these progenitor cells in a clinically relevant in vivo environment.

Animals↗

PubMed1999

Femoral strength is better predicted by finite element models than QCT and DXA.

Clinicians and patients would benefit if accurate methods of predicting and monitoring bone strength in-vivo were available. A group of 51 human femurs (age range 21-93; 23 females, 28 males) were evaluated for bone density and geometry using quantitative computed tomography (QCT) and dual energy X-ray absorptiometry (DXA). Regional bone density and dimensions obtained from QCT and DXA were used to develop statistical models to predict femoral strength ex vivo. The QCT data also formed the basis of a three-dimensional finite element (FE) models to predict structural stiffness. The femurs were separated into two groups; a model training set (n = 25) was used to develop statistical models to predict ultimate load, and a test set (n = 26) was used to validate these models. The main goal of this study was to test the ability of DXA, QCT and FE techniques to predict fracture load non-invasively, in a simple load configuration which produces predominantly femoral neck fractures. The load configuration simulated the single stance phase portion of normal gait; in 87% of the specimens, clinical appearing sub-capital fractures were produced. The training/test study design provided a tool to validate that the predictive models were reliable when used on specimens with "unknown" strength characteristics. The FE method explained at least 20% more of the variance in strength than the DXA models. Planned refinements of the FE technique are expected to further improve these results. Three-dimensional FE models are a promising method for predicting fracture load, and may be useful in monitoring strength changes in vivo.

Absorptiometry, Photon↗

PubMed1999

Elastic modulus and hardness of cortical and trabecular bone lamellae measured by nanoindentation in the human femur.

The mechanical properties of bone tissue are determined by composition as well as structural, microstructural and nanostructural organization. The aim of this study was to quantify the elastic properties of bone at the lamellar level and compare these properties among osteonal, interstitial and trabecular microstructures from the diaphysis and the neck of the human femur. A nanoindentation technique with a custom irrigation system was used for simultaneously measuring force and displacement of a diamond tip pressed 500 nm into the moist bone tissue. An isotropic elastic modulus was calculated from the unloading curve with an assumed Poisson ratio of 0.3, while hardness was defined as the maximal force divided by the corresponding contact area. The elastic moduli ranged from 6.9 +/- 4.3 GPa in trabecular tissue from the femoral neck of a 74 yr old female up to 25.0 +/- 4.3 GPa in interstitial tissue from the diaphyseal cortex of a 69 yr old female. The mean elastic modulus was found to be significantly influenced by the type of lamella (p < 10(-6)) and by donor (p < 10(-6)). The interaction between the type of lamella and the donor was also highly significant (p < 10(-6)). Hardness followed a similar distribution as elastic modulus among types of lamellae and donor, but with lower statistical contrast. It is concluded that the nanostructure of bone tissue must differ substantially among lamellar types, anatomical sites and individuals and suggests that tissue heterogeneity is of potential importance in bone fragility and adaptation.

Aged↗

PubMed1999

Perspectives on tissue engineering of bone.

The normal repair and regeneration of bone occurs through an ordered and regulated sequence of cellular events. Successful replacement of bone through tissue engineering likely will be dependent on the recapitulation of this cascade of events. This report presents some of the principles to be considered in the design of engineered bone constructs. The role of cells, a supporting matrix, and endogenous or exogenous biologic or mechanical factors are introduced. The authors' experience with a gene therapeutic approach to bone regeneration is presented as one example of using the principles discussed to promote reproducible bone formation.

Animals↗

PubMed1999

Viscoelastic characterization of mesenchymal gap tissue and consequences for tension accumulation during distraction.

Nonlinear viscoelastic analysis was used to characterize the time-dependent behavior of mesenchymal gap tissue generated during distraction osteogenesis. Six (n = 6) lengthened tibiae were harvested from New Zealand white rabbits at 18 days. This gap tissue was subjected to a series of step displacement tests of increasing magnitude, and force relaxation behavior was monitored. Isochrones in stress-strain space were fit to odd cubic functions of strain. An analytic expression, linear in both e and e3, was developed to predict stress accumulation within the gap tissue as a function of time during distraction. Stress relaxation functions were described well by two-term Prony series. The two time constants determined from mechanical testing results were consistent, suggesting the presence of two fundamental physiologic relaxation processes. Gap tissue stresses were predicted to rise considerably during early stages of lengthening when distraction magnitudes exceeded the clinical norm of 0.25 mm. These differences in tension accumulation were less pronounced by the time lengthening was completed. Specifically, these results may in part explain clinical observations of decreased bone regeneration and altered tissue proliferation and differentiation at higher distraction rates. More generally, this work provides a framework for the rigorous characterization of the viscoelastic properties of biologic tissues ordinarily exposed to step strains.

Animals↗