RESUMEN
Alterations in resorption cavities and bone remodeling events during anti-resorptive treatment are believed to contribute to reductions in fracture risk. Here, we examine changes in the size of individual remodeling events associated with treatment with a selective estrogen receptor modulator (raloxifene) or a bisphosphonate (risedronate). Adult female rats (6months of age) were submitted to ovariectomy (n=17) or sham surgery (SHAM, n=5). One month after surgery, the ovariectomized animals were separated into three groups: untreated (OVX, n=5), raloxifene treated (OVX+Ral, n=6) and risedronate treated (OVX+Ris, n=6). At 10months of age, the lumbar vertebrae were submitted to three-dimensional dynamic bone histomorphometry to examine the size (depth, breadth and volume) of individual resorption cavities and formation events. Maximum resorption cavity depth did not differ between the SHAM (23.66±1.87µm, mean±SD) and OVX (22.88±3.69µm) groups but was smaller in the OVX+Ral (14.96±2.30µm) and OVX+Ris (14.94±2.70µm) groups (p<0.01). Anti-resorptive treatment was associated with reductions in the surface area of resorption cavities and the volume occupied by each resorption cavity (p<0.01 each). The surface area and volume of individual formation events (double-labeled events) in the OVX+Ris group were reduced as compared to other groups (p<0.02). Raloxifene treated animals showed similar amounts of bone remodeling (ES/BS and dLS/BS) compared to sham-operated controls but smaller cavity size (depth, breadth and volume). The current study shows that anti-resorptive agents influence the size of resorption cavities and individual remodeling events and that the effect of anti-resorptives on individual remodeling events may not always be directly related to the degree of suppression of bone remodeling.
Asunto(s)
Conservadores de la Densidad Ósea/uso terapéutico , Remodelación Ósea/efectos de los fármacos , Resorción Ósea/tratamiento farmacológico , Ácido Etidrónico/análogos & derivados , Clorhidrato de Raloxifeno/uso terapéutico , Animales , Ácido Etidrónico/uso terapéutico , Femenino , Ovariectomía , Ratas , Ratas Sprague-Dawley , Ácido RisedrónicoRESUMEN
The number and size of resorption cavities in cancellous bone are believed to influence rates of bone loss, local tissue stress and strain and potentially whole bone strength. Traditional two-dimensional approaches to measuring resorption cavities in cancellous bone report the percent of the bone surface covered by cavities or osteoclasts, but cannot measure cavity number or size. Here we use three-dimensional imaging (voxel size 0.7×0.7×5.0 µm) to characterize resorption cavity location, number and size in human vertebral cancellous bone from nine elderly donors (7 male, 2 female, ages 47-80 years). Cavities were 30.10 ± 8.56 µm in maximum depth, 80.60 ± 22.23∗10(3) µm(2) in surface area and 614.16 ± 311.93∗10(3) µm(3) in volume (mean ± SD). The average number of cavities per unit tissue volume (N.Cv/TV) was 1.25 ± 0.77 mm(-3). The ratio of maximum cavity depth to local trabecular thickness was 30.46 ± 7.03% and maximum cavity depth was greater on thicker trabeculae (p<0.05, r(2)=0.14). Half of the resorption cavities were located entirely on nodes (the intersection of two or more trabeculae) within the trabecular structure. Cavities that were not entirely on nodes were predominately on plate-like trabeculae oriented in the cranial-caudal (longitudinal) direction. Cavities on plate-like trabeculae were larger in maximum cavity depth, cavity surface area and cavity volume than cavities on rod-like trabeculae (p<0.05). We conclude from these findings that cavity size and location are related to local trabecular microarchitecture.
Asunto(s)
Remodelación Ósea/fisiología , Resorción Ósea/fisiopatología , Anciano , Anciano de 80 o más Años , Fenómenos Biomecánicos , Femenino , Humanos , Imagenología Tridimensional , Vértebras Lumbares/fisiología , Vértebras Lumbares/fisiopatología , Masculino , Persona de Mediana EdadRESUMEN
The amount of bone turnover in the body has been implicated as a factor that can influence fracture risk and bone strength. Here we test the idea that remodeling cavities promote local tissue failure by determining if microscopic tissue damage (microdamage) caused by controlled loading in vitro is more likely to form near resorption cavities. Specimens of human vertebral cancellous bone (L4, 7 male and 2 female, age 70±10, mean±SD) were loaded in compression to the yield point, stained for microscopic tissue damage and submitted to three-dimensional fluorescent imaging using serial milling (image voxel size 0.7×0.7×5.0 µm). We found the resulting damage volume per bone volume (DV/BV) was correlated with percent eroded surface (p<0.01, r(2)=0.65), demonstrating that whole specimen measures of resorption cavities and microdamage are related. Locations of microdamage were more than two times as likely to have a neighboring resorption cavity than randomly selected sites without microdamage (relative risk 2.39, 95% confidence interval of relative risk: 2.09-2.73), indicating a spatial association between resorption cavities and microdamage at the local level. Individual microdamage sites were 48,700 (40,100; 62,700) µm(3) in size (median, 25th and 75th percentiles). That microdamage was associated with resorption cavities when measured at the whole specimen level as well as at the local level provides strong evidence that resorption cavities play a role in mechanical failure processes of cancellous bone and therefore have the potential to influence resistance to clinical fracture.
Asunto(s)
Fracturas por Compresión/fisiopatología , Traumatismos Vertebrales/fisiopatología , Columna Vertebral/fisiopatología , Anciano , Anciano de 80 o más Años , Fenómenos Biomecánicos , Remodelación Ósea/fisiología , Resorción Ósea/patología , Resorción Ósea/fisiopatología , Fuerza Compresiva , Femenino , Fracturas por Compresión/patología , Humanos , Imagenología Tridimensional , Técnicas In Vitro , Masculino , Persona de Mediana Edad , Traumatismos Vertebrales/patología , Columna Vertebral/patología , Estrés MecánicoRESUMEN
Serial block face imaging is a microscopy technique in which the top of a specimen is cut or ground away and a mosaic of images is collected of the newly revealed cross-section. Images collected from each slice are then digitally stacked to achieve 3D images. The development of fully automated image acquisition devices has made serial block face imaging more attractive by greatly reducing labour requirements. The technique is particularly attractive for studies of biological activity within cancellous bone as it has the capability of achieving direct, automated measures of biological and morphological traits and their associations with one another. When used with fluorescence microscopy, serial block face imaging has the potential to achieve 3D images of tissue as well as fluorescent markers of biological activity. Epifluorescence-based serial block face imaging presents a number of unique challenges for visualizing bone specimens due to noise generated by sub-surface signal and local variations in tissue autofluorescence. Here we present techniques for processing serial block face images of trabecular bone using a combination of non-uniform illumination correction, precise tiling of the mosaic in each cross-section, cross-section alignment for vertical stacking, removal of sub-surface signal and segmentation. The resulting techniques allow examination of bone surface texture that will enable 3D quantitative measures of biological processes in cancellous bone biopsies.