Detecting hip osteoarthritis on clinical CT: a deep learning application based on 2-D summation images derived from CT.

We developed and compared deep learning models to detect hip osteoarthritis on clinical CT. The CT-based summation images, CT-AP, that resemble X-ray radiographs can detect radiographic hip osteoarthritis and in the absence of large training data, a reliable deep learning model can be optimized by combining CT-AP and X-ray images. INTRODUCTION: In this study, we aimed to investigate the applicability of deep learning (DL) to assess radiographic hip osteoarthritis (rHOA) on computed tomography (CT). METHODS: The study data consisted of 94 abdominopelvic clinical CTs and 5659 hip X-ray images collected from Cohort Hip and Cohort Knee (CHECK). The CT slices were sequentially summed to create radiograph-like 2-D images named CT-AP. X-ray and CT-AP images were classified as rHOA if they had osteoarthritic changes corresponding to Kellgren-Lawrence grade 2 or higher. The study data was split into 55% training, 30% validation, and 15% test sets. A pretrained ResNet18 was optimized for a classification task of rHOA vs. no-rHOA. Five models were trained using (1) X-rays, (2) downsampled X-rays, (3) combination of CT-AP and X-ray images, (4) combination of CT-AP and downsampled X-ray images, and (5) CT-AP images. RESULTS: Amongst the five models, Model-3 and Model-5 performed best in detecting rHOA from the CT-AP images. Model-3 detected rHOA on the test set of CT-AP images with a balanced accuracy of 82.2% and was able to discriminate rHOA from no-rHOA with an area under the receiver operating characteristic curve (ROC AUC) of 0.93 [0.75-0.99]. Model-5 detected rHOA on the test set at a balanced accuracy of 82.2% and classified rHOA from no-rHOA with an ROC AUC of 0.89 [0.67-0.97]. CONCLUSION: CT-based summation images that resemble radiographs can be used to detect rHOA. In addition, in the absence of large training data, a reliable DL model can be optimized by combining CT-AP and X-ray images.

Effect of progressive high-impact exercise on femoral neck structural strength in postmenopausal women with mild knee osteoarthritis: a 12-month RCT.

It is uncertain whether subjects with mild knee osteoarthritis, and who may be at risk of osteoporosis, can exercise safely with the aim of improving hip bone strength. This RCT showed that participating in a high-impact exercise program improved femoral neck strength without any detrimental effects on knee cartilage composition. INTRODUCTION: No previous studies have examined whether high-impact exercise can improve bone strength and articular cartilage quality in subjects with mild knee osteoarthritis. In this 12-month RCT, we assessed the effects of progressive high-impact exercise on femoral neck structural strength and biochemical composition of knee cartilage in postmenopausal women. METHODS: Eighty postmenopausal women with mild knee radiographic osteoarthritis were randomly assigned into the exercise (n = 40) or control (n = 40) group. Femoral neck structural strength was assessed with dual-energy X-ray absorptiometry. The knee cartilage region exposed to exercise loading was measured by the quantitative MRI techniques of T2 mapping and delayed gadolinium-enhanced MRI of cartilage (dGEMRIC). Also, an accelerometer-based body movement monitor was used to evaluate the total physical activity loading on the changes of femoral neck strength in all participants. Training effects on the outcome variables were estimated by the bootstrap analysis of covariance. RESULTS: A significant between-group difference in femoral neck bending strength in favor of the trainees was observed after the 12-month intervention (4.4%, p < 0.01). The change in femoral neck bending strength remained significant after adjusting for baseline value, age, height, and body mass (4.0%, p = 0.020). In all participants, the change in bending strength was associated with the total physical activity loading (r = 0.29, p = 0.012). The exercise participation had no effect on knee cartilage composition. CONCLUSION: The high-impact training increased femoral neck strength without having any harmful effect on knee cartilage in women with mild knee osteoarthritis. These findings imply that progressive high-impact exercise is a feasible method in seeking to prevent hip fractures in postmenopausal women whose articular cartilage may also be frail.

Bone and cartilage characteristics in postmenopausal women with mild knee radiographic osteoarthritis and those without radiographic osteoarthritis.

OBJECTIVES: To evaluate the association between radiographically-assessed knee osteoarthritis and femoral neck bone characteristics in women with mild knee radiographic osteoarthritis and those without radiographic osteoarthritis. METHODS: Ninety postmenopausal women (mean age [SD], 58 [4] years; height, 163 [6] cm; weight, 71 [11] kg) participated in this cross-sectional study. The severity of radiographic knee osteoarthritis was defined using Kellgren-Lawrence grades 0=normal (n=12), 1=doubtful (n=25) or 2=minimal (n=53). Femoral neck bone mineral content (BMC), section modulus (Z), and cross-sectional area (CSA) were measured with DXA. The biochemical composition of ipsilateral knee cartilage was estimated using quantitative MRI measures, T2 mapping and dGEMRIC. The associations between radiographic knee osteoarthritis grades and bone and cartilage characteristics were analyzed using generalized linear models. RESULTS: Age-, height-, and weight-adjusted femoral neck BMC (p for linearity=0.019), Z (p for linearity=0.033), and CSA (p for linearity=0.019) increased significantly with higher knee osteoarthritis grades. There was no linear relationship between osteoarthritis grades and knee cartilage indices. CONCLUSIONS: Increased DXA assessed hip bone strength is related to knee osteoarthritis severity. These results are hypothesis driven that there is an inverse relationship between osteoarthritis and osteoporosis. However, MRI assessed measures of cartilage do not discriminate mild radiographic osteoarthritis severity.

Light physical activity determined by a motion sensor decreases insulin resistance, improves lipid homeostasis and reduces visceral fat in high-risk subjects: PreDiabEx study RCT.

OBJECTIVE: To examine physical activity (PA) thresholds affecting glucose, insulin and lipid concentrations and body fat composition in high-risk patients for type 2 diabetes (T2D). INTERVENTION: A total of 113 subjects of both genders having abnormal glucose levels in the oral glucose tolerance test were contacted. A total of 78 subjects with age 58.8±10.4 years and body mass index 31.7±5.3 kg m(-2) were randomly assigned to intervention and control groups. INTERVENTION consisted of a supervised walking (60 min three times weekly) for 3 months. All the subjects received standard care for PA and weight reduction and wore an accelerometer during the whole wakeful time. RESULTS: Over 80% of the daily steps clustered at an acceleration level of 0.3-0.7 g (2-3 km h(-1) of walking) and were 5870 in the intervention and 4434 in the control group (P<0.029). Between 0 and 3 months no significant changes were observed in fasting and 2-h glucose, body weight or maximal oxygen uptake. In contrast, changes in fasting and 2-h insulin (-3.4 mU l(-1), P=0.035 and -26.6, P=0.003, respectively), homeostasis model assessment-estimated insulin resistance (-1.0, P=0.036), total cholesterol (-0.55 mmol l(-1), P=0.041), low-density lipoprotein (LDL) cholesterol (-0.36 mmol l(-1), P=0.008) and visceral fat area (-5.5 cm(2), P=0.030) were significantly greater in the intervention than in control subjects. The overall effects of PA were analyzed by quartiles of daily steps of all subjects. There were significant reductions in total and LDL cholesterol and visceral fat area between the highest (daily steps over 6520) and the lowest quartile (1780-2810 daily steps). The changes associated with PA remained significant after adjustments of baseline, sex, age and body weight change. CONCLUSION: Habitual and structured PAs with the acceleration levels of 0.3-0.7 g and daily steps over 6520, equivalent to walking at 2-3 km h(-1) for 90 min daily, standing for the relative PA intensity of 30-35% of the maximal oxygen uptake, are clinically beneficial for overweight/obese and physically inactive individuals with a high risk for T2D.

Quantification of differences in bone texture from plain radiographs in knees with and without osteoarthritis.

OBJECTIVE: To quantify differences in bone texture between subjects with different stages of knee osteoarthritis (OA) and age- and gender-matched controls from plain radiographs using advanced image analysis methods. DESIGN: Altogether 203 knees were imaged using constant X-ray parameters and graded according to Kellgren-Lawrence (KL) grading scale (KL0: n = 110, KL1: n = 28, KL2: n = 27, KL3: n = 31, KL4: n = 7). Bone density-related and structure-related parameters were calculated from medial and lateral tibial subchondral bone plate and trabecular bone and from femur. Density-related parameters were derived from grayscale values and structure-related parameters from Laplacian- and local binary patterns (LBP)-based images. RESULTS: Reproducibilities of structure-related parameters were better than bone density-related parameters. Bone density-related parameters were significantly (P < 0.05) higher in KL2-4 groups than in control group (KL0) in medial tibial subchondral bone plate and trabecular bone. LBP-based structure parameters differed significantly between KL0 and KL2-4 groups in medial subchondral bone plate, between KL0 and KL1-4 groups in medial and lateral trabecular bone, and between KL0 and KL1-4/KL2-4 in medial and lateral femur. Laplacian-based parameters differed significantly between KL0 and KL2-4 groups in medial side regions-of-interest (ROIs). CONCLUSIONS: Our results indicate that the changes in bone texture in knee OA can be quantitatively evaluated from plain radiographs using advanced image analysis. Based on the results, increased bone density can be directly estimated if the X-ray imaging conditions are constant between patients. However, structural analysis of bone was more reproducible than direct evaluation of grayscale values, and is therefore better suited for quantitative analysis when imaging conditions are variable.

Discrimination of fractures by low-frequency axial transmission ultrasound in postmenopausal females.

SUMMARY: In this cross-sectional study, 95 postmenopausal women, with and without fracture history, were measured by low-frequency axial transmission ultrasound. The measured ultrasound velocity discriminated the fractured subjects from the nonfractured ones equally or better than peripheral quantitative computed tomography (pQCT) and dual energy x-ray absorptiometry (DXA). These results suggest that low-frequency ultrasound is suitable for bone fragility assessment. INTRODUCTION: Quantitative low-frequency axial transmission ultrasound is a promising modality for assessing mineral density and geometrical properties of long bones such as radius and tibia. The aim of the current study was to evaluate the ability of low-frequency axial transmission ultrasound to discriminate fractures retrospectively in postmenopausal women. METHODS: A cross-sectional study involved 95 female subjects aged 45-88 years, whose fracture information was gathered retrospectively. The fracture group was defined as subjects with one or more low-/moderate-energy fractures. The radius and tibial shaft were measured with a custom-made ultrasonometer to assess the velocity of the low-frequency first-arriving signal (V (LF)). Site-matched pQCT was used to measure volumetric cortical and subcortical bone mineral density (sBMD), and cortical thickness (CTh). Areal BMD (aBMD) was measured using DXA for the whole body (WB), lumbar spine, and hip. RESULTS: The majority (19/32; 59 %) of the fractures were in the upper limb. V (LF) in the radius, but not in the tibia, discriminated fractures with an age- and BMI-adjusted odds ratio (OR) of 2.06 (95 % CI 1.21-3.50, p < 0.01). In the radius, CTh and cortical BMD (CBMD) significantly discriminated fractures, as did the total, cortical, and sBMD in the tibia (adjusted OR 1.35-2.15, p < 0.05). Sensitivity and specificity were similar among all the measurements (area under the receiver operating characteristic curve 0.74-0.81, p < 0.001). CONCLUSIONS: Low-frequency axial transmission ultrasound in the radius was able to discriminate fractured subjects from the nonfractured ones. This suggests that low-frequency axial transmission ultrasound has the potential to assess bone fragility in postmenopausal women.

BMD T-score discriminates trochanteric fractures from unfractured controls, whereas geometry discriminates cervical fracture cases from unfractured controls of similar BMD.

UNLABELLED: The ability of bone mineral density (BMD) to discriminate cervical and trochanteric hip fractures was studied. Since the majority of fractures occur among people who are not diagnosed as having osteoporosis, we also examined this population to elucidate whether geometrical risk factors can yield additional information on hip fracture risk beside BMD. The study showed that the T-score criterion was able to discriminate fracture patients from controls in the cases of trochanteric fractures, whereas geometrical measures may discriminate cervical fracture cases in patients with T-score >-2.5. INTRODUCTION: Low bone mineral density (BMD) is a well-established risk factor for hip fracture. However, majority of fractures occur among people not diagnosed as having osteoporosis. We studied the ability of BMD to discriminate cervical and trochanteric hip fractures. Furthermore, we examined whether geometrical measures can yield additional information on the assessment of hip fracture risk in the fracture cases in subjects with T-score >-2.5. METHODS: Study group consisted of postmenopausal females with non-pathologic cervical (n = 39) or trochanteric (n = 18) hip fracture (mean age 74.2 years) and 40 age-matched controls. BMD was measured at femoral neck, and femoral neck axis length, femoral neck and shaft cortex thicknesses (FNC and FSC), and femoral neck-shaft angle (NSA) were measured from radiographs. RESULTS: BMD T-score threshold of -2.5 was able to discriminate trochanteric fractures from controls (p < 0.001). Seventeen out of 18 trochanteric fractures occurred in individuals with T-score -2.5. Within these non-osteoporotic cervical fracture patients (N = 19) and non-osteoporotic controls (N = 35), 83.3% were classified correctly based on a model including NSA and FNC (p < 0.001), area under the receiver operating characteristics curve being 0.85 for the model, while it was only 0.56 for BMD alone. CONCLUSIONS: The study suggests that the risk of trochanteric fractures could be discriminated based on a BMD T-score <-2.5 criterion, whereas cervical fracture cases would remain under-diagnosed if solely using this criterion. Instead, geometrical risk factors are able to discriminate cervical fracture cases even among individuals with T-score >-2.5. For cervical and trochanteric fractures combined, BMD and geometric measures independently contributed to hip fracture discrimination. Our data support changing from T-score <-2.5 to a more comprehensive assessment of hip fracture etiology, in which fracture type is also taken into account. The findings need to be confirmed with a larger sample, preferably in a prospective study.

Effect of impact exercise on bone metabolism.

SUMMARY: Regular impact exercise in premenopausal women caused positive osteogenic effects associated to low basal serum parathormone (PTH) but had no effects on bone turnover markers PINP or TRACP5b. The low serum basal PTH levels during impact exercise may be a sign of increased incorporation of calcium to bone. INTRODUCTION: This study aimed to determine the long-term effects of high-impact exercise on bone turnover and calciotropic hormones. METHODS: We performed a 12-month population-based, randomized, controlled exercise trial in 120 women (age 35-40 years) randomly assigned to an exercise group (EG; n = 60) or a control group (CG; n = 60). The exercise regimen consisted of supervised high-impact exercises three times per week. Daily impact loading was assessed by using an accelerometer. Bone turnover markers and calciotropic hormones were analyzed at 0, 6, and 12 months. RESULTS: Twelve months of impact exercise did not reveal any treatment effects in bone turnover markers PINP or TRAPC5b, whereas serum basal PTH decreased significantly more in the EG than in the CG (-11.2 vs. -2.2 pg/mL; p = 0.03). The change in PTH was dose dependent and most clearly seen in subjects with 96 to 130 daily impacts at 2.5 to 5.3 g (e.g., running or jumping). CONCLUSIONS: Regular impact exercise does not cause persistent alterations in bone turnover emphasizing necessity of continuous training to achieve bone benefits. Impact exercise training lowers the serum basal PTH levels and possibly enables greater difference between the basal PTH and transient exercise-induced PTH peaks leading to osteogenic effects.

High insulin levels are positively associated with peripheral nervous system function.

OBJECTIVE: The aim of this study was to analyze peripheral nervous system (PNS) function in overweight and obese individuals. MATERIALS AND METHODS: Forty-four adult non-diabetic overweight individuals were recruited. Peroneal motor nerve conduction and radial, sural, and medial plantar sensory nerve conduction were studied. Insulin and glucose levels were determined twice (over a 2- to 3-year period) with an oral glucose tolerance test (OGTT). Multiple stepwise linear regression models adjusted for age, height, weight, and skin temperature were used to analyze the data. RESULTS: Analysis revealed that baseline insulin levels measured 120 min after an OGTT explained 18% of the variation in peroneal F-wave minimum latency, 8% of peroneal F-wave maximum latency variation, 15% of sural sensory latency variation, 13% of sural sensory nerve conduction velocity (NCV) variation, and 10% of the variation in medial plantar sensory NCV. DISCUSSION AND CONCLUSION: Our study shows that serum insulin levels measured 120 min after an OGGT are positively associated with PNS function. High insulin levels without notably high glucose levels appear to be beneficial for the function of the PNS.

Are signs of temporomandibular disorders stable and predictable in adolescents with headache?

The aim of the present study was to study changes in signs and symptoms of temporomandibular disorders (TMD) and factors predicting TMD signs in adolescents with and without headache. A population-based sample (n = 212) of 13-year-olds with and without headache was re-examined at the age of 16. The study included a questionnaire, face-to-face interview and somatic examination. In addition, a neurological examination, a muscle evaluation and a stomatognathic examination were performed. Significant changes were seen in TMD signs during the follow-up, but TMD signs at the end of the follow-up could not be predicted by baseline headache, sleeping difficulties, depression or muscle pain. TMD signs at the age of 16 were associated with female gender and muscle pain. We conclude that considerable changes in TMD signs occur in the follow-up of adolescents with and without headache. Headache-related TMD are not predictable in adolescents with and without headache.

The phase state of NiTi implant material affects osteoclastic attachment.

In the present work, the responses of mature osteoclasts cultured on austenite and martensite phases of NiTi shape memory implant material were studied. We used the sensitivity of osteoclasts to the underlying substrate and actin ring formation as an indicator of the adequacy of the implant surface. The results showed osteoclasts with actin ring on both NiTi phases. However, significantly more osteoclasts were present on the austenitic NiTi than on the martensitic NiTi. We also analyzed the surface free energy of the samples but found no significant difference between austenite and martensite phases. The results revealed that osteoclasts tolerated well the austenite phase of NiTi. The chemically identical martensitic NiTi was not as well tolerated by osteoclasts (e.g., indicated by diminished actin ring formation). This leads to the conclusion that certain physical properties specific to the martensitic NiTi have an adverse effect to the surviving of osteoclasts on this NiTi phase. These results confirm that mature, authentic osteoclasts can act as cell probes in experiments concerning aspects of biocompatibility of bone implant materials.

Gamma irradiation and ethylene oxide in the sterilization of native reindeer bone morphogenetic protein extract.

BACKGROUND AND AIMS: For human use, it is necessary to sterilize bone morphogenetic proteins (BMPs), in order to reduce the risk of infections and associated complications. We compared the effects of ethylene oxide and gamma irradiation in the sterilization of native reindeer BMP extract with regard to bone induction in the Balb/C mouse thigh muscle pouch model. MATERIALS AND METHODS: BMP extract, sterilized with ethylene oxide gas (Steri-Vac 4XL, temperature 29 degrees C, exposure time 4 h, ethylene oxide concentration 860 mg/l), or gamma irradiation at doses of 3.15 MRad was administered in implants containing 5 or 10 mg of BMP extract with collagen carrier. Non-sterilized collagen implants served as controls. New bone formation was evaluated based on the incorporation of Ca45 and radiographically three weeks after implantation. RESULTS: The collagen was not able to induce new bone visible in radiographs. The mean Ca45 incorporation in the gamma sterilized group containing 5 mg of BMP extract was 30% (p = 0.04) and that containing 10 mg of BMP extract was 60% (p = 0.02) higher than seen in the corresponding ethylene oxide sterilized groups. The mean new bone areas were 45% higher in the gamma sterilized groups than in the corresponding ethylene oxide sterilized groups, but the differences were not significant. The mean optical density of new bone in the gamma sterilized group containing 5 mg of BMP extract was 75% (p = 0.00) and in that containing 10 mg of BMP extract was 70% (p = 0.00) higher than seen in the corresponding ethylene oxide sterilized groups. CONCLUSION: Native reindeer BMP extract is more sensitive to the effects of ethylene oxide gas sterilization than gamma irradiation. These results suggest that gamma irradiation is recommendable for the sterilization of BMP extracts.

Influence of the upper femur and pelvic geometry on the risk and type of hip fractures.

The geometry of the upper femur has been reported to associate with the hip fracture risk in postmenopausal women. However, these associations seem to be partly conflicting, probably because of differences in measurement setup. Here, we compared the upper femur and pelvic geometries of 70 hip fracture patients (46 cervical and 24 trochanteric fractures) and 40 age-adjusted controls based on plain anteroposterior radiographs, eliminating the possible sources of inaccuracy as far as possible by using a standardized patient position and calibrated dimension measurements by digital image analysis. The femoral neck/shaft angle (NSA) was larger in the fracture patients compared with the controls (p < 0.001). The fracture group had thinner cortices in the upper femur than the controls (p < 0.001). The femoral shaft diameter (FSD; p < 0.001), trochanter width (TW; p < 0.01), and the pelvic dimensions, that is, the smallest outer pelvic diameter (SOPD; p < 0.01) and the largest inner pelvic diameter (LIPD; p < 0.05) were smaller in the fracture group. Comparing the fracture types, we found NSA larger in the cervical hip fracture patients than in the patients with a trochanteric fracture (p < 0.01). The femoral neck/shaft cortex ratio was lower and the FSD was smaller in the cervical hip fracture group (p < 0.05). Acetabular width (AW) was greater and the SOPD was wider in the cervical fracture patients (p < 0.01). We concluded that the upper femur and pelvic dimensions as defined from calibrated and position-standardized plain radiographs are useful in the evaluation of hip fracture risk and fracture type.

Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on bone in two rat strains with different aryl hydrocarbon receptor structures.

Polychlorinated dibenzo-p-dioxins (PCDDs) are highly toxic environmental contaminants, and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is the most potent dioxin. Here, we studied the effects of TCDD on bone. Two rat strains, Han/Wistar (H/W) and Long-Evans (L-E), were used because they exhibit a 1000-fold sensitivity difference in acute lethality of TCDD, which difference is related to the aryl hydrocarbon receptor (AHR). TCDD inhibited the tibial growth dose dependently, the effect being manifested at lower doses in the more sensitive L-E strain. In H/W rats the effect of TCDD was seen only at the high dose of 170 microg/kg (p < 0.05), whereas in the sensitive L-E rats a significant reduction of bone growth was already seen at 1.7 microg/kg (p < 0.01). This reduction was caused by the smaller tibial size because the diaphyseal bone mineral density (BMD) did not change. The three-point bending breaking force of the tibia was significantly reduced in H/W rats at 170 microg/kg (p < 0.05), but tibial stiffness was lower already at the dose of 17 microg/kg (p < 0.05). In the sensitive L-E strain, both breaking force and stiffness were reduced at the dose of 17 microg/kg (p < 0.001). These results indicate that TCDD dose-dependently interferes with bone growth, modeling, and mechanical strength. The altered transactivation domain of AHR is associated with a lower sensitivity of bone to TCDD in H/W rats, suggesting that AHR plays a role in modulating the effects of dioxins on bone.

Femoral neck is a sensitive indicator of bone loss in immobilized hind limb of mouse.

The present study was carried out to evaluate a unilateral hind limb immobilization model in the mouse. The right legs of male mice (age 10-12 weeks) were immobilized for 3 weeks against the abdomen by an elastic bandage. Body weight decreased significantly during the immobilization. Peripheral quantitative computed tomography (pQCT) analysis showed that the cross-sectional cortical area (CSA), the bone mineral content (BMC), and the bone mineral density (BMD) of the tibial diaphysis were lower in both legs of the immobilized animals than in age-matched controls, but the difference was mainly due to weight reduction. At the tibial metaphysis, CSA, BMC, and BMD were reduced in both legs of the immobilized animals, even after weight adjustment. At the femoral neck, CSA, BMC, and BMD were significantly lower in both legs of the immobilized animals, and the difference between the hind legs of the immobilized animals was also highly significant. The findings of the pQCT study were in good agreement with the changes in mechanical strength. The tibia was a more sensitive indicator of diaphyseal bone weakening than the femur when measuring the bending breaking force of the diaphysis. The femoral neck showed significantly decreased strength, and the difference between the immobilized leg and the contralateral leg was most clearly seen in lateral loading. We conclude that 3 weeks of hind limb immobilization weakened the tibia and femur significantly compared with their contralateral counterparts. The reduction was more significantly seen in the mechanical bending strength than in the pQCT evaluation, and the femoral neck was the most sensitive indicator of bone weakening.

Expression profiles of mRNAs for osteoblast and osteoclast proteins as indicators of bone loss in mouse immobilization osteopenia model.

An experimental mouse model for disuse osteopenia was developed using unilateral cast immobilization. Analysis of the distal femurs and proximal tibias by quantitative histomorphometry revealed significant osteopenia within 10-21 days of immobilization. At 3 weeks, bone loss was also demonstrated with peripheral quantitative computed tomography as diminished bone mineral content and as concomitant reduction in the cross-sectional moment of inertia. These structural and geometrical alterations resulted in decreased strength of the distal femurs tested by cantilever bending. Analysis of the underlying cellular and molecular mechanisms of bone loss revealed a rapid increase in bone resorption within 3 days of immobilization. The mRNA levels for cathepsin K, matrix metalloproteinase-9, and tartrate resistant acid phosphatase were all significantly increased during the 21-day immobilization period, but with different expression profiles. These increases were paralleled by an increased number of osteoclasts as measured by histomorphometry. By day 6 of immobilization, the balance of bone turnover was further shifted toward net bone loss as the mRNA levels for major bone components (type I collagen and osteocalcin) were decreased. In histomorphometric analysis this was observed as reduced rates of mineral apposition and bone formation after 10 days of immobilization. The results of this study demonstrate that immobilization has a dual negative effect on bone turnover involving both depressed bone formation and enhanced bone resorption.

Mineral density and bone strength are dissociated in long bones of rat osteopetrotic mutations.

Bone mineral density (BMD) and mechanical strength generally show strong positive correlations. However, osteopetrosis is a metabolic bone disease with increased skeletal density radiographically and increased risk of fracture. We have evaluated mechanical strength and mineral density in three osteopetrotic mutations in the rat (incisors-absent [ia/ia], osteopetrosis [op/op], and toothless [tl/tl]) to test the hypothesis that reduced bone resorption in one or more of these mutations results in weaker bones in the presence of greater mineral density and skeletal mass. Peripheral quantitative computed tomography (pQCT) was used to analyze BMD and cross-sectional geometry in the tibial diaphysis and metaphysis as well as the femoral diaphysis and femoral neck. The bending breaking force of tibial and femoral midshafts was obtained using the three-point bending test and femoral neck strength was tested by axial loading. Osteopetrotic mutants were significantly smaller than their normal littermates (NLMs) in each stock. The pQCT analysis showed that BMD and bone mineral content (BMC) were higher than or equal to NLMs in all skeletal sites measured in the osteopetrotic mutants. However, the mechanical breaking force was equal to or lower than their NLMs in all sites. The cross-sectional structure of long bone shafts was markedly different in osteopetrotic mutants, having a thin cortex and a medullary area filled with primary trabecular bone. These results indicate that osteopetrotic mutations in the rat increase bone density and decrease bone strength. The tibial diaphysis was significantly weaker in tl/tl and ia/ia mutants and the tibial metaphysis showed the greatest increase in BMD in all mutants. These data are another illustration that an increased BMD does not necessarily lead to stronger bones.

Comparison of three-point bending test and peripheral quantitative computed tomography analysis in the evaluation of the strength of mouse femur and tibia.

We compared the mechanical and densitometric testing methods in evaluating the mechanical strength of mouse cortical bones. The femora and tibiae of 10 male mice (weight 32.8 +/- 4.0 g) were utilized. Volumetric cortical bone mineral density (vCtBMD), cross-sectional cortical area at midshaft (CSA), cross-sectional moment of inertia (CSMI), and strength strain index (SSI) were measured by peripheral quantitative computed tomography (pQCT). The precision of pQCT expressed as a coefficient of variation (CV) was 1.1%, 2.7%, and 6.4% for vCtBMD, CSA, and CSMI, respectively. The mechanical properties were measured by a three-point bending test. The method error measured from paired bones was 7.3%-10.1% for breaking bending force, 15.0%-15.2% for stiffness, 2.0%-2.4% for vCtBMD, 5.2%-6.4% for CSA, 13.5%-17.6% for CSMI, and 8.9%-18.1% for SSI. CSMI and CSA were found to be the best explanatory variables for the breaking force of femur and tibia, respectively, while CSA and CSMI were the best predictors for the elastic modulus of femur and tibia, respectively. CSA had a higher correlation with mechanical parameters than vCtBMD. On the basis of this study, the mechanical tests and the pQCT measurements are relevant in biomechanical studies on mouse bones and justify the use of the murine model. High-resolution pQCT gives better precision than the three-point bending test in studies of mouse bones.

The mechanical strength of bone in different rat models of experimental osteoporosis.

In order to discover good parameters for experimental osteoporosis, we measured the failure load of the femoral neck and the bending strength of the tibia in orchidectomized (ORC) (20 rats for 4 weeks), ovariectomized (OVX) (28 rats for 6 weeks), and immobilized (IMM) (33 rats for 3 weeks) rats. Each of these operations led to a significant decrease in trabecular bone volume when compared with corresponding controls (p < 0.001). The ash weight of femurs was significantly decreased in ORC (p < 0.05) and IMM (p < 0.001) rats, but not in OVX rats. Growth of the femur was somewhat slower in ORC (p < 0.05) and IMM rats (p < 0.05), but not in OVX animals. All three osteoporosis models showed significant decreases in the maximal load of the femoral neck (ORC: 23.9%, p < 0.001; OVX: 15.8%, p < 0.001; IMM: 27.7%, p < 0.001), as well as in energy absorption (ORC: 43.9%, p < 0.001; OVX: 28.3%, p < 0.001; IMM: 45.3%, p < 0.001). In tibia orchidectomy reduced maximal strength and energy absorption significantly (10%, p < 0.01; 27.8%, p < 0.01), but ovariectomy decreased only maximal load (8.7%, p < 0.01) and immobilization only energy absorption (18.0%, p < 0.01). Our results suggest that the mechanical strength of the femoral neck is a sensitive indicator of bone loss in all three osteoporosis models.

A metaphyseal defect model of the femur for studies of murine bone healing.

A bone defect model was developed in the distal metaphysis of the femur for studies on bone healing in the mouse. The circular defect involving 20% of the bone circumference resulted in a 34% reduction in the bending moment compared to intact bone. The healing process was followed using histomorphometry, peripheral quantitative computed tomography (pQCT), biomechanical testing, and molecular biological analyses. Histologically, healing of the defect was characterized by filling of the medullary cavity with trabecular new bone during the first week of healing, and by closing of the cortical window by 6 weeks. Small areas of periosteal chondrogenesis were frequently observed during defect healing. In pQCT, bone mineral content (BMC) of the defect area approached that of intact control bone already by 3 weeks, reflecting the production of trabecular bone. Similarly, the bending strength and stiffness of the healing femur reached the level of intact control femur already at 3 weeks. Bone formation and remodeling was followed by Northern analyses, which demonstrated elevated mRNA levels for bone components (type I collagen and osteocalcin), and for osteoclastic enzymes (cathepsin K, matrix metalloproteinase-9, and tartrate-resistant acid phosphatase) throughout the healing period. Finally, the applicability of the defect model for gene therapy experiments was tested using adenovirus-mediated transfer of the LacZ reporter gene. Both histochemistry and mRNA analyses demonstrated that the gene was expressed in the repair tissue with the highest expression during the first week of healing. The present model thus provides a standardized environment for studies on induction and remodeling of trabecular new bone in normal and genetically engineered mice.