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Particles released from implants cause inflammatory bone loss, which is a key factor in aseptic loosening, the most common reason for joint replacement failure. With the anticipated increased incidence of total joint replacement in the next decade, implant failure will continue to burden patients. The gut microbiome is increasingly recognized as an important factor in bone physiology, however, its role in implant loosening is currently unknown. We tested the hypothesis that implant loosening is associated with changes in the gut microbiota in a preclinical model. When the particle challenge caused local joint inflammation, decreased peri-implant bone volume, and decreased implant fixation, the gut microbiota was affected. When the particle challenge did not cause this triad of local effects, the gut microbiota was not affected. Our results suggest that cross-talk between these compartments is a previously unrecognized mechanism of failure following total joint replacement.

Biomarkers are of interest to identify patients at risk for peri-implant osteolysis and aseptic loosening. We used a rat model of particle-induced peri-implant osteolysis to investigate if early changes in biomarkers were associated with subsequent implant fixation strength. Implants were placed in rat femora, which were then challenged with intra-articular knee injections of either clean polyethylene, lipopolysaccharide-doped polyethylene, or cobalt-chromium alloy particles, with particle-free vehicle serving as control (n ≥ 8 per group). Rats were weighed weekly, blood was collected at weeks 0, 3, 5, and 6, and locomotor behavior was assessed 4 days before study conclusion. Rats were euthanized 6 weeks post surgery. Week 6 serum was analyzed for five bone remodeling markers, while longitudinal serum was assessed for osteocalcin. Bone-implant contact, peri-implant trabecular architecture, and implant fixation strength were measured. Rats challenged with cobalt-chromium particles had a significant reduction in implant fixation strength compared with the vehicle-control group (P = .034). This group also had elevated serum osteocalcin (P = .005), depressed weight gain (P = .001) and less frequent rearing behavior (P = .029). Regardless of group, change in serum osteocalcin at week 3 (r = -.368; P = .046), change in weight at week 2 (r = .586; P < .001), as well as weight change at all other time intervals were associated with fixation strength. The finding that early alterations in serum osteocalcin and body weight were predictive of subsequent implant fixation strength supports continued investigation of biomarkers for early detection of peri-implant osteolysis and implant loosening. Further, change in biomarker levels was found to be more indicative of implant fixation status than any single measurement.

BACKGROUND: Osteoporosis and Alzheimer’s disease are common diseases of aging that would seem to be unrelated, but may be linked through the influence of bone-derived signals on brain function. The aim of the current study is to investigate the relationship between circulating levels of bone-related biomarkers and cognition. METHODS: The population included 103 community-dwelling older individuals with memory concerns but without cognitive impairment. A global cognition summary measure was collected at baseline and 6, 12, and 18 months post-enrollment by converting raw scores from 19 cognitive function tests to z-scores and averaging. Baseline plasma concentrations of bone-related biomarkers, including undercarboxylated, carboxylated, and total osteocalcin, parathyroid hormone, C-terminal telopeptide of collagen 1 (CTX-1), procollagen type 1 amino-terminal propeptide, osteoprotegrin, osteopontin, Dickkopf WNT signaling pathway inhibitor 1 (Dkk1), sclerostin, and amyloid β peptides (Aβ(40) and Aβ(42)), were measured. RESULTS: Using sex, age, and education-adjusted mixed-effects models, we found that baseline levels of TNF-related apoptosis-inducing ligand (TRAIL; p < .001), Dkk1 (p = .014), and CTX-1 (p = .046) were related to the annual rate of change of global cognition over the 18 month follow-up. In cognitive domain-specific analysis, baseline TRAIL was found to be positively related to the annual rate of change in episodic (p < .001) and working memory (p = .016), and baseline Dkk1 was positively related to semantic memory (p = .027) and negatively related to working memory (p = .016). CONCLUSIONS: These results further confirm the link between bone and brain health and suggest that circulating levels of bone-related biomarkers may have diagnostic potential to predict worsening cognition.

Objective To assess the ability of sclerostin antibody therapy to blunt the negative effects of polyethylene particles on implant fixation and peri-implant bone structure in a rat implant fixation model. Methods Thirty-six adult male rats received intramedullary titanium implants; 12 rats received vehicle injections only (control), and 24 rats received intraarticular injections of lipopolysaccharide-doped polyethylene particles. Twelve of the rats that received particles also received sclerostin antibody treatment. The 3 groups of rats were maintained for 12 weeks in a pathogen-free environment, at which time mechanical, micro–computed tomography, and dynamic and static histomorphometry end points were assessed. Results Sclerostin antibody treatment completely blocked the negative effect of the lipopolysaccharide-doped polyethylene particles on implant fixation and peri-implant bone volume by increasing the bone formation rate and depressing bone resorption. Conclusion Anabolic agents targeting the Wnt signaling pathway are a promising new alternative for the prevention of periprosthetic osteolysis and aseptic loosening.

Background: Previous studies have demonstrated that sclerostin blockade is anabolic for bone. This study examined whether systemic administration of sclerostin antibody would increase implant fixation and peri-implant bone volume in a rat model. Methods: Titanium cylinders were placed in the femoral medullary canal of ninety male Sprague-Dawley rats. One-half of the rats (n = 45) received murine sclerostin antibody (Scl-Ab, 25 mg/kg, twice weekly) and the other one-half (n = 45) received saline solution. Equal numbers of rats from both groups were sacrificed at two, four, or eight weeks after the implant surgery and the femora were examined by microcomputed tomography, mechanical pull-out testing, and histology. Results: Fixation strength in the two groups was similar at two weeks but was 1.9-fold greater at four weeks (p = 0.024) and 2.2-fold greater at eight weeks (p < 0.001) in the rats treated with sclerostin antibody. At two weeks, antibody treatment led to increased cortical area, with later increases in cortical thickness and total cross-sectional area. Significant differences in peri-implant trabecular bone were not evident until eight weeks but included increased bone volume per total volume, bone structure that was more plate-like, and increased trabecular thickness and number. Changes in bone architecture in the intact contralateral femur tended to precede the peri-implant changes. The peri-implant bone properties accounted for 61% of the variance in implant fixation strength, 32% of the variance in stiffness, and 63% of the variance in energy to failure. The implant fixation strength at four weeks was approximately equivalent to the strength in the control group at eight weeks. Conclusions: Sclerostin antibody treatment accelerated and enhanced mechanical fixation of medullary implants in a rat model by increasing both cortical and trabecular bone volume. Clinical Relevance: Sclerostin antibody treatment may be useful for improving implant fixation.

Enhancement of in vivo mobilization and homing of endogenous mesenchymal stem cells (MSCs) to an injury site is an innovative strategy for improvement of bone tissue engineering and repair. The present study was designed to determine whether mobilization by AMD3100 and/or local homing by delivery of stromal cell-derived factor-1 (SDF-1) enhances recombinant human bone morphogenetic protein-2 (rhBMP-2) induced ectopic bone formation in an established rat model. Rats received an injection of either saline or AMD3100 treatment 1 h before harvesting of bone marrow for in vitro colony-forming unit-fibroblasts (CFU-F) culture or the in vivo subcutaneous implantation of absorbable collagen sponges (ACSs) loaded with saline, recombinant human bone morphogenetic protein-2 (rhBMP-2), SDF-1, or the combination of SDF-1 and rhBMP-2. AMD3100 treatment resulted in a significant decrease in CFU-F number, compared with saline, which confirmed that a single systemic AMD3100 treatment rapidly mobilized MSCs from the bone marrow. At 28 and 56 days, bone formation in the explanted ACS was assessed by microcomputed tomography (μCT) and histology. At 28 days, AMD3100 and/or SDF-1 had no statistically significant effect on bone volume (BV) or bone mineral content (BMC), but histology revealed more active bone formation with treatment of AMD3100, loading of SDF-1, or the combination of both AMD3100 and SDF-1, compared with saline-treated rhBMP-2 loaded ACS. At 56 days, the addition of AMD3100 treatment, loading of SDF-1, or the combination of both resulted in a statistically significant stimulatory effect on BV and BMC, compared with the saline-treated rhBMP-2 loaded ACS. Histology of the 56-day ACS were consistent with the μCT analysis, exhibiting more mature and mineralized bone formation with AMD3100 treatment, SDF-1 loading, or the combination of both, compared with the saline-treated rhBMP-2 loaded ACS. The present study is the first that provides evidence of the efficacy of AMD3100 and SDF-1 treatment to stimulate trafficking of MSCs to an ectopic implant site, in order to ultimately enhance rhBMP-2 induced long-term bone formation.

Bone growth and repair are under the control of biochemical and mechanical signals. Low-intensity pulsed ultrasound (LIPUS) stimulation at 30mW/cm(2) is an established, widely used and FDA approved intervention for accelerating bone healing in fractures and non-unions. Although this LIPUS signal accelerates mineralization and bone regeneration, the actual intensity experienced by the cells at the target site might be lower, due to the possible attenuation caused by the overlying soft tissue. The aim of this study was to investigate whether LIPUS intensities below 30mW/cm(2) are able to provoke phenotypic responses in bone cells. Rat bone marrow stromal cells were cultured under defined conditions and the effect of 2, 15, 30mW/cm(2) and sham treatments were studied at early (cell activation), middle (differentiation into osteogenic cells) and late (biological mineralization) stages of osteogenic differentiation. We observed that not only 30mW/cm(2) but also 2 and 15mW/cm(2), modulated ERK1/2 and p38 intracellular signaling pathways as compared to the sham treatment. After 5 days with daily treatments of 2, 15 and 30mW/cm(2), alkaline phosphatase activity, an early indicator of osteoblast differentiation, increased by 79%, 147% and 209%, respectively, compared to sham, indicating that various intensities of LIPUS were able to initiate osteogenic differentiation. While all LIPUS treatments showed higher mineralization, interestingly, the highest increase of 225% was observed in cells treated with 2mW/cm(2). As the intensity increased to 15 and 30mW/cm(2), the increase in the level of mineralization dropped to 120% and 82%. Our data show that LIPUS intensities lower than the current clinical standard have a positive effect on osteogenic differentiation of rat bone marrow stromal cells. Although Exogen™ at 30mW/cm(2) continues to be effective and should be used as a clinical therapy for fracture healing, if confirmed in vivo, the increased mineralization at lower intensities might be the first step towards redefining the most effective LIPUS intensity for clinical use.

Two methods to improve bone repair include the use of recombinant human bone morphogenetic protein-2 (rhBMP-2) and low-intensity pulsed ultrasound (LIPUS). The present study was designed to determine if LIPUS enhances the effect of rhBMP-2–induced bone formation in a well characterized ectopic implant model. Absorbable collagen sponges loaded with 0-, 1-, 2.5- or 5-μg doses of rhBMP-2 were implanted subcutaneously in 11-week-old, male Long Evans rats, followed by daily 20-min LIPUS or sham LIPUS treatment beginning 1 d after surgery. Explanted sponges were assessed for bone volume, mineral density and mineral content by microcomputed tomography (μCT). At two weeks, LIPUS had no effect on rhBMP-2–induced bone formation, but at four weeks, LIPUS increased bone volume in the 1-μg rhBMP-2–treated implants 117.7-fold (0.02 ± 0.04 mm 3 vs. 2.07 S.E.M. ± 1.67 mm 3 ; p = 0.028), and 2.3-fold in the 5-μg dose implants (5.96 ± 3.68 mm 3 vs. 13.52 ± 6.81 mm 3 ; p = 0.077) compared with sham LIPUS. Bone mineral density was not affected by LIPUS treatment. Total mineral content followed the same pattern as bone volume. Histologic staining for mineralized tissue was consistent with the μCT observations. The present study is the first to demonstrate that LIPUS enhances bone formation induced by rhBMP-2. (E-mail: robert_leven@rush.edu)

Short-term osseointegration of orthopedic implants is critical for the long-term stability of the implant–bone interface. To improve initial implant stability, one strategy under consideration involves the presentation of adhesion ligands on the implant surface to stimulate bone regeneration in the peri-implant region. To assess the relative effects of implant surface chemistry and topography on osseointegration within the rat femoral ablation implant model, a nonfouling, enzymatically degradable interpenetrating polymer network (edIPN) of poly(AAm-co-EG/AAc) amenable to presenting the cell signaling domain Arg-Gly-Asp (RGD), was developed. Moderate enhancement of peri-implant bone formation was found after 28 days using the edIPN without peptide modification (p = 0.032). However, no data supported a benefit of peptide modification, as bone–implant contact, normalized bone volume and normalized fixation strength was equivalent or poorer than dual acid-etched (DAE) treated implants after 28 days. Surface topography was determined to be the dominant factor in modulating osseointegration, as DAE implants produced equivalent roughness-normalized fixation strength versus previously reported data on plasma-sprayed hydroxyapatite/tricalcium phosphate-coated implants (Barber et al., J Biomed Mater Res A, forthcoming). An ideal osseointegrated implant will require optimization of all three aforementioned parameters, and may take the form of biomolecule delivery from thin degradable polymer networks. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007

Objective The relationship between knee pain and radiographic evidence of knee osteoarthritis (OA) is notoriously imperfect. In particular, conditions that distinguish individuals with symptoms from those with comparable radiographic involvement who remain asymptomatic are unclear. We investigated dynamic loading across the knee in individuals with mild radiographic OA who were distinguished by the presence or absence of knee pain. Methods Subjects were recruited into 3 groups: symptomatic with a Kellgren/Lawrence (K/L) grade of 2 (n = 52), asymptomatic with a K/L grade of 2 (n = 19), and asymptomatic with a K/L grade of 0 or 1 (n = 37), the latter representing a normal comparator group. Dynamic knee loading was assessed with gait analysis, and both the peak external knee adduction moment and the knee adduction angular impulse were determined. Results Peak knee adduction moment and knee adduction angular impulse were 19% and 30% higher, respectively, in symptomatic K/L grade 2 individuals than in asymptomatic individuals with the same radiographic grade (P < 0.05). Conversely, the asymptomatic K/L grade 2 group did not differ from the K/L grade 0–1 normal comparator group (P = 1.00). Conclusion Among individuals with mild radiographic knee OA (K/L grade 2), those who are symptomatic have significantly higher medial compartment loads than those who are asymptomatic, whereas those who are asymptomatic do not differ from normal controls (asymptomatic K/L grade 0 or 1). These findings suggest a biomechanical component to the distinction between asymptomatic and symptomatic radiographic OA, which may be pathophysiologically important.

We demonstrate that a biomimetic polymer network is capable of affecting bone regeneration in vivo. Starting with a foundation consisting of an environmentally responsive poly(N-isopropylacrylamide-co-acrylic acid) hydrogel, we incorporated matrix metalloproteinase-13 (MMP-13) degradable crosslinkers and peptides containing integrin-binding domains (i.e., Arg-Gly-Asp) to create a biomimetic matrix designed to encourage osteoblast migration and proliferation. We independently tuned matrix stiffness and peptide concentration to generate a response surface model of osteoblast proliferation on different types of matrices. Osteoblast proliferation was significantly influenced by matrix stiffness (i.e., its complex modulus) and peptide concentration. When implanted in a rat femoral ablation model, these matrices induced bone regeneration only when protease degradable crosslinks were used to create the network. For the matrices with MMP-13 degradable crosslinkers, the bone formed had a trabecular-like structure and was distributed throughout the marrow space. Based on the correlated effects of matrix stiffness and ligand concentration, the response surface model will facilitate improvements in the regenerative capacity of these artificial extracellular matrices.

Background: Gaps at the interface between implant and bone increase the risk of diminished implant fixation and eventual loosening. The purpose of the present study was to determine if combined use of recombinant human transforming growth factor-beta 2 (rhTGF-β2) and bone morphogenetic protein 2 (rhBMP-2) led to greater implant fixation strength in the presence of interface gaps than the use of either growth factor alone. Methods: Twenty-eight skeletally mature adult male dogs received one porous-coated titanium implant in the proximal part of each humerus, for a total of fifty-six implantation sites. Spacers were used to establish an initial 3-mm gap between the implant and the host bone at all fifty-six sites. Forty-two implants were coated with hydroxyapatite-tricalcium phosphate and were used in three growth-factor-treatment groups in which the implants placed in the left humerus were loaded with 12 μg of rhTGF-β2 (Group 1, seven animals), 25 μg of rhBMP-2 (Group 2, seven animals), or 12 μg of rhTGF-β2 combined with 25 μg of rhBMP-2 (Group 3, seven animals). In these animals, the twenty-one implants that were placed in the right humerus were loaded with buffer only to serve as contralateral controls. In Group 4 (seven animals), the implants were not coated with hydroxyapatite-tricalcium phosphate, the gap in the left humerus was lightly packed with autogenous bone graft, and the gap in the right humerus was left empty to serve as a contralateral control. All animals were killed at twenty-eight days. The primary end points included three mechanical variables: fixation strength, interface stiffness, and energy to failure. Secondary end points included bone ingrowth and bone volume and trabecular architecture in the gap and in a region located 2 mm medial to the implantation site. Results: The hydroxyapatite-tricalcium phosphate coating had no effect on implant fixation, bone ingrowth, or bone formation in the 3-mm gap. Individual growth factor treatments led to 2.3 to 3.2-fold increases in fixation strength and stiffness as compared with the values for the contralateral controls (p < 0.05). The combined growth factor treatment led to 5.7-fold increases in fixation strength and stiffness compared with the values for the contralateral controls (p < 0.01). Autogenous bone graft treatment was associated with 4.5 to 6.4-fold increases in implant fixation strength and stiffness as compared with the values for the contralateral controls (p < 0.01). Compared with the relevant contralateral controls, energy to failure was increased 3.5-fold in association with TGF-β2 alone (p < 0.05), 4.5-fold in association with TGF-β2 combined with BMP-2 (p < 0.01), and 2.5-fold in association with autogenous bone-grafting. As much as 63% of the variance in the mechanical end points was associated with variance in bone volume and architecture in the 3-mm gap and in the region of interest located 2 mm medial to the implantation site (p < 0.01). Conclusions: In this animal model, the combined use of TGF-β2 and BMP-2 led to more secure mechanical fixation of the implant than did the use of either growth factor alone and demonstrated results that were similar to those associated with the use of autogenous bone graft. Clinical Relevance: This growth factor cocktail is more useful for enhancing implant fixation than individual growth factor therapy and may provide an alternative strategy to autogenous bone-grafting.

Mandibular ramus height restoration by distraction osteogenesis (DO) is a key procedure in mandibular hypoplasia reconstruction. The objective of this study was to evaluate short-term skeletal changes in the regenerated bone after vertical mandibular ramus DO using a buried distraction device. Eight subadult beagle dogs underwent bilateral vertical mandibular ramus DO. After a 7-day latency period, distraction was performed at a rate of 0.5 mm twice a day for 12 days. Four dogs were killed at 1 month and four dogs at 2 months after the end of distraction. One intact beagle was included as an unoperated control. After sacrifice, micro computed tomography (muCT) and mechanical testing of distracted sites were used to measure bone volume (BV), total volume (TV), and mechanical peak load strength, respectively. The muCT images showed wide variation in the response, with some animals demonstrating considerable bone formation and reconstitution of the canal for the inferior alveolar nerve. Quantitatively, BV was no more than 67% and BV/TV was less than 25% of the intact control, and strength was approximately 33% of the intact control value. The 1 and 2 month values were similar. These results suggest that internal distractors can successfully reconstitute bone but that the regenerated tissue did not regain structural and mechanical characteristics of native bone within the 2 month study period.

The purpose of this study was to determine if acetabular articular cartilage damage occurs in the presence or absence of changes in subchondral plate thickness or porosity and trabecular bone architecture after hip hemiarthroplasty. Eight canines were sacrificed 6 months after receiving unilateral hemiarthroplasties in which a cobalt chrome alloy femoral head was used. The acetabular cartilage, subchondral plate, and trabecular bone were quantitatively evaluated. Although the articular cartilage in the treated hip showed gross and histologic degenerative changes, there were no differences in the treated and contralateral hips in any of the trabecular bone parameters or subchondral plate thickness. However, the subchondral plate porosity was increased 2.6-fold in the treated hip. Therefore, degradation of cartilage can occur in the absence of thickening of the subchondral plate or alterations in the supporting trabecular bone architecture. These observations provide a better understanding of the role that periarticular bone has in the degenerative process after hemiarthoplasty.

Various anabolic agents, including transforming growth factor-beta (TGF-beta), have been shown to enhance intramembranous bone regeneration and strengthen the mechanical connection between implant and host skeleton, a prerequisite for clinical success with orthopedic and dental implants. Mechanisms underlying these observations at the level of the gene have received little attention. A rat model was used to examine levels of gene transcription for 21 "osteogenic" genes by real-time polymerase chain reaction at days 1, 3, 5, 7, 10, 14, and 28 in a control group and a group in which the implant was treated with 1 microg recombinant human TGF-beta2 (n = 42, equally divided among the 2 groups and 7 time points). Genes were chosen to represent three functional categories: (1) growth factors, their receptors and antagonists; (2) bone differentiation markers; and (3) inflammation markers. Examination of the transcription profiles showed that nine genes had up-regulated or down-regulated expression levels without a change in timing and 12 genes had accelerated or delayed expression profiles with or without a concomitant change in maximal or minimal expression. The earliest changes (days 1-3) involved accelerated expression profiles for IGF-1R and VEGF and up-regulation of TGF-beta2, TbetaRI, BMP-2, BMP-7, and Cbfa1. Furthermore, principal components analyses showed that some subsets of genes were co-expressed in both groups, although the temporal relationship of these subsets was altered following growth factor treatment. Thus, in addition to changes in individual transcription profiles, the regulatory connections between sets of co-expressed genes may also be affected by exogenously delivered anabolic agents during bone regeneration.

Several previous studies of bone repair have shown 2- to 4-fold increases in bone formation following local delivery of exogenous transforming growth factor-beta (TGF-beta). Here, we use quantitative backscatter electron microscopy to test the effect of TGF-beta1 on mineralization of regenerated bone by examining tissue samples from a previously published canine study in which we found increased bone formation. In the experiment, the proximal humeri of 10 male canines were implanted bilaterally for 28 days with porous-coated implants in the presence of a 3 mm gap between the surface of the implant and the host bone. Implants placed in the left humeri were treated with TGF-beta1 at a dose of either 120 microg (n = 5) or 335 microg (n = 5), and the implants placed in the contralateral humeri served as untreated controls. Quantitative backscatter scanning electron microscopy was used to assess the volume fraction of bone and its degree of mineralization in the 3 mm gaps. The calibrated grayscale mean and median values were depressed compared to the controls in the high dose group (p = 0.048 and p = 0.041, respectively), suggesting that high dose TGF-beta delayed or inhibited mineralization of newly formed osteoid.

Rat bone marrow stromal cells were cultured on either Ra (0.14 μm) or Ra (5.8 μm) Ti6Al4V discs for 24 or 48 h. Cells on the Ra (0.14 μm) surface showed typical fibroblastic morphology, whereas cells on the Ra (5.8 μm) surface were in clusters with a more epithelial appearance. RNA was extracted from the cells at both time points, and gene expression was analyzed by using a rat gene microarray. At 24 and 48 h, a similar number of genes were both up- and down-regulated at least twofold on the Ra (5.8 μm) surface compared to the Ra (0.14 μm) surface. We analyzed the relative level of specific groups of genes related to bone and cartilage development, cell adhesion and extracellular matrix proteins, transcription factors, bone morphogenetic proteins, phospholipases, and protein kinases. Roughness did not appear to be a specific stimulator of osteogenesis because genes of both the bone and cartilage lineage were up-regulated on the Ra (5.8 μm) surface. The most prominent change among transcription factors was up-regulation of Hox 1.4 on the Ra (5.8 μm) surface. Up-regulation of phospholipase A2 and SMAD 4 indicate these genes are also involved in the response of cells to an Ra (5.8 μm) surface. Our data show surface roughness alters the expression of a large number of genes in marrow stromal cells, which are related to multiple pathways of mesenchymal cell differentiation. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 70A: 391–401, 2004

This study investigated the association between non-invasive measurements of bone mass and markers of dynamic and static hip joint loads in subjects expected to be at peak bone mass. The bone mineral density (BMD) and bone mineral content (BMC) of three proximal femoral sites (neck, greater trochanter, and total) were measured by dual energy X-ray absorptiometry, and the peak external joint moments at the hip during walking and jogging were calculated from gait analyses of 31 normal human subjects ranging in age from 30 to 49 years (18 females, 13 males). Various multiple regression analyses were performed to determine how much of the variance in BMD and BMC was explained by height, body mass, and the peak hip joint moments. In total, the models explained up to 40% of the variance in BMD and 58% of the variance in BMC. Inclusion of height or body mass did not increase the explanatory power of the models for BMD and explained no more than 8% of the total variance in BMC once the joint moments from walking were allowed to enter the models. These data support the hypothesis that variance in peak bone mass is associated with variance in dynamic hip loads largely independent of the effect of static factors such as height and body mass.

The present study demonstrates increased early mechanical fixation of titanium implants coated with a new biomimetic apatite surface in a rat model. Male Sprague- Dawley rats received unilateral femoral medullary implants for periods of 1- 4 weeks. The strength of fixation of the implant to the host bone increased more rapidly in the group receiving apatite-treated implants compared with the control group as evidenced by the apatite group's 21-fold greater fixation strength at 1 week (p 0.009), 4-fold greater fixation strength at 2 weeks (p 0.041), and 2-fold greater fixation strength at 4 weeks (p 0.093) compared with the control. Fixation strength was correlated with bone-implant contact as determined from micro computed tomography assessment of the specimens (r 2 0.338, p 0.011 in the control group and r 2 0.543, p 0.001 in the apatite group). Furthermore, for a given amount of bone-implant contact, the fixation strength was higher in the apatite group than in the control group (p 0.011), suggesting that the bone formed a stronger bond to the apatite coating than to the titanium. This difference in bonding strength accounted for the difference in mechanical behavior. © 2004 Wiley Period- icals, Inc. J Biomed Mater Res 70A: 66 -73, 2004

Joint moments are commonly used to characterize gait. Factors like height and weight influence these moments. This study determined which of two commonly used normalization methods, body mass or body weight times height, most reduced the effects of height and weight on peak hip, knee, and ankle external moments during walking. The effectiveness of each normalization method in reducing gender differences was then tested. Gait data from 158 normal subjects were analyzed using unnormalized values, body mass normalized values, and body weight times height normalized values. Without normalization, height or weight accounted for 7-82% of the variance in all 10 peak components of the moments. With normalization, height and weight accounted for at most 6% of the variance with the exception of the hip adduction moment normalized by body weight times height and the ankle dorsiflexion moment normalized by body mass. For the hip adduction moment normalized by body weight times height, height still accounted for 13% of the variance (p

Osteoarthritis is a disease of synovial joints that involves articular cartilage breakdown with accompanying bone changes, including subchondral sclerosis and osteophytosis. However, conflicting data have been reported concerning the cause-and-effect relationship, if any, between these changes. The authors studied the subchondral plate (subchondral bone plus calcified cartilage) in relation to the degree of articular cartilage degeneration on the distal articular surface of the first metatarsal, a region prone to osteoarthritis. No correlation was found between subchondral plate thickness or porosity and the degree of cartilage degeneration in the study sample of 96 metatarsals. Owing to the suggestion that initiation of cartilage fibrillation may be a result of steep stiffness gradients in the subchondral bone, the ratios of subchondral plate thickness in adjacent regions of the metatarsal head were examined in detail, but no correlation was found with subchondral degeneration. Thus increases in subchondral bone thickness are not associated with increases in cartilage degeneration on the first metatarsal, which may imply that subchondral bone changes do not cause osteoarthritis in this joint. (J Am Podiatr Med Assoc 93(2): 104-110, 2003)

Objective The purpose of this work was to test the effect of inhibition of bone remodeling, through the use of the bisphosphonate, zoledronic acid, on cartilage matrix damage in an animal model of cartilage matrix damage. Design New Zealand white rabbits were divided into four groups for treatment purposes: (1) untreated controls; (2) injected into one knee joint with the cartilage matrix degradation enzyme, chymopapain; (3) injected into one knee joint with chymopapain and also given subcutaneous injections of the bisphosphonate, zoledronic acid, three times per week until sacrifice at either day 28 or 56 post-chymopapain-injection; (4) received only the zoledronic acid injections. At sacrifice, the knee joints were examined grossly and histologically, and biochemically for proteoglycan content. Urine samples were analysed, at intervals, for levels of collagen cross-links which are biochemical markers of cartilage and bone. Results Animals receiving both intraarticular chymopapain injections and subcutaneous zoledronic acid injections displayed a significantly lower degree of grossly and histologically detectable cartilage degeneration on the tibial articular surfaces (the articular surface displaying the greatest degree of degeneration) than did animals only receiving the chymopapain injections. In addition, urinary levels of collagen cross-links for bone and cartilage were significantly higher in those animals only receiving chymopapain injections. Conclusion The bone resorption observed after chymopapain injection into the rabbit knee joint can be inhibited through the use of the bisphosphonate, zoledronic acid. Furthermore, zoledronic acid does not increase the level of cartilage degeneration and appears to provide some level of chondroprotection in this model.

Osteoarthritis (OA) is a common, disabling condition of synovial joints that can eventually lead to reduced, or lost, mobility. It is characterized by both articular cartilage degeneration and subchondral bone changes. However, a cause-and-effect relationship between the two tissues remains controversial. Increased subchondral bone density has been associated with early degenerative changes in the cartilage of knee, hip, and finger joints-joints in which progressive changes to OA are common. In contrast, the ankle joint is known to exhibit early cartilage changes, but is not prone to the development of OA. In the present study, it was found that cartilage degeneration on the talus is not associated with an increase in bone density, as assessed through peripheral quantitative computed tomography (pQCT).

The purposes of the present study were to determine if recombinant human transforming growth factor-beta-2 (rhTGF-beta2) enhances bone ingrowth into porous-coated implants and bone regeneration in gaps between the implant and surrounding host bone. The implants were placed bilaterally for four weeks in the proximal humeri of skeletally mature, adult male dogs in the presence of a 3-mm gap. In three treatment groups of animals, the test implant was treated with hydroxyapatite/tricalcium phosphate (HA/TCP) and rhTGF-beta2 in buffer at a dose per implant of 1.2 microg (n = 6), 12 microg (n = 7), or 120 microg (n = 7) and placed in the left humerus. In these same animals, an internal control implant treated only with HA/TCP and buffer was placed in the right humerus. In a non-TGF-beta treated external control group of animals (n = 7), one implant was treated with HA/TCP while the contralateral implant was not treated with the ceramic. In vitro analyses showed that approximately 15%, of the applied dose was released within 120 h with most of the release occurring in the first 24 h. The TGF-beta treated implants had significantly more bone ingrowth than the controls with the greatest effect in the 12 microg/implant group (a 2.2-fold increase over the paired internal control (P = 0.004) and a 4-fold increase over the external control (P < 0.001)). The TGF-beta treated implants had significantly more bone formation in the gap than the controls with the greatest effect in the 12 and 120 microg groups (1.8-fold increases over the paired internal controls (P = 0.003 and P = 0.012, respectively) and 2.8-fold increases over the external controls (P < 0.001 and P = 0.001, respectively)). Compared to the external controls, the internal control implants tended to have more bone ingrowth (1.9-fold increase, P = 0.066) and had significantly more bone formation in the gap (1.7-fold increase. P = 0.008). Thus, application of rhTGF-beta2 to a porous-coated implant-stimulated local bone ingrowth and gap healing in a weakly dose-dependent manner and stimulated bone regeneration in the 3-mm gap surrounding the contralateral control implant, a site remote from the local treatment with the growth factor.

The purpose of this study was to quantify the volume and density of cancellous bone available from 3 commonly used bone graft sites in upper extremity surgery: the distal radius, olecranon process, and anterior iliac crest. Sixteen cadavers (age range, 59-98 years) with no prior history of bone harvest or metabolic conditions affecting bone were used. Cancellous graft was obtained using standardized techniques designed to simulate the clinical setting. Packed cancellous bone volume was determined as the volume occupied by the harvested bone after compression packing with a uniform load. Defect volume was determined by measuring the volume of the site from which bone was harvested. The distal radius and olecranon provided similar volumes of packed cancellous bone (2.7 and 2.8 cc, respectively). The anterior iliac crest provided approximately twice this amount (5.3 cc). The packed cancellous bone volume/defect volume ratio was not different between the 3 sites studied. Male gender was associated with a significantly greater amount of packed cancellous bone volume for all sites. We believe the olecranon to be an alternative to the distal radius as a source of bone graft for upper extremity procedures. We suggest using the anterior iliac crest when a large volume of cancellous bone is required.

Bone repair is regulated by biological factors and the local mechanical environment. We hypothesize that the combined use of low-intensity pulsed ultrasound (LIPUS) and recombinant human bone morphogenetic protein-2 (rhBMP-2) will synergistically or additively enhance bone regeneration in a model simulating the more difficult scenarios in orthopaedic traumatology.Femoral defects in rats were replaced with absorbable collagen sponges carrying rhBMP-2 (0, 1.2, 6, or 12 μg; n = 30). Each group was divided equally to receive daily treatment of either LIPUS or sham stimulation. At 4 weeks, new bone formation was assessed using quantitative (radiography and microcomputed tomography), qualitative (histology), and functional (biomechanical) end points.LIPUS with 1.2 μg of rhBMP-2 significantly improved the radiographic healing as compared with its sham control starting as early as 2 weeks. Quantitatively, the use of LIPUS with 6 μg of rhBMP-2 significantly increased the bone volume. However, using LIPUS with 12 μg of rhBMP-2 indicated a reduction in callus size, without compromising the bone volume, which was also observable histologically, showing organized lamellar bone and repopulated marrow in the original defect region. Histologically, 1.2 μg of rhBMP-2 alone showed the presence of uncalcified cartilage in the defect, which was reduced with LIPUS treatment. Biomechanically, LIPUS treatment significantly increased the peak torsion and stiffness in the 6- and 12 μg rhBMP-2 groups.LIPUS enhances rhBMP-2-induced bone formation at lower doses (1.2 and 6 μg) and callus maturation at 12-μg dose delivered on absorbable collagen sponge for bone repair in a rat critical-sized femoral segmental defect.

Changes in long bone cross-sectional geometry during growth can be influenced by biological and mechanical factors. Here, we assess relationships between cross-sectional geometric properties and length of the human humerus and femur during postnatal growth to test the hypothesis that loading history plays an important role in the development of adult bone morphology. A skeletal sample including 83 paired humeri and femora from individuals between birth and age 30 was examined. Midshaft cross-sectional geometric properties were determined based on computed tomographic scans and the two bones were compared by examining growth trajectories and scaling relationships between the cross-sectional properties and bone length. The growth trajectories for both bones were similar in many respects and showed that increase in length ceased by age 20, whereas increase in cross-sectional properties continued into the third decade of life. When compared to bone length, the cross-sectional geometric properties of the femur and humerus were similar early in postnatal life, but increased at a greater rate in the femur particularly during the first decade of life, leading to divergent adult morphologies. A beam model was developed to predict maximum midshaft strains in each bone as a function of age. The moment acting on the femur was estimated from an analysis of gait in children and the moment acting on the humerus was chosen so that the magnitude of the maximum midshaft strains in the two bones was equivalent in adulthood. With this model, the maximum midshaft strains for the femur were predicted to be higher than for the humerus during the first decade of life. These data support the concept that load history plays an important role in accretion of bone mass during postnatal growth.

Enhancement of bone ingrowth with transforming growth factor-beta was evaluated in a canine model. Ten dogs had bilateral implantation of a titanium-fiber-metal-coated rod in the proximal part of the humerus. A three-millimeter gap between the outer surface of the porous coating and the surrounding cancellous bone was created to impair bone ingrowth. All of the implants were plasma-flame-sprayed with hydroxyapatite and tricalcium phosphate. In each animal, one implant was also treated with recombinant transforming growth factor-beta 1 while the other implant, which was not so treated, served as a paired control. Two doses of transforming growth factor-beta 1 were used: 335 micrograms in five animals and 120 micrograms in the other five. At four weeks, the amount of bone ingrowth in the implants that had been treated with 120 micrograms of transforming growth factor-beta 1 was threefold higher than that in the paired controls (p = 0.009), but with the numbers available there was no significant increase in bone ingrowth with the higher dose. The amount of new-bone formation in the three-millimeter gaps adjacent to the treated implants was twice that in the gaps of the paired controls, regardless of the dose. The differences between the treated and control implants with regard to the architecture of the new bone in the gap indicate that the mechanism of action of transforming growth factor-beta 1 may include both proliferation of osteoprogenitor cells and production of matrix by committed osteoblasts. Compared with the findings in a previous study in which this canine model was used, the data from the present investigation indicate that enhancement of bone ingrowth in implants that have been treated with a combination of a hydroxyapatite-tricalcium phosphate coating and transforming growth factor-beta 1 may exceed that obtainable with grafting of the gap with autogenous cancellous bone.

Bone ingrowth and the distribution of wear debris within the porous coating of 13 primary cementless porous-coated tibial components removed for reasons unrelated to fixation or infection were quantitatively described. The average length of implantation was 15.3 months (range, 3–30 months). The implants were all of the same design, made from Ti6A14V with a commercially pure titanium fiber-metal porous coating, which covered the undersurface of the tray and the four fixation pegs. In all but one component, supplemental screw fixation was used. The average extent of bone ingrowth within the tray was 27.1 ± 16.1%, and the average volume fraction was 9.5 ± 7.5%. There was significantly more bone ingrowth within the fixation pegs than within the tray and also more bone ingrowth in the anterior hall of the tray than posteriorly. There was no correlation between the amount of bone ingrowth and the length of implantation, age, or sex of the patient; however, the depth and orientation of the resection plane were found to correlate with the topographic distribution of bone ingrowth. Particulate debris appeared to gain access to the interface via soft tissue pathways both at the periphery and through the holes for adjuvant screw fixation.

SummaryObjectiveThe current study was undertaken to adapt Equilibrium Partitioning of an Ionic Contrast agent via microcomputed tomography (EPIC-μCT) to mouse articular cartilage (AC), which presents a particular challenge because it is thin (∼30 μm) and has a small volume (0.2–0.4 mm3), meaning there is only approximately 2–4 μg of chondroitin sulfate (CS) glycosaminoglycan per joint surface cartilage.DesignUsing 6 μm isotropic voxels and the negatively charged contrast agent ioxaglate (Hexabrix), we optimized contrast agent concentration and incubation time, assessed two methods of tissue preservation (formalin fixation and freezing), examined the effect of ex vivo chondroitinase ABC digestion on X-ray attenuation, assessed accuracy and precision, compared young and skeletally mature cartilage, and determined patterns of degradation in a murine cartilage damage model induced by treadmill running.ResultsThe optimal concentration of the contrast agent was 15%, formalin fixation was preferred to freezing, and 2 h of incubation was needed to reach contrast agent equilibrium with formalin-fixed specimens. There was good agreement with histologic measurements of cartilage thickness, although μCT over-estimated thickness by 13% (∼5 μm) in 6-week-old mice. Enzymatic release of 0.8 μg of chondrotin sulfate (about 40% of the total) increased X-ray attenuation by ∼17%. There was a 15% increase in X-ray attenuation in 14-week-old mice compared to 6-week-old mice (P

One hundred and thirty-eight consecutive revisions of an acetabular component were performed in 132 patients between 1983 and 1986. The revision prosthesis was a hemispherical component that was coated with porous titanium mesh and was secured to the pelvis with a variable number of screws. There were seventy-five women and fifty-seven men, and the mean age at the time of the revision was fifty-two years (range, twenty to seventy-nine years). Due to defects in the acetabulum, 80 per cent of the hips were treated with bone grafts, usually a mixture of local autogenous graft and freeze-dried allograft. One hundred and twenty-four patients (129 hips) were available for follow-up, at a mean of forty-four months (range, thirty-six to eighty months). Seven hips (5 per cent) were revised again: four because of infection and three because of instability. There were no revisions for loosening without infection, and none of the components migrated in the absence of infection. Radiolucent lines were common and usually corresponded to regions in which allograft had been used. All bone grafts united by twelve months. Non-contained medial grafts underwent a phenomenon of resorption, consolidation, and remodeling to a sclerotic rim by twenty-four months. There were no complications related to placement of the screws. The results in this series were superior to results of revisions of acetabular components with the use of cement, which were associated with rates of failure of almost 10 per cent after a similar duration of follow-up. The technique of maximization of host-bone coverage and of packing of all defects with cancellous autogenous graft or allograft, or both, was successful for all classes of acetabular deficiencies.

The authors examined 11 cementless acetabular components of one design retrieved at autopsy and made observations concerning tissue ingrowth and local tissue reaction, radiographic-histologic correlation, and the distribution of particulate wear debris. The cups were hemispherical in design with a commercially pure titanium fibermetal porous coating. All of the prostheses were implanted with screws. The implants were in place for an average of 41 months (range, 5 weeks to 75 months). Ten of the cups had bone ingrowth, with the average volume fraction being 12.1±8.2%. There were no differences in the amount of bone ingrowth when the component was partitioned into nine anatomic regions. However, there was more bone adjacent to screw holes through which screws were inserted compared with empty screw holes. As the number of radiolucent zones increased on the clinical radiographs less bone ingrowth was observed histologically. The amount of metal debris in holes with screws and holes without screws was similar. In the longest term cases, polyethylene debris was noted within empty screw holes, but no granulomatous reactions or osteolytic processes were observed.

25 hemispherical acetabular components retrieved from patients who had a total hip arthroplasty (THA) were studied histologically. All of the components had titanium fiber metal porous coatings and were implanted with adjuvant screw fixation, 18 in primary THA and 7 in revision THA. None of the cups had been removed because of infection or failure of fixation. The components had been in place an average of 30 (1-140) weeks. 18 cups had bone ingrowth into the bone-implant interface with bone ingrowth was over 80 percent. Bone ingrowth was more often observed at the dome and in the vicinity of sites of screw fixation than elsewhere. Peripherally, bone ingrowth was less consistent, and in isolated areas fibrocartilage and mineralized fibrocartilage were occasionally observed. The longer-term cases had more bone ingrowth than the shorter-term cases. Apparent metallic debris was observed within histiothe porous coating. Up to one third of the available void space within the porous coating was occupied by bone, while the maximal proportion of the area of Departments of Orthopedics, 'Rush-Presbyterian-St. Luke's Medical Center, 1653 West Congress Parkway, Chicago, IL 6061 2, and *Massachussetts General Hospital, Boston, MA 02114, U.S.A. Tel+1-312 942 8151 or 5850. Fax -312 942 2040 Submitted 92-10-1 8. Accepted 93-05-04 cytes adjacent to screw holes, and the amount of this material increased with duration of implantation. - ___-~ ~ ~~ _~__

Low-intensity pulsed ultrasound (LIPUS) is an established therapy for fracture repair and has been used widely in the clinics, but its underlying mechanism of action remains unclear. The aim of the current research was to determine the effect of LIPUS on gap junctional cell-to-cell intercellular communication in rat bone marrow stromal cells (BMSC) in vitro and to determine whether the ability of BMSCs to communicate by gap junctions would affect their response to LIPUS. Single or daily-multiple LIPUS treatment at 1.5 MHz, 30 mW/cm2, for 20 min was applied to BMSC. We demonstrated that BMSC form functional gap junctions and single LIPUS treatment significantly increased the intracellular dye transfer between BMSC. In addition, activated phosphorylation of ERK1/2 and p38 by LIPUS stimulation was diminished when cells were treated with a gap junction inhibitor 18β-glycyrrhetinic acid (18β). We further demonstrated that 18β diminished the significant increase in alkaline phosphatase activity following LIPUS stimulation. These results suggest a potential role of gap junctional cell-to-cell intercellular communication on the effects of LIPUS in BMSC.

The canine is often used as a model to study functional bone adaptation after total hip replacement. To improve our understanding of the model, we defined the central tendencies and statistical variations in the cross-sectional geometry, angle of anteversion, and cervicodiaphyseal angle of the proximal femur in 15 adult male mongrel dogs and compared the results with published reports of the human femur. Numerous similarities in the cross-sectional geometry of the canine and the human femur were noted, supporting the use of the canine as a model. The two species differed in that the orientation of the principal axes in the proximal cross sections was not related to the angle of anteversion in the canine femur, whereas these angles are related in humans. In addition, the canine medullary canal is larger than the human medullary canal relative to the external dimensions of the femur, and hence the canine has relatively thinner cortical bone. This difference in femoral cross-sectional geometry may explain, in part, why the canine provides an accentuated model of bone loss in hip arthroplasty experiments.

Angiogenesis is essential for bone formation and several bone morphogenetic proteins (BMPs) have been shown to induce angiogenesis through osteoblast-derived vascular endothelial growth factor (VEGF)-A. Growth differentiation factor-5 (GDF-5) is a member of the BMP family expressed in bone and known to induce angiogenesis in vivo. In this study, the effects of GDF-5 on osteogenic differentiation and expression of VEGF-related genes were determined using rat bone marrow stromal cells. GDF-5 stimulated osteogenic differentiation. It also upregulated the expression of VEGF-A after 3 hr, accompanied by a trend of decrease in its receptor VEGFR-2 at 6 and 24 hr. VEGF-D and its receptor VEGFR-3 showed peak expression at later time points. This regulation may be further controlled by neuropilin 2 that exhibited a parallel profile to VEGF-D. These observations indicate that GDF-5 stimulates osteogenic differentiation and has a potential to induce angiogenesis through osteoblast-derived VEGF-A in bone.

Interpenetrating polymer networks (IPNs) of poly (acrylamide-co-ethylene glycol/acrylic acid) functionalized with an -Arg-Gly-Asp- (RGD) containing 15 amino acid peptides, derived from rat bone sialoprotein (bsp-RGD(15), were grafted to titanium implants in an effort to modulate bone formation in the peri-implant region in the rat femoral ablation model. Bone-implant contact (BIC) and bone formation within the medullary canal were determined using microcomputed tomography at 2 and 4 weeks postimplantation. BIC for bsp-RGD(15)-IPN implants was enhanced relative to hydroxyapatite tricalcium phosphate (HA-TCP) coated implants, but was similar to all other groups. Aggregate bone formation neither indicated a dose-dependent effect of bsp-RGD(15) nor a meaningful trend. Mechanical testing of implant fixation revealed that only the HA-TCP coated implants supported significant (>1 MPa) interfacial shear strength, despite exhibiting lower overall BIC, an indication that bone ingrowth into the rougher coating was the primary mode of implant fixation. While no evidence was found to support the hypothesis that bsp-RGD(15)-modified IPN coated implants significantly impacted bone-implant bonding, these results point to the lack of correlation between in vitro studies employing primary osteoblasts and in vivo wound healing in the peri-implant region.

Based on the premise that bone mass and bone geometry are related to load history and that subchondral bone may play a role in osteoarthritis (OA), we sought to determine if static and dynamic markers of knee joint loads explain variance in the medial-to-lateral ratio of proximal tibial bone mineral density (BMD) in subjects with mild and moderate medial knee OA. We utilized two surrogate markers of dynamic load, the peak knee adduction moment and the knee adduction angular momentum, the latter being the time integral of the frontal plane knee joint moment. BMD for medial and lateral regions of the proximal tibial plateau and one distal region in the tibial shaft was measured in 84 symptomatic subjects with Kellgren and Lawrence radiographic OA grades of 2 or 3. Utilizing gait analysis, the peak knee adduction moment (the external adduction moment of greatest magnitude) and the time integral of the frontal plane knee joint moment (the angular momentum) over the entire stance phase as well as for each of the four subdivisions of stance were calculated. The BMD ratio was not significantly different in grade 2 (1.32 +/- 0.27) and grade 3 knees (1.47 +/- 0.40) (P = 0.215). BMD of the tibial shaft was not correlated with any loading parameter or static alignment. Of all the surrogate gait markers of dynamic load, the knee adduction angular momentum in terminal stance explained the most variance (20%) in the medial-to-lateral BMD ratio (adjusted r(2) = 0.196, P < 0.001). The knee adduction angular momentum for the entire stance phase explained 18% of the variance in the BMD ratio (adjusted r(2) 0.178, P < 0.001), 10% more variance than explained by the overall peak knee adduction moment (adjusted r(2) = 0.081, P < 0.001). 18% of the variance in the BMD ratio was also explained by the knee alignment angle (adjusted r(2) = 0.183, P < 0.001), and the total explanatory power was increased to 22% when the knee adduction angular momentum in terminal stance was added (change in r(2) = 0.041, P < 0.05, total adjusted r(2) = 0.215, P < 0.001). The BMD ratio and its relationship to dynamic and static markers of loading were independent of height, weight, and the body mass index, demonstrating that both dynamic markers of knee loading as well as knee alignment explained variance in the tibial BMD ratio independent of body size. (c) 2006 Elsevier Inc. All rights reserved.

Tissue formation and repair are dependent upon cascades of biological events, but the signals involved and the possible gene coexpression patterns during intramembranous bone repair are only poorly understood. We sought to place this mode of regeneration in context by profiling quantitative gene expression for a panel of 39 genes between days 1 and 14 following rat femoral marrow ablation. In situ hybridization was employed to localize a subset of genes. Additionally, principal components analysis was conducted to identify underlying factors suggestive of coexpression patterns. During inflammation (days 1-5), several genes, including cyclooxygenase-1 and -2, showed downregulation. Other proinflammatory cytokines, tumor necrosis factor-alpha and interleukin-1beta, exhibited increasing levels around day 5. During repair (days 3-10), growth factors, receptors, and inhibitor genes for transforming growth factor- beta; basic fibroblast growth factor; bone morphogenetic proteins 2, 4, and 7; vascular endothelial growth factor; and insulin-like growth factor-I were upregulated. In addition, the gene for core binding factor-alpha1 and markers of osteoblast function such as alkaline phosphatase, collagen type I, osteonectin, osteopontin, and osteocalcin had peak expression at day 5 or 7. The remodeling phase (days 10-14) was characterized by peaks for cytokines associated with osteoclastic activity including receptor activator of nuclear factor-kappaB, receptor activator of nuclear factor-kappaB ligand (RANKL), cathepsin K, tumor necrosis factor-alpha, interleukin-6, and cyclooxygenase-2. In situ hybridization showed that the most common sites of increased signal were within osteoblastic cells on trabecular and endosteal surfaces. Principal components analysis identified eight underlying factors that together explained over 80% of the variance in the data.

Orthopedic and dental implant fixation depends upon bone regeneration. Growth factors such as transforming growth factor-beta (TGF-beta) have been shown to enhance bone repair and strengthen the mechanical connection between implant and host skeleton in canine models. To provide a platform for studying molecular mechanisms of growth factor stimulated bone regeneration and implant fixation, the present study examined peri-implant bone volume as a response to TGF-beta treatment in a rodent model. The rat femoral ablation model in which an implant is placed in the medullary cavity of the femur was used to examine the dose response to TGF-beta2 applied to the implant (0, 0.1, 1.0, or 10 microg). The study included a total of 40 rats (10 per dose) examined at 28 days. Peri-implant bone volume and bone-implant contact were assessed through microcomputed tomography and implant fixation strength was determined by a mechanical pullout test. Treatment of the implant with 10 microg TGF-beta2 led to a 2-fold increase in bone volume (P

To determine how low intensity pulsed ultrasound alters gene expression in rat bone marrow stromal cells and to see if combining this stimulation with BMP-2, cells were pre-cultured for eight days in the presence of 50 microg/ml ascorbic acid and then exposed to either low intensity US or 100 ng/ml BMP-2 or both combined, beginning on the first, third fifth or seventh day of culture so that cells were exposed to the stimuli for one, three, five or seven days. Real time PCR was used to determine the effect of these treatments on gene expression of several genes associated with osteogenesis. The expression of some of the genes (Cbfa-1/Runx2, IGF-receptor, Alk-3, alkaline phosphatase, osteopontin, TGF-beta1, BMP-7) was increased compared to untreated controls. Combination of US and BMP-2 treatment did not lead to synergy of the two stimuli. Cbfa-1 stimulation occurred more quickly with US than with BMP-2. Increases in gene expression were greatest after 3 days exposure to US, with similar results for BMP-2 treatment implying that there may be a time dependence for the stimulus of osteogenic gene expression in stromal cells.

A prospective, randomized, pilot study comparing 500 cGy (group A) versus 1,000 cGy (group B) radiation treatment for the prevention of heterotopic bone in a consecutive group of high-risk patients undergoing total hip arthroplasty was performed. Treatment was initiated within ninety-six hours and given in equal, divided doses: 2 doses for group A and 5 doses for group B. No statistically significant difference was found in the demographics between the 2 groups. At follow-up evaluation, the distribution of heterotopic bone according to the Brooker classification was: group A, 9 class 0, 17 class 1, one class 2, and 2 class 3. Group B: 17 class 0, 10 class 1, 2 class 2, and one class 3. This difference was not statistically significant (P=.086). Only 3 cases were considered treatment failures, for a success rate of 93% in group A and 97% in group B. As a result of this pilot study, 500 cGy radiation treatment appears to be effective in the prevention of clinically significant heterotopic ossification after total hip arthroplasty.

Concentrations of the bone resorption markers pyridinoline and deoxypyridinoline and the bone formation marker osteocalcin were measured in 24-h urine collections from 30 subjects who underwent unilateral total hip replacements for monoarticular symptomatic osteoarthrosis and 10 controls. The patient groups were divided based on the femoral implant type (cemented cobalt alloy stem, cementless porous coated cobalt alloy stem, and cementless porous coated titanium alloy stem). Urine collections were performed before surgery and then at 3, 6, 12, 24, and 36 months. There were significant changes over time in the three patient groups for pyridinoline, deoxypyridinoline, and the ratio of osteocalcin to deoxypyridinoline (p < or = 0.01), but the control group values did not change over time. The resorption markers tended to peak at 3 months and the osteocalcin to deoxypyridinoline ratio was more variable, having depressed values in the cementless cobalt alloy group and elevated values in the other two groups compared with baseline. The cementless cobalt alloy group had higher resorption marker levels than the cemented cobalt alloy group at 6, 12, 24, and 36 months and higher levels than the cementless titanium alloy group at all postoperative times (p < 0.05). The osteocalcin to deoxypyridinoline ratio was lower in the cementless cobalt alloy group than in the cemented cobalt alloy group at 3, 6, and 24 months and the cementless titanium alloy group at 6, 12, and 24 months (p < 0.05). For the cemented cobalt chrome group, the baseline-normalized resorption marker values at 3 months and 6 months were correlated with the severity of radiographically assessed bone loss at 36 months (0.749 < r < 0.840; p < 0.05). For the cementless titanium alloy group, baseline-normalized osteocalcin/ deoxypyridinoline ratios at 3 months and 6 months were related inversely to radiographic bone loss at 36 months (0.687 < r < 0.749; p < 0.05). Thus, body fluid markers of bone metabolism change after total hip replacement. In addition, the changes in the marker concentrations were sensitive to implant design and were correlated with subsequent stress-shielding-induced bone loss.

In osteoarthritis, one postulate is that changes in the mechanical properties of the subchondral bone layer result in cartilage damage. The goal of this study was to examine changes in subchondral trabecular bone properties at the calcified tissue level in the early stages of cartilage damage. Finite element models were constructed from microCT scans of trabectilar bone from the proximal tibia of donors with mild cartilage damage and from normal donors. In the donors with cartilage damage, macroscopic damage was present only in the medial compartment. The effective tissue elastic moduli were determined using a combination of finite element models and mechanical testing. The bone tissue modulus was reduced by 60% in the medial condyle of the cases with cartilage damage compared to the control specimens. Neither the presence of cartilage damage nor the anatomic site (medial vs. lateral) affected the elastic modulus at the apparent level. The volume fraction of trabecular bone was higher in the medial compartment compared to the lateral compartment of tibiae with cartilage damage (but not the controls), suggesting that mechanical properties were preserved in part at the apparent level by an increase in the bone volume fraction. It seems likely that the normal equilibrium between cartilage properties, bone tissue properties and bone volume fraction is disrupted early in the development of osteoarthritis.

We investigated, at the whole bone level, the contribution of bone density and geometry to the fracture load of the second metatarsal, a bone that is prone to stress fracture. Dual-energy X-ray absorptiometry (DXA) was used to determine the areal bone mineral density (BMD), projected area of bone, and bone mineral content. Peripheral quantitative computed tomography (pQCT) was used to determine the volumetric cortical bone mineral density (vCtBMD) and cross-sectional moment of interia. Various metatarsal linear dimensions were also measured. The load at failure in cantilever bending was determined. The only linear dimension that had a significant correlation with load at failure was the height of the metatarsal base (r(2) = 0.30, p = 0.008). Utilizing all of the information provided by DXA gave no greater indication of whole bone strength than just BMD alone (adjusted r(2) = 0.40, p = 0.001). Using all of the information provided by pQCT gave no greater indication of whole bone strength than just vCtBMD alone (r(2) = 0. 46, p < 0.001). Volumetric cortical density and BMD were strongly correlated (r(2) = 0.81, p < 0.001). Our data suggest that, in the human second metatarsal, a variable such as material strength (as inferred from cortical density), and not geometry, may be the major factor in determining cantilever load to failure.

The present study examined the loads at the hip joint during gait and the bone mineral density of the proximal femur in 25 patients with end-stage hip osteoarthritis. Dual energy X-ray absorptiometry was used to determine the bone mineral density of the greater trochanter, femoral neck and Ward's triangle of the osteoarthritic group. The bone mineral density was normalized for the patient's age, gender, weight and ethnic origin (Z score). Gait analysis was used to determine the external hip joint moments and motion during walking for the osteoarthritic group and a control group of 21 normal subjects. The gait parameters of the osteoarthritic group which were significantly diminished compared to the normal group (p0.001) accounted for as much as 42% (p0.001) of the variation in the normalized bone mineral density. Specifically, the dynamic sagittal plane hip motion during gait (maximum flexion minus maximum extension) and peak external rotation and adduction moments were significantly correlated with greater trochanter (R = 0.429-0.648, p = 0.032-0.0001) and Ward's triangle (R = 0.418-0.532, p = 0.038-0.006) normalized bone mineral density while the adduction moment was also significantly correlated with the femoral neck normalized bone mineral density (R = 0.5394, p = 0.005). The normalized bone mineral density of the femoral neck and Ward's triangle was elevated while that of the greater trochanter was decreased as compared to normal reference values. The significant correlation between the hip joint moments during gait and femoral bone mineral density indicate that hip joint loads need to be included when explaining local variation in bone mineral density in hip osteoarthritis.

The purpose of the present study was to test the hypothesis that cortical bone loss, trabecular bone density and the amount of bone ingrowth vary as a function of stem stiffness in a canine cementless hip replacement model. The study was motivated by the problem of cortical bone atrophy in the proximal femur following cementless total hip replacement. Two stem stiffnesses were used and both designs were identical in external geometry and porous coating placement. The high stiffness stem caused approximately 26% cortical bone stress-shielding and the low stiffness stem caused approximately 7.5% stress-shielding, as assessed by beam theory. Each group included nine adult, male canines who received unilateral arthroplasties for a period of six months. The animals with the low stiffness stems tended to lose less proximal cortical bone than the animals with high stiffness stems (4% +/- 9 as opposed to 11% +/- 14), but the difference was not statistically significant (p = 0.251). However, the patterns of bone ingrowth into the implant and change in medullary bone density adjacent to the implant were fundamentally different as a function of stem stiffness (p < 0.01). Most importantly, while the high stiffness group had peaks in these variables at the distal end of the stem, the low stiffness group had peak values proximally. These different patterns of functional adaptation are consistent with the idea that reduced stem stiffness enhances proximal load transfer.

It is well known that bone changes occur in relationship to the articular cartilage damage seen in osteoarthritis. Numerous studies have shown a relationship between bone density and articular cartilage damage in knee and hip joints, but it is controversial as to whether or not an increased subchondral bone density results in greater cartilage damage. We studied 49 pairs of cadaveric first metatarsals in an attempt to correlate the level of cartilage damage seen on the distal articular surface with areal and volumetric density and other parameters such as bone geometry. A positive correlation was detected between articular cartilage damage and bone density by peripheral quantitative computed tomography in five of eight frontal slices in the head of the left metatarsal in the male sample only. The more extensive subchondral cysts found in the right male metatarsal may have contributed to the lack of correlation between the two variables on the right side. In addition, females displayed significantly greater osteophytosis than males at a given grade of cartilage damage, thus, possibly leading to a painful, apropulsive gait and a disuse osteopenia which would neutralize any subchondral bone density increase associated with cartilage damage. Microscopic investigations are, therefore, warranted to detect bone changes on a more discreet level, particularly within the subchondral plate.

Dual energy x-ray absorptiometry was used to measure periprosthetic, distal femoral, and proximal tibial bone mass in the affected and contralateral limbs of eight patients 10 years after unilateral total hip arthroplasty with a cementless, porous-coated titanium alloy femoral stem. Gait analyses to assess the presence of asymmetries in loading of the lower extremities were also performed 10 years postoperatively. The patients had excellent clinical results and no other significant lower extremity pathology. On the basis of comparison of the affected and unaffected proximal femora, bone loss adjacent to the proximal medial aspect of the femoral stem was determined to be 34% (p < 0.001). However, the patients also had 16% less bone in the ipsilateral proximal tibia (p = 0.003) and 15% less bone in the ipsilateral femur 3 cm distal to the prosthesis (p = 0.007) compared with the contralateral limb. When normalized to the asymmetry in tibial bone mineral content, the estimated proximal medial periprosthetic bone loss was still statistically significant, but the magnitude was reduced from 34 to 17% (p = 0.009). The gait analyses indicated that several measures that influence the loads at the hip and knee joints were reduced in the involved limb compared with the contralateral limb. Furthermore, the bilateral difference in the vertical component of the external force acting on the proximal tibia was correlated with the bilateral difference in tibial bone mineral content (r = 0.80, p = 0.02). These data suggest that two mechanical factors, the local stress-shielding effect of the prosthesis and the global effect of decreased loading of the limb, can both make significant contributions to periprosthetic bone loss. It is apparent that the magnitude of the periprosthetic bone loss related to stress-shielding has been overestimated by as much as 50% in retrospective studies.

The tibial component of a canine cementless total knee replacement model was used to determine the degree to which pegs and screws contributed to the initial in vitro stability of the device. Three implant designs were investigated: (1) a four-peg implant in which cortical bone screws passed through the pegs, (2) the four-peg implant without adjuvant screw fixation, and (3) a flat implant with screws placed in the same positions as in the first design. For measuring the interface motion, the tibial component and proximal tibia were modeled as rigid bodies and an experimental method was developed which permitted all six degrees of freedom of the motion between these two objects to be determined. In tests performed to validate this methodological approach, the potential confounding influences of tibial deformation and differential amounts of tibial deformation with the use of screws or pegs were shown to be minimal, supporting the use of the rigid-body method. In general, the areas of greatest motion were at the periphery of the bone-implant interface, regardless of whether or not screws or pegs were used. The components secured with screws had up to five-fold reductions in interface motion compared to components which had pegs but lacked screw fixation. Components with pegs and screws and components with screws only had the same amount of interface motion. Thus, in the presence of screw fixation, the addition of pegs did not increase the stability of the tibial component.