The consequences of growth hormone-releasing hormone receptor haploinsufficiency for bone quality and insulin resistance

Objective Growth hormone (GH)/insulin-like growth factor (IGF) axis and insulin are key determinants of bone remodelling. Homozygous mutations in the GH-releasing hormone receptor (GHRHR) gene (GHRHR) are a frequent cause of genetic isolated GH deficiency (IGHD). Heterozygosity for GHRHR mutation causes changes in body composition and possibly an increase in insulin sensitivity, but its effects on bone quality are still unknown. The objective of this study was to assess the bone quality and metabolism and its correlation with insulin sensitivity in subjects heterozygous for a null mutation in the GHRHR. Patients and methods A cross-sectional study was performed on 76 normal subjects (68.4% females) (N/N) and 64 individuals (64.1% females) heterozygous for a mutation in the GHRHR (MUT/N). Anthropometric features, quantitative ultrasound (QUS) of the heel, bone markers [osteocalcin (OC) and CrossLaps], IGF-I, glucose and insulin were measured, and homeostasis model assessment of insulin resistance (HOMAIR) was calculated. Results There were no differences in age or height between the two groups, but weight (P = 0.007) and BMI (P = 0.001) were lower in MUT/N. There were no differences in serum levels of IGF-I, glucose, T-score or absolute values of stiffness and OC, but insulin (P = 0.01), HOMAIR (P = 0.01) and CrossLaps (P = 0.01) were lower in MUT/N. There was no correlation between OC and glucose, OC and HOMAIR in the 140 individuals as a whole or in the separate MUT/N or N/N groups. Conclusions This study suggests that one allele mutation in the GHRHR gene has a greater impact on energy metabolism than on bone quality.

Pamidronate for the treatment of osteoporosis secondary to chronic cholestatic liver disease in Wistar rats

Osteoporosis is a major complication of chronic cholestatic liver disease (CCLD). We evaluated the efficacy of using disodium pamidronate (1.0 mg/kg body weight) for the prevention (Pr) or treatment (Tr) of cholestasis-induced osteoporosis in male Wistar rats: sham-operated (Sham = 12); bile duct-ligated (Bi = 15); bile duct-ligated animals previously treated with pamidronate before and 1 month after surgery (Pr = 9); bile duct-ligated animals treated with pamidronate 1 month after surgery (Tr = 9). Rats were sacrificed 8 weeks after surgery. Immunohistochemical expression of IGF-I and GH receptor was determined in the proximal growth plate cartilage of the left tibia. Histomorphometric analysis was performed in the right tibia and the right femur was used for biomechanical analysis. Bone material volume over tissue volume (BV/TV) was significantly affected by CCLD (Sham = 18.1 ± 3.2 vs Bi = 10.6 ± 2.2%) and pamidronate successfully increased bone volume. However, pamidronate administered in a preventive regimen presented no additional benefit on bone volume compared to secondary treatment (BV/TV: Pr = 39.4 ± 12.0; Tr = 41.2 ± 12.7%). Moreover, the force on the momentum of fracture was significantly reduced in Pr rats (Sham = 116.6 ± 23.0; Bi = 94.6 ± 33.8; Pr = 82.9 ± 22.8; Tr = 92.5 ± 29.5 N; P < 0.05, Sham vs Pr). Thus, CCLD had a significant impact on bone histomorphometric parameters and pamidronate was highly effective in increasing bone mass in CCLD; however, preventive therapy with pamidronate has no advantage regarding bone fragility.

Exploring anorganic bovine bone granules as osteoblast carriers for bone bioengineering: a study in rat critical-size calvarial defects

It is known that current trends on bone bioengineering seek ideal scaffolds and explore innovative methods to restore tissue function. In this way, the objective of this study was to evaluate the behavior of anorganic bovine bone as osteoblast carrier in critical-size calvarial defects. MC3T3-E1 osteoblast cells (1x10(5) cells/well) were cultured on granules of anorganic bovine bone in 24-well plates and after 24 h these granules were implanted into rat critical-size calvarial defects (group Biomaterial + Cells). In addition, other groups were established with different fillings of the defect: Blood Clot (negative control); Autogenous Bone (positive control); Biomaterial (only granules) and Cells (only MC3T3-E1 cells). After 30 days, the animals were euthanized and the calvaria were technically processed in order to allow histological and morphometric analysis. It was possible to detect blood vessels, connective tissue and newly formed bone in all groups. Particularly in the Biomaterial + Cells group, it was possible to observe a profile of biological events between the positive control group (autogenous bone) and the group in which only anorganic bovine granules were implanted. Altogether, the results of the present study showed that granules of anorganic bovine bone can be used as carrier to osteoblasts and that adding growth factors at the moment of implantation should maximize these results.

Assessment of cumulative damage by using ultrasonic C-scan on carbon fiber/epoxy composites under thermal cycling

In recent years, structural composites manufactured by carbon fiber/epoxy laminates have been employed in large scale in aircraft industries. These structures require high strength under severe temperature changes of -56° until 80 °C. Regarding this scenario, the aim of this research was to reproduce thermal stress in the laminate plate developed by temperature changes and tracking possible cumulative damages on the laminate using ultrasonic C-scan inspection. The evaluation was based on attenuation signals and the C-scan map of the composite plate. The carbon fiber/epoxy plain weave laminate underwent temperatures of -60° to 80 °C, kept during 10 minutes and repeated for 1000, 2000, 3000 and 4000 times. After 1000 cycles, the specimens were inspected by C-scanning. A few changes in the laminate were observed using the inspection methodology only in specimens cycled 3000 times, or so. According to the found results, the used temperature range did not present enough conditions to cumulative damage in this type of laminate, which is in agreement with the macro - and micromechanical theory.

Biochemical, bone and renal patterns in hyperparathyroidism associated with multiple endocrine neoplasia type 1

Primary hyperparathyroidism associated with multiple endocrine neoplasia type I (hyperparathyroidism/multiple endocrine neoplasia type 1) differs in many aspects from sporadic hyperparathyroidism, which is the most frequently occurring form of hyperparathyroidism. Bone mineral density has frequently been studied in sporadic hyperparathyroidism but it has very rarely been examined in cases of hyperparathyroidism/multiple endocrine neoplasia type 1. Cortical bone mineral density in hyperparathyroidism/multiple endocrine neoplasia type 1 cases has only recently been examined, and early, severe and frequent bone mineral losses have been documented at this site. Early bone mineral losses are highly prevalent in the trabecular bone of patients with hyperparathyroidism/multiple endocrine neoplasia type 1. In summary, bone mineral disease in multiple endocrine neoplasia type 1related hyperparathyroidism is an early, frequent and severe disturbance, occurring in both the cortical and trabecular bones. In addition, renal complications secondary to sporadic hyperparathyroidism are often studied, but very little work has been done on this issue in hyperparathyroidism/multiple endocrine neoplasia type 1. It has been recently verified that early, frequent, and severe renal lesions occur in patients with hyperparathyroidism/multiple endocrine neoplasia type 1, which may lead to increased morbidity and mortality. In this article we review the few available studies on bone mineral and renal disturbances in the setting of hyperparathyroidism/multiple endocrine neoplasia type 1. We performed a meta-analysis of the available data on bone mineral and renal disease in cases of multiple endocrine neoplasia type 1-related hyperparathyroidism.

Exercise and bone mass in adults

[EN] There is a substantial body of evidence indicating that exercise prior to the pubertal growth spurt stimulates bone growth and skeletal muscle hypertrophy to a greater degree than observed during growth in non-physically active children. Bone mass can be increased by some exercise programmes in adults and the elderly, and attenuate the losses in bone mass associated with aging. This review provides an overview of cross-sectional and longitudinal studies performed to date involving training and bone measurements. Cross-sectional studies show in general that exercise modalities requiring high forces and/or generating high impacts have the greatest osteogenic potential. Several training methods have been used to improve bone mineral density (BMD) and content in prospective studies. Not all exercise modalities have shown positive effects on bone mass. For example, unloaded exercise such as swimming has no impact on bone mass, while walking or running has limited positive effects.

Evaluation of bone strength: microtomographic tecniques

This thesis is a part of a larger study about the characterization of mechanical and histomorphometrical properties of bone. The main objects of this study were the bone tissue properties and its resistance to mechanical loads. Moreover, the knowledge about the equipment selected to carry out the analyses, the micro-computed tomography (micro-CT), was improved. Particular attention was given to the reliability over time of the measuring instrument. In order to understand the main characteristics of bone mechanical properties a study of the skeletal, the bones of which it is composed and biological principles that drive their formation and remodelling, was necessary. This study has led to the definition of two macro-classes describing the main components responsible for the resistance to fracture of bone: quantity and quality of bone. The study of bone quantity is the current clinical standard measure for so-called bone densitometry, and research studies have amply demonstrated that the amount of tissue is correlated with its mechanical properties of elasticity and fracture. However, the models presented in the literature, including information on the mere quantity of tissue, have often been limited in describing the mechanical behaviour. Recent investigations have underlined that also the bone-structure and the tissue-mineralization play an important role in the mechanical characterization of bone tissue. For this reason in this thesis the class defined as bone quality was mainly studied, splitting it into two sub-classes of bone structure and tissue quality. A study on bone structure was designed to identify which structural parameters, among the several presented in the literature, could be integrated with the information about quantity, in order to better describe the mechanical properties of bone. In this way, it was also possible to analyse the iteration between structure and function. It has been known for long that bone tissue is capable of remodeling and changing its internal structure according to loads, but the dynamics of these changes are still being analysed. This part of the study was aimed to identify the parameters that could quantify the structural changes of bone tissue during the development of a given disease: osteoarthritis. A study on tissue quality would have to be divided into different classes, which would require a scale of analysis not suitable for the micro-CT. For this reason the study was focused only on the mineralization of the tissue, highlighting the difference between bone density and tissue density, working in a context where there is still an ongoing scientific debate.

Generierung und Charakterisierung von in-vitro- und in- vivo-Modellsystemen zur Untersuchung des Skelettsystems

Die Entstehung und Aufrechterhaltung von Knorpel- und Knochengewebe wird durch eine Vielzahl von hemmenden oder fördernden Faktoren hoch komplex reguliert, wobei die dabei involvierten physiologischen Prozesse bisher nur teilweise verstanden werden. Auch die Ursachen sowohl degenerativer Erkrankungen, aber auch durch Mutationen im FGFR3-Gen verursachter Chondrodysplasien sind in ihrer Ätiopathogenese noch nicht vollständig erforscht. In dieser Arbeit wurden verschiedene experimentelle Ansätze verfolgt, die zur weiteren Aufklärung der Pathophysiologie zweier unterschiedlicher Skeletterkrankungen beitragen sollten.rnEin relevantes Charakteristikum der degenerativen Gelenkserkrankung Osteoarthrose ist der Verlust an Aggrekan, hauptverantwortlich verursacht durch die Aggrekanase ADAMTS5. Es wurde ein Tiermodell generiert, bei dem gezielt mittels des Tet-ON-Systems die Aggrekanase mAdamts-5 überexprimiert werden kann. Nach Konstruktherstellung und Generierung als auch Charakterisierung des in vitro-Modells wurde das Tiermodell hergestellt, um die Folgen der Überexpression im Hinblick auf einen verstärkten Aggrekanabbau im Knorpel der Mäuse zu analysieren. Nach initialer Charakterisierung auf Induzierbarkeit zeigte eine Gründerlinie eine induzierbare transgene mAdamts5-Expression. Die Überprüfung auf Knorpelspezifität zeigte, sowohl embryonal als auch im adulten Tier, dass sich der verwendete, zusammengesetzte Kollagen-Typ II Promotor wie der endogene verhielt und somit funktional war. Nach Doxyzyklininduktion wurde bei der optimalen Dosis von 1 mg/ml im Vergleich zum induzierten Wildtyp-Tier eine 15%ige Abnahme des Gesamt-Glykosamino-glykan(GAG)-Gehaltes und eine um 120% erhöhte GAG-Abgabe ins Medium detektiert, was eine verstärkte Spaltung von Aggrekan bedeutete. Die transgene Aggrekanase wurde überexprimiert und spaltete verstärkt Aggrekan. Da aufgrund der histologischen Untersuchungen jedoch keine Knorpelerosionen feststellbar waren, konnte im Umkehrschluss gefolgert werden, dass der Knorpel einen Verlust an Glykosaminoglykanen bis zu einer gewissen Grenze tolerieren kann. Mit dem generierten und charakterisierten Tiermodell konnte mit dem Verlust an GAG eine Osteoarthrose-ähnliche Situation simuliert werden, insbesondere im Hinblick auf frühe Stadien der Erkrankung, bei denen noch keine makroskopisch eindeutig sichtbare Knorpelerosionen vorliegen. rnIm zweiten Teil der Arbeit wurden Zellkulturexperimente zur weiteren Aufklärung FGFR3-regulierter Prozesse durchgeführt. Nach Generierung und Verifizierung der stabilen Zelllinien, die mittels des Tet-ON-Systems das FGFR3-Gen mit jeweils einer Chondrodysplasie-assoziierten Mutation (Achondroplasie-Mutation G380R, Thanatophore Dysplasie Typ II-Mutation K650E) induzierbar überexprimieren, wurden die Auswirkungen der zwei verschiedenen Mutationen anhand bereits beschriebener Signalwege untersucht. Über die Rekrutierung des ERK-Signalweges konnte bei beiden Zelllinien die Funktionalität nachgewiesen werden, wobei die Zelllinie mit der einen schwereren Phänotyp beim Menschen verursachenden TDII-Mutation eine stärkere Aktivierung zeigte. Bei der Aktivierung von STAT1 wies nur die TDII-Zelllinie eine Phosphorylierung auf, nicht jedoch die ACH-Zelllinie; dies deckte sich mit bereits publizierten Untersuchungen. Beide Kaskaden zeigten eine unterschiedliche Signalantwort aufgrund der verschiedenen Mutationen. Des Weiteren konnte eine unterschiedliche MMP13-Zielgenexpression nachgewiesen werden, wobei lediglich die ACH-Zelllinie eine erhöhte MMP13-Expression (6-fach) zeigte. Zur Identifizierung neuer involvierter FGFR3-Zielgene wurde die differentielle Genexpression der TDII-Zelllinie im Vergleich induziert/nicht induziert mittels Microarray-Hybridisierung untersucht. Als interessantes Zielgen fiel STC1 auf, welches ebenfalls eine Rolle in der Chondrogenese spielt und bislang nicht mit FGFR3 in Verbindung gebracht wurde. Es konnte jedoch nur auf RNA-Ebene eine Regulation nachgewiesen werden. Nachfolgend durchgeführte transiente Experimente zeigten, dass die Wildtyp-Variante von FGFR3 möglicherweise eine Funktion in der Sekretion des Proteins STC1 hat und dass durch die beiden eingefügten Mutationen (ACH, TDII) diese aufgehoben ist. Der Einfluss von FGFR3 auf die Sekretion von STC1 stellt ein neues Ergebnis dar, insbesondere auch die Auswirkungen der beiden für die unterschiedlichen Krankheitsbilder stehenden Mutationen. Welche Relevanz allerdings die STC1-Sekretion im Rahmen FGFR3-assoziierter Erkrankungen hat, kann nicht eindeutig beurteilt werden. Weitere Faktoren aus dem hoch komplexen Zusammenspiel während der Knorpel/Knochenentwicklung müssen untersucht werden, um eine definitive Einordnung zu ermöglichen.

Characterisation of human co-culture systems for applications in bone tissue engineering

For the successful integration of bone tissue engineering constructs into patients, an adequate supply with oxygen and nutrients is critical. Therefore, prevascularisation of bone tissue engineering constructs is desirable for bone formation, remodelling and regeneration. Co-culture systems, consisting of human endothelial cells and primary osteoblasts (pOB) as well as osteosarcoma cell lines, represent a promising method for studying the mechanisms involved in the vascularisation of constructs in bone tissue en- gineering and could provide new insights into the molecular and cellular mechanisms that control essential processes during angiogenesis. The present study demonstrated the im- portant components of co-culture systems with a focus on bone tissue replacement and the angiogenic effects of pOB and osteosarcoma cell lines on human endothelial cells. Furthermore, the studies emphasised an overall approach for analysis of signal molecules that are involved in the angiogenic activation of human endothelial cells by the regulation of VEGF-related pathways at the transcriptional and translational levels. The osteosarcoma cell lines Cal-72, MG-63 and SaOS-2, as well as pOB from several donors, differed in their angiogenesis-inducing potential in 2-D and 3-D co-culture systems. SaOS-2 cells appeared to have a high osteogenic differentiation level with no detectable angiogenesis-inducing potential in co-culture with human endothelial cells. The angiogenic potential of the osteoblast-like cells is mainly correlated with the upregulation of essential angiogenic growth factors, such as VEGF, bFGF and HGF and the downregulation of the angiogenesis inhibitor, endostatin. However, other factors involved in angiogenic regulation were found to differ between SaOS-2 cells, compared to Cal-72 and MG-63. The present study focuses on VEGF pathway-effecting genes as key players in the regulation of angiogenesis. The levels of VEGF and VEGF-effecting genes, such as TGF-α and TIMP-2 are down-regulated in SaOS-2 cells. In contrast, direct regulators of VEGF, such as IL6, IL8 and TNF are strongly upregulated, which indicates disruptions in growth factor regulating pathways in SaOS-2 cells. Potential pathways, which could be involved include MEK, PI3K, MAPK, STAT3, AKT or ERK. Additional treatment of co-cultures with single growth factors did not accelerate or improve the angiogenesis-inducing potential of SaOS-2 cells. Knowledge of the detailed molecular mechanisms involved in angiogenesis control will hopefully allow improved approaches to be developed for prevascularisation of bone tissue engineering constructs.

DensiProbe Spine: a novel instrument for intraoperative measurement of bone density in transpedicular screw fixation

STUDY DESIGN.: Cadaver study. OBJECTIVE.: To determine bone strength in vertebrae by measuring peak breakaway torque or indentation force using custom-made pedicle probes. SUMMARY OF BACKGROUND DATA.: Screw performance in dorsal spinal instrumentation is dependent on bone quality of the vertebral body. To date no intraoperative measuring device to validate bone strength is available. Destructive testing may predict bone strength in transpedicular instrumentations in osteoporotic vertebrae. Insertional torque measurements showed varying results. METHODS.: Ten human cadaveric vertebrae were evaluated for bone mineral density (BMD) measurements by quantitative computed tomography. Peak torque and indentation force of custom-made probes as a measure for mechanical bone strength were assessed via a transpedicular approach. The results were correlated to regional BMD and to biomechanical load testing after pedicle screw implementation. RESULTS.: Both methods generated a positive correlation to failure load of the respective vertebrae. The correlation of peak breakaway torque to failure load was r = 0.959 (P = 0.003), therewith distinctly higher than the correlation of indentation force to failure load, which was r = 0.690 (P = 0.040). In predicting regional BMD, measurement of peak torque also performed better than that of indentation force (r = 0.897 [P = 0.002] vs. r = 0.777 [P = 0.017]). CONCLUSION.: Transpedicular measurement of peak breakaway torque is technically feasible and predicts reliable local bone strength and implant failure for dorsal spinal instrumentations in this experimental setting.

Abnormal bone geometry at the metacarpal bone shaft of rheumatoid arthritis patients with maintained muscle-bone relationship

OBJECTIVES:: Metacarpal juxta-articular bone is altered in Rheumatoid Arthritis (RA). However, a detailed analysis of disease related geometrical adaptations of the metacarpal shaft is missing. The aim of the present study was to assess the role of RA disease, forearm muscle cross-sectional area (CSA), age and sex on bone geometry at the metacarpal shaft. METHODS:: In 64 RA patients and 128 control subjects geometric properties of the third metacarpal bone mid-shaft and forearm muscle CSA were measured by peripheral quantitative computed tomography (pQCT). Linear models were performed for cortical CSA, total bone CSA, polar stress-strain Index (polar SSI, a surrogate for bone's resistance to bending and torsion), cortical thickness and Metacarpal Index (MI=cortical CSA/total CSA) with explanatory variables muscle CSA, age, RA status and sex. RESULTS:: Forearm muscle CSA was associated with cortical and total metacarpal CSA, and polar SSI. RA group status was associated with all bone parameters except cortical CSA. There was a significant interaction between RA status and age, indicating that the RA group had a greater age-related decrease in cortical CSA, cortical thickness and MI. CONCLUSIONS:: Bone geometry of the metacarpal shaft is altered in RA patients compared to healthy controls. While bone mass of the metacarpal shaft is adapted to forearm muscle mass, cortical thickness and MI are reduced but outer bone shaft circumference and polar SSI increased in RA patients. These adaptations correspond to an enhanced aging pattern in RA patients.

[Examination reports on survived strangulation cases]

Clinical-forensic examination of strangulation victims is an increasing part of the routine of many forensic pathology institutes. The cases examined between 2004 and 2008 at the Institute of Legal Medicine of the Hanover Medical School were retrospectively analysed. In total, the study material comprised 218 victims (175 females and 43 males). In 80.7 %, the clinical-forensic examination was performed within 24 hours after the incident. In the overwhelming number of cases, the alleged perpetrator was no stranger. 128 victims (58.7 %) had strangulation marks, 32 victims (14.7 %) ligature marks and 65 victims (29.8 %) nail marks. Four victims showed injuries of the laryngeal and pharyngeal structures (reddening, hematomas, swelling and in one case a fracture of the cricoid cartilage on both sides). Extensive petechiae were predominantly seen in the conjunctivae, the buccal mucosa and the skin of the face in cases where the victims suffered a loss of consciousness. 87 cases (39.9% were classified as potentially life-threatening and 30 cases (13.8 %) as acute life-threatening events. This classification is of legal relevance for the penalty. In addition, 60 victims experienced sexual violence. These results suggest that early clinical-forensic examination is crucial for documenting forensic evidence in support of police investigations and may deliver significant details relevant in court.

Osteogenic potential of autogenous bone grafts harvested with four different surgical techniques

The osteogenic potential of autogenous bone grafts is superior to that of allografts and xenografts because of their ability to release osteoinductive growth factors and provide a natural osteoconductive surface for cell attachment and growth. In this in vitro study, autogenous bone particles were harvested by four commonly used techniques and compared for their ability to promote an osteogenic response. Primary osteoblasts were isolated and seeded on autogenous bone grafts prepared from the mandibles of miniature pigs with a bone mill, piezo-surgery, bone scraper, and bone drill (bone slurry). The osteoblast cultures were compared for their ability to promote cell attachment, proliferation, and differentiation. After 4 and 8 hrs, significantly higher cell numbers were associated with bone mill and bone scraper samples compared with those acquired by bone slurry and piezo-surgery. Similar patterns were consistently observed up to 5 days. Furthermore, osteoblasts seeded on bone mill and scraper samples expressed significantly elevated mRNA levels of collagen, osteocalcin, and osterix at 3 and 14 days and produced more mineralized tissue as assessed by alizarin red staining. These results suggest that the larger bone graft particles produced by bone mill and bone scraper techniques have a higher osteogenic potential than bone slurry and piezo-surgery.

Relative Contributions of Osteogenic Tissues to New Bone Formation in Periosteal Distraction Osteogenesis: Histological and Histomorphometrical Evaluation in a Rat Calvaria

Background: The relative contributions of different, potential factors to new bone formation in periosteal distraction osteogenesis are unknown. Purpose: The aim of the present study was to assess the influence of original bone and periosteum on bone formation during periosteal distraction osteogenesis in a rat calvarial model by means of histology and histomorphometry. Methods: A total of 48 rats were used for the experiment. The contribution of the periosteum was assessed by either intact or incised periosteum or an occlusive versus a perforated distraction plate. The cortical bone was either left intact or perforated. Animals were divided in eight experimental groups considering the three possible treatment modalities. All animals were subjected to a 7-day latency period, a 10-day distraction period and a 7-day consolidation period. The newly formed bone was analyzed histologically and histomorphometrically. Results: New, mainly woven bone was found in all groups. Differences in the maximum height of new bone were observed and depended on location. Under the distraction plate, statistically significant differences in maximum bone height were found between the group with perforations in both cortical bone and distraction plate and the group without such perforations. Conclusions: If the marrow cavities were not opened, the contribution to new bone formation was dominant from the periosteum. If the bone perforations opened the marrow cavities, a significant contribution to new bone formation originated from the native bone.

In vitro cytotoxicity of silver nanoparticles on osteoblasts and osteoclasts at antibacterial concentrations

Abstract Nanoparticulate silver coatings for orthopaedic implants promise to decrease postoperative infection rates. However, silver-induced cytotoxicity on bone cells has not been investigated in detail. This study investigated the cytotoxic effects of silver nano- and microparticles and Ag(+) on osteoblasts (OBs) and osteoclasts (OCs) and correlated their effects with the antibacterial efficacy on Staphylococcus epidermidis. Silver nanoparticles (50 nm) exhibited strong cytotoxic effects on OBs and OCs. Weak cytotoxic effects were observed for silver microparticles (3 μm). The cytotoxicity was primarily mediated by a size-dependent release of Ag(+). Antibacterial effects occurred at Ag(+) concentrations that were 2-4 times higher than those inducing cytotoxic effects. Such adverse effects on OB and OC survival may have deleterious effects on the biocompatibility of orthopaedic implants. Our study represents an important step toward the detailed investigation of orthopaedic implant with nanoparticulate silver coatings prior to their widespread clinical usage.