Implant retention in a rabbit model of fracture-related infection.

Aims: Fracture-related infection (FRI) is commonly classified based on the time of onset of symptoms. Early infections (< two weeks) are treated with debridement, antibiotics, and implant retention (DAIR). For late infections (> ten weeks), guidelines recommend implant removal due to tolerant biofilms. For delayed infections (two to ten weeks), recommendations are unclear. In this study we compared infection clearance and bone healing in early and delayed FRI treated with DAIR in a rabbit model. Methods: Staphylococcus aureus was inoculated into a humeral osteotomy in 17 rabbits after plate osteosynthesis. Infection developed for one week (early group, n = 6) or four weeks (delayed group, n = 6) before DAIR (systemic antibiotics: two weeks, nafcillin + rifampin; four weeks, levofloxacin + rifampin). A control group (n = 5) received revision surgery after four weeks without antibiotics. Bacteriology of humerus, soft-tissue, and implants was performed seven weeks after revision surgery. Bone healing was assessed using a modified radiological union scale in tibial fractures (mRUST). Results: Greater bacterial burden in the early group compared to the delayed and control groups at revision surgery indicates a retraction of the infection from one to four weeks. Infection was cleared in all animals in the early and delayed groups at euthanasia, but not in the control group. Osteotomies healed in the early group, but bone healing was significantly compromised in the delayed and control groups. Conclusion: The duration of the infection from one to four weeks does not impact the success of infection clearance in this model. Bone healing, however, is impaired as the duration of the infection increases.

A Hyaluronic Acid Hydrogel Loaded with Gentamicin and Vancomycin Successfully Eradicates Chronic Methicillin-Resistant Staphylococcus aureus Orthopedic Infection in a Sheep Model.

Implantable orthopedic devices have had an enormously positive impact on human health; however, despite best practice, patients are prone to developing orthopedic device-related infections (ODRI) that have high treatment failure rates. One barrier to the development of improved treatment options is the lack of an animal model that may serve as a robust preclinical assessment of efficacy. We present a clinically relevant large animal model of chronic methicillin-resistant Staphylococcus aureus (MRSA) ODRI that persists despite current clinical practice in medical and surgical treatment at rates equivalent to clinical observations. Furthermore, we showed that an injectable, thermoresponsive, hyaluronic acid-based hydrogel loaded with gentamicin and vancomycin outperforms current clinical practice treatment in this model, eliminating bacteria from all animals. These results confirm that local antibiotic delivery with an injectable hydrogel can dramatically increase treatment success rates beyond current clinical practice, with efficacy proven in a robust animal model.

Cortical parameters predict bone strength at the tibial diaphysis, but are underestimated by HR-pQCT and µCT compared to histomorphometry.

Cortical bone and its microstructure are crucial for bone strength, especially at the long bone diaphysis. However, it is still not well-defined how imaging procedures can be used as predictive tools for mechanical bone properties. This study evaluated the capability of several high-resolution imaging techniques to capture cortical bone morphology and assessed the correlation with the bone's mechanical properties. The microstructural properties (cortical thickness [Ct.Th], porosity [Ct.Po], area [Ct.Ar]) of 11 female tibial diaphysis (40-90 years) were evaluated by dual-energy X-ray absorptiometry (DXA), high-resolution peripheral-quantitative-computed-tomography (HR-pQCT), micro-CT (µCT) and histomorphometry. Stiffness and maximal torque to failure were determined by mechanical testing. T-Scores determined by DXA ranged from 0.6 to -5.6 and a lower T-Score was associated with a decrease in Ct.Th (p ≤ 0.001) while the Ct.Po (p ≤ 0.007) increased, and this relationship was independent of the imaging method. With decreasing T-Score, histology showed an increase in Ct.Po from the endosteal to the periosteal side (p = 0.001) and an exponential increase in the ratio of osteons at rest to those after remodelling. However, compared to histomorphometry, HR-pQCT and µCT underestimated Ct.Po and Ct.Th. A lower T-Score was also associated with significantly reduced stiffness (p = 0.031) and maximal torque (p = 0.006). Improving the accuracy of Ct.Po and Ct.Th did not improve prediction of the mechanical properties, which was most closely related to geometry (Ct.Ar). The ex-vivo evaluation of mechanical properties correlated with all imaging modalities, with Ct.Th and Ct.Po highly correlated with the T-Score of the tibial diaphysis. Cortical microstructural changes were underestimated with the lower resolution of HR-pQCT and µCT compared to the histological 'gold standard'. The increased accuracy did not result in an improved prediction for local bone strength in this study, which however might be related to the limited number of specimens and thus needs to be evaluated in a larger collective.

Single-stage revision of MRSA orthopedic device-related infection in sheep with an antibiotic-loaded hydrogel.

Local antimicrobial therapy is an integral aspect of treating orthopedic device-related infection (ODRI), which is conventionally administered via polymethyl-methacrylate (PMMA) bone cement. PMMA, however, is limited by a suboptimal antibiotic release profile and a lack of biodegradability. In this study, we compare the efficacy of PMMA versus an antibiotic-loaded hydrogel in a single-stage revision for chronic methicillin-resistant Staphylococcus aureus (MRSA) ODRI in sheep. Antibiofilm activity of the antibiotic combination (gentamicin and vancomycin) was determined in vitro. Swiss alpine sheep underwent a single-stage revision of a tibial intramedullary nail with MRSA infection. Local gentamicin and vancomycin therapy was delivered via hydrogel or PMMA (n = 5 per group), in conjunction with systemic antibiotic therapy. In vivo observations included: local antibiotic tissue concentration, renal and liver function tests, and quantitative microbiology on tissues and hardware post-mortem. There was a nonsignificant reduction in biofilm with an increasing antibiotic concentration in vitro (p = 0.12), confirming the antibiotic tolerance of the MRSA biofilm. In the in vivo study, four out of five sheep from each treatment group were culture-negative. Antibiotic delivery via hydrogel resulted in 10-100 times greater local concentrations for the first 2-3 days compared with PMMA and were comparable thereafter. Systemic concentrations of gentamicin were minimal or undetectable in both groups, while renal and liver function tests were within normal limits. This study shows that a single-stage revision with hydrogel or PMMA is equally effective, although the hydrogel offers certain practical benefits over PMMA, which make it an attractive proposition for clinical use.

Development of bone seeker-functionalised microspheres as a targeted local antibiotic delivery system for bone infections.

OBJECTIVE: Bone infections are challenging to treat because of limited capability of systemic antibiotics to accumulate at the bone site. To enhance therapeutic action, systemic treatments are commonly combined with local antibiotic-loaded materials. Nevertheless, available drug carriers have undesirable properties, including inappropriate antibiotic release profiles and nonbiodegradability. To alleviate such limitations, we aim to develop a drug delivery system (DDS) for local administration that can interact strongly with bone mineral, releasing antibiotics at the infected bone site. METHODS: Biodegradable polyesters (poly (ε-caprolactone) or poly (D,l-lactic acid)) were selected to fabricate antibiotic-loaded microspheres by oil in water emulsion. Antibiotic release and antimicrobial effects on Staphylococcus aureus were assessed by zone of inhibition measurements. Microsphere bone affinity was increased by functionalising the bisphosphonate drug alendronate to the microsphere surface using carbodiimide chemistry. Effect of bone targeting microspheres on bone homeostasis was tested by looking at the resorption potential of osteoclasts exposed to the developed microspheres. RESULTS: In vitro, the antibiotic release profile from the microspheres was shown to be dependent on the polymer used and the microsphere preparation method. Mineral binding assays revealed that microsphere surface modification with alendronate significantly enhanced interaction with bone-like materials. Additionally, alendronate functionalised microspheres did not differentially affect osteoclast mineral resorption in vitro, compared with nonfunctionalised microspheres. CONCLUSION: We report the development and characterisation of a DDS which can release antibiotics in a sustained manner. Surface-grafted alendronate groups enhanced bone affinity of the microsphere construct, resulting in a bone targeting DDS. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: The DDS presented can be loaded with hydrophobic antibiotics, representing a potential, versatile and biodegradable candidate to locally treat bone infection.

Current Concepts of Osteomyelitis: From Pathologic Mechanisms to Advanced Research Methods.

Osteomyelitis is an inflammation of the bone and bone marrow that is most commonly caused by a Staphylococcus aureus infection. Much of our understanding of the underlying pathophysiology of osteomyelitis, from the perspective of both host and pathogen, has been revised in recent years, with notable discoveries including the role played by osteocytes in the recruitment of immune cells, the invasion and persistence of S. aureus in submicron channels of cortical bone, and the diagnostic role of polymorphonuclear cells in implant-associated osteomyelitis. Advanced in vitro cell culture models, such as ex vivo culture models or organoids, have also been developed over the past decade, and have become widespread in many fields, including infectious diseases. These models better mimic the in vivo environment, allow the use of human cells, and can reduce our reliance on animals in osteomyelitis research. In this review, we provide an overview of the main pathologic concepts in osteomyelitis, with a focus on the new discoveries in recent years. Furthermore, we outline the value of modern in vitro cell culture techniques, with a focus on their current application to infectious diseases and osteomyelitis in particular.

Biomechanical investigation of four different fixation techniques in sacrum Denis type II fracture with low bone mineral density.

With increasing life expectancy, fragility fractures of the pelvic ring are seen more frequently. Although their osteosynthesis can be very challenging, specific biomechanical studies for investigation of the fixation stability are still lacking. The aim of this study was to biomechanically evaluate four different fixation methods for sacrum Denis type II fractures in osteoporotic bone. Unstable Denis type II vertical sacrum fractures were created in 16 human pelves. Their osteosynthesis was performed with one sacro-iliac screw, posterior sacral plating, triangular fixation, or spino-pelvic fixation. For that purpose, each pelvis was randomly assigned to two paired groups for treatment with either SI-screw/posterior sacral plating or triangular fixation/spino-pelvic fixation. Each hemi-pelvis was cyclically tested under progressively increasing axial compression. Relative interfragmentary movements were investigated via optical motion tracking analysis. Axial stiffness of triangular fixation was significantly higher versus posterior sacral plating and spino-pelvic fixation (p ≤ 0.022), but not significantly different in comparison to SI-screw fixation (p = 0.337). Cycles to 2, 3, 5, and 8 mm fracture displacement, as well as to 3°, 5°, and 8° gap angle at the fracture site were significantly higher for triangular fixation compared to all other groups (p ≤ 0.041). Main failure mode for all osteosynthesis techniques was screw cutting through the bone, leading loss of fixation stability. From a biomechanical point of view, triangular fixation in sacrum Denis type II fractures demonstrated less interfragmentary movements and should be considered in unstable fragility fractures of the sacrum. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1624-1629, 2018.

High-Resolution Tomography-Based Quantification of Cortical Porosity and Cortical Thickness at the Surgical Neck of the Humerus During Aging.

Fractures of the proximal humerus are highly related to age and osteoporotic bone remodeling. Previous studies have highlighted the cortex as a major side of the bone loss, but the microstructural changes of the humerus have not been evaluated entirely. Sixty-four (n = 64) humeri of a representative collective (18-100 years) were scanned with high-resolution peripheral quantitative computed tomography (82 µm). Bone mineral density (BMD), trabecular bone volume fraction (Tb.BV/TV), cortical thickness (Ct.Th), and cortical porosity (Ct.Po) were determined with respect to four age groups. The BMD (r = -0.42), Ct.Th (r = 0.57), and Tb.BV/TV (r = 0.68) showed an age group-related decrease, while the Ct.Po increased (r = -0.55). The oldest group (80-100 years) revealed an extensively higher Ct.Po of +87% compared to the youngest group (18-44 years), while the Ct.Th and Tb.BV/TV were significantly lower by -35 and -49% (p < 0.05). The main cortical bone loss occurred after 65 years with the Ct.Th (-34%) and Tb.BV/TV (-40%) being clearly lower and the Ct.Po (+93%) clearly higher compared to the youngest group. In summary, osteoporosis leads to an age-related higher Ct.Po and reduced Ct.Th at the humeral cortex of the surgical neck. The bone loss of the cortex predominantly occurs around the age of 65 years and is very likely to reduce the mechanical strength and highly increases the fracture risk.

Antimicrobial delivery systems for local infection prophylaxis in orthopedic- and trauma surgery.

Infectious complications occur in a minor but significant portion of the patients undergoing joint replacement surgery or fracture fixation, particularly those with severe open fractures, those undergoing revision arthroplasty or those at elevated risk because of poor health status. Once established, infections are difficult to eradicate, especially in the case of bacterial biofilm formation on implanted hardware. Local antibiotic carriers offer the prospect of controlled delivery of antibiotics directly in target tissues and implant, without inducing toxicity in non-target organs. Polymeric carriers have been developed to optimize the release and targeting of antibiotics. Passive polymeric carriers release antibiotics by diffusion and/or upon degradation, while active polymeric carriers release their antibiotics upon stimuli provided by bacterial pathogens. Additionally, some polymeric carriers gelate in-situ in response to physiological stimuli to form a depot for antibiotic release. As antibiotic resistance has become a major issue, also other anti-infectives such as silver and antimicrobial peptides have been incorporated in research. Currently, several antibiotic loaded biomaterials for local infection prophylaxis are available for use in the clinic. Here we review their advantages and limitations and provide an overview of new materials emerging that may overcome these limitations.

A combined biomaterial and cellular approach for annulus fibrosus rupture repair.

Recurrent intervertebral disc (IVD) herniation and degenerative disc disease have been identified as the most important factors contributing to persistent pain and disability after surgical discectomy. An annulus fibrosus (AF) closure device that provides immediate closure of the AF rupture, restores disc height, reduces further disc degeneration and enhances self-repair capacities is an unmet clinical need. In this study, a poly(trimethylene carbonate) (PTMC) scaffold seeded with human bone marrow derived mesenchymal stromal cells (MSCs) and covered with a poly(ester-urethane) (PU) membrane was assessed for AF rupture repair in a bovine organ culture annulotomy model under dynamic load for 14 days. PTMC scaffolds combined with the sutured PU membrane restored disc height of annulotomized discs and prevented herniation of nucleus pulposus (NP) tissue. Implanted MSCs showed an up-regulated gene expression of type V collagen, a potential AF marker, indicating in situ differentiation capability. Furthermore, MSCs delivered within PTMC scaffolds induced an up-regulation of anabolic gene expression and down-regulation of catabolic gene expression in adjacent native disc tissue. In conclusion, the combined biomaterial and cellular approach has the potential to hinder herniation of NP tissue, stabilize disc height, and positively modulate cell phenotype of native disc tissue.

Osseointegration of machined, injection moulded and oxygen plasma modified PEEK implants in a sheep model.

Machined and injection moulded polyetheretherketone (PEEK) implants with and without an oxygen plasma modification were prepared and implanted in sheep cancellous and cortical bone. After 4, 12 and 26 weeks, osseointegration was evaluated through mechanical push-out tests and histomorphometry. In the cancellous bone, push-out force increased with time, a trend toward higher force was observed for machined compared to moulded, and oxygen plasma modified compared to unmodified. On-going remodelling of the bone was detected in the periphery of the implants at 4 weeks. Minimal or no inflammation was observed with all the implants at all locations and time-points. Bone-implant contact (BIC) was quantified at all-time points and locations for all the four PEEK implant surfaces. The BIC values ranged from 15 to 75% with an average of 29 ± 13% in the cancellous bone and 25-65% with an average of 50 ± 12% in the cortical bone. In the cortical bone the BIC increased significantly from 4 to 26 weeks. This in vivo study has identified that surface topography of PEEK implants influences osseointegration. In addition, oxygen plasma has the potential to increase bone-implant interface stability. This study provides a unique reference for further modifications and in vivo assessment of PEEK implants.

Assessment of intraosseous femoral head pressures during cement augmentation of the perforated proximal femur nail antirotation blade.

OBJECTIVES: The benefits of cement augmentation with fixation of osteoporotic pertrochanteric fractures have been previously demonstrated. The objective of this study was 3-fold: (1) To quantify the intraosseous pressure produced during cement augmentation of the perforated proximal femoral nail antirotation (PFNA) blades; (2) To assess whether the pressure generated is influenced by the injection rate; and (3) To assess the amount of force applied during the injection. METHODS: Six pairs of human cadaveric femurs were used in the study. A basicervical osteotomy was performed, and the heads were instrumented with the PFNA blade. Each pair was randomly assigned into 1 of 2 groups: slow versus rapid injection with polymethylmethacrylate (PMMA) cement. In the slow group, the augmentation was performed using 6 consecutive 1 mL injections, each over 10 seconds. In the rapid group, each 1 mL injection was performed over 5 seconds. For intraosseous pressure measurements, transmitters were inserted to a depth of 5 mm at both the superior and inferior apices of the head. The reaction forces on the syringe were measured as well. RESULTS: There were no significant differences between the slow and rapid injection rates with respect to the peak pressures measured at the 6 time points immediately after cement injection. In both groups, elevations in pressure were transient and returned to baseline values within 30 seconds. The highest pressure recorded in the slow group was 37.3 and 30.7 mm Hg in the rapid group. The force required after each sequential injection increased in both groups; however, significantly higher forces were required to inject cement over 5 than 10 seconds (P = 0.036). CONCLUSIONS: This in vitro model is the first one to demonstrate that femoral head cement augmentation is associated with a small transient increase in intraosseous pressure with sequential fast and slow 1 mL injections of up to 6 mL PMMA. We conclude that cement augmentation of the perforated PFNA blade carries a low risk of pressure-induced avascular necrosis.

A biomechanical study on proximal plate fixation techniques in periprosthetic femur fractures.

INTRODUCTION: Proximal plate fixation is a crucial factor in osteosynthesis of periprosthetic femur fractures. Stability and strength of different fixation concepts for proximal plate fixation were compared. MATERIALS AND METHODS: Twelve fresh frozen, bone mineral density matched human femora, instrumented with cemented hip endoprosthesis were osteotomized simulating a Vancouver B1 fracture. Specimens were instrumented with locking compression plates, fixed proximally with either locking attachment plate (LAP), monocortical screws, cerclage plus monocortical screws (1cerclage) or cerclages only (4cerclages). Cyclic testing was performed with monotonically increasing load until failure. Relative movements at the proximal plate-femur interface were registered by motion tracking. RESULTS: The LAP construct exhibited a significantly longer cumulative survival (failure criterion 1mm separation at the proximal plate fixation) compared to the monocortical (p=0.048) and 4cerclages constructs (p=0.012) but not to 1cerclage constructs. CONCLUSION: Bicortical screw anchorage improves proximal plate fixation in periprosthetic fractures. The cerclage-screw combination is a valuable alternative especially in osteoporotic bone.

The locking attachment plate for proximal fixation of periprosthetic femur fractures--a biomechanical comparison of two techniques.

PURPOSE: Mechanical properties of a locking attachment plate construct (LAP-LCP), allowing bicortical screw placement laterally to the prosthesis stem, are compared to a cerclage-LCP construct. METHODS: Eight right synthetic femora with implanted uncemented hip endoprosthesis were cut distally and fixed with LCP, monocortical locking screws and either LAP (n = 4) or cerclage (n = 4). Cyclic testing was performed with monotonically increasing sinusoidal load until failure. Relative movements at the plate-femur interface were registered by motion tracking. Statistical differences were detected by unpaired t-test and general linear model repeated measures. RESULTS: Stiffness of the LAP-LCP was significantly higher at the beginning (875.4 N/mm ± 29.8) and after 5000 cycles (1213.0 N/mm ± 101.1) compared to the cerclage-LCP (644.96 N/mm ± 50.1 and 851.9 N/mm ± 81.9), with p = 0.013. Relative movements for AP-bending (B) and axial translation (T) of the LAP-LCP at the beginning (0.07° ± 0.02, 0.20 mm ± 0.08), after 500 cycles (0.16° ± 0.10, 0.26 mm ± 0.07) and after 5000 cycles (0.26° ± 0.11, 0.31 mm ± 0.07) differed significantly from the cerclage-LCP (beg.: 0.26° ± 0.04, 0.28 mm ± 0.05; 500 cyc: 0.47° ± 0.03, 0.53 mm ± 0.07; 5000 cyc.: 0.63° ± 0.18, 0.79 mm ± 0.13), with B: p = 0.02, T: p = 0.04. Relative movements for medial bending were not significantly different between the two constructs. Cycles to failure (criterion 1 mm axial translation) differed significantly between LAP-LCP (19,519 ± 1,758) and cerclage-LCP (11,265 ± 2,472), with p = 0.035. CONCLUSIONS: Biomechanically, the LAP-LCP construct improves proximal fixation of periprosthetic fractures compared to the cerclage-LCP construct.

Potential of polymethylmethacrylate cement-augmented helical proximal femoral nail antirotation blades to improve implant stability--a biomechanical investigation in human cadaveric femoral heads.

BACKGROUND: Cement augmentation may improve fixation stability and reduce cut-out rate in the treatment of intertrochanteric hip fractures. The aim of this study was to compare the number of cycles to failure of polymethylmethacrylate (PMMA)-augmented helical blades with nonaugmented ones in human cadaveric femoral heads. METHODS: Six pairs of cadaveric femoral heads were instrumented with a perforated proximal femoral nail antirotation blade. Within each pair, one blade was augmented using 3 mL of PMMA. All specimens underwent cyclic axial loading under physiologic conditions.Starting at 1,000 N, the load was monotonically increased by 0.1 N/cycle until construct failure occurred. To monitor the migration of the blade, anteroposterior radiographs were taken at 250 cycle increments. Nonparametric test statistics were done to calculate correlations and identify differences between study groups. RESULTS: Inducing failure required a significantly higher number of cycles in the augmented group (p = 0.028). Bone mineral density was significantly related with the number of cycles to failure in nonaugmented specimens (p 0.001, R2 = 0.97), but not in the augmented group (p = 0.91, R2 = 0.34). CONCLUSION: Implant augmentation with small amounts of PMMA enhances the cut-out resistance in proximal femoral fractures. Especially in osteoporotic bone, the procedure may improve patient care.

In vivo evaluation of the effect of intramedullary nail microtopography on the development of local infection in rabbits.

BACKGROUND AND AIM: Fractures of the tibia and femoral diaphysis are commonly repaired by intramedullary (IM) nails, which are currently composed of either electropolished stainless steel (EPSS) or standard, non-polished titanium-aluminum-niobium (TAN). Once the fracture has fully healed, removal of IM nails is common, but the strong adhesion of bone to standard TAN complicates removal. Polishing the surface of TAN IM nails has been shown to reduce bony adhesion and ease implant removal without compromising fixation. Polished TAN nails are, therefore, expected to have significant clinical benefit in situations where the device is to be removed. The aim of the present study was to determine the effect of polishing TAN IM nails on susceptibility to infection in an animal model. MATERIALS AND METHODS: Solid IM nails (Synthes, Betlach, Switzerland) composed of standard TAN were compared with polished equivalents and also to clinically available EPSS nails. The surface chemical and topographical properties of the materials were assessed by X-ray photon spectroscopy (XPS), white light profilometry, and scanning electron microscopy (SEM). An in vivo infection study was performed using a clinical isolate of Staphylococcus aureus that was characterized with respect to various virulence factors. RESULTS: Polishing TAN IM nails caused no significant change to the chemistry of the nails, but the topography of the polished TAN nails was significantly smoother than standard TAN nails. In the infection study, the rank order based on descending infectious dose 50 (ID(50)) was: standard TAN, polished TAN, and finally EPSS. The ID(50) values did not differ greatly between any of the groups. CONCLUSIONS: Polishing the surface TAN IM nails was not found to influence the susceptibility to infection in our animal model.

Influence of material and microtopography on the development of local infection in vivo: experimental investigation in rabbits.

Polishing the surface of internal fracture fixation (IFF) implant materials can ease implant removal and reduce irritation to gliding tissues by reducing soft tissue adhesion and bony overgrowth. Thus, polishing the surface of these implants is expected to have significant clinical benefit in certain situations. The aim of the present study was to determine if polishing the surface of an IFF device influences susceptibility to infection. The local infection rate associated with 4-hole 2.0 mm Synthes locking compression plates (LCPs) composed of clinically available commercially pure titanium (cpTi) and titanium aluminium niobium (TAN) in their standard microrough form was compared with that of their test polished equivalents and also to clinically available electropolished stainless steel (EPSS). The LCPs were fixed in locking mode onto the tibia of mature, female New Zealand White rabbits and a clinical strain of Staphylococcus aureus was added to the implantation site. Twenty eight days after surgery the rabbits were euthanized and assessed for infection. The rank order based on descending ID50 was; polished TAN, standard TAN, standard cpTi, EPSS and finally polished cpTi, however, the ID50 values did not differ greatly between the groups with the same material. Using the LCP model in locking mode, polishing the surface of both cpTi and TAN was not found to influence the susceptibility to infection in our animal model.

TGFbeta3 and loading increases osteocyte survival in human cancellous bone cultured ex vivo.

The goal of this study was to assess the effect of the addition of TGFbeta(3), alone or in combination with loading, on the survival of osteocytes in 3D human explant cancellous bone during long-term culture in an ex vivo loading bioreactor. Human cancellous bone explants were cultured for up to 14 days with or without TGFbeta(3) (15 ng ml(-1)) and with or without loading (300 cycles, at 1 Hz, producing 4000 microstrain). Bone core response was visualized using undecalcified histology with morphological methods after embedding with Technovit 9100 New resin. Histological examination revealed normal gross level bone structure with or without the application of load or the addition of TGFbeta(3). The viability of the osteocytes within the bone was assessed by lactate dehydrogenase (LDH) activity. We demonstrate that this ex vivo loading bioreactor is able to maintain a high percentage (over 50%) of viable osteocytes throughout the bone explants after 14 days in ex vivo culture. Further to this, the combination of daily loading and TGFbeta(3) administration produced superior osteocyte survival at the core centres when compared to loading or TGFbeta alone.

Microtopography of metal surfaces influence fibroblast growth by modifying cell shape, cytoskeleton, and adhesion.

Stainless Steel (SS), titanium (cpTi), and Ti-6Al-7Nb (TAN) are frequently used metals in fracture fixation, which contact not only bone, but also soft tissue. In previous soft tissue cytocompatibility studies, TAN was demonstrated to inhibit cell growth in its "standard" micro-roughened state. To elucidate a possible mechanism for this inhibition, cell area, shape, adhesion, and cytoskeletal integrity was studied. Only minor changes in spreading were observed for cells on electropolished SS, cpTi, and TAN. Cells on "standard" cpTi were similarly spread in comparison with electropolished cpTi and TAN, although the topography influenced the cell periphery and also resulted in lower numbers and shorter length of focal adhesions. On "standard" microrough TAN, cell spreading was significantly lower than all other surfaces, and cell morphology differed by being more elongated. In addition, focal adhesion numbers and mean length were significantly lower on standard TAN than on all other surfaces, with 80% of the measured adhesions below a 2-microm threshold. Focal adhesion site location and maturation and microtubule integrity were compromised by the presence of protruding beta-phase microspikes found solely on the surface of standard TAN. This led us to propose that the impairment of focal adhesion numbers, maturation (length), and cell spreading to a possibly sufficient threshold observed on standard TAN blocks cell cycle progress and eventually cell growth on the surface. We believe, as demonstrated with standard cpTi and TAN, that a difference in surface morphology is influential for controlling cell behavior on implant surfaces.