Layer-by-layer polyelectrolyte encapsulation of Mycoplasma pneumoniae for enhanced Raman detection.

Mycoplasma pneumoniae is a major cause of respiratory disease in humans and accounts for as much as 20% of all community-acquired pneumonia. Existing mycoplasma diagnosis is primarily limited by the poor success rate at culturing the bacteria from clinical samples. There is a critical need to develop a new platform for mycoplasma detection that has high sensitivity, specificity, and expediency. Here we report the layer-by-layer (LBL) encapsulation of M. pneumoniae cells with Ag nanoparticles in a matrix of the polyelectrolytes poly(allylamine hydrochloride) (PAH) and poly(styrene sulfonate) (PSS). We evaluated nanoparticle encapsulated mycoplasma cells as a platform for the differentiation of M. pneumoniae strains using surface enhanced Raman scattering (SERS) combined with multivariate statistical analysis. Three separate M. pneumoniae strains (M129, FH and II-3) were studied. Scanning electron microscopy and fluorescence imaging showed that the Ag nanoparticles were incorporated between the oppositely charged polyelectrolyte layers. SERS spectra showed that LBL encapsulation provides excellent spectral reproducibility. Multivariate statistical analysis of the Raman spectra differentiated the three M. pneumoniae strains with 97-100% specificity and sensitivity, and low (0.1-0.4) root mean square error. These results indicated that nanoparticle and polyelectrolyte encapsulation of M. pneumoniae is a potentially powerful platform for rapid and sensitive SERS-based bacterial identification.

Chemical generation of nitric oxide in the mouth from the enterosalivary circulation of dietary nitrate.

High concentrations of nitrite present in saliva (derived from dietary nitrate) may, upon acidification, generate nitrogen oxides in the stomach in sufficient amounts to provide protection from swallowed pathogens. We now show that, in the rat, reduction of nitrate to nitrite is confined to a specialized area on the posterior surface of the tongue, which is heavily colonized by bacteria, and that nitrate reduction is absent in germ-free rats. We also show that in humans increased salivary nitrite production resulting from nitrate intake enhances oral nitric oxide production. We propose that the salivary generation of nitrite is accomplished by a symbiotic relationship involving nitrate-reducing bacteria on the tongue surface, which is designed to provide host defence against microbial pathogens in the mouth and lower gut. These results provide further evidence for beneficial effects of dietary nitrate.

The local and global geometry of trabecular bone.

The organization and shape of the microstructural elements of trabecular bone govern its physical properties, are implicated in bone disease, and serve as blueprints for biomaterial design. To devise fundamental structure-property relationships and design truly bone-mimicking biomaterials, it is essential to characterize trabecular bone structure from the perspective of geometry, the mathematical study of shape. Using micro-CT images from 70 donors at five different sites, we analyze the local and global geometry of human trabecular bone in detail, respectively by quantifying surface curvatures and Minkowski functionals. We find that curvature density maps provide distinct and sensitive shape fingerprints for bone from different sites. Contrary to a common assumption, these curvature maps also show that bone morphology does not approximate a minimal surface but exhibits a much more intricate curvature landscape. At the global (or integral) perspective, our Minkowski analysis illustrates that trabecular bone exhibits other types of anisotropy/ellipticity beyond interfacial orientation, and that anisotropy varies substantially within the trabecular structure. Moreover, we show that the Minkowski functionals unify several traditional morphometric indices. Our geometric approach to trabecular morphometry provides a fundamental language of shape that could be useful for bone failure prediction, understanding geometry-driven tissue growth, and the design of bone-mimicking tissue scaffolds. STATEMENT OF SIGNIFICANCE: The architecture of trabecular bone is key in determining bone properties, and is often a starting point for the design of bone-substitutes. Despite the substantial history of bone morphometry, a fundamental characterization of trabecular bone geometry is still lacking. Therefore, we introduce a robust framework to quantify local and global trabecular bone geometry, which we apply to hundreds of micro-CT scans. Our approach relies on quantifying surface curvatures and Minkowski functionals, which are the most fundamental local and global shape quantifiers. Our results show that these shape metrics are sensitive to differences between bone types and unify traditional metrics within a single mathematical framework. This geometrical framework could also be useful to design bone-mimicking scaffolds and understand geometry-driven tissue growth.

Spatio-temporal characterization of fracture healing patterns and assessment of biomaterials by time-lapsed in vivo micro-computed tomography.

Thorough preclinical evaluation of functionalized biomaterials for treatment of large bone defects is essential prior to clinical application. Using in vivo micro-computed tomography (micro-CT) and mouse femoral defect models with different defect sizes, we were able to detect spatio-temporal healing patterns indicative of physiological and impaired healing in three defect sub-volumes and the adjacent cortex. The time-lapsed in vivo micro-CT-based approach was then applied to evaluate the bone regeneration potential of functionalized biomaterials using collagen and bone morphogenetic protein (BMP-2). Both collagen and BMP-2 treatment led to distinct changes in bone turnover in the different healing phases. Despite increased periosteal bone formation, 87.5% of the defects treated with collagen scaffolds resulted in non-unions. Additional BMP-2 application significantly accelerated the healing process and increased the union rate to 100%. This study further shows potential of time-lapsed in vivo micro-CT for capturing spatio-temporal deviations preceding non-union formation and how this can be prevented by application of functionalized biomaterials. This study therefore supports the application of longitudinal in vivo micro-CT for discrimination of normal and disturbed healing patterns and for the spatio-temporal characterization of the bone regeneration capacity of functionalized biomaterials.

High-throughput synthesis and screening of hydrogen-storage alloys.

Libraries of mixed-metal hydride materials are synthesized on a silicon microfabricated array of "hot-plate" MEMS devices, which allow high-throughput screening using temperature programmed desorption and infrared thermography. The heating plate of the MEMS device is a membrane with low heat capacity, allowing fast and localized temperature control and the extraction of calorimetric data from thermography. The combination of the synthetic method and screening chip allows a fast determination of the desorption temperature and hydrogen content of the materials. Mixed metal hydrides are synthesized directly. The potential of the method is exemplified by presenting results for the sorption properties of Mg xNi 1- x hydride thin-film materials. The results are consistent with the literature, showing the highest hydrogen capacity and desorption temperature for the MgH 2 phase in Mg-rich compositions and the promotion of a lower temperature desorption from the Mg 2NiH 4 phase, with a concomitant reduction in hydrogen capacity.

Revision for adverse local tissue reaction following metal-on-polyethylene total hip arthroplasty is associated with a high risk of early major complications.

Aims: Fretting and corrosion at the modular head/neck junction, known as trunnionosis, in total hip arthroplasty (THA) is a cause of adverse reaction to metal debris (ARMD). We describe the outcome of revision of metal-on-polyethylene (MoP) THA for ARMD due to trunnionosis with emphasis on the risk of major complications. Patients and Methods: A total of 36 patients with a MoP THA who underwent revision for ARMD due to trunnionosis were identified. Three were excluded as their revision had been to another metal head. The remaining 33 were revised to a ceramic head with a titanium sleeve. We describe the presentation, revision findings, and risk of complications in these patients. Results: The patients presented with pain, swelling, stiffness, or instability and an inflammatory mass was confirmed radiologically. Macroscopic material deposition on the trunnion was seen in all patients, associated with ARMD. Following revision, six (18.2%) dislocated, requiring further revision in four. Three (9.1%) developed a deep infection and six (18.2%) had significant persistent pain without an obvious cause. One developed a femoral artery thrombosis after excision of an iliofemoral pseudotumor, requiring a thrombectomy. Conclusion: The risk of serious complications following revision MoP THA for ARMD associated with trunnionosis is high. In the presence of extensive tissue damage, a constrained liner or dual mobility construct is recommended in these patients. Cite this article: Bone Joint J 2018;100-B:720-4.

Cement penetration and primary stability of the femoral component after impaction allografting. A biomechanical study in the cadaveric femur.

This study explored the relationship between the initial stability of the femoral component and penetration of cement into the graft bed following impaction allografting. Impaction allografting was carried out in human cadaveric femurs. In one group the cement was pressurised conventionally but in the other it was not pressurised. Migration and micromotion of the implant were measured under simulated walking loads. The specimens were then cross-sectioned and penetration of the cement measured. Around the distal half of the implant we found approximately 70% and 40% of contact of the cement with the endosteum in the pressure and no-pressure groups, respectively. The distal migration/micromotion, and valgus/varus migration were significantly higher in the no-pressure group than in that subjected to pressure. These motion components correlated negatively with the mean area of cement and its contact with the endosteum. The presence of cement at the endosteum appears to play an important role in the initial stability of the implant following impaction allografting.

Carvable calcium phosphate bone substitute material.

This study investigated the use of partially set hydroxyapatite forming calcium phosphate cement as a carvable and mechanically stable bone substitute material. Hydroxyapatite-forming cements were made of either mechanically activated alpha-tricalcium phosphate or a mixture of tetracalcium phosphate and dicalcium phosphate anhydrous and setting was arrested up to 4 h post setting. The study showed that these partially set rigid samples of defined geometry could be carved into a desired shape when the degree of reaction was 30-40% and the relative porosity between 40 and 50%; samples are then expected to set completely after implantation in the presence of water or serum, having the same compressive strength as a continuously set calcium phosphate cement (up to 36 MPa). The development of compressive strength, phase composition, and crystallinity when varying production parameters of these partially "preset" bone substitute materials are presented for both cement systems.

Mandibular osteopathy in a Hagerman horse, Equus simplicidens (Equidae, Mammalia), from Hagerman Fossil Beds National Monument (Idaho, USA).

Multimodality imaging, including computed tomography (CT) and digital radiography, was utilized to examine a fossilized hemimandible of a probable female Hagerman horse (Equus simplicidens) with a large, ventrally located, osseous deformation. Utilizing comparative pathology to the modern day horse, it was determined that the most likely etiology of the pathologic bony swelling along the ventral hemimandible was abnormal tooth development that led to chronic osteomyelitis and subsequent osseous proliferation, sclerosis and deformation.

Delayed dislocation following metal-on-polyethylene arthroplasty of the hip due to 'silent' trunnion corrosion.

AIMS: We present a case series of ten metal-on-polyethylene total hip arthroplasties (MoP THAs) with delayed dislocation associated with unrecognised adverse local tissue reaction due to corrosion at the trunnion and pseudotumour formation. METHODS: The diagnosis was not suspected in nine of the ten patients (six female/four male; mean age 66 years), despite treatment in a specialist unit (mean time from index surgery to revision was 58 months, 36 to 84). It was identified at revision surgery and subsequently confirmed by histological examination of resected tissue. Pre-operative assessment and culture results ruled out infection. A variety of treatment strategies were used, including resection of the pseudotumour and efforts to avoid recurrent dislocation. RESULTS: The rate of complications was high and included three deep infections, two patients with recurrent dislocation, and one recurrent pseudotumour. CONCLUSION: This series (mean follow-up of 76 months following index procedure and 19 months following revision THA) demonstrates that pseudotumour is an infrequent but important contributor to delayed instability following MoP THA. It is easy to overlook in the differential diagnosis, especially if the alignment of the components is less than optimal, leading to an assumption that malalignment is the cause of the dislocation. The instability is likely to be multifactorial and the revision surgery is complex. TAKE HOME MESSAGE: Due to the high complication rate associated with revision in this cohort, the diagnosis should be borne in mind when counselling patients regarding the risks of revision surgery.

Cortical bone screw fixation in ionically modified apatite cements.

Hydroxyapatite cements are used in reconstruction of the face; usually in well-defined cavities where the cement can be stabilized without the need for internal fixation. A hydroxyapatite cement that could enable screw fixation and some loading therefore has considerable potential in maxillofacial reconstruction. It has been demonstrated recently that water demand of calcium phosphate cements can be reduced by ionically modifying the liquid component. This study investigated the capacity of an ionically modified precompacted apatite cement to retain self-tapping cortical bone screws. Screw pullout forces were determined in the direction of the screw long axis and perpendicular to it, using cortical bone and polymethylmethacrylate cement as a control. In bending pullout tests, measured forces to remove screws from ionically modified precompacted cement were insignificantly different from cortical bone. However, pullout forces of bone screws from hydroxyapatite cement decreased with aging time in vitro.

Adverse local tissue reactions in metal-on-polyethylene total hip arthroplasty due to trunnion corrosion: the risk of misdiagnosis.

Adverse reaction to wear and corrosion debris is a cause for concern in total hip arthroplasty (THA). Modular junctions are a potential source of such wear products and are associated with secondary pseudotumour formation. We present a consecutive series of 17 patients treated at our unit for this complication following metal-on-highly cross-linked polyethylene (MoP) THA. We emphasise the risk of misdiagnosis as infection, and present the aggregate laboratory results and pathological findings in this series. The clinical presentation was pain, swelling or instability. Solid, cystic and mixed soft-tissue lesions were noted on imaging and confirmed intra-operatively. Corrosion at the head-neck junction was noted in all cases. No bacteria were isolated on multiple pre- and intra-operative samples yet the mean erythrocyte sedimentation rate was 49 (9 to 100) and C-reactive protein 32 (0.6 to 106) and stromal polymorphonuclear cell counts were noted in nine cases. Adverse soft-tissue reactions can occur in MoP THA owing to corrosion products released from the head-neck junction. The diagnosis should be carefully considered when investigating pain after THA. This may avoid the misdiagnosis of periprosthetic infection with an unidentified organism and mitigate the unnecessary management of these cases with complete single- or two-stage exchange.

Treatment of infection associated with segmental bone loss in the proximal part of the femur in two stages with use of an antibiotic-loaded interval prosthesis.

Treatment of an infection at the site of a total hip replacement associated with extensive loss of the proximal part of the femur is a challenging problem. In the present preliminary report, we describe the results after use of a prosthesis of antibiotic-loaded acrylic cement (PROSTALAC) in thirty such hips. The purpose of the prosthesis, which acts as an internal splint, is to maintain the length of the femur as well as the range of motion of the joint and the mobility of the patient between stages. A local level of antibiotics is maintained by the antibiotic-coated surface. A PROSTALAC with a cement-on-cement articulation was used in the first fifteen hips (Group I) in the study, and a custom metal-on-polyethylene articulating PROSTALAC was inserted in the subsequent fifteen hips (Group II). One patient who had a recurrent infection was managed with a second two-stage exchange and was included in both groups. Between stages, the average limb-length discrepancy was twenty-five millimeters despite a loss of more than 25 per cent of the femur in nineteen limbs. Sixteen patients were discharged home and seven, to a community hospital between stages. Six patients in Group I and only one in Group II were hospitalized for the entire course of treatment. The total duration of hospitalization for both stages averaged thirty-eight days. Twenty-eight patients were mobile even though they did not bear weight on the involved limb between stages: three patients used a cane, fifteen used crutches, and ten used a walker. Twenty-six patients reported no, slight, or moderate pain in the thigh, groin, or buttock between stages. The average Harris hip score before the first stage of the operation was 23 points (range, 0 to 63 points), which improved to 74 points (range, 40 to 91 points) at an average of forty-seven months (range, twenty-four to 114 months) postoperatively. Two patients died of unrelated causes before two years (the minimum follow-up period) had elapsed and were excluded from the final analysis; they had no evidence of recurrent infection. Of the remaining twenty-eight hips, twenty-seven (96 per cent) had no evidence of infection at the most recent follow-up examination.

The cement mantle in femoral impaction allografting. A comparison of three systems from four centres.

An analysis of the cement mantle obtained with the Exeter impaction allografting system at one centre showed that it was either deficient or absent in almost 47% of Gruen zones. We therefore examined the mantle obtained using this system at another hospital and compared the results with those from the CPT and Harris Precoat Systems at other centres. The surgical indications for the procedure and the patient details were broadly similar in all four hospitals. There was some variation in the frequency of use of cortical strut allografts, cerclage wires and wire mesh to supplement the impaction allograft. Analysis of the cement mantles showed that when uncertain Gruen zones were excluded, the incidence of zones with areas of absence or deficiency of the cement was 47% and 50%, respectively, for the two centres using the Exeter system, 21% for the CPT system and 18% for the Harris Precoat system. We measured the difference in size between the proximal allograft impactors and the definitive prosthesis for each system. The Exeter system impactors are shorter than the definitive prosthesis and taper sharply so that the cavity created is inadequate, especially distally. The CPT proximal impactors are considerably longer than the definitive prosthesis and are designed to give a mantle of approximately 2 mm medially and laterally and 1.5 mm anteriorly and posteriorly. The Harris Precoat proximal impactors allow for a mantle with a circumference of 0.75 mm in the smaller sizes and 1 mm in the larger. Many reports link the longevity of a cemented implant to the adequacy of the cement mantle. For this reason, femoral impaction systems require careful design to achieve a cement mantle which is uninterrupted in its length and adequate in its thickness. Our results suggest that some current systems require modification.

Allograft impaction and cement penetration after revision hip replacement. A histomorphometric analysis in the cadaver femur.

We studied various aspects of graft impaction and penetration of cement in an experimental model. Cancellous bone was removed proximally and local diaphyseal lytic defects were simulated in six human cadaver femora. After impaction grafting the specimens were sectioned and prepared for histomorphometric analysis. The porosity of the graft was lowest in Gruen zone 4 (52%) and highest in Gruen zone 1 (76%). At the levels of Gruen zones 6 and 2 the entire cross-section was almost filled with cement. Cement sometimes reached the endosteal surface in other Gruen zones. The mean peak impaction forces exerted with the impactors were negatively correlated with the porosity of the graft.

The PROSTALAC functional spacer in two-stage revision for infected knee replacements. Prosthesis of antibiotic-loaded acrylic cement.

The PROSTALAC functional spacer is made of antibiotic-loaded acrylic cement but has a small metal-on-polythene articular surface. We have used it as an interim spacer in two-stage exchange arthroplasty for infected total knee replacement. PROSTALAC allows continuous rehabilitation between stages as it maintains good alignment and stability of the knee and a reasonable range of movement. It also helps to maintain the soft-tissue planes, which facilitates the second-stage procedure. We reviewed 45 consecutive patients, treated over a period of nine years. The mean follow-up was for 48 months (20 to 112). At final review, there was no evidence of infection in 41 patients (91%); only one had a recurrent infection with the same organism. There was improvement in the Hospital for Special Surgery knee score between stages and at final review. The range of movement was maintained between stages. Complications were primarily related to the extensor mechanism and stability of the knee between stages. Both of these problems decreased with refinement of the design of the implant. The rate of cure of the infection in our patients was similar to that using other methods. Movement of the knee does not appear to hinder control of infection.

Surgical approaches in revision hip replacement.

Revision hip arthroplasty will be performed with frequency in the future. A successful outcome depends on careful preoperative planning, and a key component of that plan is the surgical approach. The choice of the approach should be based on the indication for revision, the particular implant to be removed, the presence of acetabular or femoral bone loss, previous surgical approaches used, and the preferences and training of the surgeon. For simple revision procedures, one of the standard approaches used in primary hip arthroplasty may be adequate. More complex cases may necessitate an extended exposure or one of the techniques developed specifically for revision arthroplasty. No single approach is suitable for all revision procedures, and the surgeon must be familiar with a range of exposures if the clinical result is to be optimized.

Use of reconstruction rings for the management of acetabular bone loss during revision hip surgery.

Aseptic loosening remains the leading cause of failure after total hip replacement. Extensive bone loss and acetabular bone stock deficiency are frequently encountered. Simple autografting techniques are often not possible, and copious amounts of allograft may be required to reconstruct the defect. The mechanically stable environment that is a prerequisite for successful graft incorporation cannot be achieved with routine acetabular fixation techniques alone. A reconstruction ring that is secured to the surrounding pelvis provides a more rigid construct. Several types of reconstruction rings are available for the management of acetabular bone loss during revision hip surgery. Early results suggest that these devices may prove to be a useful alternative for treatment of a difficult problem.

Impaction allografting with cement for the management of femoral bone loss.

Impaction allografting with cement is the only technique currently available which reverses the diminution of bone stock that occurs in a revision hip arthroplasty, and as such, has great potential. It is particularly appropriate in the younger patient, though older patients may also benefit from the technique. Although the short term results are encouraging, there is a need for further basic science research to determine the optimal graft material and prosthesis design. Refinements in surgical instrumentation and technique will continue to improve the predictability of the clinical result and expand the indications for this important addition to the available options in revision hip arthroplasty.