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Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXXVII | Pages 139 - 139
1 Sep 2012
Topolovec M Milosev I Coer A Bloebaum R
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Background. Wear particles are considered to be the major culprit for the aseptic loosening. Their characterization is thus crucial for the understanding of their bioreactivity and contribution to the development of aseptic loosening. Methods. Metal wear debris particles were analyzed directly in periprosthetic tissue resins by scanning electron microscopy (SEM) combined with back-scattered electron imaging (BSE) and energy dispersive X-ray spectroscopy (EDS). Four groups of tissue samples retrieved at revision operations of loosened hip implants with different bearing surfaces (metal-on-metal, ceramic-on-polyethylene and metal-on-polyethylene), and different material of the femoral stem (Ti alloy, CoCrMo and polymer combined with stainless steel) were investigated. Tissue samples were first analyzed histologicaly. Sections from the same paraffin blocks were then carbon coated and analyzed using SEM/BSE/EDS method. Results. Metal particles were detected in all samples. Their composition corresponded to the composition of the implant components. The gradation of metal particles ranged from +1 to +3. A considerable number of big metal particles were actually agglomerates of submicron particles visible only at higher magnification. The clustering of particles was observed primarily for CoCrMo and, to a lesser extent, for stainless steels particles. The median sizes of CoCrMo clusters in two groups of samples were 2.9 1.8 m (range, 0.5 to 7.6 m) and 3.2 1.0 m (range, 1.9 to 5.4 m). The effect of clustering was not observed for Ti particles. The median sizes of individual Ti particles determined in two groups of samples were 2.5 3.6 m (range, 0.4 to 17.3 m) and 4.3 2.8 m (range, 0.8 to 11.0 m). Conclusion. Scanning electron microscopy combined with back-scattered electron imaging is an appropriate and selective method to recognize metal particles in tissue sections, without being destructive to specimens. When the size of the particles is considered, however, it should be differed between the size of individual particles and size of clusters of particles. Besides its benefits, this study has some limitations: the detection of particles smaller than 0.4 m is difficult, and this method cannot be used to identify polyethylene particles


Orthopaedic Proceedings
Vol. 87-B, Issue SUPP_II | Pages 205 - 205
1 Apr 2005
Fadda M Manunta A Milia F Zirattu F Zirattu G
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With the plasma–spray technique of applying a hydrox-ylapatite (HA) coating bone ingrowth can be enhanced and early migration of hip prostheses reduced. The significance of coating resorption is controversial. In this study the bone growth and the degradation of the HA coatings were evaluated and compared by SEM.

Premature loosening was identified in four cups with an Ha coating over a porous-coated surface 3 years post-operatively.The Ha coating has a thickness of up to 50 μm. The cup specimens were soaked in 6% sodium hypochlorite to render them anorganic, dehydrated, and sputter-coated with gold-palladium. Secondary electron images of all specimens were obtained by field emission SEM (Zeiss:DSM.962).

Ultrastructural analysis showed that all porous-coated Ha-coated cups had bridges of lamellar bone in direct contact with the implant surface (30% bone in-on growth). Different types of coating degradation were observed. Delamination between the coating and implant surface releases numerous particles or fragments; the resorption by osteoclasts of the amorphous phase was shown to expose the crystalline phase of the coating grains.

This study suggests that resorption disintegrates the Ha coating and reduces the bonding strength between implant and bone and the strength of the coating-implant interface, which might lead to implant loosening,coating delamination and acceleration of third-body wear processes.


Orthopaedic Proceedings
Vol. 86-B, Issue SUPP_III | Pages 367 - 367
1 Mar 2004
Fadda M Zirattu G Manunta A Fadda G Delrio A
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Aims: Hydroxylapatite (HA) coating is able to enhance bone ingrowth and to reduce early migration of hip prostheses. The optimum coating quality and surface texture is still a matter of debate. Moreover, the signiþ-cance of coating resorption is controversial. In this study the degradation of the coatings HA was evaluated and comparate by SEM. Materials and methods: Four cups with HA coating over a porous-coated surface was iden-tiþed with premature loosening at 2–3 years post-operatively. The HA coating has a thickness of up to 50 μ. The cup was stored in formalin before the SEM analysis. The cup specimens was soaked in 6% sodium hypochlorite to render them anorganic, dehydrated, sputter Ð coated with gold-palladium. Secondary electron images of all specimens were obtained by þeld-emission SEM (Zeiss: DSM.962). Results: Ultrastructural analysis showed that all porous-coated HA coated cups had bridges of bone in direct contact with the implant surface (30% bone on-growth). Different types of coating degradation were observed. Delamination between the coating and implant surface; release of numerous particles or fragments ranging from a few to several dozens of microns. Under high magniþcation resorption of the amorphous phase is shown to be exposing the crystalline phase of the coating grains so that the grain boundaries become fragile and easily to be phagocytosed by osteoclasts. Conclusions: This study suggested that resorption disintegrates the HA Ð coating and reduces the bonding strength between implant and bone and the strength of the coatingÐimplant interface, which might lead to implant loosening, coating delamination and acceleration of third body wear processes.


Bone & Joint Research
Vol. 8, Issue 6 | Pages 246 - 252
1 Jun 2019
Liddle A Webb M Clement N Green S Liddle J German M Holland J

Objectives. Previous studies have evidenced cement-in-cement techniques as reliable in revision arthroplasty. Commonly, the original cement mantle is reshaped, aiding accurate placement of the new stem. Ultrasonic devices selectively remove cement, preserve host bone, and have lower cortical perforation rates than other techniques. As far as the authors are aware, the impact of ultrasonic devices on final cement-in-cement bonds has not been investigated. This study assessed the impact of cement removal using the Orthosonics System for Cemented Arthroplasty Revision (OSCAR; Orthosonics) on final cement-in-cement bonds. Methods. A total of 24 specimens were manufactured by pouring cement (Simplex P Bone Cement; Stryker) into stainless steel moulds, with a central rod polished to Stryker Exeter V40 specifications. After cement curing, the rods were removed and eight specimens were allocated to each of three internal surface preparation groups: 1) burr; 2) OSCAR; and 3) no treatment. Internal holes were recemented, and each specimen was cut into 5 mm discs. Shear testing of discs was completed by a technician blinded to the original grouping, recording ultimate shear strengths. Scanning electron microscopy (SEM) was completed, inspecting surfaces of shear-tested specimens. Results. The mean shear strength for OSCAR-prepared specimens (33.6 MPa) was significantly lower than for the control (46.3 MPa) and burr (45.8 MPa) groups (p < 0.001; one-way analysis of variance (ANOVA) with Tukey’s post hoc analysis). There was no significant difference in shear strengths between control and burr groups (p = 0.57). Scanning electron microscopy of OSCAR specimens revealed evidence of porosity undiscovered in previous studies. Conclusion. Results show that the cement removal technique impacts on final cement-in-cement bonds. This in vitro study demonstrates significantly weaker bonds when using OSCAR prior to recementation into an old cement mantle compared with cement prepared with a burr or no treatment. This infers that care must be taken in surgical decision-making regarding cement removal techniques used during cement-in-cement revision arthroplasty, suggesting that the risks and benefits of ultrasonic cement removal need consideration. Cite this article: A. Liddle, M. Webb, N. Clement, S. Green, J. Liddle, M. German, J. Holland. Ultrasonic cement removal in cement-in-cement revision total hip arthroplasty: What is the effect on the final cement-in-cement bond? Bone Joint Res 2019;8:246–252. DOI: 10.1302/2046-3758.86.BJR-2018-0313.R1


Aims. This study investigated vancomycin-microbubbles (Vm-MBs) and meropenem (Mp)-MBs with ultrasound-targeted microbubble destruction (UTMD) to disrupt biofilms and improve bactericidal efficiency, providing a new and promising strategy for the treatment of device-related infections (DRIs). Methods. A film hydration method was used to prepare Vm-MBs and Mp-MBs and examine their characterization. Biofilms of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli were treated with different groups. Biofilm biomass differences were determined by staining. Thickness and bacterial viability were observed with confocal laser scanning microscope (CLSM). Colony counts were determined by plate-counting. Scanning electron microscopy (SEM) observed bacterial morphology. Results. The Vm-MBs and Mp-MBs met the experimental requirements. The biofilm biomass in the Vm, Vm-MBs, UTMD, and Vm-MBs + UTMD groups was significantly lower than in the control group. MRSA and E. coli biofilms were most notably damaged in the Vm-MBs + UTMD group and Mp-MBs + UTMD group, respectively, with mean 21.55% (SD 0.08) and 19.73% (SD 1.25) remaining in the biofilm biomass. Vm-MBs + UTMD significantly reduced biofilm thickness and bacterial viability (p = 0.005 and p < 0.0001, respectively). Mp-MBs + UTMD could significantly decrease biofilm thickness and bacterial viability (allp < 0.001). Plate-counting method showed that the numbers of MRSA and E. coli bacterial colonies were significantly lower in the Vm-MBs + UTMD group and the Mp, Mp-MBs, UTMD, Mp-MBs + UTMD groups compared to the control group (p = 0.031). SEM showed that the morphology and structure of MRSA and E. coli were significantly damaged in the Vm-MBs + UTMD and Mp-MBs + UTMD groups. Conclusion. Vm-MBs or Mp-MBs combined with UTMD can effectively disrupt biofilms and protectively release antibiotics under ultrasound mediation, significantly reducing bacterial viability and improving the bactericidal effect of antibiotics. Cite this article: Bone Joint Res 2024;13(9):441–451


Bone & Joint Research
Vol. 7, Issue 5 | Pages 357 - 361
1 May 2018
Shin T Lim D Kim YS Kim SC Jo WL Lim YW

Objectives. Laser-engineered net shaping (LENS) of coated surfaces can overcome the limitations of conventional coating technologies. We compared the in vitro biological response with a titanium plasma spray (TPS)-coated titanium alloy (Ti6Al4V) surface with that of a Ti6Al4V surface coated with titanium using direct metal fabrication (DMF) with 3D printing technologies. Methods. The in vitro ability of human osteoblasts to adhere to TPS-coated Ti6Al4V was compared with DMF-coating. Scanning electron microscopy (SEM) was used to assess the structure and morphology of the surfaces. Biological and morphological responses to human osteoblast cell lines were then examined by measuring cell proliferation, alkaline phosphatase activity, actin filaments, and RUNX2 gene expression. Results. Morphological assessment of the cells after six hours of incubation using SEM showed that the TPS- and DMF-coated surfaces were largely covered with lamellipodia from the osteoblasts. Cell adhesion appeared similar in both groups. The differences in the rates of cell proliferation and alkaline phosphatase activities were not statistically significant. Conclusions. The DMF coating applied using metal 3D printing is similar to the TPS coating, which is the most common coating process used for bone ingrowth. The DMF method provided an acceptable surface structure and a viable biological surface. Moreover, this method is automatable and less complex than plasma spraying. Cite this article: T. Shin, D. Lim, Y. S. Kim, S. C. Kim, W. L. Jo, Y. W. Lim. The biological response to laser-aided direct metal-coated Titanium alloy (Ti6Al4V). Bone Joint Res 2018;7:357–361. DOI: 10.1302/2046-3758.75.BJR-2017-0222.R1


Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_7 | Pages 128 - 128
4 Apr 2023
Li M Wu G Liu Y
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Miniscrew implants (MSIs) are widely used to provide absolute anchorage for the orthodontic treatment. However, the application of MSIs is limited by the relatively high failure rate (22.86%). In this study, we wished to investigate the effects of amorphous and crystalline biomimetic calcium phosphate coating on the surfaces of MSIs with or without the incorporated BSA for the osteointegration process with an aim to facilitate the early loading of MSIs. Amorphous and crystalline coatings were prepared on titanium mini-pin implants. Characterizations of coatings were examined by Scanning electron microscopy (SEM), Confocal laser-scanning dual-channel-fluorescence microscopy (CLSM) and Fourier-transform infrared spectroscopy (FTIR). The loading and release kinetics of bovine serum albumin (BSA) were evaluated by Enzyme linked immunosorbent assay (ELISA). Activity of alkaline phosphate (ALP) was measured by using the primary osteoblasts. In vivo, a model of metaphyseal tibial implantation in rats was used (n=6 rats per group). We had 6 different groups: no coating no BSA, no coating but with surface adsorption of BSA and incorporation of BSA in the biomimetic coating in the amorphous and crystalline coatings. Time points were 3 days, 1, 2 and 4 weeks. Histological and histomorphometric analysis were performed and the bone to implant contact (BIC) of each group was compared. In vitro, the incorporation of BSA changed the crystalline coating from sharp plates into curly plates, and the crystalline coating showed slow-release profile. The incorporation of BSA in crystalline coating significantly decreased the activity of ALP in vitro. In vivo study, the earliest significant increase of BIC appeared in crystalline coating group at one week. The crystalline coating can serve as a carrier and slow release system for the bioactive agent and accelerate osteoconductivity at early stage in vivo. The presence of BSA is not favorable for the early establishment of osteointegration


Orthopaedic Proceedings
Vol. 106-B, Issue SUPP_16 | Pages 11 - 11
19 Aug 2024
Hamilton JL Gianotti S Impergre A AbuAlia M Markovics A Jacobs JJ Valle CJD Wimmer MA
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We created TiO. 2. nanotubes (TNTs) on the surface of titanium (Ti) implants with subsequential loading with gentamicin and chitosan, acting as a control release agent, by electrophoretic deposition (EPD). We hypothesized femoral implants with TNTs loaded with gentamicin and chitosan would localize antibiotic to the implant and surgical site and prevent PJI in a mouse model. Ti-6Al-4V ELI wires underwent TNT surface modification by two-step anodization. EPD was then used to load gentamicin and chitosan onto the Ti wire with surface TNTs. Control Ti wires contained TNTs with EPD of chitosan only. 12-week-old male C57BL/6J mice underwent received a right femoral intramedullary implant followed by inoculation at the surgical site with 1×10. 3. CFUs of bioluminescent Xen36 Staphylococcus aureus (S. aureus). Mice were randomly divided into two implant groups: 1) Gentamicin + Chitosan Group (n=7; experimental group); 2) Chitosan Group (n=7; control group). Outcomes included: 1) Relative S. aureus abundance by bioluminescence imaging; 2) Quantification of S. aureus at the implant and surrounding tissue by colony forming unit (CFU) analysis; 3) Scanning electron microscopy (SEM) for implant bacterial biofilm; 4) Radiographic signs of PJI. Over 14 days assessment following wire implantation and inoculation with S. aureus, the experimental group had no evidence of infection based on i) no increased Xen36 S. aureus bioluminescence signal and ii) no CFUs present. All control had increased bioluminescence signal, above baseline, at all time-points and presence of CFUs. Ti femoral implants modified with surface TNTs and coated with gentamicin and chitosan through EPD prevented PJI in all mice through 14 days. In comparison, all control mice demonstrated evidence of PJI over 14 days. Implants with TNTs and EPD of gentamicin were highly effective in this mouse PJI model


Orthopaedic Proceedings
Vol. 91-B, Issue SUPP_II | Pages 289 - 289
1 May 2009
Mirza SB Greenwood M Blunn G
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Introduction There are 1 million cases of major skeletal defects :that occur worldwide each year that lead to significant morbidity and disability and currently require bone grafting as the main mode of treatment. Limitations of bone-grafting include donor site morbidity, reduced osseoinductivity and risk of pathogen transmission to the host. There is considerable interest in finding ways of differentiating mesenchymal stem cells down the osteoblastic lineage to form bone tissue. We hypothesized that there is an optimum strain that promotes differentiation of mesenchymal stem cells into osteoblasts. Methods: A bioreactor was developed that was capable of applying tensional forces across a culture strip in a graduated manner within a range of 1-4373me. Mesenchymal stem cells were grown on these strips and subjected to cyclical tensile strain at 1Hz. Cell morphology using Scanning Electron Microscopy, mineralization using specialized stains and expression of core binding factor1 (Cbfa1) was studied at various strain levels. Results: Scanning Electron Microscopy revealed classic osteoblastic cells in the regions subjected to tensile force, especially in the region where average strain was 1312me. X-ray microanalysis revealed calcium deposits on the strip, indicating osteoblastic differentiation. Cbfa1 expression was greatest in the region with an average strain 1312 me followed by a region on the strip subjected to just fluid shear without any tension. Cbfa1 expression was significantly greater in cells subjected to tensile forces than unstrained controls at all levels of strain tested (p< 0.05). Cbfa1 expression was further enhanced significantly by the addition of osteogenic factors (p< 0.05). Significantly greater mineralization (p< 0.05) occurred in the regions subject to tension with the greatest being in the region with an average strain of 1312 me. Conclusions: Mechanical tensile forces especially in the range of up to 2173me promote differentiation of Mesenchymal Stem Cells into osteoblasts and encourage expression of the Cbfa1 gene. Tensile strain also promotes mineralization. Chemical factors in form of osteogenic media accelerate the differentiation of MSCs and encourages earlier production of osteoblast specific markers. Fluid shear appears to have a beneficial effect in stimulating differentiation into the osteoblast phenotype and, combined with tensile strain, may offer an even greater osteogenic stimulus


Orthopaedic Proceedings
Vol. 106-B, Issue SUPP_2 | Pages 132 - 132
2 Jan 2024
Rau J
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Over the last decades, biodegradable metals emerged as promising materials for various biomedical implant applications, aiming to reduce the use of permanent metallic implants and, therefore, to avoid additional surgeries for implant removal. However, among the important issue to be solved is their fast corrosion - too high to match the healing rate of the bone tissue. The most effective way to improve this characteristic is to coat biodegradable metals with substituted calcium phosphates. Tricalcium phosphate (β-TCP) is a resorbable bioceramic widely used as synthetic bone graft. In order to modulate and enhance its biological performance, the substitution of Ca2+ by various metal ions, such as strontium (Sr2+), magnesium (Mg2+), iron (Fe2+) etc., can be carried out. Among them, copper (Cu2+), manganese (Mn2+), zinc (Zn2+) etc. could add antimicrobial properties against implant-related infections. Double substitutions of TCP containing couples of Cu2+/Sr2+ or Mn2+/Sr2+ ions are considered to be the most perspective based on the results of our study. We established that single phase Ca3−2x(MˊMˊˊ)x(PO4)2 solid solutions are formed only at x ≤ 0.286, where Mˊ and Mˊˊ—divalent metal ions, such as Zn2+, Mg2+, Cu2+, Mn2+, and that in case of double substitutions, the incorporation of Sr2+ ions allows one to extend the limit of solid solution due to the enlargement of the unit cell structure. We also reported that antimicrobial properties depend on the substitution ion occupation of Ca2+ crystal sites in the β-TCP structure. The combination of two different ions in the Ca5 position, on one side, and in the Ca1, Ca2, Ca3, and Ca4 positions, on another side, significantly boosts antimicrobial properties. In the present work, zinc-lithium (Zn-Li) biodegradable alloys were coated with double substituted Mn2+/Sr2+ β-TCP and double substituted Cu2+/ Sr2+ β-TCP, with the scope to promote osteoinductive effect (due to the Sr2+ presence) and to impart antimicrobial properties (thanks to Cu2+ or Mn2+ ions). The Pulsed Laser Deposition (PLD) method was applied as the coating's preparation technique. It was shown that films deposited using PLD present good adhesion strength and hardness and are characterized by a nanostructured background with random microparticles on the surface. For coatings characterization, Fourier Transform Infrared Spectroscopy, X-ray Diffraction, and Scanning Electron Microscopy coupled with Energy Dispersive X-ray and X-ray Photoelectron Spectroscopy were applied. The microbiology tests on the prepared coated Zn-Li alloys were performed with the Gram-positive (Staphylococcus aureus, Enterococcus faecalis) and Gram-negative (Salmonella typhimurium, Escherichia coli) bacteria strains and Candida albicans fungus. The antimicrobial activity tests showed that Mn2+/Sr2+ β-TCP -coated and Cu2+/Sr2+ β-TCP coated Zn-Li alloys were able to inhibit the growth of all five microorganisms. The prepared coatings are promising in improving the degradation behavior and biological properties of Zn-Li alloys, and further studies are necessary before a possible clinical translation


Orthopaedic Proceedings
Vol. 95-B, Issue SUPP_15 | Pages 360 - 360
1 Mar 2013
Wang T Pelletier M Bertollo N Crosky A Walsh W
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Introduction. Implant contamination prior to cement application has the potential to affect the cement-implant bond. the consequences of implant contamination were investigated in vitro using static shear loading with bone cement and titanium dowels of differing surface roughness both with, and without contamination by substances that are likely to be present during surgery. Namely; saline, fat, blood and oil, as a negative control. Methods. Fifty Titanium alloy (Ti-6Al-4V) dowels were prepared with two surface finishes comparable to existing stems. The roughness (Ra and Rq) of the dowel surface was measured before and after the pushout test. Four contaminants (Phosphate Buffered Saline (PBS), ovine marrow, ovine blood, olive oil) were prepared and heated to 37°C. Each contaminant was smeared on the dowel surface completely and uniformly approximately 4 minutes prior to implantation. Samples were separated into ten groups (n=5 per group) based on surface roughness and contaminant. Titanium alloy dowels was placed in the center of Polyvinyl chloride (PVC) tubes with bone cement, and equilibrated at 37°C in PBS for 7 days prior to mechanical testing. The push out test was performed at 1 mm per minute. The dowel surface and cement mantel were analyzed using a Scanning Electron Microscopy (SEM) to determine the distribution and composition of any debris and contaminates on the surface. Results. All contaminants decreased stem-bone cement interfacial shear strength. Saline produced the greatest decrease, followed by blood. The effect of fat was less pronounced and similar to that of oil likely due to the strong lipid solvent properties of the methacrylate monomer. For rough dowels, there were differences in ultimate shear strength between control and contaminated groups (p<0.001). Blood and saline groups had lower ultimate shear strength compared to fat and oil (p<0.05) (fig. 1). The ultimate shear strength for smooth samples was not significantly affected by contamination. Increasing surface roughness increased the interfacial bonding strength, even in the presence of contaminants. In control, fat and oil groups, the effect of roughness are significant (p<0.001, p<0.05 and p<0.001 respectively) (fig. 1). Scanning Electron Microscopy (SEM) showed that contaminants influence the interfacial bond by different mechanisms. Although rough surfaces were associated with higher bond strength, they also generated more debris, which could negatively affect the longevity of the implant bond (fig. 2 and fig. 3). Conclusion. The results of this study underscores the importance of keeping an implant free from contamination, and that if contamination does occur, a saline rinse may further decrease the stability of an implant. Contaminants did not significantly affect the bond strength between bone cement and smooth Ti stem, although a trend of improved properties was seen in the presence of lipid based contaminants. Therefore, the influence of contaminants is more important to the shape-closed type stem. Increasing surface roughness dramatically improved the load carrying capability of the implant-cement interface even with contaminants


Orthopaedic Proceedings
Vol. 96-B, Issue SUPP_11 | Pages 86 - 86
1 Jul 2014
Spriano S Ferraris S Miola M
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Summary Statement. The problem facing this research is to promote rapid osteointegration of titanium implants and to minimise the risks of infections by the functionalization with different agents, each designed for a specific action. A patented process gives a multifunctional titanium surface. Introduction. A patented process of surface modification is described. It gives a multifunctional surface with a multiscale roughness (micro and nano topography), that is excellent for osteoblast adhesion and differentiation. It has a high degree of hydroxylation, that is relevant for inorganic bioactivity (apatite-HA precipitation) and it is ready for a functionalization with biological factors. A direct grafting of ALP has been obtained. Moreover, the growth of an antibacterial agent within the surface oxide layer can be useful in order to combine the osteoinduction ability to antimicrobial effects. The selection of an inorganic agent (metal nanoparticles) has the advantage to avoid an eventual development of antibiotic resistance by bacteria. Experimental Methods. Ti-cp and Ti6Al4V samples were polished or blasted, etched in diluted hydrofluoric acid (step 1a), oxidised in hydrogen peroxide (step 1b), incubated in Tresyl chloride (step 2a) and Alkaline phosphatase (ALP) enzyme (step 2b) [1, 2]. A water solution, containing a salt of the metal to be added to the surface as an inorganic antibacterial agent, can be introduced during the oxidation in hydrogen peroxide. Surface morphology and chemical composition were investigated by Scanning Electron Microscopy (SEM) and Field Emission Scanning Electron Microscopy (FESEM) equipped with Energy Dispersive Spectroscopy (EDS). The composition of the outermost surface layer and the chemical state of elements were analyzed by X-Ray Photoelectron Spectroscopy (XPS). The activity of grafted enzyme was studied by an enzymatic activity test. In vitro bioactivity was evaluated by soaking the samples in simulated body fluid and SEM observation to verify hydroxyapatite (HA) precipitation. Antibacterial activity has been determined by inhibition halo test against S aureus. Results and Discussion. A peculiar multi-scale topography, with spongy-like nanometric features, was obtained after the inorganic treatment (step 1a-1b). This morphology can be superimposed on the micro-or macro roughness deriving from acid etching or blasting, by properly optimizing the process parameters. Moreover, the treated surfaces present a high density of hydroxyl groups (XPS data) and they are bioactive (HA precipitation after soaking in SBF for 15 days). Metal (Ag, Cu, Zn) nanoparticles can be grown within the surface oxide layer and they are effective as antimicrobial inorganic agents. The amount of the metal nanoparticles can be tailored in order to have an antibacterial or a bacteriostatic surface. The effective grafting of ALP (step 2a-2b) has been shown by XPS because of the appearance of characteristic peaks in the carbon region. Moreover, it has been observed that ALP maintains its activity after grafting by an enzymatic activity test. ALP grafting improves HA precipitation kinetics. Conclusions. An innovative process was applied to titanium surfaces in order to obtain a better bone integration ability and antibacterial activity. A multi scale surface topography (micro and nano features) was successfully obtained together with an high hydroxylation degree. Modified surfaces are able to induce hydroxyapatite precipitation in vitro and to graft ALP, maintaining its activity and improving bioactivity. Metal nanoparticles embedded in the surface oxide layer have an antibacterial effect


Orthopaedic Proceedings
Vol. 104-B, Issue SUPP_12 | Pages 19 - 19
1 Dec 2022
Eltit F Wang Q Xu S Satra M Liu D Wang R Charest-Morin R Cox M
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One out of nine Canadian males would suffer prostate cancer (PC) during his lifetime. Life expectancy of males with PC has increased with modern therapy and 90% live >10 years. However, 20% of PC-affected males would develop incurable metastatic diseases. Bone metastases (BM) are present in ~80% of metastatic PC patients, and are the most severe complication of PC, generating severe pain, fractures, spinal cord compression, and death. Interestingly, PC-BMs are mostly osteoblastic. However, the structure of this newly formed bone and how it relates to pain and fracture are unknown. Due to androgen antagonist treatment, different PC phenotypes develop with differential dependency on androgen receptor (AR) signaling: androgen-dependent (AR+), double negative (AR-) and neuroendocrine. How these phenotypes are related to changes in bone structure has not been studied. Here we show a state-of-the-art structural characterization of PCBM and how PC phenotypes are associated to abnormal bone formation in PCBM. Cadaveric samples (n=14) obtained from metastases of PC in thoracic or lumbar vertebrae (mean age 74yo) were used to analyze bone structure. We used micro-computed tomography (mCT) to analyze the three-dimensional structure of the bone samples. After imaging, the samples were sectioned and one 3mm thick section was embedded in epoxy-resin, ground and polished. Scanning electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDS) and quantitative backscattering electron (qBSE) imaging were used to determine mineral morphology and composition. Another section was used for histological analysis of the PC-affected bone. Collagen structure, fibril orientation and extracellular matrix composition were characterized using histochemistry. Additionally, we obtained biopsies of 3 PCBM patients undergoing emergency decompression surgery following vertebral fracture and used them for immunohistological characterization. By using mCT, we observed three dysmorphic bone patterns: osteolytic pattern with thinned trabecula of otherwise well-organized structures, osteoblastic pattern defined as accumulation of disorganized matrix deposited on pre-existing trabecula, and osteoblastic pattern with minimum residual trabecula and bone space dominated by accumulation of disorganized mineralized matrix. Comparing mCT data with patho/clinical parameters revealed a trend for higher bone density in males with larger PSA increase. Through histological sections, we observed that PC-affected bone, lacks collagen alignment structure, have a higher number of lacunae and increased amount of proteoglycans as decorin. Immunohistochemistry of biopsies revealed that PC-cells inside bone organize into two manners: i) glandular-like structures where cells maintain their polarization in the expression of prostate markers, ii) diffuse infiltrate that spreads along bone surfaces, with loss of cell polarity. These cells take direct contact with osteoblasts in the surface of trabecula. We define that PCBM are mostly composed by AR+ with some double negative cells. We did not observe neuroendocrine phenotype cells. PCBMs generate predominantly osteoblastic lesions that are characterized by high lacunar density, lack of collagen organization and elevated proteoglycan content. These structural changes are associated with the infiltration of PC cells that are mostly androgen-dependent but have lost their polarization and contact directly with osteoblasts, perhaps altering their function. These changes could be associated with lower mechanical properties that led to fracture and weakness of the PCBM affected bone


Orthopaedic Proceedings
Vol. 102-B, Issue SUPP_11 | Pages 76 - 76
1 Dec 2020
Akdogan EK Baysan G Erkul G Cankurt U Havitcioglu H
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Meniscus has many important functions in the knee joint such as load bearing, shock absorption, joint stability, joint lubrication and proprioception. In the recent years, meniscus injuries have been the focus of orthopaedic surgeons and musculoskeletal tissue engineering applications because of its avascular nature. In this study, we aimed to compare the regeneration capacities of two composite scaffolds in a New Zealand Rabbit meniscal defect model. The first scaffold consists Poly-Lactic Acid (PLA) + chitosan + loofah and the second PLA + Hydroxyapatite (HAp) + loofah. In order to produce these scaffolds; 4% chitosan, 4% PLA and 4% HAp solutions were seperately prepared. The loofah pieces were saturated with these solutions and vacuum-dried for 14 days and sterilized with ethylene oxide. There were several characterizations performed such as Fourier Transform Infrared Spectroscopy (FTIR) for the investigation of chemical structure, Scanning Electron Microscopy (SEM) for morphological analysis, thermogravimetric differential thermal analysis (TGA/DTA) for thermal properties, mechanical compression and swelling ratio analysis. Moreover, in order to investigate biocompatibility of the scaffolds, WST-1 colorimetric assay at days 3, 7, 10, 14 and 21 was conducted. After these biocompatibility analysis, a 1.5-mm cylindrical defect was created in the avascular portion of the anterior horn of the medial meniscus in 14 New Zealand rabbits (2.5–3 kg weight) which were randomly grouped in two. The scaffolds were implanted at the defect site with the help of a freshly prepared fibrin glue. 8 weeks after the operation, the rabbits were sacrificed and their tissues were kept for further mechanical, radiological and histological analysis. In conclusion, we succeeded to produce a new meniscus scaffold. The proliferation ability of PLA + chitosan + loofah scaffold is higher than PLA + HAp + loofah scaffold. However, there was no statistically significant difference among them


Orthopaedic Proceedings
Vol. 102-B, Issue SUPP_11 | Pages 1 - 1
1 Dec 2020
Gögele CL Kerling V Lenhart A Wiltzsch S Schäfer-Eckart K Minnich B Weiger TM Schulze-Tanzil G
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Cartilage injuries often represent irreversible tissue damage because cartilage has only a low ability to regenerate. Thus, cartilage loss results in permanent damage, which can become the starting point for osteoarthritis. In the past, bioactive glass scaffolds have been developed for bone replacement and some of these variants have also been colonized with chondrocytes. However, the hydroxylapaptite phase that is usually formed in bioglass scaffolds is not very suitable for cartilage formation (chondrogenesis). This interdisciplinary project was undertaken to develop a novel slowly degrading bioactive glass scaffold tailored for cartilage repair by resembling the native extracellular cartilage matrix (ECM) in structure and surface properties. When colonized with articular chondrocytes, the composition and topology of the scaffolds should support cell adherence, proliferation and ECM synthesis as a prerequisite for chondrogenesis in the scaffold. To study cell growth in the scaffold, the scaffolds were colonized with human mesenchymal stromal cells (hMSCs) and primary porcine articular chondrocytes (pACs) (27,777.8 cells per mm. 3. ) for 7 – 35 d in a rotatory device. Cell survival in the scaffold was determined by vitality assay. Scanning electron microscopy (SEM) visualized cell ultramorphology and direct interaction of hMSCs and pACs with the bioglass surface. Cell proliferation was detected by CyQuant assay. Subsequently, the production of sulphated glycosaminoglycans (sGAGs) typical for chondrogenic differentiation was depicted by Alcian blue staining and quantified by dimethylmethylene blue assay assay. Quantitative real-time polymerase chain reaction (QPCR) revealed gene expression of cartilage-specific aggrecan, Sox9, collagen type II and dedifferentiation-associated collagen type I. To demonstrate the ECM-protein synthesis of the cells, the production of collagen type II and type I was determined by immunolabelling. The bioactive glass scaffold remained stable over the whole observation time and allowed the survival of hMSCs and pACs for 35 days in culture. The SEM analyses revealed an intimate cell-biomaterial interaction for both cell types showing cell spreading, formation of numerous filopodia and ECM deposition. Both cell types revealed initial proliferation, decreasing after 14 days and becoming elevated again after 21 days. hMSCs formed cell clusters, whereas pACs showed an even distribution. Both cell types filled more and more the pores of the scaffold. The relative gene expression of cartilage-specific markers could be proven for hMSCs and pACs. Cell associated sGAGs deposition could be demonstrated by Alcian blue staining and sGAGs were elevated in the beginning and end of the culturing period. While the production of collagen type II could be observed with both cell types, the synthesis of aggrecan could not be detected in scaffolds seeded with hMSCs. hMSCs and pACs adhered, spread and survived on the novel bioactive glass scaffolds and exhibited a chondrocytic phenotype


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_6 | Pages 31 - 31
1 May 2019
Dall'Ava L Hothi H Di Laura A Henckel J Shearing P Hart A
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Introduction. Three-dimensional (3D) printing of porous titanium implants marks a revolution in orthopaedics, promising enhanced bony fixation whilst maintaining design equivalence with conventionally manufactured components. No retrieval study has investigated differences between implants manufactured using these two methods. Our study was the first to compare these two groups using novel non-destructive methods. Materials and methods. We investigated 16 retrieved acetabular cups divided into ‘3D printed’ (n = 6; Delta TT) and ‘conventional’ (n = 10; Pinnacle Porocoat). The groups were matched for age, time to revision, size and gender (Table 1). Reasons for revision included unexplained pain, aseptic loosening, infection and ARMD. Visual inspection was performed to evaluate tissue attachment. Micro-CT was used to assess clinically relevant morphometric features of the porous structure, such as porosity, depth of the porous layer, pore size and strut thickness. Scanning electron microscopy (SEM) was applied to evaluate the surface morphology. Results. Significant differences (p = 0.0002) were found for all morphometric parameters (Table 2). Microscopic analysis revealed uniform beads over the backside of conventional implants, due to the manufacturing technique (Figure 1a). Conversely, beads of random size were found on 3D printed implants, representing a by-product of the manufacturing process, where some starting powder particles are not completely fused together (Figure 1b). The two groups showed similar tissue attachment (3D printed 76.9 ± 27.1%; conventional 73.8 ± 12.2%; p = 0.2635). Conclusion. This was the first study to analyse retrieved 3D printed orthopaedic implants. Differences were found between these and conventional implants, but both literature and registry data do not suggest a short-mid-term clinical issue with 3D printed components. Similar tissue on growth suggested a comparable behaviour with bone in situ. The key difference is the presence of the particles on 3D printed implants, whose clinical significance needs to be investigated. For any figures or tables, please contact the authors directly


Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_12 | Pages 57 - 57
1 Oct 2019
Gil D Grindy S Hugard S Muratoglu OK Oral E
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Introduction. Ultra-high molecular weight polyethylene (UHMWPE) can provide local sustained delivery of therapeutics. 1,2. For example, it can deliver analgesics to address post-arthroplasty pain. 2. Given that several analgesics, such as bupivacaine (anesthetic) and tolfenamic acid (NSAID), were shown to possess antibacterial activity against Staphylococci, we hypothesize that analgesic-loaded UHMWPE can also yield antimicrobial effects, preventing the development of periprosthetic joint infections. Methods. Bupivacaine and tolfenamic acid were incorporated into UHMWPE via phase-separated compression molding. Drug release from the prepared samples was measured using high-performance liquid chromatography. Antibacterial studies of the obtained materials were conducted against methicillin-sensitive, and methicillin-resistant S. aureus, as well as S. epidermidis. Time-kill curves were obtained to characterize antimicrobial activity against planktonic bacteria. The dynamics of bacterial adhesion were assessed to characterize antibiofilm activity. Scanning electron microscopy (SEM) was used to visualize adherent bacteria. Anticolonizing activity of the tested materials was characterized using the “daughter cell” method as outlined elsewhere. 3. Cytotoxicity profile of drug-loaded UHMWPEs was evaluated using MG-63 osteoblast cell line. Results. The bupivacaine release rate generally increased with increasing drug loading (e.g. a model knee implant loaded with bupivacaine would release ca. 15–500 mg over 24 hours). While also proportional, drug release from UHMWPE loaded with tolfenamic acid was much lower. The bacterial viability curves showed that bupivacaine-loaded UHMWPE possessed moderate antibacterial activity against planktonic MSSA, MRSA, and S. epidermidis, slowing bacteria proliferation by up to 70%. Bupivacaine-loaded UHMWPE also mitigated biofilm formation and development during the initial culture period. SEM images confirmed the observed antibiofilm effect (Fig. 1). Tolfenamic acid-loaded UHMWPE allowed proliferation of planktonic bacteria. At the same time, these materials showed pronounced dose-dependent anticolonizing activity against tested strains, providing 3-log reduction of “daughter” cells. Bupivacaine- and tolfenamic acid-loaded UHMWPEs showed little-to-no cytotoxicity against osteoblasts. Discussion & Conclusions. We demonstrated for the first time that bupivacaine-loaded UHMWPE possesses dose-dependent antibacterial properties against planktonic and adherent MSSA, MRSA, and S. epidermidis – pathogens commonly associated with periprosthetic joint infections. Pronounced anticolonizing activity was evident for tolfenamic acid-loaded UHMWPE. Due to the low solubility of tolfenamic acid, the material's antibacterial effect against planktonic bacteria was lower. These results demonstrate that analgesic-loaded UHMWPE, used as a tool in multimodal pain management, can also yield antibacterial effects, opening an entirely new avenue for providing post-arthroplasty antibacterial prophylaxis. This pioneering approach has a potential to reduce patients' morbidity and mortality after arthroplasty. For any tables or figures, please contact the authors directly


Orthopaedic Proceedings
Vol. 98-B, Issue SUPP_16 | Pages 44 - 44
1 Oct 2016
Stratton-Powell A Tipper JL Williams SD Redmond A Brockett CL
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Total ankle replacement (TAR) has a mean survivorship of 77% at 10 years which is poor compared to other types of joint arthroplasty. Osteolysis and aseptic loosening are commonly cited TAR failure modes, the mechanisms of which are unknown. Retrieval analyses of TAR devices may reveal mechanisms of failure similar or dissimilar to other joint replacements. This study investigated whether TAR explants exhibit similar damage modes to those recognised in other total joint replacements. 22 Ankle Evolution System TARs (Transystème, Nimes, France) were implanted and retrieved by the same surgeon. Mean implantation time was 7.8 yrs (5.3 to 12.1 range). Pain and/or loosening were the indications for revision. Macro photography, an Alicona Infinite microscope and the Hood/Wasielewski scale were used to classify damage modes on the polyethylene insert. Scanning electron microscopy with energy dispersive X-ray spectroscopy was used to determine the composition of third body debris and to image the fixation surface of the tibial components. Mean damage score was 185.4 (± 40.0 SD). Damage modes common to total knee replacements were identified on both the superior and inferior insert surfaces, these included: burnishing, scratching, pitting and abrasion. Titanium particles, hydroxyapatite fragments and bone debris were embedded in the insert surfaces. Fixation surface delamination was identified by the ongrowth of tissue between the cobalt chromium substrate and titanium alloy coating. Damage modes indicative of high levels of wear and deformation were evident. Pitting caused by third body debris was abundant and suggested fixation surface wear and failure


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_13 | Pages 1 - 1
1 Jun 2017
Panagiotopoulou V Davda K Hothi H Henckel J Cerquiglini A Goodier W Skinner J Hart A Calder P
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Introduction. The Precice nail is the latest intramedullary lengthening nail with excellent early outcomes. Implant complications have led to modification of the nail design. The aim of this study was to perform a retrieval study of Precice nails following lower limb lengthening. To assess macroscopic and microscopic changes to the implants and assess differences following design modification, with identification of potential surgical, implant and patient risk factors. Method. 15 nails were retrieved from 13 patients following lower limb lengthening. Macroscopic and microscopic surface damage to the nails were identified. Further analysis included radiology and micro-CT prior to sectioning. The internal mechanism was then analysed with Scanning Electron Microscopy and Energy Dispersive X-ray Spectroscopy to identify corrosion. Results. 7 male and 3 females underwent 12 femoral lengthenings, 9 antegrade and 3 retrograde. 3 females underwent tibial lengthening. All patients obtained the desired length with no implant failure and full regenerate consolidation. Surface degradation was noted on the telescopic part of every nail design, less on the latest implants. Microscopic analysis confirmed fretting and pitting corrosion. Following sectioning black debris was noted in all implants. The early designs were found to have fractured actuator pins and the pin and bearings had evidence of corrosive debris. The latest designs had evidence of biological deposits suggestive of fluid ingress within the nail. Conclusion. This study suggests fluid ingress occurs with every generation of Precice nail despite modifications. The presence of biological fluid could be an early warning sign of potential corrosion. This in theory could lead to actuator pin fracture and implant failure. The clinical relevance is the potential re-use of a “dormant” nail in patients requiring secondary limb segment lengthening. Retraction of the nail in-situ and re-use for further lengthening requires careful consent for potential implant failure


Orthopaedic Proceedings
Vol. 95-B, Issue SUPP_13 | Pages 50 - 50
1 Mar 2013
HAIDER A KARPHUKINA N MOHSIN S
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Introduction. Millions of bone graft surgeries have improved the quality of the life of millions, which cost about £ 2.5 billion annually worldwide. Current focus of the researcher is to synthesis of a bioactive bone scaffold which will reduce the cost and number of the bone graft surgeries. Hypothesis To synthesis novel, bioactive, porous, mechanical stable bone scaffold using bio glass and alginate. Methods. Sr releasing glass was made using by melt derived technique. Composite bone scaffold was made using bio-glass and alginate. Freeze dry technique was used to make the porous bone scaffold. Bone scaffold was characterized using XRD, DSC, FTIR, Particle size analyser, SEM, ICP, Mechanical testing, Cell culture with osteoblast and fibroblast and NMR techniques. RESULTS. Fig: Graph showing the FTIR spectra of bone scaffold. Discussion and conclusion. FTIR shows the Si-o-Si bonds of the glass and shifting of the bonding after adding the alginate and during cross linked with Cacl2 and Srcl2. Scanning electron microscopy result shows the average pore size of around 100µm which is desirable for the growth of the cell. Release of desirable Calcium and phosphate ions can be seen from the ICP-OES measurement. Compression test allows deciding that the composite material with alginate and bioactive glass shows much better result than the simple alginate scaffolds. Cell culture showed round cells on the surface of the scaffold. And the NMR shows the interaction between bio glass and alginate. Significance We were able to produce a new novel bone scaffold which can reduce the economical burden of bone grafting