A Polymer for Application as a Matrix Phase in a Concept of In Situ Curable Bioresorbable Bioactive Load-Bearing Continuous Fiber Reinforced Composite Fracture Fixation Plates.

The use of bioresorbable fracture fixation plates made of aliphatic polyesters have good potential due to good biocompatibility, reduced risk of stress-shielding, and eliminated need for plate removal. However, polyesters are ductile, and their handling properties are limited. We suggested an alternative, PLAMA (PolyLActide functionalized with diMethAcrylate), for the use as the matrix phase for the novel concept of the in situ curable bioresorbable load-bearing composite plate to reduce the limitations of conventional polyesters. The purpose was to obtain a preliminary understanding of the chemical and physical properties and the biological safety of PLAMA from the prospective of the novel concept. Modifications with different molecular masses (PLAMA-500 and PLAMA-1000) were synthesized. The efficiency of curing was assessed by the degree of convergence (DC). The mechanical properties were obtained by tensile test and thermomechanical analysis. The bioresorbability was investigated by immersion in simulated body fluid. The biocompatibility was studied in cell morphology and viability tests. PLAMA-500 showed better DC and mechanical properties, and slower bioresorbability than PLAMA-1000. Both did not prevent proliferation and normal morphological development of cells. We concluded that PLAMA-500 has potential for the use as the matrix material for bioresorbable load-bearing composite fracture fixation plates.

Local delivery of a selective androgen receptor modulator failed as an anabolic agent in a rat bone marrow ablation model.

BACKGROUND AND PURPOSE: Selective androgen receptor modulators (SARMs) have been developed to have systemic anabolic effects on bones and muscles without the adverse effects of steroidal androgens. One unexplored therapeutic option is the targeted application of SARMs for the enhancement of local new bone formation. We evaluated the osteogenic efficacy of a locally released SARM (ORM-11984). METHODS: ORM-11984 was mixed with a copolymer of L-lactide and ɛ-caprolactone (PLCL). An in vitro dissolution test confirmed the sustainable release of ORM-11984 from the matrix. A bone marrow ablation model was used in female Sprague-Dawley rats. Implants containing 10%, 30%, or 50% ORM-11984 by weight or pure PLCL were inserted into the medullary canal of the ablated tibia. At 6 and 12 weeks, the volume of intramedullary new bone and the perimeter of bone-implant contact were measured by micro-computed tomography and histomorphometry. RESULTS: Contrary to our hypothesis, there was a negative correlation between the amount of new bone around the implant and the dose of ORM-11984. There was only a mild (and not statistically significant) enhancement of bone formation in ablated bones subjected to the lowest dose of the SARM (10%). INTERPRETATION: This study suggests that intramedullary/endosteal osteogenesis had a negative, dose-dependent response to locally released SARM. This result highlights the complexity of androgenic effects on bones and also suggests that there are biological limits to the targeted local application of SARMs.

Load-bearing composite fracture-fixation devices with tailored fibre placement for toy-breed dogs.

Fibre reinforced composites are attractive materials for hard tissue reconstructions, due to the high strength and low flexural modulus. However, lack of contourability in the operation theatre inhibits their clinical applications. The study presents a novel in situ contourable composite implant system for load-bearing conditions. The implant system consists of a thin bioresorbable shell with several cavities, much like bubble-wrap. The central cavity contains a semi-flexible glass fibre preform prepared using Tailored Fibre Placement method. The preform is either pre-impregnated with a light curable resin, or the resin is injected into the cavity during the surgical procedure, followed by light curing. The semi-flexible glass fibre preforms were also examined as separate devices, "miniplates". Two types of miniplates were scrutinized, a simplified pilot design and a spatially refined, "optimized" design. The optimized miniplates were implemented as biostable and bioresorbable versions. The feasibility of the in situ contourable composite implant system was demonstrated. The potential of Tailored Fibre Placement for the semi-flexible glass fibre preforms and miniplates was confirmed in a series of biomechanical tests. However, structural optimization is required. Antebrachial fractures in toy-breeds of dogs are exemplar veterinary applications of the devices; further applications in veterinary and human patients are foreseen.

Biological response to an experimental implant for tibial tuberosity advancement in dogs: A pre-clinical study.

In this study, we propose a novel bioresorbable bioactive implant for tibial tuberosity advancement (TTA). The implant consists of a gradually resorbing load-bearing shell which encompasses rapidly resorbing small casings loaded with silica-based bioactive glass (BG) particulates which promote bone formation and reduce the risk of infection. The shell and the casings are manufactured by 3D printing from two medical grade bioresorbable polymers (a polyglycolide/lactide based and a polydioxanone based) that have different degradation rates. The casings are expected to resorb within days after surgery to expose the BG particulates while the shell would retain the load-bearing properties of the implant for the time required by bone healing. Unlike the currently used metallic devices, the novel implant is resorbed and excreted from the body once its purpose is fulfilled. This study presents a logical progression from the in vitro characterisation of the materials and implants to the in vivo investigation of the experimental implants. This included mechanical testing of the materials, finite element analysis of a preliminary design of the novel TTA implant, assessment of the degradation behaviour of the polymers and the ion exchange of BG in simulated body fluid, and investigation of the biological response to the novel implants after implantation in rabbits. The osteointegration of the novel implants was comparable to the osteointegration of Ti6Al4V implants in the control group; the biological efficacy and safety were confirmed. The biological response was in line with the expectations. The proof of concept for the novel TTA implants was demonstrated.

Biostable glass fibre-reinforced dimethacrylate-based composites as potential candidates for fracture fixation plates in toy-breed dogs: Mechanical testing and finite element analysis.

In toy-breed dogs (bodyweight <5 kg), the fractures of the radius and ulna are particularly common and can be caused by minimal trauma. While fracture fixation using metallic plates is a feasible treatment modality, the excessive stiffness of these devices produces the underloading of the bone which may result in the adverse bone remodelling and complications in the healing of the fracture. In this study, we investigated bisphenol A glycidylmethacrylate -based glass fibre reinforced composites as potential alternatives to metals in the devices intended for the fracture fixation of the distal radius in toy-breed dogs. Four composites with different glass fibre reinforcements were prepared as rectangular specimens and as fracture fixation plates. These were mechanically tested in three-point and four-point bending. There were two controls: polyether etherketone reinforced with short carbon fibres (specimens and plates) and commercially available stainless-steel plates. Finite element simulations were used for the assessment of the behaviour of the plates. For the control stainless steel plate, the bending strength was 1.358 N*m, superior to that of any of the composite plates. The composite plate with the matrix reinforced with continuous unidirectional glass fibres had the bending strength of 1.081 N*m, which is sufficient in this clinical context. For the plates made of polyether etherketone reinforced with carbon fibres, the strength was 0.280 N*m. Similar conclusions on the biomechanical behaviour of the plates could be made solely based on the results of the finite element simulations, provided the geometries and the material properties are well defined.

Hydroxyapatite and bioactive glass surfaces for fiber reinforced composite implants via surface ablation by Excimer laser.

In skeletal reconstructions, composites, such as bisphenol-A-glycidyldimethacrylate resin reinforced with glass fibers, are potentially useful alternatives to metallic implants. Recently, we reported a novel method to prepare bioactive surfaces for these composites. Surface etching by Excimer laser was used to expose bioactive glass granules embedded in the resin. The purpose of this study was to analyze two types of bioactive surfaces created by this technique. The surfaces contained bioactive glass and hydroxyapatite granules. The selected processing parameters were adequate for the creation of the surfaces. However, the use of porous hydroxyapatite prevented the complete exposure the granules. In cell culture, for bioactive glass coatings, the pattern of proliferation of MG63 cells was comparable to that in the positive control group (Ti6Al4V) while inferior cell proliferation was observed on the surfaces containing hydroxyapatite granules. Scanning electron microscopy revealed osteointegration of implants with both types of surfaces. The technique is suitable for the exposure of solid bioactive glass granules. However, the long-term performance of the surfaces needs further assessment.

Bioactive glass surface for fiber reinforced composite implants via surface etching by Excimer laser.

Biostable fiber-reinforced composites (FRC) prepared from bisphenol-A-glycidyldimethacrylate (BisGMA)-based thermosets reinforced with E-glass fibers are promising alternatives to metallic implants due to the excellent fatigue resistance and the mechanical properties matching those of bone. Bioactive glass (BG) granules can be incorporated within the polymer matrix to improve the osteointegration of the FRC implants. However, the creation of a viable surface layer using BG granules is technically challenging. In this study, we investigated the potential of Excimer laser ablation to achieve the selective removal of the matrix to expose the surface of BG granules. A UV-vis spectroscopic study was carried out to investigate the differences in the penetration of light in the thermoset matrix and BG. Thereafter, optimal Excimer laser ablation parameters were established. The formation of a calcium phosphate (CaP) layer on the surface of the laser-ablated specimens was verified in simulated body fluid (SBF). In addition, the proliferation of MG63 cells on the surfaces of the laser-ablated specimens was investigated. For the laser-ablated specimens, the pattern of proliferation of MG63 cells was comparable to that in the positive control group (Ti6Al4V). We concluded that Excimer laser ablation has potential for the creation of a bioactive surface on FRC-implants.

Osteointegration of PLGA implants with nanostructured or microsized ß-TCP particles in a minipig model.

Bioresorbable suture anchors and interference screws have certain benefits over equivalent titanium-alloy implants. However, there is a need for compositional improvement of currently used bioresorbable implants. We hypothesized that implants made of poly(l-lactide-co-glycolide) (PLGA) compounded with nanostructured particles of beta-tricalcium phosphate (ß-TCP) would induce stronger osteointegration than implants made of PLGA compounded with microsized ß-TCP particles. The experimental nanostructured self-reinforced PLGA (85L:15G)/ß-TCP composite was made by high-energy ball-milling. Self-reinforced microsized PLGA (95L:5G)/ß-TCP composite was prepared by melt-compounding. The composites were characterized by gas chromatography, Ubbelohde viscometry, scanning electron microscopy, laser diffractometry, and standard mechanical tests. Four groups of implants were prepared for the controlled laboratory study employing a minipig animal model. Implants in the first two groups were prepared from nanostructured and microsized PLGA/ß-TCP composites respectively. Microroughened titanium-alloy (Ti6Al4V) implants served as positive intra-animal control, and pure PLGA implants as negative control. Cone-shaped implants were inserted in a random order unilaterally in the anterior cortex of the femoral shaft. Eight weeks after surgery, the mechanical strength of osteointegration of the implants was measured by a push-out test. The quality of new bone surrounding the implant was assessed by microcomputed tomography and histology. Implants made of nanostructured PLGA/ß-TCP composite did not show improved mechanical osteointegration compared with the implants made of microsized PLGA/ß-TCP composite. In the intra-animal comparison, the push-out force of two PLGA/ß-TCP composites was 35-60% of that obtained with Ti6Al4V implants. The implant materials did not result in distinct differences in quality of new bone surrounding the implant.

(68)Ga-DOTA-Siglec-9 PET/CT imaging of peri-implant tissue responses and staphylococcal infections.

BACKGROUND: Staphylococcus epidermidis (S. epidermidis) has emerged as one of the leading pathogens of biomaterial-related infections. Vascular adhesion protein-1 (VAP-1) is an inflammation-inducible endothelial molecule controlling extravasation of leukocytes. Sialic acid-binding immunoglobulin-like lectin 9 (Siglec-9) is a leukocyte ligand of VAP-1. We hypothesized that (68)Ga-labeled 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-conjugated Siglec-9 motif containing peptide ((68)Ga-DOTA-Siglec-9) could detect inflammatory response due to S. epidermidis peri-implant infection by positron emission tomography (PET). METHODS: Thirty Sprague-Dawley rats were randomized into three groups. A sterile catheter was implanted into the medullary canal of the left tibia. In groups 1 and 2, the implantation was followed by peri-implant injection of S. epidermidis or Staphylococcus aureus (S. aureus) with adjunct injections of aqueous sodium morrhuate. In group 3, sterile saline was injected instead of bacteria and no aqueous sodium morrhuate was used. At 2 weeks after operation, (68)Ga-DOTA-Siglec-9 PET coupled with computed tomography (CT) was performed with the measurement of the standardized uptake value (SUV). The presence of the implant-related infection was verified by microbiological analysis, imaging with fluorescence microscope, and histology. The in vivo PET results were verified by ex vivo measurements by gamma counter. RESULTS: In group 3, the tibias with implanted sterile catheters showed an increased local uptake of (68)Ga-DOTA-Siglec-9 compared with the intact contralateral bones (SUVratio +29.5%). (68)Ga-DOTA-Siglec-9 PET detected inflammation induced by S. epidermidis and S. aureus catheter-related bone infections (SUVratio +58.1% and +41.7%, respectively). The tracer uptake was significantly higher in the S. epidermidis group than in group 3 without bacterial inoculation, but the difference between S. epidermidis and S. aureus groups was not statistically significant. The difference between the S. aureus group and group 3 was neither statistically significant. CONCLUSION: PET/CT imaging with novel (68)Ga-DOTA-Siglec-9 tracer was able to detect inflammatory tissue response induced by catheter implantation and staphylococcal infections.

Biofilm medium leads to apatite formation on bioactive surfaces.

PURPOSE: When investigating apatite formation on biomaterial surfaces, simulated body fluid (SBF) is used as an in vitro solution, however, it does not provide an appropriate environment for the growth of bacterial biofilm. The aim of the present study was to compare the bioactivity in terms of apatite formation on two bioactive glass (BAG) composite surfaces using both SBF and bacterial-biofilm growing medium (BM). METHODS: Polymer composite substrates with different percentages of BAG-particles (50% and 75% by weight) were prepared. Plain resin substrates were used as a negative control. The substrates were immersed in SBF and BM for 3 days. The surface and, subsequently, the cross-sections of the substrates were examined with scanning electron microscopy (SEM) and x-ray energy dispersive spectroscopy (EDS). RESULTS: All the investigated BAG-composite surfaces showed apatite formation after immersion in SBF and BM liquid media. CONCLUSIONS: The use of BM is a promising method for studies involving simultaneous biofilm growth and apatite formation on bioactive surfaces.