In Vitro and In Vivo Evaluation of Ultrasound-Triggered Release From Novel Spinal Device.

OBJECTIVES: Bacterial infection following spinal fusion is a major clinical concern with up to 20% incidence. An ultrasound-triggered bulk-release system to combat postsurgical bacterial survival was designed and evaluated. METHODS: Polylactic acid (PLA) clips were loaded with vancomycin (VAN) and microbubbles (Sonazoid, GE HealthCare) in vitro. Stability was determined over 14 days. VAN-loaded clips were submerged in water and insonated using a Logiq E10 scanner (GE HealthCare) with a curvilinear C6 probe. Doppler-induced VAN release was quantified using spectrophotometry. For in vivo testing, clips were loaded with methylene blue (MeB) solution and Sonazoid. These clips were implanted into a rabbit along the spine at L2 and L5, as well as a pig at L1 and L3, then insonated in Doppler mode using the C6 probe. RESULTS: Sonazoid microbubbles were better preserved when incubated in VAN compared with distilled water at 4°C, 25°C, and 37°C incubation temperatures (P = .0131). Contrast enhancement was observed from both solutions when incubated at 4°C storage conditions. Insonated clips achieved average cumulative VAN release of 101.8 ± 2.8% (81.4 ± 2.8 mg) after 72 hours. Uninsonated clips had only 0.3 ± 0.1% (0.3 ± 0.1 mg) average cumulative VAN release (P < .0001). Clips retrieved from the rabbit did not rupture with insonation nor produce MeB staining of surrounding tissues. In the pig, the PLA film was visibly ruptured and MeB tissue was observed following insonation, whereas the uninsonated clip was intact. CONCLUSION: These results demonstrate ultrasound-triggered release of an encapsulated prophylactic solution and provide an important proof-of-concept for continuing large animal evaluations for translational merit.

Fixation strength of swelling copolymeric anchors in artificial bone.

Fixation with suture anchors and metallic hardware for osteosynthesis is common in orthopedic surgeries. Most metallic commercial bone anchors achieve their fixation to bone through shear of the bone located between the threads. They have several deficiencies, including stress-shielding due to mechanical properties mismatch, generation of acidic by-products, poor osteointegration, low mechanical strength and catastrophic failure often associated with large bone defects that may be difficult to repair. To overcome these deficiencies, a swelling porous copolymeric material, to be used as bone anchors with osteointegration potential, was introduced. The purpose of this study was to investigate the fixation strength of these porous, swelling copolymeric bone anchors in artificial bone of various densities. The pull-out and subsidence studies indicate an effective fixation mechanism based on friction including re-fixation capabilities, and minimization of damage following complete failure. The study suggests that this swelling porous structure may provide an effective alternative to conventional bone anchors, particularly in low-density bone.

A porous swelling copolymeric material for improved implant fixation to bone.

Most metallic commercial bone anchors, such as screws and suture anchors achieve their fixation to bone through shear of the bone located between the threads. They have several deficiencies, potentially leading to failure, which are particularly evident in low-density bone. These include stress-shielding resulting from mechanical properties mismatch; lack of mechanically induced remodeling and osteointegration; and when the pullout force on the anchor, during functional activities, exceeds their pullout strength, catastrophic failure occurs leaving behind large bone defects that may be hard to repair. To overcome these deficiencies, we introduced in this study a porous swelling co-polymeric material and studied its swelling and compressive mechanical characteristics as bone anchor under different configurations. Porosity was achieved by adding a non-dissolvable agent (NaCl) during the process of polymerization, which was later dissolved in water, leaving behind a porous structure with adequate porosity for osteointegration. Three different groups of cylindrical samples of the swelling co-polymer were investigated. Solid, fully porous, and partially porous with a solid core and a porous outer layer. The results of the swelling and simple compression study show that the partially porous swelling co-polymer maintains excellent mechanical properties matching those of cancellous bone, quick swelling response, and an adequate porous outer layer for mechanically induced osteointegration. These suggest that this material may present an effective alternative to conventional bone anchors particularly in low-density bone.

Numerical analysis of the mechanical response of novel swelling bone implants in polyurethane foams.

In this study, a numerical framework was developed in order to analyze the swelling properties, mechanical response and fixation strength of swelling bone anchors. Using this framework, fully porous and solid implants, along with a novel hybrid design (consisting of a solid core and a porous sleeve), were modeled and studied. Free swelling experiments were conducted to investigate their swelling characteristics. The finite element model of swelling was validated using the conducted free swelling. Compared with the experimental data, results obtained from the finite element analysis proved the reliability of this frame-work. Afterwards, the swelling bone anchors were studied embedded in artificial bones with different densities with two different interface properties: considering frictional interface between the bone anchors and artificial bones (simulating the stages prior to osteointegration, when the bone and implant are not fully bonded and the surface of the implant can slide along the interface), and perfectly bonded (simulating the stages subsequent to osteointegration, when the bone and implant are fully bonded). It was observed that the swelling considerably decreases while the average radial stress on the lateral surface of the swelling bone anchor surges in the denser artificial bones. Ultimately, the pull-out experiments and simulations of the swelling bone anchors from the artificial bones were conducted to look into the fixation strength of the swelling bone anchors. It was found that the hybrid swelling bone anchor exhibits mechanical and swelling properties close to those of solid bone anchors, while also bone in-growth is expected to happen, which is an integral factor to these bone anchors.

Influence of a resilient, hard-carbon thin film on drilling efficiency and thermogenesis.

OBJECTIVE: To determine changes in drill bit performance attributable to application of a triaxially resilient, hard-carbon thin film. STUDY DESIGN: In vitro mechanical study. METHODS: Five matched pairs of control bits and bits with a carbon nanofilm applied were tested in equine cortical bone and a synthetic bone substrate. Thirty sequential holes were drilled with each bit. Drilling time was recorded for all holes. Maximum substrate temperature was measured with infrared thermography for holes 1, 15, and 30. Drilling time, prolongation of drilling time over successive uses, and maximum substrate temperature were compared between control and test bits in each substrate. RESULTS: Drilling time was significantly reduced with test bits in both substrates. Drilling time over successive osteotomies increased more slowly with test bits than with control bits. Test bits generated significantly lower substrate temperatures during drilling. CONCLUSIONS: Bits with the carbon nanofilm completed osteotomy faster and generated less heat than control bits. Test bits also had less degradation of drilling performance with repeated use.

Injectable radiopaque and bioactive polycaprolactone-ceramic composites for orthopedic augmentation.

The aim of this study was to develop and characterize an injectable bone void filler by incorporating baghdadite (Ca3 ZrSi2 O9 ) particles (average size of 1.7 µm) into polycaprolactone (PCL). A series of PCL composites containing different volume percentages of baghdadite [1 (PCL-1%Bag), 5 (PCL-5%Bag), 10 (PCL-10%Bag), 20 (PCL-20%Bag), and 30 (PCL-30%Bag)] were prepared, and their injectability, setting time, mechanical properties, radiopacity, degradation, and cytocompatibility were investigated. PCL, PCL-1%Bag, PCL-5%Bag, and PCL-10%Bag were able to be injected through a stainless steel syringe (Length: 9.0 mm, nozzle diameter: 2.2 mm) at 75°C at injection forces of below 1.5 kN. The core temperature of the injected material at the nozzle exit ranged between 55 and 60°C and was shown to set after 2.5-3.5 min postinjection in a 37°C environment. Injection force, melt viscosity, and radiopacity of the composites increased with increasing baghdadite content. Incorporation of 10-30 vol % baghdadite into PCL increased the compressive strength of the composites from 36 to 47.1 MPa, compared with that for pure PCL (31.4 MPa). Similar trend was found for the compressive modulus of the composites, which increased from 203.8 to 741 MPa, compared with that for pure PCL (205 MPa). Flexural strain of PCL, PCL-5%Bag, and PCL-10%Bag exceeded 30%, and PCL-10%Bag showed the highest flexural strength (29.8 MPa). Primary human osteoblasts cultured on PCL-10%Bag showed a significant upregulation of osteogenic genes compared with pure PCL. In summary, our results demonstrated that PCL-10%Bag could be a promising injectable material for orthopedic and trauma application.

Repair of dense connective tissues via biomaterial-mediated matrix reprogramming of the wound interface.

Repair of dense connective tissues in adults is limited by their intrinsic hypocellularity and is exacerbated by a dense extracellular matrix (ECM) that impedes cellular migration to and local proliferation at the wound site. Conversely, healing in fetal tissues occurs due in part to an environment conducive to cell mobility and division. Here, we investigated whether the application of a degradative enzyme, collagenase, could reprogram the adult wound margin to a more fetal-like state, and thus abrogate the biophysical impediments that hinder migration and proliferation. We tested this concept using the knee meniscus, a commonly injured structure for which few regenerative approaches exist. To focus delivery and degradation to the wound interface, we developed a system in which collagenase was stored inside poly(ethylene oxide) (PEO) electrospun nanofibers and released upon hydration. Through a series of in vitro and in vivo studies, our findings show that partial digestion of the wound interface improves repair by creating a more compliant and porous microenvironment that expedites cell migration to and/or proliferation at the wound margin. This innovative approach of targeted manipulation of the wound interface, focused on removing the naturally occurring barriers to adult tissue repair, may find widespread application in the treatment of injuries to a variety of dense connective tissues.

Hydrophobic polycationic coatings that inhibit biofilms and support bone healing during infection.

Adhesion of microorganisms to biomaterials with subsequent formation of biofilms on such foreign bodies as orthopedic trauma hardware is a critical factor in implant-associated infections; once a biofilm has been established, its microorganisms become recalcitrant to the host's immune surveillance and markedly resistant to drugs. We have previously reported that painting with the hydrophobic polycation N,N-dodecyl,methyl-PEI (PEI = polyethylenimine) renders solid surfaces bactericidal in vitro. Herein we observe that N,N-dodecyl,methyl-PEI-derivatized titanium and stainless steel surfaces resist biofilm formation by Staphylococcus aureus compared to the untreated ones. Using imaging, microbiology-, histopathology-, and scanning electron microscopy (SEM) experiments in a clinically relevant large-animal (sheep) trauma model, we subsequently demonstrate in vivo that orthopedic fracture hardware painted with N,N-dodecyl,methyl-PEI not only prevents implant colonization with biofilm but also promotes bone healing. Functionalizing orthopedic hardware with hydrophobic polycations thus holds promise in supporting bone healing in the presence of infection in veterinary and human orthopedic patients.

Vancomycin-modified implant surface inhibits biofilm formation and supports bone-healing in an infected osteotomy model in sheep: a proof-of-concept study.

BACKGROUND: Implant-associated infections contribute to patient morbidity and health care costs. We hypothesized that surface modification of titanium fracture hardware with vancomycin would support bone-healing and prevent bacterial colonization of the implant in a large-animal model. METHODS: A unilateral transverse mid-diaphyseal tibial osteotomy was performed and repaired with a titanium locking compression plate in nine sheep. Four control animals were treated with an unmodified plate and five experimental animals were treated with a vancomycin-modified plate. The osteotomy was inoculated with 2.5 × 106 colony-forming units of Staphylococcus aureus. The animals were killed at three months postoperatively, and implants were retrieved aseptically. Microbiologic and histologic analyses, scanning electron and confocal microscopy, and microcomputed tomography were performed. RESULTS: All animals completed the study. Compared with the treatment cohort, control animals exhibited protracted lameness in the operatively treated leg. Gross findings during necropsy were consistent with an infected osteotomy accompanied by a florid and lytic callus. Microcomputed tomography and histologic analysis of the tibiae further supported the presence of septic osteomyelitis in the control cohort. Thick biofilms were also evident, and bacterial cultures were positive for Staphylococcus aureus in three of four control animals. In contrast, animals treated with vancomycin-treated plates exhibited a healed osteotomy site with homogenous remodeling, there was no evidence of biofilm formation on the retrieved plate, and bacterial cultures from only one of five animals were positive for Staphylococcus aureus. CONCLUSIONS: Vancomycin-derivatized plate surfaces inhibited implant colonization with Staphylococcus aureus and supported bone-healing in an infected large-animal model.

Evaluation of a fiber reinforced drillable bone cement for screw augmentation in a sheep model--mechanical testing.

We evaluated the mechanical properties of a novel fiber reinforced calcium phosphate at time zero and after 12 weeks in vivo using a sheep long bone osteotomy model. Time zero data were obtained and compared by pullout testing of 4.5 mm bone screws from bone proper and overdrilled defects of 4.5 and 8 mm diameter. Defects were augmented with: polymethylmethacrylate (PMMA), calcium phosphate, and fiber reinforced calcium phosphate using cadaveric sheep tibiae. Twelve-week data were obtained from explanted tibiae of sheep that underwent unilateral tibial osteotomy surgery repaired with a locking compression plate. The most distal hole was overdrilled to 4.5 or 8 mm diameter, filled with fiber reinforced cement, drilled, tapped and a 4.5 mm screw was placed. Screw holding strength at t= 0 was significantly higher for reinforced when compared to nonreinforced cement, but not different from bone or PMMA in 4.5 mm defects. There was no difference in pullout strength for the 8 mm defect data. After 12 weeks fiber reinforced pullout strength increased by 45% and 8.9% for 4.5 and 8 mm defects, respectively, when compared to t= 0 testing. Fiber reinforced calcium phosphate bone cement can be drilled and tapped to support orthopedic hardware for trauma applications.