Introduction of the Anspach drill as a novel surgical driller for creating calvarial defects in animal models.

Standardized and reproducible animal models are required for the assessment of bone healing mediated by biomaterials, cells, and drugs. Among the available bone-fractured models, calvarial defect is a simple and adequate option when researchers investigate intra-membranous bone formation and the influence of their regenerative solutions. However, the conventional surgical tools required to perform calvaria osteotomies (i.e., trephine bur mounted on a dental handpiece, DS) can affect the subjacent tissues of the skull, which are the dura mater and the brain. We hypothesized that the quality of the calvaria defect and the preservation of underlying tissues can be improved using a novel Anspach high-speed drill with a Codman Neuro Disposable Perforator handheld (ACP). Using a rabbit cadaveric models, we performed calvarial defects with either conventional DS or the ACP system, and showed that both techniques allowed to create standardized defects with circular geometry. We demonstrated that the ACP had clear benefices in terms of DM preservation and absence of exothermic reaction upon drilling. Even though this comparative work was conducted on cadaver, it is of substantial importance as it introduces a novel technique, easily applicable to investigate calvaria bone healing, which brings clear advantages compared to the conventional dental station. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 9999:1-9, 2019.

Infection burden and immunological responses are equivalent for polymeric and metallic implant materials in vitro and in a murine model of fracture-related infection.

The development of an infection is a major complication for some patients with implanted biomaterials. Whether the material or surface composition of the used biomaterial influences infection has not been directly compared for key biomaterials currently in use in human patients. We conducted a thorough in vitro and in vivo investigation using titanium (Ti) and polyether-ether-ketone (PEEK) as both commercially available and as modified equivalents (surface polished Ti, and oxygen plasma treated PEEK). Complement activation and cytokine secretion of cell of the immune system was assessed in vitro for all materials in the absence and presence of bacterial stimulants. In a follow-up in vivo study, we monitored bacterial infection associated with clinically available and standard Ti and PEEK inoculated with Staphylococcus aureus. Complement activation was affected by material choice in the absence of bacterial stimulation, although the material based differences were largely lost upon bacterial stimulation. In the in vivo study, the bacterial burden, histological response and cytokine secretion suggests that there is no significant difference between both PEEK and Ti. In conclusion, the underlying material has a certain impact in the absence of bacterial stimulation, however, in the presence of bacterial stimulation, bacteria seem to dictate the responses in a manner that overshadows the influence of material surface properties. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1095-1106, 2019.

Comparison of intraperitoneal ropivacaine and bupivacaine for postoperative analgesia in dogs undergoing ovariohysterectomy.

OBJECTIVE: To compare postoperative analgesia following either intraperitoneal (IP) ropivacaine or bupivacaine in dogs undergoing ovariohysterectomy (OVH) in the scope of multimodal analgesia. STUDY DESIGN: Prospective, randomized, blinded clinical study. ANIMALS: A total of 45 privately owned dogs undergoing OVH, aged 37 ± 28 months and weighing 11.3 ± 4.5 kg. METHODS: Dogs were premedicated with acepromazine (0.05 mg kg-1) and morphine (0.5 mg kg-1) intramuscularly (IM). Anaesthesia was induced with alfaxalone and maintained with isoflurane in oxygen. Carprofen (4 mg kg-1) was injected subcutaneously after intubation. Dogs were randomly assigned to receive either bupivacaine (group B; 3 mg kg-1) or ropivacaine (group R; 3 mg kg-1) IP prior to complete closure of the linea alba. At 0.5, 1, 2, 4, 6 and 8 hours after extubation, sedation and postoperative pain were assessed, using the short form of the Glasgow Composite Pain scale (GCPS-SF), a dynamic interactive visual analogue scale (DIVAS), and mechanical nociceptive threshold (MNT) measurement. Rescue morphine (0.2 mg kg-1) was administered in case of ≥ 5/20 or ≥ 6/24 in the GCPS-SF and/or >40 mm in the DIVAS. Parametric data were compared using the t test; nonparametric data were analysed with the two-sample Wilcoxon test (p < 0.05). RESULTS: The GCPS-SF score was significantly higher in group R at 8 hours. There was no other significant difference regarding sedation or analgesia between the groups. Rescue analgesia was administered to 15 dogs (R: 9/22; B: 6/22), with no significant difference between the groups. MNT values decreased in both groups at all time points when compared to baseline. No adverse effects were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Ropivacaine or bupivacaine IP in combination with morphine IM and carprofen SC provided comparable postoperative analgesia in dogs after OVH for 6 hours. However, the anaesthetic protocol used did not prevent the administration of rescue analgesia in 41% of animals.

Clinical evaluation of intranasal medetomidine-ketamine and medetomidine-S(+)-ketamine for induction of anaesthesia in rabbits in two centres with two different administration techniques.

OBJECTIVE: The aim was to compare efficacy and side effects of induction with medetomidine-ketamine or medetomidine-S(+)-ketamine by intranasal (IN) instillation in rabbits and to evaluate both protocols during subsequent isoflurane anaesthesia. STUDY DESIGN: Prospective, blinded, randomized experimental study in two centres. ANIMALS: Eighty-three healthy New Zealand White rabbits undergoing tibial or ulnar osteotomy. METHODS: Medetomidine (0.2 mg kg-1) with 10 mg kg-1 ketamine (MK) or 5 mg kg-1 S(+)-ketamine (MS) was administered IN to each rabbit in a randomized fashion. In Centre 1 (n = 42) rabbits were held in sternal recumbency, and in Centre 2 (n = 41) in dorsal recumbency, during drug instillation. Adverse reactions were recorded. If a rabbit swallowed during endotracheal intubation, half of the initial IN dose was repeated and intubation was re-attempted after 5 minutes. Anaesthesia was maintained with isoflurane. Heart rate, blood pressure, endtidal carbon dioxide concentration and blood gases were recorded. Data were analysed using Student's t-test, Mann-Whitney test and Fisher's exact test. RESULTS: In all, 39 animals were assigned to the MK group and 44 to the MS group. Two rabbits in the MS group held in dorsal recumbency died after instillation of the drug. Eight (MK) and 11 rabbits (MS) were insufficiently anaesthetized and received a second IN dose. One rabbit in MK and three in MS required an isoflurane mask induction after the second IN dose. There were no significant differences between treatments for induction, intraoperative data, blood gas values and recovery data. CONCLUSION AND CLINICAL RELEVANCE: This study indicated that medetomidine-ketamine and medetomidine-S(+)-ketamine were effective shortly after IN delivery, but in dorsal recumbency IN administration of S(+)-ketamine led to two fatalities. Nasal haemorrhage was noted in both cases; however, the factors leading to death have not been fully elucidated.

Salter-Harris type II metacarpal and metatarsal fracture in three foals. Treatment by minimally-invasive lag screw osteosynthesis combined with external coaptation.

OBJECTIVES: To describe minimally-invasive lag screw osteosynthesis combined with external coaptation for the treatment of Salter-Harris type II third metacarpal and third metatarsal bone fractures. METHODS: Three foals aged two weeks to four months with a Salter-Harris type II third metacarpal or third metatarsal fracture. Surgery was carried out under general anaesthesia in lateral recumbency. After fracture reduction, the metaphyseal fragment was stabilized with two cortical screws placed in lag fashion under fluoroscopic control. A cast was applied for at least two weeks. RESULTS: All foals had a good outcome with complete fracture healing and return to complete soundness without any angular limb deformity. All foals had moderate transient digital hyperextension after cast removal. CLINICAL SIGNIFICANCE: Internal fixation of Salter-Harris type II third metacarpal or third metatarsal fractures with two cortical screws in lag fashion, combined with external coaptation provided good stabilization and preserved the longitudinal growth potential of the injured physis.

Cardiopulmonary effects and anaesthesia recovery quality in horses anaesthetized with isoflurane and low-dose S-ketamine or medetomidine infusions.

OBJECTIVES: To evaluate cardiopulmonary effects and anaesthesia recovery quality in horses anaesthetized with isoflurane receiving medetomidine or S-ketamine infusions. STUDY DESIGN: Randomized, blinded, prospective clinical trial. ANIMALS: Fifty horses undergoing elective surgery. METHODS: After acepromazine and flunixin meglumine premedication, horses received medetomidine (7 µg kg-1 ) intravenously (IV). Anaesthesia was induced with midazolam and racemic ketamine (Med treatment group; 2.2 mg kg-1 ; n = 25) or S-ketamine (S-ket treatment group; 1.1 mg kg-1 ; n = 25) IV and maintained with isoflurane in oxygen/air and medetomidine (Med; 3.5 µg kg-1 hour-1 ) or S-ketamine (S-ket; 0.5 mg kg-1 hour-1 ). All horses were mechanically ventilated. Cardiopulmonary variables were evaluated. Isoflurane end-tidal concentrations (Fe'Iso), dobutamine requirements and thiopental boli were recorded. Plasma samples were collected in six horses to evaluate S-ketamine and S-norketamine concentrations. After surgery, medetomidine 2 µg kg-1 was administered IV. Four independent observers scored recovery using a visual analogue scale and a numerical rating scale. RESULTS: Both groups required similar mean Fe'Iso (1%). However, S-ket horses needed more thiopental boli. Median intraoperative cardiac index values were higher with S-ket (4.5 L minute-1  m-2 ) than Med (3.9 L minute-1  m-2 ). Overall, there were no differences in heart rate, blood pressure or dobutamine requirements; however, horses in S-ket showed higher heart rate values at 30 minutes after anaesthesia induction. Compared with Med horses, S-ket horses showed decreased PaO2 and increased pulmonary venous admixture values estimated with the Fshunt calculation. Recoveries were shorter and of poorer quality with S-ket. During infusion, S-ketamine and S-norketamine plasma concentrations lay in the ranges of 0.209-0.917 µg mL-1 and 0.250-0.723 µg mL-1 , respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Despite the higher intraoperative cardiac index with S-ket, both protocols were considered to provide acceptable cardiovascular function. However, recovery quality was significantly better in the Med group.

Monitoring immune responses in a mouse model of fracture fixation with and without Staphylococcus aureus osteomyelitis.

Post-traumatic bone fractures are commonly fixed with implanted devices to restore the anatomical position of bone fragments and aid in the healing process. Bacterial infection in this situation is a challenge for clinicians due to the need for aggressive antibiotic therapy, debridement of infected tissues, and the need to maintain fracture stability. The aim of this study was to monitor immune responses that occur during healing and during Staphylococcus aureus infection, in a clinically relevant murine model of fracture fixation. Skeletally mature C57bl/6 mice received a transverse osteotomy of the femur, which was treated with commercially available titanium fracture fixation plates and screws. In the absence of infection, healing of the fracture was complete within 35days and was characterized by elevated Interleukin (IL)-4 and Interferon-gamma secretion from bone-derived cells and expression of these same genes. In contrast, mice inoculated with S. aureus could not heal the fracture within the observation period and were found to develop typical signs of implant-associated bone infection, including biofilm formation on the implant and osteolysis of surrounding bone. The immune response to infection was characterized by a TH17-led bone response, and a pro-inflammatory cytokine-led Tumor necrosis factor (TNF)-α, Interleukin (IL)-1ß) soft tissue response, both of which were ineffectual in clearing implant related bone and soft tissue infections respectively. In this murine model, we characterize the kinetics of pro-inflammatory responses to infection, secondary to bone trauma and surgery. A divergent local immune polarization is evident in the infected versus non-infected animals, with the immune response ultimately unable to clear the S. aureus infection.

The use of BoneWelding® technology in spinal surgery: an experimental study in sheep.

The innovative BoneWelding(®) technology, where ultrasound energy bonds bioresorbable implants to bone, was tested for its feasibility in spine surgery and its local thermal effects. The three tested concepts consisted of implementation of a resorbable plating system, two converging polymer pins and suture anchors to the cervical vertebral bodies. Bioresorbable polylactide implants (PLDLLA 70/30) were inserted ventrally into the third and fourth vertebral body of seven sheep, of which six were sacrificed at 2 months and one sheep immediately after temperature measurements during implant insertion. Polymer screws were used as controls. Qualitative, semi-quantitative histological, and quantitative histomorphometrical evaluation showed excellent anchorage of the implants, new mineralized bone at the implant-bone interface, no inflammatory cell reaction or thermal damage to the adjacent bone in response to the novel insertion technology. The application of two converging pins, parallel inserted polymer pins, or fusion of the implant to the polymer plates did not affect the overall excellent tissue tolerance of the technology. Temperature increase during insertion was noticed but never exceeded 47°C for less than 1 s. The BoneWelding(®) technology was proven to be safe and easy to apply.

Sequestration of extracellular hemoglobin within a haptoglobin complex decreases its hypertensive and oxidative effects in dogs and guinea pigs.

Release of hemoglobin (Hb) into the circulation is a central pathophysiologic event that contributes to morbidity and mortality in chronic hemolytic anemias and severe malaria. These toxicities arise from Hb-mediated vasoactivity, possibly due to NO scavenging and localized tissue oxidative processes. Currently, there is no established treatment that targets circulating extracellular Hb. Here, we assessed the role of haptoglobin (Hp), the primary scavenger of Hb in the circulation, in limiting the toxicity of cell-free Hb infusion. Using a canine model, we found that glucocorticoid stimulation of endogenous Hp synthesis prevented Hb-induced hemodynamic responses. Furthermore, guinea pigs administered exogenous Hp displayed decreased Hb-induced hypertension and oxidative toxicity to extravascular environments, such as the proximal tubules of the kidney. The ability of Hp to both attenuate hypertensive responses during Hb exposure and prevent peroxidative toxicity in extravascular compartments was dependent on Hb-Hp complex formation, which likely acts through sequestration of Hb rather than modulation of its NO- and O2-binding characteristics. Our data therefore suggest that therapies involving supplementation of endogenous Hb scavengers may be able to treat complications of acute and chronic hemolysis, as well as counter the adverse effects associated with Hb-based oxygen therapeutics.