Three-dimensional volume rendering planning, surgical treatment, and clinical outcomes for femoral and tibial detorsional osteotomies in dogs.

OBJECTIVES: To describe a computed tomographic (CT) methodology for planning the correction of femoral and tibial torsion and report the clinical outcomes after femoral (FDO) and tibial (TDO) detorsional osteotomy in dogs affected by torsion malalignment and patellar luxation (PL). STUDY DESIGN: Multicenter retrospective study. ANIMALS: Eighteen client-owned dogs. METHODS: Dogs underwent CT to measure femoral (FTA) and tibial torsion angle (TTA). Abnormal femoral external torsion was defined when FTA <20°, abnormal femoral internal torsion if FTA >35°; abnormal tibial external torsion was defined when TTA < -10°, and abnormal tibial internal torsion when TTA >2°. The cortical arch length (CAL) was measured with CT and used intraoperatively to determine the magnitude of correction. The medical records and radiographs were reviewed and used to report clinical and radiographic outcomes. Radiographs were reviewed to evaluate postoperative limb alignment, patellar position, and bone healing. RESULTS: Twenty-two detorsional osteotomies were performed. Mean preoperative FTA was 14° for medial-PL and 45.2° for lateral-PL. Mean preoperative TTA was 11° for medial-PL. Physiological patellar tracking was restored in 22/22 of cases. CAL measurement allowed for correction of abnormal torsion in 19/22 of cases. Seventeen out 18 dogs had full or acceptable functional outcome. The median radiographic follow-up was 3 months. Major complications occurred in 2/22 cases, which suffered an iatrogenic abnormal femoral internal torsion and a persistent hindlimb lameness. CONCLUSIONS: CAL can be measured with CT and used intraoperatively to guide the correction of abnormal torsion in dogs. CLINICAL RELEVANCE: Abnormal femoral and tibial torsion are predisposing factors for PL. A higher complication rate is expected when FDO and TDO are performed in the same hindlimb.

Active Materials for 3D Printing in Small Animals: Current Modalities and Future Directions for Orthopedic Applications.

The successful clinical application of bone tissue engineering requires customized implants based on the receiver's bone anatomy and defect characteristics. Three-dimensional (3D) printing in small animal orthopedics has recently emerged as a valuable approach in fabricating individualized implants for receiver-specific needs. In veterinary medicine, because of the wide range of dimensions and anatomical variances, receiver-specific diagnosis and therapy are even more critical. The ability to generate 3D anatomical models and customize orthopedic instruments, implants, and scaffolds are advantages of 3D printing in small animal orthopedics. Furthermore, this technology provides veterinary medicine with a powerful tool that improves performance, precision, and cost-effectiveness. Nonetheless, the individualized 3D-printed implants have benefited several complex orthopedic procedures in small animals, including joint replacement surgeries, critical size bone defects, tibial tuberosity advancement, patellar groove replacement, limb-sparing surgeries, and other complex orthopedic procedures. The main purpose of this review is to discuss the application of 3D printing in small animal orthopedics based on already published papers as well as the techniques and materials used to fabricate 3D-printed objects. Finally, the advantages, current limitations, and future directions of 3D printing in small animal orthopedics have been addressed.

Colloidal Iron Oxide Formulation for Equine Hoof Disinfection.

The presence of bacteria of various origins on horse hoofs enables the onset of infections following trauma or even post-surgical wounds. Thus, the analysis of new antibacterial substances is of fundamental importance. In this study, the antibacterial efficacy of Iron Animals (IA), a stable colloidal suspension of iron oxide, organic acids, and detergents, was tested in vitro and in vivo. In vitro assays were performed to test the unspecific inhibitory effect of IA on both gram-positive and gram-negative bacteria monitoring the microorganism growth by spectrophotometry (optical density OD600) at 37 °C for 24 h. In vivo test consists on the quantification of the bacterial load in colony forming units per gram (CFU/g) of specimens collected from the frog region of the anterior hooves of 11 horses. Sampling followed the application of four disinfectant protocols consisting of two consecutive 3 min scrubs with 50 mL of 10% Povidone-iodine (PI) or 4% Chlorhexidine (CHx), with or without an additional application for 15 min of 10 mL of Iron Animals (PI+IA and CHx+IA). In vitro, IA completely suppressed the bacterial growth of all the tested microorganisms, resulting in effectiveness also against CHx-resistant bacteria, such as Staphylococcus aureus. In vivo, PI emerged as an ineffective protocol; CHx was effective in 18% of cases, but with the addition of IA (CHx + IA) its use emerged as the best disinfectant protocol for horse hoof, achieving the lowest bacterial load in 55% of cases. The addition of IA, after PI or CHx, improves the effectiveness of both disinfectants leading to the highest bactericidal activity in 82% of cases.

Osteogenic Properties of 3D-Printed Silica-Carbon-Calcite Composite Scaffolds: Novel Approach for Personalized Bone Tissue Regeneration.

Carbon enriched bioceramic (C-Bio) scaffolds have recently shown exceptional results in terms of their biological and mechanical properties. The present study aims at assessing the ability of the C-Bio scaffolds to affect the commitment of canine adipose-derived mesenchymal stem cells (cAD-MSCs) and investigating the influence of carbon on cell proliferation and osteogenic differentiation of cAD-MSCs in vitro. The commitment of cAD-MSCs to an osteoblastic phenotype has been evaluated by expression of several osteogenic markers using real-time PCR. Biocompatibility analyses through 3-(4,5-dimethyl- thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lactate dehydrogenase (LDH) activity, hemolysis assay, and Ames test demonstrated excellent biocompatibility of both materials. A significant increase in the extracellular alkaline phosphatase (ALP) activity and expression of runt-related transcription factor (RUNX), ALP, osterix (OSX), and receptor activator of nuclear factor kappa-Β ligand (RANKL) genes was observed in C-Bio scaffolds compared to those without carbon (Bio). Scanning electron microscopy (SEM) demonstrated excellent cell attachment on both material surfaces; however, the cellular layer on C-Bio fibers exhibited an apparent secretome activity. Based on our findings, graphene can improve cell adhesion, growth, and osteogenic differentiation of cAD-MSCs in vitro. This study proposed carbon as an additive for a novel three-dimensional (3D)-printable biocompatible scaffold which could become the key structural material for bone tissue reconstruction.

A three-dimensional computed tomographic volume rendering methodology to measure the tibial torsion angle in dogs.

OBJECTIVE: To describe a three-dimensional (3D) computed tomographic (CT) methodology to measure the tibial torsion angle (TTa) and to evaluate intrarater and interrater agreements and accuracy through comparison with anatomic measurements. STUDY DESIGN: Ex vivo cadaveric study. SAMPLE POPULATION: Thirty-six tibiae from 18 dogs. METHODS: Tibial torsion angle of each tibia was measured by using two CT techniques (axial and 3D volume rendering) by three raters who blindly measured TTa in duplicate. A semitransparent bone filter was used to enhance the visibility of the target anatomical landmarks for the 3D volume rendering CT technique. Tibial torsion angle was also quantitated in tibial specimens. Intrarater and interrater agreements were analyzed by using intraclass coefficients (ICC). Accuracy was evaluated by using adjusted R2 coefficients (R2 > 80% was considered acceptable). RESULTS: The 3D volume rendering CT technique had excellent intrarater and interrater agreements (ICC > 0.94) and an R2 value of 97%. The axial CT technique had good to excellent intrarater and interrater agreements (0.8 < ICC < 0.95) and an R2 of 86%. No difference was found between axial and 3D CT techniques. A mean internal TT angle of approximately -6° was found with CT and anatomic measurements. CONCLUSION: The 3D volume rendering and axial CT techniques were precise and accurate for measuring TTa in dogs unaffected by patellar luxation. CLINICAL RELEVANCE: Combining 3D bone manipulation with application of a semitransparent filter allows simultaneous visualization of anatomic landmarks, which may facilitate the evaluation of complex bone deformations. Internal tibial torsion may be present in nonchondrodystrophic dogs without patella luxation.

Computed tomographic localization of the deepest portion of the femoral trochlear groove in healthy dogs.

OBJECTIVE: To validate a computed tomographic (CT) method to measure the femoral trochlear groove depth (FTGD). STUDY DESIGN: Cadaveric study. SAMPLE POPULATION: Fifteen dogs, 26 femoral trochleae. METHODS: Five points were identified from proximal to distal (proximal point [PP], P25, P50, P75, and distal point [DP]) along the trochlea via three-dimensional volume-rendering function on the sagittal plane and measured on multiplanar reconstruction images. Each rater repeated measurements in duplicate, unaware of the identity of the joint. The FTGD was quantitated on the anatomical specimens and statistically compared with CT measurements. Intrarater and interrater agreements were analyzed by using intraclass coefficients. Accuracy was evaluated by using either adjusted R2 coefficients (R2 > 80% was considered acceptable) or Student's t test. The ratio of the patellar and the trochlear width and the ratio of the patellar craniocaudal thickness inside the trochlear groove were calculated at three different patellar locations. RESULTS: Good to excellent intrarater and interrater agreements were observed in four of five trochlear points (P25, P50, P75, and DP), and accuracy was acceptable for these points (R2 > 80%). Computed tomographic measurements differed from the mean anatomical measurements at three of five points (PP, P50, and P75; P < .01), overestimating the FTGD by an overall mean of 0.18 mm (range, 0.02-0.3). P25 and P50 were the deepest points measured. CONCLUSION: Computed tomography allowed precise measurements of trochlear groove depth except for the most proximal point. The deepest trochlear points were P25 and P50. P25 was the most precise and accurate point measured, while PP was the least consistent. CLINICAL SIGNIFICANCE: The deepest portion of the trochlea groove may be located between P25 and P50. Evaluation of this CT method in dogs with patellar luxation is recommended.

Accuracy of an automated three-dimensional technique for the computation of femoral angles in dogs.

AIMS: The purpose of the study was to evaluate the accuracy of a three-dimensional (3D) automated technique (computer-aided design (aCAD)) for the measurement of three canine femoral angles: anatomical lateral distal femoral angle (aLDFA), femoral neck angle (FNA) and femoral torsion angle. METHODS: Twenty-eight femurs equally divided intotwo groups (normal and abnormal) were obtained from 14 dogs of different conformations (dolicomorphic and chondrodystrophicCT scans and 3D scanner acquisitions were used to create stereolithographic (STL) files , which were run in a CAD platform. Two blinded observers separately performed the measurements using the STL obtained from CT scans (CT aCAD) and 3D scanner (3D aCAD), which was considered the gold standard method. C orrelation coefficients were used to investigate the strength of the relationship between the two measurements. RESULTS: A ccuracy of the aCAD computation was good, being always above the threshold of R2 of greater than 80 per cent for all three angles assessed in both groups. a LDFA and FNA were the most accurate angles (accuracy >90 per cent). CONCLUSIONS: The proposed 3D aCAD protocol can be considered a reliable technique to assess femoral angle measurements in canine femur. The developed algorithm automatically calculates the femoral angles in 3D, thus considering the subjective intrinsic femur morphology. The main benefit relies on a fast user-independent computation, which avoids user-related measurement variability. The accuracy of 3D details may be helpful for patellar luxation and femoral bone deformity correction, as well as for the design of patient- specific, custom-made hip prosthesis implants.

Haemodynamic changes during propofol induction in dogs: new findings and approach of monitoring.

BACKGROUND: Propofol is one of the most widely used injectable anaesthetic agents in veterinary practice. Cardiovascular effects related to propofol use in dogs remain less well defined. The main objective of this study was to evaluate the haemodynamic changes during induction of general anaesthesia with propofol in healthy dogs, by a beat-to-beat continuous monitoring. All dogs were premedicated with intramuscular acepromazine (0.015 mg/kg) and methadone (0.15 mg/kg). Transthoracic echocardiography was used to measure the velocity time integral (VTI) of the left ventricular outflow tract. A syringe driver, programmed to deliver propofol 5 mg/kg over 30 s followed by a continuous infusion of 25 mg/kg/h, was used to induce and maintain anaesthesia. From the initiation of propofol administration, heart rate (HR) and mean invasive arterial blood pressure (MAP) were recorded every 5 s for 300 s, while aortic blood flow was continuously recorded and stored for 300 S. maximum cardiovascular depression was defined the lowest MAP (MAP_Tpeak) recorded during the monitored interval. VTI and VTI*HR were calculated at 0, 30, 90, 120, 150 and 300 s post administration of propofol, and at MAP_Tpeak. Haemodynamic effects of propofol in relation to plasma and biophase concentrations were also evaluated by pharmacokinetics simulation. RESULTS: The median (range) HR was significantly higher (p = 0.006) at the moment of maximum hemodynamic depression (Tpeak) [105(70-148) bpm] compared with pre-induction values (T0) [65(50-120) bpm]. The median (range) MAP was significantly lower (p < 0.001) at Tpeak [61(51-69) mmHg] compared with T0 [88(72-97) mmHg]. The median (range) VTI and VTI*HR were similar at the two time points [11.9(8.1-17.3) vs 13,3(9,4-16,5) cm, and 1172(806-1554) vs 1002(630-1159) cm*bpm, respectively]. CONCLUSIONS: Induction of anaesthesia with propofol causes a drop of arterial pressure in healthy dogs, however cardiac output is well maintained by compensatory chronotropic response. The magnitude of MAP_Tpeak may be strictly related with propofol plasma concentration.

Treatment by Therapeutic Magnetic Resonance (TMR™) increases fibroblastic activity and keratinocyte differentiation in an in vitro model of 3D artificial skin.

This study investigated the effect of extremely low-frequency pulsed electromagnetic fields (PEMFs) on skin wound healing in an in vitro dermal-like tissue. In this study, fibroblast and endothelial cells were utilized for the in vitro reconstruction of dermal-like tissues treated for various times up to 21 days with PEMFs. The effects of PEMFs on cell proliferation (MTT test), cell ageing (ß-galactosidase test, ROS production), gene expression, the quality of the extracellular matrix and the amount of fibroblast growth factors were analysed. The high quality of the dermis products in the presence of PEMFs at the end of the study was confirmed through the high degree of organization of keratinocytes, which were subsequently seeded on the aforementioned in vitro reconstructed dermis. The cells organized themselves in well-defined multi-layers and were better organized compared with the epidermis present on the dermis that was obtained without PEMF treatment. Copyright © 2015 John Wiley & Sons, Ltd.

Computation of Femoral Canine Morphometric Parameters in Three-Dimensional Geometrical Models.

OBJECTIVE: To define and validate a method for the measurement of 3-dimensional (3D) morphometric parameters in polygonal mesh models of canine femora. STUDY DESIGN: Ex vivo/computerized model. SAMPLE POPULATION: Sixteen femora from 8 medium to large-breed canine cadavers (mean body weight 28.3 kg, mean age 5.3 years). METHODS: Femora were measured with a 3D scanner, obtaining 3D meshes. A computer-aided design-based (CAD) software tool was purposely developed, which allowed automatic calculation of morphometric parameters on a mesh model. Anatomic and mechanical lateral proximal femoral angles (aLPFA and mLPFA), anatomic and mechanical lateral distal femoral angles (aLDFA and mLDFA), femoral neck angle (FNA), femoral torsion angle (FTA), and femoral varus angle (FVA) were measured in 3D space. Angles were also measured onto projected planes and radiographic images. RESULTS: Mean (SD) femoral angles (degrees) measured in 3D space were: aLPFA 115.2 (3.9), mLPFA 105.5 (4.2), aLDFA 88.6 (4.5), mLDFA 93.4 (3.9), FNA 129.6 (4.3), FTA 45 (4.5), and FVA -1.4 (4.5). Onto projection planes, aLPFA was 103.7 (5.9), mLPFA 98.4 (5.3), aLDFA 88.3 (5.5), mLDFA 93.6 (4.2), FNA 132.1 (3.5), FTA 19.1 (5.7), and FVA -1.7 (5.5). With radiographic imaging, aLPFA was 109.6 (5.9), mLPFA 105.3 (5.2), aLDFA 92.6 (3.8), mLDFA 96.9 (2.9), FNA 120.2 (8.0), FTA 30.2 (5.7), and FVA 2.6 (3.8). CONCLUSION: The proposed method gives reliable and consistent information about 3D bone conformation. Results are obtained automatically and depend only on femur morphology, avoiding any operator-related bias. Angles in 3D space are different from those measured with standard radiographic methods, mainly due to the different definition of femoral axes.

Decellularization and Delipidation Protocols of Bovine Bone and Pericardium for Bone Grafting and Guided Bone Regeneration Procedures.

The combination of bone grafting materials with guided bone regeneration (GBR) membranes seems to provide promising results to restore bone defects in dental clinical practice. In the first part of this work, a novel protocol for decellularization and delipidation of bovine bone, based on multiple steps of thermal shock, washes with detergent and dehydration with alcohol, is described. This protocol is more effective in removal of cellular materials, and shows superior biocompatibility compared to other three methods tested in this study. Furthermore, histological and morphological analyses confirm the maintenance of an intact bone extracellular matrix (ECM). In vitro and in vivo experiments evidence osteoinductive and osteoconductive properties of the produced scaffold, respectively. In the second part of this study, two methods of bovine pericardium decellularization are compared. The osmotic shock-based protocol gives better results in terms of removal of cell components, biocompatibility, maintenance of native ECM structure, and host tissue reaction, in respect to the freeze/thaw method. Overall, the results of this study demonstrate the characterization of a novel protocol for the decellularization of bovine bone to be used as bone graft, and the acquisition of a method to produce a pericardium membrane suitable for GBR applications.

Tissue Engineering Strategies as Tools for Personalized Meningioma Treatment.

Pharmacogenomics, the science of how genetic makeup influences an individual's reaction to drugs, is an innovative tool for providing critical insights into how a patient will respond to a particular treatment. In the present work, we constructed cancer-like tissues to be used as tools for determining the most effective drug for an individual patient. Using tissue engineering strategies, we generated two different solid tumor-like tissues in vitro, a neuronal tumor (meningioma) and a nonmelanoma skin cancer. Samples were tested by both histological and genetic approaches (using a comparative genomic hybridization array, and the relative World Health Organization classification of the samples was compared with the results obtained by the molecular analyses. Our data confirmed the ability of the cells to maintain their phenotype in three-dimensional scaffolds as well as the strong relationship between chromosomal alterations and histological malignancy grades. We then validated the in vitro construction of tumor-like tissues as a potential tool for developing personalized drug treatments.

Pharmacokinetics and efficacy of intravenous and extradural tramadol in dogs.

Tramadol is a synthetic opioid agonist used extensively in human and, to a lesser extent, veterinary medicine throughout the world. The clinical efficacy and pharmacokinetic profile of intravenous (i.v.) and extradural (e.d.) tramadol (2 mg/kg) and its o-desmethyl metabolite were studied in dogs undergoing tibial plateau levelling osteotomy (TPLO). Intra-operative cardiorespiratory variables were monitored and post-operative pain was assessed using the short form of the Glasgow Composite Pain Scale. A rapid (<5 min) and effective production of o-desmethyl tramadol was recorded. The pharmacokinetic profile was similar for tramadol and its metabolite irrespective of the route of administration. E.d. tramadol provided sufficient intra- and post-operative analgesia without significant clinical side-effects, but the post-operative analgesia was comparable to that following i.v. administration and the e.d. route could therefore not be considered a practical alternative to the i.v. route.