Ladislav Stehlík

The pig as an experimental model for clinical craniofacial research

The pig represents a useful, large experimental model for biomedical research. Recently, it has been used in different areas of biomedical research. The aim of this study was to review the basic anatomical structures of the head region in the pig in relation to their use in current research. Attention was focused on the areas that are frequently affected by pathological processes in humans: the oral cavity with teeth, salivary gland, orbit, nasal cavity and paranasal sinuses, maxilla, mandible and temporomandibular joint. Not all of the structures have an equal morphology in the pig and human, and these morphological dissimilarities must be taken into account before choosing the pig as an experimental model for regenerative medicine.

Quantification of Humeroulnar Incongruity in Labrador Retrievers with and without Medial Coronoid Disease

OBJECTIVE To quantify humeroulnar incongruity on elbow radiographs in Labrador Retrievers with or without medial coronoid disease (MCD). STUDY DESIGN Retrospective study of 92 elbows. SAMPLE POPULATION Radiographic projections of elbow joints from Labrador Retrievers with MCD (n = 42 elbows; 26 dogs) and without MCD (n = 50 elbows; 25 dogs). PROCEDURE The congruity of the humeroulnar joint was measured using an index of subluxation (SI) for each elbow. SI was defined as the distance between the centers of 2 circles drawn along the margins of the incisura trochlearis and the trochlea of humerus on mediolateral digital radiographic projections, normalized by the radius of the circle circumscribing the humeral trochlea. SI was compared between right and left elbows with and without pathology using a Wilcoxon test for paired data, and between normal and abnormal groups with a Wilcoxon test for unpaired data. Mismatch between ulnar curvature and curvature of humeral trochlea and radioulnar incongruency were also noted (Wilcoxon test). The intraobserver repeatability, correlation between SI and radioulnar incongruency, and between SI and mismatch elbow curvature were estimated with a Pearsons correlation coefficient. RESULTS Intraobserver repeatability of SI measurement was high (r = 0.97). Mean ± SD humeroulnar incongruity (SI) was greater in elbows with MCD (18.5 ± 6.6) than in the normal elbows (1.7 ± 2.0, P < 0.001). The difference between the diameters of the curvatures of the ulnar and humeral trochlea was greater in elbows with MCD (12.5 ± 4.4) than in the normal group (10.7 ± 4.1, P < 0.05). A moderate correlation was found between the degree of humeroulnar incongruity and a radioulnar step (r = 0.63); however, no correlation was identified between SI and the difference between the diameters of the curvatures of the ulnar and humeral trochleae (r = 0.14). CONCLUSION We propose a radiographic index to measure humeroulnar incongruity on mediolateral digital radiographic projections. This index (SI) supports the presence of humeroulnar incongruity in Labrador Retrievers with MCD. Further evaluation of its reproducibility and clinical importance are warranted. Although there is a moderate correlation between humeroulnar incongruity and radioulnar incongruency, causation has not been established.Objective To quantify humeroulnar incongruity on elbow radiographs in Labrador Retrievers with or without medial coronoid disease (MCD). Study Design Retrospective study of 92 elbows. Sample Population Radiographic projections of elbow joints from Labrador Retrievers with MCD (n = 42 elbows; 26 dogs) and without MCD (n = 50 elbows; 25 dogs). Procedure The congruity of the humeroulnar joint was measured using an index of subluxation (SI) for each elbow. SI was defined as the distance between the centers of 2 circles drawn along the margins of the incisura trochlearis and the trochlea of humerus on mediolateral digital radiographic projections, normalized by the radius of the circle circumscribing the humeral trochlea. SI was compared between right and left elbows with and without pathology using a Wilcoxon test for paired data, and between normal and abnormal groups with a Wilcoxon test for unpaired data. Mismatch between ulnar curvature and curvature of humeral trochlea and radioulnar incongruency were also noted (Wilcoxon test). The intraobserver repeatability, correlation between SI and radioulnar incongruency, and between SI and mismatch elbow curvature were estimated with a Pearsons correlation coefficient. Results Intraobserver repeatability of SI measurement was high (r = 0.97). Mean ± SD humeroulnar incongruity (SI) was greater in elbows with MCD (18.5 ± 6.6) than in the normal elbows (1.7 ± 2.0, P < 0.001). The difference between the diameters of the curvatures of the ulnar and humeral trochlea was greater in elbows with MCD (12.5 ± 4.4) than in the normal group (10.7 ± 4.1, P < 0.05). A moderate correlation was found between the degree of humeroulnar incongruity and a radioulnar step (r = 0.63); however, no correlation was identified between SI and the difference between the diameters of the curvatures of the ulnar and humeral trochleae (r = 0.14). Conclusion We propose a radiographic index to measure humeroulnar incongruity on mediolateral digital radiographic projections. This index (SI) supports the presence of humeroulnar incongruity in Labrador Retrievers with MCD. Further evaluation of its reproducibility and clinical importance are warranted. Although there is a moderate correlation between humeroulnar incongruity and radioulnar incongruency, causation has not been established.

Effect of medetomidine-butorphanol and dexmedetomidine-butorphanol combinations on intraocular pressure in healthy dogs.

OBJECTIVE To assess the effects of intravenous (IV) medetomidine-butorphanol and IV dexmedetomidine-butorphanol on intraocular pressure (IOP). STUDY DESIGN Prospective, randomized, blinded clinical study. ANIMALS Forty healthy dogs. Mean ± SD body mass 37.6 ± 6.6 kg and age 1.9 ± 1.3 years. METHODS Dogs were allocated randomly to receive an IV combination of dexmedetomidine, 0.3 mg m(-2), combined with butorphanol, 6 mg m(-2), (group DEX) or medetomidine 0.3 mg m(-2), combined with butorphanol 6 mg m(-2), (group MED). IOP and pulse (PR) and respiratory (f(R) ) rates were measured prior to (baseline) and at 10 (T10), 20 (T20), 30 (T30) and 40 (T40) minutes after drug administration. Oxygen saturation of hemoglobin (SpO(2)) was monitored following sedation. Data were analyzed by anova followed by Dunnetts tests for multiple comparisons. Changes were considered significant when p < 0.05. RESULTS Following drug administration, PR and f(R) were decreased significantly at all time points but did not differ significantly between groups. Baseline IOP in mmHg was 14 ± 2 for DEX and 13 ± 2 for MED. With both treatments, at T10, IOP increased significantly (p < 0.001), reaching 20 ± 3 and 17 ± 2 for DEX and MED respectively. This value for DEX was significantly higher than for MED. There were no significant differences in IOP values between groups at any other time points. At T30 and T40, IOP in both groups was below baseline (DEX, 12 ± 2 and 11 ± 2: MED 12 ± 2 and 11 ± 2) and this was statistically significant, for DEX. CONCLUSIONS AND CLINICAL RELEVANCE At the documented doses, both sedative combinations induced a transient increase and subsequent decrease of IOP relative to baseline, which must be taken into consideration when planning sedation of animals in which marked changes in IOP would be undesirable.

Comparative morphology of normal and cleft minipigs demonstrates dual origin of incisors

OBJECTIVE The incisors of the mammalian dental arch develop from tissues arising from separated facial prominences. These primordial craniofacial structures undergo complex morphogenetic processes as they merge and fuse in a time and space dependent fashion. However, local contributions of precursor facial prominences to the incisors that develop subsequently remain unknown. The purpose of this study was to characterize the development of all three deciduous upper rostral teeth in the pig (Sus scrofa f. domestica) for the identification of the likely facial prominence contributions to the incisors based on normal and pathological developmental relationships. DESIGN Embryonic minipigs were collected between gestational days 20-36 (E20-36), processed for histological analysis and subjected to computerized 3D modelling. The location and morphology of the incisors (i) in these specimens were characterized and compared between developmental stages. A second set of neonatal minipigs displaying cleft lip and/or cleft palate defects were also obtained and incisor locations and eruption patterns were morphologically examined. RESULTS Palate formation begins during the third week of gestation (E20) in the minipig with ossification of the premaxilla initiating soon afterwards (E24). The third incisor (i3) develops caudally to the contact seam formed by the fusion of the primary and secondary palates in normal embryos. All cleft animals displayed normal i3 and canine, on other hand, development of i1 and i2 was often disrupted similar to human. CONCLUSIONS Our observations suggest a dual embryonic origin of the incisors in minipigs with the first and second incisors originating from the frontonasal prominence whilst the third incisor forms from tissues derived from the maxillary prominence.

CONGENITAL ABNORMALITIES OF THE VERTEBRAL COLUMN IN FERRETS

Vertebral column pathologies requiring surgical intervention have been described in pet ferrets, however little information is available on the normal vertebral formula and congenital variants in this species. The purpose of this retrospective study was to describe vertebral formulas and prevalence of congenital vertebral anomalies in a sample of pet ferrets. Radiographs of 172 pet ferrets (96 males and 76 females) were included in this retrospective study. In 143 ferrets (83.14%), five different formulas of the vertebral column were recorded with normal morphology of vertebrae (rib attachment included) but with a variable number of thoracic (Th), lumbar (L), and sacral (S) vertebrae. The number of cervical (C) vertebrae was constant in all examined animals. Observed vertebral formulas were C7/Th14/L6/S3 (51.74%), C7/Th14/L6/S4 (22.10%), C7/Th14/L7/S3 (6.98%), C7/Th15/L6/S3 (1.74%), and C7/Th15/L6/S4 (0.58%). Formula C7/Th14/L6/S4 was significantly more common in males than in females (P < 0.05). Congenital spinal abnormalities were found in 29 ferrets (16.86%), mostly localized in the thoracolumbar and lumbosacral regions. The cervical region was affected in only one case. Transitional vertebrae represented the most common congenital abnormalities (26 ferrets) in the thoracolumbar (13 ferrets) and lumbosacral regions (10 ferrets) or simultaneously in both regions (three ferrets). Other vertebral anomalies included block (two ferrets) and wedge vertebra (one ferret). Spina bifida was not detected. Findings from the current study indicated that vertebral formulas may vary in ferrets and congenital abnormalities are common. This should be taken into consideration for surgical planning.

Radiography, Computed Tomography and Magnetic Resonance Imaging of Craniofacial Structures in Pig

The pig has recently become popular as a large animal experimental model in many fields of biomedical research. The aim of this study is to evaluate the basic anatomical structures in the head region of the pig to lay the groundwork for its practical clinical usage or pre‐clinical research in the future. We used three different diagnostic imaging methods: radiography, computed tomography (CT) and magnetic resonance imaging (MRI). The analysis showed that radiographic imaging is suitable only for general evaluation of the facial area of the pig skull. CT images showed excellent spatial definition of bony structures of the whole craniofacial area, and MRI images revealed fine soft tissue details. Radiography is preferentially suited to general assessment of bone structures of the facial skeleton; however, the thick layer of adipose tissue in the craniofacial region of the pig makes the imaging of some parts difficult or even impossible. CT is useful for revealing morphological details of mineralized tissues, whereas MRI is more suitable for soft tissue analysis and the detection of subtle pathologic changes in both bone and soft tissues. Therefore, before using pigs as an experimental model in craniofacial research, it is necessary to evaluate the suitability and disadvantages of potential imaging methods and how appropriate they are for accurate visualization of desired structures.

Computed tomography-guided cementoplasty combined with radiation therapy for an aneurysmal bone cyst in a dog: a case report

Aneurysmal bone cysts are expansile osteolytic bone lesions that can manifest clinically as lameness, pain and swelling. In humans, aneurysmal bone cysts are considered benign tumours that have been classified as primary or secondary to a previous coexisting lesion. Local recurrence after treatment or malignant transforma - tion has been reported in both humans and in a dog. Different treatment options have been proposed. This report describes the treatment of an 11-month-old Irish wolfhound with an aneurysmal bone cyst with a combination of computed tomography-guided cementoplasty and radiation therapy, 30 Gy delivered in 10 fractions. The dog recovered completely, with no lameness observed 30 months after treatment.

Comparison of the Resistance to Bending Forces of the 4.5 LCP Plate-rod Construct and of 4.5 LCP Alone Applied to Segmental Femoral Defects in Miniature Pigs

The study deals with the determination of mechanical properties, namely resistance to bending forces, of flexible buttress osteosynthesis using two different bone-implant constructs stabilizing experimental segmental femoral bone defects (segmental ostectomy) in a miniature pig ex vivo model using 4.5 mm titanium LCP and a 3 mm intramedullary pin (“plate and rod” construct) (PR-LCP), versus the 4.5 mm titanium LCP alone (A-LCP). The “plate and rod” fixation (PR-LCP) of the segmental femoral defect is significantly more resistant ( p < 0.05) to bending forces (200 N, 300 N, and 500 N) than LCP alone (A-LCP). Stabilisation of experimental segmental lesions of the femoral diaphysis in miniature pigs by flexible bridging osteosynthesis 4.5 mm LCP in combination with the “plate and rod” construct appears to be a suitable fixation of non-reducible fractures where considerable strain of the implants by bending forces can be assumed. These findings will be used in upcoming in vivo experiments in the miniature pig to investigate bone defect healing after transplantation of mesenchymal stem cells in combination with biocompatible scaffolds. Fracture fixation, comminuted fracture, ostectomy, buttress osteosynthesis, implant failure

Vertebral formula and congenital abnormalities of the vertebral column in rabbits

The aim of this retrospective study of 330 rabbits (164 males, 166 females) was to determine different vertebral formulas and prevalence of congenital vertebral anomalies in rabbits from radiographs of the cervical (C), thoracic (Th), lumbar (L) and sacral (S) segments of the vertebral column. The number of vertebrae in each segment of vertebral column, position of anticlinal vertebra and localisation and type of congenital abnormalities were recorded. In 280/330 rabbits (84.8%) with normal vertebral morphology, seven vertebral formulas were identified: C7/Th12/L7/S4 (252/330, 76.4%), C7/Th12/L6/S4 (11/330, 3.3%), C7/Th13/L7/S4 (8/330, 2.4%), C7/Th12/L7/S5 (4/330, 1.2%), C7/Th12/L8/S4 (3/330, 0.9%), C7/Th12/L7/S6 (1/330, 0.3%) and C7/Th11/L7/S4 (1/330, 0.3%). The anticlinal vertebra was identified as Th10 in 56.4% of rabbits and Th11 in 42.4% of rabbits. Congenital spinal abnormalities were identified in 50/330 (15.2%) rabbits, predominantly as a single pathology (n=44). Transitional vertebrae represented the most common abnormalities (n=41 rabbits) in the thoracolumbar (n=35) and lumbosacral segments (n=6). Five variants of thoracolumbar transitional vertebrae were identified. Cervical butterfly vertebrae were detected in three rabbits. One rabbit exhibited three congenital vertebral anomalies: cervical block vertebra, thoracic hemivertebra and thoracolumbar transitional vertebra. Five rabbits exhibited congenital vertebral abnormalities with concurrent malalignment, specifically cervical kyphosis/short vertebra (n=1), thoracic lordoscoliosis/thoracolumbar transitional vertebrae (n=1), thoracic kyphoscoliosis/wedge vertebrae (n=2) and thoracolumbar lordoscoliosis/thoracolumbar transitional vertebrae/lumbosacral transitional vertebrae (n=1). These findings suggest that vertebral columns in rabbits display a wide range of morphologies, with occasional congenital malformations.

A modified technique for radiographic measurement of the tibial plateau angle in dogs

This prospective study was aimed at testing a modified method for the measurement of the tibial plateau angle on radiographs of the stifle joint and the proximal part of the tibia. Forty-four stifle joints of 32 clientowned dogs were included in this study. Dogs were presented for hindlimb lameness and suspected cranial cruciate ligament rupture. The diagnostic procedures included radiographic examination. The mediolateral projection of the stifle joint including the tibia and the tarsal joint was used in this study. We tested the most widely used gold standard method as well as three additional methods for tibial plateau angle measurement on each joint. The tibial plateau angle, the positioning of the stifle joint and the presence of osteoarthrosis were recorded. Only 29 (66%) joints had correct position on the radiograph and were used for further study. Repeated-measures ANOVA identified significant differences in mean tibial plateau angle between the different measuring methods. Dunnett’s post-hoc test identified a significant difference between the TA-2 and TA-2i methods and TA-0. No significant differences in tibial plateau angle were identified between joints with and without osteoarthrosis. The positioning of the limb significantly influenced the tibial plateau angle. One of the modified methods was found to not differ significantly from the gold standard method; thus, it can be recommended for further testing.