Female Sheep Research Articles

Occurrence of squamous cell carcinoma in Milchschaf sheep in Uruguay

ABSTRACT: In this study, the epidemiological and pathological features of an outbreak of squamous cell carcinoma (SCC) in adult female sheep and a young ram, Friesian Milchschaf breed or crossbreed is described. Seven adult females belonging to an original flock nucleus of 20 animals were affected, as well as a young ram belonging to the same nucleus was affected in other establishment. Multiple verrucous neoplastic masses of several months of evolution were evident in the head region, with local invasion and regional lymph node metastases. Histological studies revealed a cancerous stage of invasion of the dermis and neoplastic proliferation characteristic of invasive SCC, and a pre-cancerous stage with solar elastosis and chronic solar keratosis lesions induced by sunlight. Decrease in latitude, increased level of solar radiation with long exposure to sunlight, and the phenotypic characteristics of the breed are the main factors responsible for the high prevalence of SCC, showing the susceptibility of the Friesian Milchschaf breed and crossbreed in temperate zones as Uruguay.

Wool-derived keratin hydrogel enhances implant osseointegration in cancellous bone.

This study investigated whether a keratin hydrogel derived from wool enhances ossointegration of dental implants, using a cancellous bone model in sheep. Ten female sheep received one dental implant per hind leg in the distal femoral condyles. Test implants were coated with 1 mL of keratin hydrogel containing sulphonated keratin and hydroxyapatite; control implants remained uncoated; implants were not functionally loaded axially. Biomechanical stability was tested with resonance frequency analysis (RFA) at placement and euthanasia. Animals were sacrificed after 4 weeks and resin-embedded histomorphometry performed. Bone appeared denser and more mature around the keratin-treated test implants. Average % bone-implant contact was significantly increased for the test (58.1 ± 4.6 [SD]%) compared to control implants (34.4 ± 7.5%) (p = 0.01). RFA values were lower for control (76.5 ± 4.2 ISQ) than test implants (80.4 ± 1.4) after 4 weeks (p = 0.04). the increase in osseointegration found in this model suggests that keratin hydrogel has the potential to enhance the successful osseointegration of dental implants placed into poor quality bone. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2447-2454, 2018.

Radiographic measurements related with the cardiac size in young female Bergamasca sheep

ABSTRACT: In thoracic radiographic examination, routinely used in cases which cardiac evaluation is indicated, quantitative assessment of the heart is a useful role to be used in combination with subjective analysis. Numerous studies about objective assessment of the cardiac size have been performed in several species, including sheep; however, there is scarce information regarding cardiac parameters of young Bergamasca sheep. Therefore, the purpose of this study was to determine the average results and suggest the range of expected normal values for parameters related to the heart size of young female Bergamasca sheep by radiographic evaluation. Fifteen healty 8 months-old female Bergamasca sheep (mean weight: 41.13±4.71kg) were submitted to right lateral recumbency thoracic radiography. The length of the fourth and third to fifth thoracic vertebrae, cardiac height and width, vertebral heart size, cardiophrenic contact, caudal vena cava height, aorta caliber and tracheal angle were measured and the mean results found were, respectively: 2.46±0.11cm (95% CI 2.41-2.52), 7.53±0.30cm (95% CI 7.38-7.68), 13.83±0.57cm, (95% CI 13.54-14.12), 8.99±0.37cm (8.80-9.17), 8.99±0.27 vertebrae (95% 8.85-9.13), 4.55±0.70cm (95% CI 4.19-4.90), 1.88±0.19cm (95% CI 1.79-1.97), 2.05±0.11cm (95% CI 2.00-2.11) and 14.36±2.73° (95% CI 12.98-15.75). Cardiac height and width and the sum of these parameters were compared to the length of third to fifth thoracic vertebrae, resulting in the respective mean values: 1.84±0.08 (95% IC 1.80-1.88), 1.20±0.05 (1.17-1.22) and 3.04±0.11 (95% IC 2.98-3.09). Ratios of cardiophrenic contact to cardiac height and caudal vena cava height to length of fourth thoracic vertebra were also evaluated and the mean values obtained were 0.33±0.05 (95% IC 0.30-0.35) and 0.76±0.08 (95% IC 0.72-0.81), respectively. Authors suggest that the values available in this study may be used as reference for normal heart size in young female Bergamasca sheep and as basis for further studies.

A tissue-based approach to selection of reference genes for quantitative real-time PCR in a sheep osteoporosis model

BackgroundIn order to better understand the multifactorial nature of osteoporosis, animal models are utilized and compared to healthy controls. Female sheep are well established as a model for osteoporosis induced by ovariectomy, calcium and vitamin D low diet, application of steroids, or a combination of these treatments. Transcriptional studies can be performed by applying quantitative real time PCR (RT-qPCR). RT-qPCR estimates mRNA-levels of target genes in relation to reference genes. A chosen set of reference genes should not show variation under experimental conditions. Currently, no standard reference genes are accepted for all tissue types and experimental conditions. Studies examining reference genes for sheep are rare and only one study described stable reference in mandibular bone. However, this type of bone differs from trabecular bone where most osteoporotic fractures occur. The present study aimed at identifying a set of reference genes for relative quantification of transcriptional activity of ovine spine bone and ovine in vitro differentiated mesenchymal stromal cells (MSC) for reliable comparability.MethodsTwelve candidate reference genes belonging to different functional classes were selected and their expression was measured from cultured ovMSCs (n = 18) and ovine bone samples (n = 16), respectively. RefFinder was used to rank the candidate genes.ResultsWe identified B2M, GAPDH, RPL19 and YWHAZ as the best combination of reference genes for normalization of RT-qPCR results for transcriptional analyses of these ovine samples.ConclusionThis study demonstrates the importance of applying a set of reference genes for RT-qPCR analysis in sheep. Based on our data we recommend using four identified reference genes for relative quantification of gene expression studies in ovine bone or for in vitro experiments with osteogenically differentiated ovine MSCs.

R-100 improves pulmonary function and systemic fluid balance in sheep with combined smoke-inhalation injury and Pseudomonas aeruginosa sepsis

BackgroundSeptic shock is a major cause of death in intensive care units around the world . The aim of the study was to investigate whether the novel drug R-100 (a superoxide degradation catalyst and nitric oxide donor) improves pulmonary function in a sheep model of septic shock caused by Pseudomonas aeruginosa and smoke inhalation.MethodsEleven female sheep were prepared surgically and randomly assigned to a treatment group (n = 5) or a control group (n = 6) after inhalation of cooled cotton smoke and airway instillation of live P. aeruginosa (2.5 × 1011 CFU) by bronchoscope under deep anesthesia and analgesia. The treatment group received an intravenous infusion of a total of 80 mg/kg of R-100 diluted in 500 mL of 5% dextrose. The control group was given 500 mL of 5% dextrose. All animals received intravenous lactated Ringer’s solution to maintain a hematocrit level at baseline ± 3%. Blood gas and hemodynamics were measured at baseline and then analyzed every 3 h during the 24-h study period. Results are expressed as mean ± SEM.ResultsThe treated animals showed significant improvement in their pulmonary gas exchange (PaO2/FiO2 ratio at 24 h: 246 ± 29 vs. 90 ± 40 mmHg control, P < 0.05). Pulmonary arterial pressures were reduced in the treated group (24 h: 26 ± 1 vs. 30 ± 2 cm mmHg control, P < 0.05). The treated animals also had an improved total fluid balance after 24 h (190 ± 45/24 h mL vs. 595 ± 234/24 h mL control, P < 0.05).ConclusionsTreatment with R-100 improves pulmonary gas exchange and blood oxygenation, and prevents a fluid imbalance in sheep subjected to smoke inhalation and P. aeruginosa.

Esmolol Administration to Control Tachycardia in an Ovine Model of Peritonitis.

Excessive adrenergic signaling may be harmful in sepsis. Using β-blockers to reduce sympathetic overactivity may modulate sepsis-induced cardiovascular, metabolic, immunologic, and coagulation alterations. Using a randomized ovine fecal peritonitis model, we investigated whether administration of a short-acting β-blocker, esmolol, could control tachycardia without deleterious effects on hemodynamics, renal perfusion, cerebral perfusion, cerebral metabolism, or outcome. After induction of fecal peritonitis, 14 anesthetized, mechanically ventilated, and hemodynamically monitored adult female sheep were randomly assigned to receive a continuous intravenous infusion of esmolol to control heart rate between 80 and 100 bpm (n = 7) or a saline infusion (control group, n = 7). Esmolol was discontinued when the mean arterial pressure decreased below 60 mm Hg. Fluid resuscitation was titrated to maintain pulmonary artery occlusion pressure at baseline values. Left renal blood flow and cerebral cortex perfusion and metabolism were monitored in addition to standard hemodynamic variables. Esmolol was infused for 11 (9-14) hours; the target heart rate (80-100 bpm) was achieved between 3 and 8 hours after feces injection. In the first 5 hours after the start of the infusion, the decrease in heart rate was compensated by an increase in stroke volume index; later, stroke volume index was not statistically significantly different in the 2 groups, so that the cardiac work index was lower in the esmolol than in the control group. Hypotension (mean arterial pressure <60 mm Hg) occurred earlier (10 [8-12] vs 14 [11-20] hours; P= .01) in the esmolol group than in the control animals. Renal blood flow decreased earlier in the esmolol group, but there were no differences in urine output, cerebral cortex perfusion, metabolism, or survival between the groups. In this ovine model of abdominal sepsis, early control of tachycardia by esmolol was associated with a transient increase in stroke volume, followed by earlier hypotension. There were no significant effects of esmolol on cerebral perfusion, metabolism, urine output, or survival.

Efecto nematicida de la infusión de ajo (Allium sativum) comparada con albendazol administrados vía oral en ovinos

La carne de las ovejas de pelo es muy aceptada por su característico sabor, sin embargo, este ganado está muy expuesto a las parasitosis gastrointestinales limitando su producción cárnica. El uso de productos medicinales naturales es una alternativa a los tratamientos convencionales. Al Ajo se le atribuyen propiedades medicinales principalmente por sus compuestos órganos ulfurados. En la presente investigación se evaluó el efecto nematicida de la infusión de ajo (Allium sativum) al 10%, determinar si existe diferencia significativa con una prueba de medias T studenty efecto residual del efecto nematicida de la infusión de ajo al 10% comparado con albendazol al 15% administrados por vía oral en ovinos. Se seleccionaron 30 hembras ovinas híbridas cruces de razas Merina, Pelibuey, Blackbelly y Katahdine comprendidas entre 1 año a 2½ años de edad, divididas en 2 grupos de 15 animales cada uno. Grupo 1:se le administró la infusión de ajo a una dosis de 5ml/100kg, por 3 días consecutivos; Grupo 2: recibió un producto comercial a base de albendazol a dosis única de 5mg/kg. Se colectaron muestras fecales al día 0 antes del tratamiento,24 horas, 7, 14, 21 y 28 días después del tratamiento, usando como métodos diagnósticos Flotación y McMaster (huevos/g heces). La infusión de ajo administrada vía oral en ovinos, fue efectiva como nematicida en el primer díapost tratamiento obteniéndose el 47% de las muestras sin carga parasitaria en relación del 40% de albendazol. El grupo tratado con la infusión de ajo fue disminuyendo su efecto nematicida a los 7, 14, 21 y 28 días a diferencia de lalbendazol que fue a los 21 días. Se determinó que no existe diferencia significativa (P ≥0.05) del efecto nematicida dela infusión de ajo comparada con el albendazol administrado por vía oral en ovinos, por lo que es una alternativa para disminuir la carga de nematodos en ovinos.

Serological survey of antibodies against Toxoplasma gondii in organic sheep and goat farms in Greece

Toxoplasmosis is a parasitic infection with global distribution caused by the protozoon Toxoplasma gondii. The present study is divided into the preliminary and the basic research. In the preliminary research, 413 serum samples (184 sheep and 229 goats) from organic farms of Péloponnèse and Western Sterea Hellas were tested for IgG antibodies against Τ gondii, by using enzyme-linked immunosorbent assay (ELISA). Blood sampling took place during spring of 2005 and samples were taken from male animals and female that had aborted during the reproductive period of 2004-2005. Significant prevalence of antibodies was the reason that further research of toxoplasmosis in organic farms was conducted. For the basic research, small ruminants' blood samples from organic farms of Péloponnèse, Western Sterea Hellas and the county of Ioannina, were collected during summer of 2006. Blood samples were collected from flocks with cases of abortions during the reproductive period of 2005-2006. Animals were separated into 3 groups: males, females that had abortions during the reproductive period of 2005-2006 and females that did not have abortions during the same period. A total number of 182 sheep sera and 167 goat sera were tested by using the enzyme-linked immunosorbent assay (ELISA). The results showed that antibodies were detected in 38.46% of male sheep, 60.87% of female sheep that had abortions during the reproductive period 2005-2006 and in 54.21% of female sheep that did not have abortions during the same period. Significant difference was determined only between male and female animals (p&lt;0.05). In addition, antibodies were detected in 12.12% of male goats, 14.29% of female goats that had abortions and in 22.35% of female goats that did not have abortions in the reproductive period 2005-2006. There was no significant difference between the results (p&gt;0.05). Seroprevalence in goats was significandy lower than that of the sheep (p&lt;0.05). In animals, aged less than 4 years old, seroprevalence was significantly lower (39.58% for sheep and 13.04% for goats) than that for animals aged more than 4 years old (61.97% for sheep and 24% for goats) (p&lt;0.05). The results of the survey suggest that there is high prevalence of toxoplasmosis both in animals that had aborted and in animals that had not aborted and that there are different species and gender susceptibility. Results also indicate that toxoplasmosis is more common in older animals.

Mitral valve replacement with the novel TRIBIO and an established stented bioprosthesis in a sheep model

Heart valve replacement with a bioprosthesis is one of the most frequently performed procedures in cardiac surgery and represents a highly effective therapy to relieve diseased heart valves. Nevertheless, as postoperatively elevated transvalvular gradients and prosthesis-patient mismatch are reported as shortcomings of the procedure or of the currently used devices, there is a need for novel bioprostheses with improved haemodynamics. This study presents preclinical haemodynamic results after mitral valve replacement with the novel TRIBIO bioprosthesis (TRIBIO) compared with an established bioprosthesis in a sheep model. Six female sheep had their mitral valves replaced with the TRIBIO and 3 with the Carpentier-Edwards Perimount, both sized 19 mm. The TRIBIO consists of a flexible valve-bearing crown, a force-decoupled interface and an intra-annular base ring. Mean and peak transvalvular gradients as well as an effective orifice area were monitored in both groups using transthoracic echocardiography over the course of the 90-day study. In both groups, haemodynamic performance diminished over time. The TRIBIO demonstrated overall superior haemodynamics, i.e. lower transvalvular gradients and a larger effective orifice area, although the results were not statistically significant. On Day 90, the mean values for the mean and peak transvalvular gradients and the effective orifice area were 6 mmHg, 10.2 mmHg and 1.2 cm2 for the TRIBIO and 10 mmHg, 15.8 mmHg and 0.8 cm2 for the Carpentier-Edwards Perimount, respectively. This study demonstrates a trend towards improved preclinical haemodynamic performance following mitral valve replacement with the TRIBIO compared to that with an established bioprosthesis in a sheep model.

A case of kidney infection by Corynebacterium pseudotuberculosis in sheep

A three year-old female sheep was presented to the Clinic of Farm Animals, School of Veterinary Medicine, Aristotle University of Thessaloniki, with mild nasal seromucous discharge and progressive weight loss, while rectal temperature was normal. Hematological examination revealed mild leukocytosis, while traces of hemoglobin and protein were found in urinalysis. The animal was euthanized and forwarded for necropsy. An extremely enlarged, nodular right kidney, whose renal parenchyma was completely replaced by multiple abscesses, was the remarkable finding of the necropsy. Microbiological and molecular examinations were performed to establish the diagnosis and rule out Maedi/Visna infection. Tissue samples from the kidney and other organs were, also, examined histologically. Corynebacterium pseudotuberculosis was isolated in microbiological tests. A kidney infection by C. pseudotuberculosis, presented as a clinical case with scant symptoms, is discussed in this case report.

Ecotypic variation in population dynamics of reintroduced bighorn sheep

ABSTRACTSelection of bighorn sheep (Ovis canadensis) for translocation historically has been motivated by preservation of subspecific purity rather than by adaptation of source stocks to similar environments. Our objective was to estimate cause‐specific, annual, and age‐specific mortality of introduced bighorn sheep that originated at low elevations in southern British Columbia, Canada (BC ecotype), or in the Missouri River Breaks region of central Montana, USA (MT ecotype). In North Dakota, USA, mortality was similar and typically low for adult female bighorn sheep from Montana (0.09 ± 0.029 [SE]) and British Columbia (0.08 ± 0.017) during 2000–2016. Median life expectancy was 11 years for females that reached adulthood (2 yrs old); however, mortality accelerated with age and reached 86% by age 16. Mortalities resulted primarily from low rates of predation, disease, accidents, and unknown natural causes (&lt;0.04 [upper 90% CI]). Similar survival rates of female bighorn sheep from female bighorn sheep from British Columbia and Montana, coupled with greater recruitment of bighorn sheep from Montana, resulted in a greater projected rate of increase for the MT ecotype (λ = 1.21) than for the BC ecotype (1.02), and a more youthful age structure. These results support translocation of bighorn sheep from areas that are environmentally similar to areas that will be stocked. Potential benefits include more rapid population growth, greater resilience to and more rapid recovery from density‐independent losses, an increased possibility that rapidly growing populations will expand into adjacent habitat, increased hunter opportunity, increased connectivity among herds, and a more complete restoration of ecosystem processes. © 2017 The Wildlife Society.

Continuous Kisspeptin Restores Luteinizing Hormone Pulsatility Following Cessation by a Neurokinin B Antagonist in Female Sheep.

Pulsatile secretion of the gonadotropin-releasing hormone (GnRH) drives pulsatile secretion of the luteinizing hormone (LH), with evidence that this depends on kisspeptin (Kiss) input to GnRH neurons. Kiss administration causes acute GnRH/LH secretion, and electrophysiological data suggest that Kiss neurons may act in a phasic manner to drive GnRH secretion, but there is not definitive evidence for this. The product of the Kiss-1 gene is proteolytically cleaved to smaller products, and the 10 amino acid C-terminal product (Kiss-10) displays full bioactivity. We have shown previously that continuous delivery of Kiss-10 to anestrous ewes can cause a surge in GnRH secretion and ovulation and increases LH pulse frequency in humans. Here, we tested the hypothesis that continuous Kiss-10 delivery can support pulsatile GnRH/LH secretion in the sheep. Neurokinin B (NKB) provides positive drive to Kiss neurons, so we therefore infused an NKB antagonist (ANT-08) intracerebroventricularly to induce cessation of pulsatile GnRH/LH secretion, with or without concomitant continuous Kiss-10 infusion. ANT-08 suppressed GnRH/LH pulsatility, which was immediately restored with continuous Kiss-10 infusion. These data support the notion that Kiss-10 action is downstream of NKB signaling and that continuous Kiss-10 stimulation of GnRH neurons is sufficient to support a pulsatile pattern of GnRH/LH secretion. This offers further support to the theory that GnRH pulse generation is intrinsic to GnRH neurons and that pulsatile GnRH release can be affected with continuous stimulation by Kiss-10.

* Six-Year-Old Sheep as a Clinically Relevant Large Animal Model for Aortic Valve Replacement Using Tissue-Engineered Grafts Based on Decellularized Allogenic Matrix.

Tissue-engineered (TE) grafts based on decellularized grafts have shown very promising results in preclinical and clinical studies. However, in animal models valves have either been tested in juvenile models or in the clinically less relevant pulmonary valve position. In this study, we tested the grafts in the aortic valve (AV) position of 6-year-old sheep, as geriatric patients in need of an AV substitute due to calcification are the largest patient group benefiting from TE grafts. Decellularized AV (DAV; n = 4) and DAV additionally re-endothelialized with autologous cells (n = 3) were implanted in the AV position of 6-year-old female sheep. Function was investigated at implantation and explantation 12 months later. Regeneration capacity was analyzed by the repopulation degree of the graft with recipient's cells, by the generation of a new endothelial layer and by intracellular staining against pro-collagen type I. DAV and re-endothelialized AV demonstrated excellent function with only two valves developing mild insufficiencies (1°). Of the repopulating cells only few cells were identified as inflammation cells, while the majority was found to be interstitial cells producing procollagen type I. Endothelial coverage was found, but seemed to be reduced. The regenerative capacity of decellularized matrix is not only a feature exhibited when implanted in juvenile individuals but also is evident when implanted in the high-pressure AV position of older sheep, revealing the potential of TE grafts in age-advanced patients.

Obestatin stimulates the somatotrophic axis activity in sheep

The effects of obestatin (an anorexigenic peripheral peptide) on somatotrophic axis activity in ruminants have not yet been determined. The aim of this study was to investigate the consequence of intracerebroventricular infusions of obestatin on the activity of the somatotrophic axis in peripubertal female sheep. Animals were randomly divided into two groups: control group received intracerebroventricular infusions of the vehicle, and the obestatin group was infused with obestatin (25 µg/120 µL h−1). The series of four hourly infusions on three consecutive days were performed. The blood samples were collected on day 0 and on day 3. Immediately after the end of experiment sheep were slaughtered. Parts of the brains were fixed in situ for further immunohistochemical analysis, while the remaining brains were frozen for Real Time RT-qPCR analysis.Substantial changes in the activity of the somatotrophic axis were observed in obestatin-infused sheep. In those animals obestatin evoked an increase in growth hormone-releasing hormone (GHRH) mRNA expression and a decrease in somatostatin mRNA expression in the anterior hypothalamic area. Moreover, a decrease in somatostatin immunoreactivity in the periventricular nucleus and an increase in somatostatin immunopositive fibers in the median eminence were noted. Changes in the GHRH and somatostatin activity are associated with an increase in growth hormone (GH) gene expression and in the amount of GH immunoreactive material stored in the somatotrophic pituitary cells. Consequently, an increase in GH concentration in the peripheral blood, due to an increase in the number of pulses was observed.It was revealed that obestatin affects the somatostatin/GHRH/GH system at the level of protein synthesis, accumulation and release. It is suggested that obestatin participates in the mechanism modulating somatotrophic axis activity at the central level by stimulating GH release through suppression of somatostatin output. Thereby, it can be concluded that obestatin may be involved in the modulation of growth processes in sheep.

Freeze-Dried Chitosan-Platelet-Rich Plasma Implants for Rotator Cuff Tear Repair: Pilot Ovine Studies.

Rotator cuff tears are a very common shoulder pathology. Different suturing techniques have been used for surgical cuff repair, but failure of healing remains a significant clinical challenge. The objective of this study was to establish and compare chronic and acute ovine rotator cuff tear models in our laboratory and investigate the feasibility of using chitosan (CS)-platelet-rich plasma (PRP) implants in conjunction with suture anchors to treat rotator cuff tears in large animal models. Repair with suture anchors only was used as control. In two preliminary pilot studies, unilateral full-thickness tears were created in the infraspinatus (ISP) tendon of mature female Texel-cross sheep. In the chronic model (n = 4 sheep), the tendons were capped with silicon and allowed to retract for 6 weeks, leading to degenerative changes, whereas the tendons were immediately repaired in the acute model (n = 4 sheep). Transected ISP tendons were reattached with suture anchors and, in the case of treated shoulders, implants composed of freeze-dried CS solubilized in autologous PRP were additionally applied to the tendon-bone interface and on top of the repaired site. The chronic defect model induced significant tendon degeneration and retraction, which made repair more challenging than in the acute defect model. Half the tendons in the chronic repair model were found to be irrepairable at 6 weeks. In the other half, the tendons could not be reattached to the footprint due to significant retraction, which made this a model of tissue formation in a gap. In contrast, the acute tendon repair model was executed easily. Extensive bone remodeling and tissue ingrowth at the tendon-bone interface were observed in the case of treatment with anchors + CS-PRP in both models, suggesting that CS-PRP implants could potentially modulate rotator cuff healing processes in large animal models.

The GDF5 mutant BB-1 enhances the bone formation induced by an injectable, poly(l-lactide-co-glycolide) acid (PLGA) fiber-reinforced, brushite-forming cement in a sheep defect model of lumbar osteopenia

Background ContextTargeted delivery of osteoinductive bone morphogenetic proteins (eg, GDF5) in bioresorbable calcium phosphate cement (CPC), potentially suitable for vertebroplasty and kyphoplasty of osteoporotic vertebral fractures, may be required to counteract augmented local bone catabolism and to support complete bone regeneration. The biologically optimized GDF5 mutant BB-1 may represent an attractive drug candidate for this purpose. PurposeThe aim of the current study was to test an injectable, poly(l-lactide-co-glycolide) acid (PLGA) fiber-reinforced, brushite-forming CPC containing low-dose BB-1 in a sheep lumbar osteopenia model. Study Design/ SettingThis is a prospective experimental animal study. MethodsBone defects (diameter 5 mm) were generated in aged, osteopenic female sheep and were filled with fiber-reinforced CPC alone (L4; CPC+fibers) or with CPC containing different dosages of BB-1 (L5; CPC+fibers+BB-1; 5, 100, and 500 µg BB-1; n=6 each). The results were compared with those of untouched controls (L1). Three and 9 months after the operation, structural and functional effects of the CPC (±BB-1) were analyzed ex vivo by measuring (1) bone mineral density (BMD); (2) bone structure, that is, bone volume/total volume (BV/TV) (assessed by micro-CT and histomorphometry), trabecular thickness (Tb.Th), and trabecular number (Tb.N); (3) bone formation, that is, osteoid volume/bone volume (OV/BV), osteoid surface/bone surface (OS/BS), osteoid thickness, mineralizing surface/bone surface (MS/BS), mineral apposition rate, and bone formation rate/bone surface; (4) bone resorption, that is, eroded surface/bone surface; and (5) compressive strength. ResultsCompared with untouched controls (L1), CPC+fibers (L4) and/or CPC+fibers+BB-1 (L5) significantly improved all parameters of bone formation, bone resorption, and bone structure. These effects were observed at 3 and 9 months, but were less pronounced for some parameters at 9 months. Compared with CPC without BB-1, additional significant effects of BB-1 were demonstrated for BMD, bone structure (BV/TV, Tb.Th, and Tb.N), and bone formation (OS/BS and MS/BS). The BB-1 effects on bone formation at 3 and 9 months were dose dependent, with 100 µg as the potentially optimal dosage. ConclusionsBB-1 significantly enhanced the bone formation induced by a PLGA fiber-reinforced CPC in sheep lumbar osteopenia. A single local dose as low as 100 µg BB-1 was sufficient to augment middle- to long-term bone formation. A CPC containing the novel GDF5 mutant BB-1 may thus represent an alternative to the bioinert, supraphysiologically stiff polymethylmethacrylate cement presently used to treat osteoporotic vertebral fractures by vertebroplasty and kyphoplasty.

Effects of age and sex on blood biochemistry of dorper lambs

The purpose of this study was to evaluate the physiological variations and influence of age group and sex on biochemical variables of 32 male and female Dorper sheep at the age of 15 to 121 days. Four blood samples were collected from each animal, at monthly intervals, totaling 128 samples. Biochemical variables determined in each sample were total protein (TP), albumin (ALB), globulin (GLB), albumin/globulin ratio (A/G), glucose (GLU), triglycerides (TG), total cholesterol (tCHOL), high density cholesterol carrying lipoproteins (HDL-C), low density cholesterol carrying lipoproteins (LDL-C), very low density cholesterol carrying lipoproteins (VLDL-C), urea, creatinine (CRE), total calcium (tCa), ionized calcium (iCa), phosphorus (P), calcium/phosphorus ratio (Ca/P), magnesium (Mg), sodium (Na), potassium (K), chloride (Cl), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT) and alkaline phosphatase (ALP). The results were obtained in four moments according to age: M1 - sheep from 15 to 30 days, M2 - sheep from 45 to 61 days, M3 - sheep from 74 to 90 days and M4 - sheep from 105 to 121 days. Comparing the values of the biochemical variables among the four age groups studied a significant difference (p &lt; 0.05) was observed in the concentrations of all the variables evaluated except for Na, Cl and ALT. Sex did not influence (p &gt; 0.05) the values of the biochemical variables analyzed. Based on the results, it is concluded that the age groups exert a significant influence on the majority of blood biochemical variables of Dorper sheep from 15 to 121 days of age.

A New Clinically Relevant T-Score Standard to Interpret Bone Status in a Sheep Model.

BackgroundOsteoporosis is diagnosed by bone loss using a radiological parameter called T-score. Preclinical studies use DXA to evaluate bone status were the T-score is referenced on bone mineral density (BMD) values of the same animals before treatment. Clinically, the reference BMD represents values of an independent group of healthy patients around 30 years old. The present study established a clinically similar T-score standard to diagnose osteoporosis in a sheep model.Material/MethodsWe used 31 female merino land sheep (average 5.5 years old) to study osteoporosis. The following groups were compared using DXA measurement: 1) control; 2) ovariectomized (OVX); 3) OVX combined with a deficient diet (OVXD); and 4) OVXD combined with methylprednisolone administration (OVXDS). Further, an independent group of 32 healthy sheep (4–6 years old) were measured as an independent baseline. BMD was measured at 0 months, 3 months, and 8 months after treatment.ResultsThe same significance pattern between the treated groups and either baseline groups was seen. However, using an independent baseline changed the “clinical” interpretation of the data from an osteoporotic bone status (T-score <−2.5) after 3 months of OXDS treatment into an osteopenic bone status (T-score <−1.5 to −2.4).ConclusionsUsing an independent baseline enhanced the statistical significance and showed the clinical relevance. Furthermore, an independent baseline is a reliable alternative to use of a new control group for future experiments and thus reduces the number of animals needed by eliminating the need for a control and corresponding to clinical practice.

A description of the sequence of long-term behavior of autogenous boneblock in maxillary sinus augmentation in sheep: Additional morphologic and histomorphometric evidence

To document the behavior of autogenous bone block in sinus lift and surgical consequences. Twelve sinus lifts with autogenous hip bone blocks and simultaneous insertion of two implants in 6 adult female sheep. Polychrome sequential labelling and histologic and histomorphometric evaluation after 6, 16, and 26 weeks. Augmentation material in the apical third was almost fully resorbed after 26 weeks (P = .00388). Percentage of bone tissue increased 0.5-1.0 mm from the implant in crestal region (15.3 ± 7.5% to 16.2 ± 10.1%), whereas it vanished in the apical region from 16 to 26 weeks (4.2 ± 10.4% to 0%) (P = .363). Autogenous bone block leaves an apical thin but functionally crucial layer covering implants in a form follows function way. Denial of animal-originated biomaterials and prion diseases remain a rarely discussed issue. The use of an implant length-adapted autogenous transplant with osseoinductive advantages should be taken into consideration.

Developmental Morphology and Topography of the Components of the Cervical Sympathetic Trunk in Sheep (Ovis aries) During the Fetal Period.

The objective of this study was to clarify the typical architecture and morphological variations of cervical sympathetic trunk (CST) in sheep during fetal period. Components of CST were examined on both sides of 40 male and female sheep fetuses aged from 60 to 140 days under a stereomicroscope. Skeletotopy and frequency of presence of cranial cervical ganglion (CCG), syntopy of cervical ganglia, and composition and topography of vagosympathetic trunk were consistent among specimens whereas the shape of cervical ganglia, the skeletotopy and number of three middle cervical ganglia (MG), and the frequency of communicating branches of CCG to the first cervical spinal nerve exhibited differences during fetal period. A reduction in the number of MG and the caudal movement of main MG were noted by increasing fetal age. Based on these detailed findings, comparative and developmental anatomy and evolutionary changes are discussed and compared with previous studies. The number of MG, skeletotopy of CCG and main MG, the number and range of communicating branches of CCG to spinal nerves, and the association of vagus and sympathetic nerves in fetal sheep were fundamentally different from those of mostly reported species. These results suggest that data obtained from CST of fetal sheep are significantly different from those obtained from humans, and it is problematic to apply them to humans because of the more cranial position of CCG, very narrow contribution of CCG to spinal nerve, absence of the vertebral ganglion, existence of multiple MG, and no communicating branches from MG to spinal nerves. Anat Rec, 300:2250-2262, 2017. © 2017 Wiley Periodicals, Inc.