Mesenchymal stem cells (MSCs) constitute a great promise for regenerative therapy, but these cells are difficultly recovered in large amounts. A potent alternative is the stromal vascular fraction (SVF), non-cultured MSCs, separated from adipose tissue (AT). We aim to evaluate AT harvesting site effect on the SVF cells’ quantity and quality in dogs. Subcutaneous abdominal fat, falciform ligament and peri-ovarian fat were sampled. After SVF isolation, the trypan blue exclusion test and a hemocytometer were used to assess the cell viability and cellular yield. SVF cells were labeled for four surface antigenic markers, clusters of differentiation CD90, CD44, CD29, and CD45, and then examined by flow cytometry. Semi-quantitative RT-PCR was used to evaluate the gene expression of the former markers in addition to OCT-4 and CD34. SVF cells in the peri-ovarian AT recorded the highest viability% (99.63 ± 0.2%), as well as a significantly higher cellular yield (36.87 ± 19.6 × 106 viable cells/gm fat, p < 0.001) and a higher expression of adipose-derived mesenchymal stem cells AD-MSCs surface markers than that of other sites. SVF cells from the peri-ovarian site revealed a higher expression of MSC markers (CD90, CD44, and CD29) and OCT-4 compared to the other sites, with weak CD45 and CD34 expressions. The positive OCT-4 expression demonstrated the pluripotency of SVF cells isolated from different sites. To conclude, the harvesting site is a strong determinant of SVF cells’ quantity and quality, and the peri-ovarian site could be the best AT sampling site in dogs.

Mitral valve regurgitation is a common canine heart disease. Transcatheter Edge-to-Edge Repair (TEER) is a transcatheter, edge-to-edge mitral repair device that uses a hybrid approach. No detailed information has been published on the hemodynamic effect of TEER on cardiac function. The aim of this report is to provide a longitudinal observation of the cardiac functional changes observed after TEER implantation in normal dogs using traditional, two-dimensional speckle tracking, and color M-mode echocardiographic methods. In the current report, TEER was implanted into two healthy dogs under general anesthesia. An echocardiographic examination was performed at baseline and weekly postoperative follow-ups were conducted until the fourth week. Successful TEER implantation was achieved with a short operation time (98 and 63 min) in the two dogs. Functional mitral valve regurgitation, elevated E/e’ ratio, elevated radial strain, and stable intraventricular pressure gradients (IVPG) were observed after the operation in the dogs. Mild non progressive mitral valve stenosis was observed in both dogs. TEER is a minimally invasive method for mitral valve surgery that necessitates more clinical trials. With longitudinal observation of heart function using novel approaches, better outcomes will be expected.

BackgroundUremic cardiomyopathy (UC), the main cause of death in progressive chronic kidney disease (CKD), is characterized by diastolic dysfunction. Intraventricular pressure gradients (IVPG) derived from color m-mode echocardiography (CMME) and two-dimensional speckle tracking echocardiography (2DSTE) were established as novel echocardiographic approaches for non-invasive and repeatable assessment of cardiac function. Previously, salvianolic acid B (Sal B) showed the potential to alleviate concentric LV hypertrophy in the pressure overload model. The purpose of this study was to evaluate the changes in cardiac function in UC and assess the efficacy of Sal B therapy using IVPG and 2DSTE techniques.Materials and MethodsTwenty-four rats underwent subtotal nephrectomy to produce progressive renal failure and were allocated equally into UC (n = 12) and Sal B-UC (n = 12) groups and monitored for 8 weeks. A sham-operated group was also included in this study (n = 12). Sal B was injected from weeks 4 to 8 in the Sal B-UC group. Conventional echocardiography, 2DSTE, and CMME were performed every 2 weeks post-operation, concomitantly with an evaluation of renal function. Histopathological and immunohistochemistry analyses were carried out to confirm the echocardiography findings.ResultsRenal failure and myocardial dysfunction were confirmed in the UC group from weeks 2 through 8. Eccentric and concentric hypertrophy was observed in the UC group, while the Sal B-UC group showed only eccentric hypertrophy. IVPG analysis did not reveal any significant differences between the groups. Edema, inflammation, fibrosis, and immunohistochemical expression of CD3 infiltration were higher in the UC group compared with sham and Sal B-UC groups.Conclusion2DSTE and IVPG explored the pathophysiology during the development of UC and indicated the incidence of myocardial dysfunction before ventricular morphological changes without intracardiac flow changes. This study confirmed increased ventricular stiffness and fibrosis in UC rats which was potentially treated by Sal B via decreasing edema, inflammation, and fibrosis.

Mesenchymal stem cells (MSCs) derived from adipose tissue are evolved into various cell-based regenerative approaches. Adipose-derived stem cells (ASCs) isolated from rats are commonly used in tissue engineering studies. Still, there is a gap in knowledge about how the harvest locations influence and guide cell differentiation. This study aims to investigate how the harvesting site affects stem-cell-specific surface markers expression, pluripotency, and differentiation potential of ASCs in female Sprague Dawley rats. ASCs were extracted from the adipose tissue of the peri-ovarian, peri-renal, and mesenteric depots and were compared in terms of cell morphology. MSCs phenotype was validated by cell surfaces markers using flow cytometry. Moreover, pluripotent gene expression of Oct4, Nanog, Sox2, Rex-1, and Tert was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR). ASCs multipotency was evaluated by specific histological stains, and the results were confirmed by quantitative polymerase chain reaction (RT-qPCR) expression analysis of specific genes. There was a non-significant difference detected in the cell morphology and immunophenotype between different harvesting sites. ASCs from multiple locations were significantly varied in their capacity to differentiate into adipocytes, osteoblastic cells, and chondrocytes. To conclude, depot selection is a critical element that should be considered when using ASCs in tissue-specific cell-based regenerative therapies research.

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Silk fibroin (SF) is a suitable material for vascular prostheses for small arteries. SF is useful not only as a base material for artificial vascular grafts but also as a coating material. This study prepared three types of transgenic SF (vascular endothelial growth factor (VEGF), Arg-Glu-Asp-Val (REDV), and Tyr-Ile-Gly-Ser-Arg (YIGSR)) incorporating expression factors that are thought to be effective for endothelialization as coating materials. We compared the contribution of these materials to early endothelialization in vivo when using them as a porous transgenic SF coating. A porous coating of transgenic SF containing VEGF, REDV, and YIGSR was applied to a silk small-diameter artificial vascular graft base with a diameter of 1.5 mm and a length of 3 cm. Two and four weeks after implantation of these artificial grafts into the abdominal aorta of rats, they were removed and evaluated by histologic examination. Transgenic SF coating incorporating VEGF and REDV demonstrated higher tissue infiltration and continuous endothelialization in the center of the graft compared to YIGSR at 4 weeks after implantation. VEGF and REDV are capable of endothelialization as early as 4 weeks after implantation, confirming the usefulness of transgenic SF when used as a porous coating.

Funding Acknowledgements Type of funding sources: None. Background/Introduction: Congestive heart failure (CHF), the main reason for morbidity and mortality, is considered a serious consequence of myocardial infarction (MI). The use of left ventricular end-diastolic pressure (LVEDP) as a chief indicator of CHF becomes limited because of the possible impairment of cardiac function and induced aortic valve damage during its recording. Echocardiography is the gold standard approach to diagnose structural myocardial dysfunction. However, its ability to predict chronic CHF following MI is still limited. Recently, intraventricular pressure gradient (IVPG) was presented as a non-invasive, highly sensitive preload-independent diastolic function parameter to assess cardiac function, especially during cardiomyopathy. However, there have not been any investigations demonstrating the feasibility of IVPG in the evaluation of post-infarction chronic CHF. Purpose This study aimed to investigate the utility of IVPG to assess the heart function in a rat model with chronic CHF following MI with evaluating its capacity to predict these changes. Methods Fifty male rats were included. MI was induced via ligation of the left anterior descending artery (LAD) at the level of the atrioventricular junction (MI animals, n = 35). Sham animals were subjected to the same left thoracotomy procedure without LAD ligation (Sham animals, n = 15). Transthoracic conventional echocardiography and colour M-mode echocardiography (CMME) for IVPG were performed in all animals 6 months post-surgery. The next day, animals were anesthetized, ventilated, and euthanized after the recording of hemodynamics. The heart weight, and lung and liver wet-to-dry weight ratios were recorded. J-tree cluster-analysis was performed based on ten echocardiographic variables indicative of CHF. Results Based on the cluster analysis, animals were joined into two clusters; CHF+ (n = 22) and named MI/HF+, and CHF- (n = 28) that was joined from sham (n = 15), and MI/HF- (n = 13). MI/HF+ presented the most severe anatomical and echocardiographic changes indicative of CHF with significant reduction of all IVPG indices and impairment of the hemodynamics. The IVPG indices were significantly (P&lt; 0.0001) correlated with the anatomical and echocardiographic findings, LVDP, LVEDP, HR, -dP/dtmin, and Tau. Meanwhile, LVSP was only significantly correlated with apical IVPG (R = 0.677, P = 0.022). dP/dtmax was significantly correlated with total IVPG, basal IVPG, and apical IVPG (R = 0.797, P = 0.017, R = 0.724, P = 0.003, and R = 0.652, P = 0.026 ; respectively). Moreover, total, basal, mid-to-apical, mid-, and apical IVPG were significant (P&lt; 0.0001) predictors of chronic CHF following MI. Conclusion Compared to the structural, and functional indices of conventional echocardiography, IVPG derived from CMME could provide a substantial non-invasive tool to diagnose and predict CHF after long-term MI.

In all organs, control of blood flow is important but might be particularly critical for testicular functions. This is because of the very low oxygen concentration and high metabolic rate of the seminiferous tubules, the physiological temperature of the testis, and its location outside the abdominal cavity. Many factors affect the characteristics of TBF in farm and companion animals, such as environment (thermal and seasonal effects) and physiological (species, breeds, age, body weight, and sexual maturity). Thermal environment stress has detrimental effects on spermatogenesis and consequently has more serious impacts on both human and animal fertility. Numerous studies have been performed to assess TBF in different animal species including bulls, rams, bucks, alpacas, stallions, and dogs with varied results. Hence, assessment of TBF by Doppler ultrasonography is of great importance to estimate the effect of high environment temperature on testicular functions. Also, differences observed in the TBF may result from different technical aspects such as the identification of the segment of the testicular artery to be examined. In the current review, we focused on the imperative roles of TBF in various animal species. Besides, we discussed in detail various factors that could affect TBF. These factors can significantly modify the TBF and thus should be considered when establishing reference values in farm animals for better clinical diagnosis. The information provided in this review is valuable for researchers and veterinarians to help them a better understanding of testicular hemodynamics for the proper evaluation of breeding soundness examination in males.