Search

Background: Granulosa cell tumours (GCT) are the most common ovarian tumours in mares. The classical presentation of a GCT is a unilaterally enlarged ovary appearing as a multicystic honeycomb mass. In rare cases, GCTs cause haemoperitoneum as a result of rapid growth of the tumour. The clinical diagnosis of GCT is usually based on history, rectal examination, ultrasonographic examination and serum hormone analysis, and surgical removal of the affected ovary is the treatment of choice. The different surgical approaches are based on the dimension of the GCT. Case description: A 7-year-old mare was referred to the department for horses due to suspicion of a large colon impaction. The mare presented with clinical signs of colic, fever and signs of hypovolemic shock. Rectal and ultrasonographic examination showed haemoperitoneum and a honey-comb mass within the abdomen, and a granulosa cell tumour (GCT) as the cause of an acute haemoperitoneum was diagnosed based on the serum level of Anti-Mullerian hormone. After stabilization of the mare, the GCT was removed through a ventral midline incision. Because of the enormous dimensions of the GCT, intra-abdominal partial resection of the tumour using a tenotomy knife was necessary to exteriorize the ovarian pedicle. At 3 months follow-up, the mare was ridden for her intended use. Conclusion: This report provides an approach to an uncommon case of a very large and heavy GCT. [Open Vet J 2023; 13(9.000): 1212-1218]

Osteoarthritis (OA) of the temporomandibular joint (TMJ) occurs spontaneously in humans and various animal species, including horses. In humans, obtaining tissue samples is challenging and clinical symptoms appear late in the disease progression. Therefore, genetically modified, induced, and naturally occurring animal models play a crucial role in understanding the pathogenesis and evaluating potential therapeutic interventions for TMJ OA. Among the naturally occurring models, the equine TMJ OA model is characterized by slow, age-related progression, a wide range of clinical examinations, and imaging modalities that can be performed on horses, as well as easy tissue and synovial fluid collection. The morphological and functional similarities of TMJ structures in both species make the equine model of TMJ OA an excellent opportunity to track disease progression and response to treatment. However, much work remains to be carried out to determine the utility of human TMJ OA biomarkers in horses. Among the main TMJ OA biomarkers, IL-1, IL-6, TGF-β, TNF-α, and PGE2 have been recently investigated in the equine model. However, the majority of biomarkers for cartilage degradation, chondrocyte hypertrophy, angiogenesis, and TMJ overload—as well as any of the main signaling pathways—have not been studied so far. Therefore, it would be advisable to focus further research on equine specimens, considering both mediators and signaling.

The utilization of Autologous Conditioned Serum (ACS) for treating osteoarthritis (OA) in horses has seen a notable increase in recent years. In vitro studies have consistently identified ACS as a promising therapy for OA joints, contributing to its growing popularity in OA treatment. Despite this, there is a noticeable absence of systematic reviews focused solely on the clinical data of OA patients treated with ACS, excluding the in vitro perspective. This study aims to address this gap by systematically reviewing the latest literature, concentrating solely on clinical data in in vivo studies to evaluate the efficacy of ACS in OA lesions. All clinical studies involving ACS treatments for horses with OA were included in the assessment. Surprisingly, only six trials met the inclusion criteria for this systematic review. The results indicate that the majority of the considered articles support the use of ACS as a treatment for horses, albeit with a control group provided in only one study. However, the absence of a control group and the exclusion of histological evaluation diminish the validity of the majority of clinical research. While several studies suggest a beneficial effect of ACS on OA horses without significant adverse effects, this systematic review affirms that there is no definitive evidence for its effectiveness. Therefore, further investigation of the efficacy of ACS products as a treatment for OA is warranted, emphasizing the need for more controlled trials. Poorly designed and biased studies, lacking blinding or control and adopting inadequate outcome measures, may favor positive results and, thus, necessitate a more rigorous approach to validate the efficacy of ACS in OA treatment.

Abstract Background Lameness in cattle is a major health problem and causes great economic losses. Carpal injury is a common cause of forelimb lameness in cattle. Radiography and/or ultrasonography of the carpus is a challenge due to complex anatomy of the joint. Additional imaging using computed tomography (CT) or magnetic resonance imaging (MRI) may be indispensable for reaching a decisive diagnosis. Precise evaluation of the clinical CT and MRI images necessitates an in-depth knowledge of the normal CT and MRI tissue variants. Therefore, our purpose was to provide a detailed description of the normal CT and MRI appearance of the osseous and soft tissue structures of twelve cadaveric bovine carpi using CT and 3 Tesla MRI. Carpi were frozen, transected in sagittal, dorsal and transverse planes then adjoined to their corresponding CT and MRI images. Results The clinically significant articular and peri-articular structures of the bovine carpus were identified and characterized on the CT and MRI images. CT images provided a remarkable delineation of the cortical, subchondral, and cancellous bone. The high-field 3 Tesla MRI offered high definition and distinction of the delicate soft tissues of the bovine carpus. Conclusions 3 Tesla high-field MRI offers new opportunities in soft tissue tomography but cannot be compared with CT in terms of bone imaging. Clinicians have to determine whether CT, MRI or both imaging techniques are required in clinical situations.

Equine veterinarians face challenges in treating horses with osteoarthritic joint pain in routine veterinary practice. All common treatment options aim to reduce the clinical consequences of osteoarthritis (OA) characterized by persistent synovitis and progressive degradation of articular cartilage. A range of joint-associated cell types and extracellular matrices are involved in the not yet entirely understood chronic inflammatory process. Regeneration of articular tissues to re-establish joint hemostasis is the future perspective when fundamental healing of OA is the long-term goal. The use of intra-articular applied biologic therapeutics derived from blood or mesenchymal stroma cell (MSC) sources is nowadays a well-accepted treatment option. Although this group of therapeutics is not totally consistent due to the lack of clear definitions and compositions, they all share a potential regenerative effect on articular tissues as described in in vivo and in vitro studies. However, the current stage of science in regenerative medicine needs to be supported by clinical reports as in fact, in vitro studies as well as studies using induced OA models still represent a fragment of the complex pathomechanism of naturally occurring OA. This systemic review aims to determine the long-term effect of orthobiologic therapeutics in horses suffering naturally occurring OA. Thereby, a meta-analysis of randomized controlled trials (RCTs) is conducted to describe the efficiency and safety of intra-articular applied orthobiologics in terms of lameness reduction in the long-term. Using the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines, thirteen studies met the inclusion criteria for the systemic review. Four of those studies have further been evaluated by the meta-analysis comparing the long-term effect in lameness reduction. Each study was examined for risk of bias. For data evaluation, a random-effects model was used, describing the overall outcome in a forest plot. The I2 statistic was used to assess heterogeneity. Results indicate, that orthobiologic therapies represent an effective long-term and safe OA treatment option. Due to the inhomogeneity of included studies, no statements are provided addressing specific orthobiologic therapies, affected joints, OA stage and horse's intended use. Future clinical trials should follow standardized study designs to provide comparable data.

The treatment of tendinopathies with multipotent mesenchymal stromal cells (MSCs) is a promising option in equine and human medicine. However, conclusive clinical evidence is lacking. The purpose of this study was to gain insight into clinical treatment efficacy and to identify suitable outcome measures for larger clinical studies. Fifteen horses with early naturally occurring tendon disease were assigned to intralesional treatment with allogeneic adipose-derived MSCs suspended in serum or with serum alone through block randomization (dosage adapted to lesion size). Clinicians and horse owners remained blinded to the treatment during 12 months (seven horses per group) and 18 months (seven MSC-group and five control-group horses) of follow-up including clinical examinations and diagnostic imaging. Clinical inflammation, lameness, and ultrasonography scores improved more over time in the MSC group. The lameness score difference significantly improved in the MSC group compared with the control group after 6 months. In the MSC group, five out of the seven horses were free of re-injuries and back to training until 12 and 18 months. In the control group, three out of the seven horses were free of re-injuries until 12 months. These results suggest that MSCs are effective for the treatment of early-phase tendon disease and provide a basis for a larger controlled study.

ObjectivesIndividuals with irregular physical activity (PA) participation are defined as fluctuators. This study aimed to comprehend how fluctuators’ perceived barriers and motivators in their subjective theories are exhibited and cognitively represented in relation to their everyday PA practices and lapses.MethodsThe design of “Research Program Subjective Theories” was used to explore and present fluctuators’ cognition concerning PA participation. Thirty fluctuators were invited to a semi-structured interview. By inductive and deductive coding, fluctuators’ verbal data were converted into word categories for extracting commonalities and comparing differences. By retaining the remaining word categories of high frequency and exploring the interrelationships among the remained word categories using statistical analyses, a superstructure (i.e., visualized representation of fluctuators’ overall cognition) including fluctuators’ main PA motivators, barriers, and behavioral outcomes was compiled.ResultsFluctuators face common motivators as barriers, such as lack of time, lack of willpower, lack of social support, and physical reasons (discomfort, injuries, or diseases). Fluctuators’ subjective theories primarily differed in motivational configurations. The physically motivated fluctuators (i.e., those predominantly motivated by physically related motivators) were more linked with low PA level, while the mixed motivated fluctuators (i.e., motivated by both physically related and emotionally or socially related motivators) were more likely associated with moderate PA level. The exemplars of the three typical fluctuators were also demonstrated to reveal their real experiences and situations in the daily life context.ConclusionDue to the fact that fluctuation research is still in its infancy, this study represents a significant opportunity to promote knowledge growth in this area. Future studies are recommended to convert findings of the present study into interventions that benefit fluctuators in overcoming perceived barriers and enhancing motivations to eventually participate in regular PA.

Mesenchymal stromal cells (MSC) represent a promising treatment option for tendon disorders and joint diseases, primarily osteoarthritis. Since MSC are highly context-sensitive to their microenvironment, their therapeutic efficacy is influenced by their tissue-specific pathologically altered targets. These include not only cellular components, such as resident cells and invading immunocompetent cells, but also components of the tissue-characteristic extracellular matrix. Although numerous in vitro models have already shown potential MSC-related mechanisms of action in tendon and joint diseases, only a limited number reflect the disease-specific microenvironment and allow conclusions about well-directed MSC-based therapies for injured tendon and joint-associated tissues. In both injured tissue types, inflammatory processes play a pivotal pathophysiological role. In this context, MSC-mediated macrophage modulation seems to be an important mode of action across these tissues. Additional target cells of MSC applied in tendon and joint disorders include tenocytes, synoviocytes as well as other invading and resident immune cells. It remains of critical importance whether the context-sensitive interplay between MSC and tissue- and disease-specific targets results in an overall promotion or inhibition of the desired therapeutic effects. This review presents the authors’ viewpoint on disease-related targets of MSC therapeutically applied in tendon and joint diseases, focusing on the equine patient as valid animal model.

Abstract Background Human obesity is linked with systemic inflammation. However, it is still controversial if equines produce more inflammatory cytokines with increasing body weight and if the production of those show breed type specific patterns. The main objective of this study was to determine if diet induced obesity is associated with increased inflammatory signatures in adipose tissue of equines and if a breed predisposition exists between ponies and horses. Additionally, we aimed to identify adipose tissue depot differences in inflammatory cytokine expression. Nineteen healthy, non-overweight and metabolically healthy equines received a hypercaloric diet for 2 years. Body weight, body condition score and cresty neck score were assessed weekly throughout the study. At three time points, insulin sensitivity was determined by a combined glucose-insulin test. Adipose tissue samples were collected from two intra-abdominal and two subcutaneous depots under general anesthesia at each time point after an endotoxin trigger. In the adipose tissue samples levels of CD68 mRNA (a marker of macrophage infiltration) and pro-inflammatory cytokine mRNA (IL-1β, IL-6 and TNFα) were analyzed with RT-qPCR. As markers of lipid metabolism mRNA levels of lipoprotein lipase (LPL) and fatty acid binding protein 4 (FABP4) were determined with RT-qPCR. Results CD68 mRNA levels increased with body weight gain in several adipose tissue (AT) depots (Wilcoxon signed rank test with Bonferroni correction; retroperitoneal AT horses: P = 0.023, mesocolonial AT horses: P = 0.023, subcutaneous tail head AT ponies: P = 0.015). In both abdominal depots CD68 mRNA levels were higher than in subcutaneous adipose tissue depots (Kruskal–Wallis-ANOVA with Bonferroni correction: P

Abstract Background Urban parks play an important role in promoting physical activity (PA) among adults and especially among older city residents. According to the socioecological approach the association of physical environments and psychosocial factors in the context of park-based PA of elderly have not been systematically examined until now, let alone the relevance of the city (urban area) on a cross-cultural level. This study investigated selected aspects of (1) the association of psychosocial and park environmental factors with park-based physical activity (PBPA) of older people; and (2) the moderating effect of city on the association of these factors with PBPA. Methods A face-to-face survey was conducted of a mixed-culture sample from different urban surroundings in Hong Kong (HK) and Leipzig (L). In six parks of each city physically active elderly (> = 60 years; HK: n = 306; L: n = 311) were recruited. Multiple linear regressions were used to analyse the association between psychosocial factors and perceived environmental factors with PBPA and the moderating effect of city. Results Controlled for demographic variables, all other psychosocial factors were significantly related to PBPA, except social support. In terms of environmental factors, PBPA was positively associated with safety, attractiveness, features and negatively associated with park time distance. Controlled for demographic variables, psychosocial and environmental factors, the moderating effect of city on the associations of park features and park time distance with PBPA was not significant in HK. In contrast, there was a significant positive relationship for park features and a negative relationship for park time distance with PBPA in L. Conclusions Psychosocial and perceived environmental factors significantly influence PBPA of older people. City moderates the associations of these factors and independently contributes to park-based PA of the elderly. The different interactions of environmental factors and urban area for PBPA of elderly can support policy makers on the municipal level in choosing adequate strategies for promoting PA of older people in parks.

Abstract Background Despite appropriate medical therapy, many horses with equine recurrent uveitis continue to suffer from recurrent bouts of inflammation. Surgical intervention via the pars plana vitrectomy or suprachoroidal cyclosporine implant placement may control and/or prevent recurrences, however, these procedures may be contraindicated, unavailable, or declined by an owner. Thus, an effective adjunctive treatment option may help to improve the clinical outcomes in those situations. There are several anecdotal reports on the use of intravitreal gentamicin injections, but to date, no data evaluating the complication rate and/or treatment effect following this treatment have been published. Thus, the aim of this prospective study was to describe the intravitreal gentamicin injection technique, describe the associated peri-injection (within 24 h) and post-injection (30 to 780 days) complications, and to report the effects of the injection on the clinical signs of uveitis. Additionally, evaluation of the systemic and ocular Leptospira-status, and its effect on the treatment outcome was performed. A total of 86 horses of various ages, breeds, and gender presenting with recurrent or persistent uveitis were treated via intravitreal injection of 4 mg of undiluted gentamicin (0.04 ml, Genta 100, 100 mg/ml in 35 horses) or preservative-free gentamicin (0.05 ml, 80 mg/ml in 52 horses) under sedation and local anesthesia. All 86 horses were observed for immediate peri-injection and post-injection complications. Response to therapy was evaluated in 59 of the 86 horses (follow-up: 30 to 780 days). Results Peri-injection complications consisted of subconjunctival (26/86; 30.2%) or intracameral hemorrhage (4/86; 4.7%); both of which completely resolved within 5 days. Post-injection complications consisted of cataract formation/maturation (5/59 horses, 8.5%) and diffuse retinal degeneration (3/59 eyes 5.1%). The majority of horses 52/59 (88.1%) with a minimum follow-up period of 30 days were controlled (absence of recurrent or persistent inflammation) at their last recheck examination. Recurrent inflammation was documented in 5/59 (8.5%) horses and persistent inflammation was diagnosed in 2/59 (3.4%) horses. Conclusions Intravitreal injection of low-dose gentamicin shows promise at controlling different types and stages of uveitis. The ability of intravitreal injections of low-dose gentamicin (4 mg) to control persistent and recurrent inflammation warrants further investigation.

BackgroundThere are conflicting reports about the fitness status of European adults, partly due to the lack of a standardized fitness test battery used across Europe. The European Fitness Badge (EFB) was developed in 2017 as an online-based tool to assess the health-related fitness of persons aged ≥ 18 years residing in European countries. We examined the demographic characteristics and fitness status of persons who completed the EFB between June 2017 and May 2019.MethodsWe conducted a multinational study in eight European countries. Participants completed the EFB which includes 11 validated motor tests to measure endurance, strength, coordination, and flexibility performance, under the supervision of an EFB instructor in different settings (e.g., sports club sessions, public events). Two different test batteries [test profiles (TPs)] are available to distinguish between less active (TP1) and active individuals (TP2). We calculated descriptive statistics and conducted analyses of variance to examine sample characteristics and a potential impact of sex, age, body mass index (BMI), physical activity, and posture on fitness as assessed by the EFB.ResultsThe sample included 6,019 adults (68.7% females; mean age 52.7 years; age range 18–89 years). Participants who completed TP1 were older (TP1: 61.4 years; TP2: 44.2 years; p = 0.00), reported a lower level of physical activity (TP1: 3.8; TP2: 4.0; p = 0.00), had a higher BMI (TP1: 25.7; TP2: 24.3; p = 0.00) and a higher frequency of postural abnormalities (TP1: 43%; TP2: 33%; p = 0.00) than TP2 participants. Among 3,034 participants who completed TP2, males had higher performance in endurance, strength, and overall fitness, whereas females performed better in coordination and flexibility tests. In addition, younger age, lower BMI, and higher level of physical activity engagement were associated with better EFB test performance.ConclusionThe EFB can be used to assess the health-related fitness status of individuals aged ≥ 18 years. Our results show that TP1 and TP2 were completed by persons from the respective target groups (i.e., less active vs. active), and also confirm findings from previous studies on potential determinants of fitness such as sex or age.

Strong evidence exists that fitness is a physical health resource, which serves to protect one’s health. There is still uncertainty about which fitness level provides the best health outcome and which measurements can be used for analyzing this question. This cross-sectional study analyzed 462 (64.07% female) German middle-aged adults regarding their fitness status, physical activity (Non-Exercise test), body composition (Body Mass Index) and heart-related health status. Motor tests were used to measure the health-related fitness status. The heart-related health status was surveyed by questionnaire and diagnosis was done in part by a physician. Relationships between risk factors and fitness factors are visible during the correlative analysis. They are substantially more visible in the differentiation of people with and without risks. People with low fitness show noticeable risks in activity, Body Mass Index and heart-related health. People with high fitness show health resources for activity, Body Mass Index and heart-related health. This study points out that all fitness dimensions influence one’s heart-related health in a positive way. Fitness is measured objectively and includes all health-related fitness dimensions such as endurance, strength, coordination and flexibility. Apart from this standardization, we ask for more longitudinal studies and more objective health measurements.

OBJECTIVES:Fluctuation is a common but neglected phenomenon of physical activity (PA) behavior. This study aimed to explore the psychosocial profiles of PA fluctuation in office employees, and to examine the association of latent profiles with demographics and PA level. METHOD:434 Chinese office employees who were identified as PA fluctuators (M = 32.4 years, SD = 6.9, 55.5% female) completed a cross-sectional online survey covering demographics, PA behavior, and six psychosocial indicators (self-efficacy, planning, action control, affective attitude, social support, and perceived barriers). Latent profile analysis was used to determine PA fluctuators' psychosocial profiles. Associated factors of profile membership were identified with multinomial logistic regression. RESULTS:The two-profile model (uncommitted vs. moderately committed) was selected as the best solution. The moderately committed group (n = 346, 79.7%) possessed a more active mindset by reporting significantly higher scores of self-efficacy (t = 9.42 p < .001), planning (t = 16.33 p < .001), action control (t = 14.55 p < .001), affective attitude (t = 13.33 p < .001), and social support (t = 11.50 p < .001) compared with the uncommitted group (n = 88, 20.3%). Results from a multinomial logistic regression showed that the moderately committed profile was associated with normal weight status (OR = 2.00, p< .05), having a medium managerial position (OR = 2.54, p< .01), and high level of moderate to vigorous PA behavior (OR = 4.85, p< .001). CONCLUSIONS:These findings demonstrate the variability of PA fluctuators' mindsets. Future tailored interventions are recommended to promote PA behavior for this population based on the categorization from the present study.

Presurgical hand asepsis is part of the daily routine in veterinary medicine. Nevertheless, basic knowledge seems to be low, even among specialised veterinary surgeons. The major objectives of our study were to assess current habits for presurgical hand preparation (phase 1) among personnel in a veterinary hospital and their effectiveness in reducing bacteria from hands in comparison to a standardised protocol (phase 2). Assessment of individual habits focused on time for hand washing and disinfection, the amount of disinfectant used, and the usage of brushes. The standardised protocol defined hand washing for 1 min with liquid neutral soap without brushing and disinfection for 3 min. All participants (2 surgeons, 8 clinic members, 32 students) used Sterillium®. Total bacterial counts were determined before and after hand washing, after disinfection, and after surgery. Hands were immersed in 100 ml sterile sampling fluid for 1 min and samples were inoculated onto Columbia sheep blood agar using the spread-plate method. Bacterial colonies were manually counted. Glove perforation test was carried out at the end of the surgical procedure. Differences in the reduction of relative bacterial numbers between current habits and the standardised protocol were investigated using Mann-Whitney-Test. The relative increase in bacterial numbers as a function of operation time (≤60 min, >60 min) and glove perforation as well as the interaction of both was investigated by using ANOVA. Forty-six and 41 preparations were carried out during phase 1 and phase 2, respectively. Individual habits differed distinctly with regard to time (up to 8 min) and amount of disinfectant (up to 48 ml) used both between participants and between various applications of a respective participant. Comparison of current habits and the standardised protocol revealed that the duration of hand washing had no significant effect on reducing bacteria. Contrary, the reduction in bacterial numbers after disinfection by the standardised protocol was significantly higher (p

Background Non-alcoholic fatty liver disease is known as determining part of human obesity. The impact of body weight (BW) gain on liver metabolism has not been extensively investigated yet. Objectives To investigate hepatic alterations caused by increasing BW in ponies and horses. Animals A total of 19 non-obese equines (10 Shetland ponies, geldings; nine Warmblood horses, geldings). Methods Animals received 200% of their metabolizable maintenance energy requirements for 2 years. Serum alkaline phosphatase, glutamate dehydrogenase (GLDH), aspartate aminotransferase (AST), and gamma-glutamyl transferase activities and bile acids were analyzed several times during 2 years of hypercaloric diet. Hepatic lipid content and hepatic levels of the interleukin (IL)-6, tumor necrosis factor α (TNFα), cluster of differentiation (CD) 68, IL-1β, lipoprotein lipase (LPL), fatty acid-binding protein 1, chemerin and nuclear factor-κB mRNAs were assessed at the start of the study and after 1 and 2 years of excess energy intake. Results The mean (±SD) BW gain recorded during 2 years of excess energy intake was 29.9 ± 19.4% for ponies and 17 ± 6.74% for horses. The hepatic lipid content was not profoundly affected by increasing BW. Levels of the IL-6, TNFα, CD68 and IL-1β mRNAs did not change during BW gain. Levels of the chemerin mRNA increased significantly in both breeds (ponies: P = 0.02; horses: P = 0.02) in response to BW gain. Significant differences in serum GLDH and AST activities, serum bile acid concentrations and hepatic levels of the LPL mRNA were observed between ponies and horses at the end of the study. Conclusions Chemerin might represent an interesting marker for future equine obesity research. Interestingly, steatosis caused by increasing BW may occur later in the development of obesity in equines than in humans. Additionally, the hepatic metabolism exhibits differences between ponies and horses, which may explain in part the greater susceptibility of ponies to obesity-associated metabolic dysregulations.

The immunomodulatory potential of multipotent mesenchymal stromal cells (MSC) provides a basis for current and future regenerative therapies. In this study, we established an approach that allows to address the effects of pro-inflammatory stimulation and co-culture with MSC on different specific leukocyte subpopulations. Equine peripheral blood leukocyte recovery was optimized to preserve all leukocyte subpopulations and leukocyte activation regimes were evaluated. Allogeneic labeled equine adipose-derived MSC were then subjected to direct co-culture with either non-stimulated, concanavalin A (ConA)-activated or phosphate 12-myristate 13-acetate and ionomycin (PMA/I)-activated leukocytes. Subsequently, production of the cytokines interferon-γ (IFN- γ), interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) and presence of FoxP3 were determined in specific cell populations using multicolor flow cytometry. Prostaglandin E2 (PGE2) was measured in the supernatants. ConA-stimulation induced mild activation of leukocytes, whereas PMA/I-stimulation led to strong activation. In T cells, PMA/I promoted production of all cytokines, with no distinct suppressive effects of MSC. However, increased numbers of CD25/FoxP3-positive cells indicated that MSC supported regulatory T cell differentiation in PMA/I-activated leukocyte cultures. MSC also reduced numbers of cytokine-producing B cells and granulocytes, mostly irrespective of preceding leukocyte activation, and reversed the stimulatory effect of ConA on IFN-γ production in monocytes. Illustrating the possible suppressive mechanisms, higher numbers of MSC produced IL-10 when co-cultured with non-stimulated or ConA-activated leukocytes. This was not observed in co-culture with PMA/I-activated leukocytes. However, PGE2 concentration in the supernatant was highest in the co-culture with PMA/I-activated leukocytes, suggesting that PGE2 could still mediate modulatory effects in strongly inflammatory environment. These context- and cell type-specific modulatory effects observed give insight into the interactions between MSC and different types of immune cells and highlight the roles of IL-10 and PGE2 in MSC-mediated immunomodulation. The approach presented could provide a basis for further functional MSC characterization and the development of potency assays.

Fatty acids, as key components of cellular membranes and complex lipids, may play a central role in endocrine signalling and the function of adipose tissue and liver. Thus, the lipid fatty acid composition may play a role in health status in the equine. This study aimed to investigate the fatty acid composition of different tissues and liver lipid classes by comparing Warmblood horses and Shetland ponies under defined conditions. We hypothesized that ponies show different lipid patterns than horses in adipose tissue, liver and plasma. Six Warmblood horses and six Shetland ponies were housed and fed under identical conditions. Tissue and blood sampling were performed following a standardized protocol. A one-step lipid extraction, methylation and trans-esterification method with subsequent gas chromatography was used to analyse the total lipid content and fatty acid profile of retroperitoneal, mesocolon and subcutaneous adipose tissue, liver and plasma. Fatty acids were grouped according to their degree of saturation and their conjugated double bond into the respective lipid classes. In the adipose tissues, saturated fatty acids (SFAs) and n-9 monounsaturated fatty acids (n-9 MUFAs) were most present in ponies and horses. N-6 polyunsaturated fatty acids (n-6 PUFAs), followed by SFAs, were most frequently found in liver tissue and plasma in all animals. Horses, in comparison to ponies, had significantly higher n-6 PUFA levels in all tissues and plasma. In liver tissue, horses had significantly lower hepatic iso-branched-chain fatty acids (iso-BCFAs) than ponies. The hepatic fatty acid composition of selected lipid classes was different between horses and ponies. In the polar PL fraction, horses had low n-9 MUFA and n-3 PUFA contents but higher n-6 PUFA contents than ponies. Furthermore, iso-BCFAs are absent in several hepatic lipid fractions of horses but not ponies. The differences in fatty acid lipid classes between horses and ponies provide key information on the species- and location-specific regulation of FA metabolism, thus affecting health status such as inflammatory responses.

The magic angle effect increases the MRI signal of healthy tendon tissue and could be used for more detailed evaluation of tendon structure. Furthermore, it could support the discrimination of hypointense artefacts induced by contrast agents such as superparamagnetic iron oxide used for cell tracking. However, magic angle MRI of the equine superficial digital flexor tendon has not been accomplished in vivo in standing low-field MRI so far. The aim of this in vivo study was to evaluate the practicability of this magic angle technique and its benefit for tracking superparamagnetic iron oxide-labelled multipotent mesenchymal stromal cells. Six horses with induced tendinopathy in their forelimb superficial digital flexor tendons were injected locally either with superparamagnetic iron oxide-labelled multipotent mesenchymal stromal cells or serum. MRI included standard and magic angle image series in T1- and T2∗-weighted sequences performed at regular intervals. Image analysis comprised blinded evaluation and quantitative assessment of signal-to-noise ratio. The magic angle technique enhanced the tendon signal-to-noise ratio (P

Background Development of incisional complications following ventral median celiotomy might depend on suture pattern for skin closure. Methods In this prospective study, 21 healthy male horses underwent celiotomy. Skin closure was either performed via a continuous percutaneous pattern (CO group; 5 warmbloods/5 ponies) or an intradermal pattern (ID group; 5 warmbloods/6 ponies). Follow-up examination of the incisional site included daily monitoring for edema, dehiscence, and drainage. Transcutaneous ultrasound was performed at Days 3, 6, and 10 as well as on Week 8 and 12 to evaluate size of edema and presence or absence of sinus formation, and hernia formation. Prevalence of incisional infection on base of positive microbiological analysis at any time up to Day 10 was evaluated and compared between ID and CO group. Furthermore, edema size was analysed by a linear mixed-effect model for group and time dependency. Results Observed incisional complications included edema (9/10 in CO, 10/11 in ID), suture sinus formation (2/10 in CO, 1/11 in ID), surgical site infection (2/10 in CO, 0/11 in ID), and incisional hernia (1/10 in CO, 0/11 in ID). The overall prevalence of incisional infection was 9.5% without significant differences between both groups (20% in CO, 0% in ID; p = 0.214). Edema size was not dependent on time or group (p = 0.545 and p = 0.627, respectively). Discussion CO and ID suture pattern are appropriate for skin closure following ventral median celiotomy in horses. None of the animals in the continuous ID group developed surgical site infections, even without the use of antibiotics.

Abstract Background The study examined the measurement quality of a stage algorithm measuring the Four steps from Inactivity to activity Theory (FIT Model). Methods In a cross-sectional study, stages were assessed in 1012 Chinese university students in terms of physical activity, social-cognitive variables and health outcomes. Main outcome measures were stages of change, self-reported physical activity, perceived barriers, intrinsic motivation, plans, fitness and health satisfaction. Misclassification, sensitivity, specificity, receiver operating characteristic (ROC) curves, nonlinear trends, and planned comparison were computed. Results Compared to previous studies, sensitivity was at the average level (64 %-71 %), and specificity was comparably higher (76%-89%). When using higher PA intensity criteria (moderate and strenuous intensities), sensitivity was higher, whereas specificity was lower in comparison to the lower PA intensity criteria (also including mild activity). After running contrast and trend analyses, nonlinear trends for all indicative variables across the stages and a match of 77 % of predictions of stage differences were confirmed. Conclusion The measurement quality of the stage algorithm was supported in a young adult sample.

Age-related degenerative changes in tendon tissue represent a common cause for acute tendon pathologies. Although the regenerative potential of multipotent mesenchymal stromal cells (MSC) was reported to restore functionality in injured tendon tissue, cellular mechanisms of action remain partly unclear. Potential tenogenic differentiation of applied MSC is affected by various intrinsic and extrinsic factors. The current study presents an in vitro model to evaluate the combined extrinsic effects of decellularized equine tendon matrix, transforming growth factor beta 3 (TGFβ3) and bone morphogenetic protein 12 (BMP12) on the tenogenic fate of equine adipose tissue-derived MSC. Monolayer MSC cultures supplemented with TGFβ3 and BMP12 as well as MSC cultured on tendon matrix scaffolds preloaded with the growth factors were incubated for 3 and 5 days. Histological evaluation and real time reverse transcription polymerase chain reaction (RT-PCR) revealed that growth factor-mediated tenogenic induction of MSC was modified by the conditions of the surrounding microenvironment. While the gene expression pattern in monolayer cultures supplemented with TGFβ3 or TGFβ3 and BMP12 revealed an upregulation for collagen 1A2, collagen 3A1, tenascin c, scleraxis and mohawk ( p < 0.05 ) , the presence of tendon matrix led to an upregulation of decorin and osteopontin as well as to a downregulation of smad8 ( p < 0.05). Preloading of scaffolds with either TGFβ3, or with TGFβ3 and BMP12 promoted a tenocyte-like phenotype and improved cell alignment. Furthermore, gene expression in scaffold culture was modulated by TGFβ3 and/or BMP12, with downregulation of collagen 1A2, collagen 3A1, decorin, scleraxis, smad8 and osteopontin, whereas gene expression of tenascin c was increased. This study shows that growth factor-induced tenogenic differentiation of equine MSC is markedly altered by topographical constraints of decellularized tendon tissue in vitro. While TGFβ3 represents an effective mediator for tenogenic induction, the role of BMP12 in tenogenesis may be of modulatory character and needs further evaluation.

Transforming growth factor beta 3 (TGFβ3) promotes tenogenic differentiation and may enhance tendon regeneration in vivo. This study aimed to apply TGFβ3 absorbed in decellularized equine superficial digital flexor tendon scaffolds, and to investigate the bioactivity of scaffold-associated TGFβ3 in an in vitro model. TGFβ3 could effectively be loaded onto tendon scaffolds so that at least 88% of the applied TGFβ3 were not detected in the rinsing fluid of the TGFβ3-loaded scaffolds. Equine adipose tissue-derived multipotent mesenchymal stromal cells (MSC) were then seeded on scaffolds loaded with 300 ng TGFβ3 to assess its bioactivity. Both scaffold-associated TGFβ3 and TGFβ3 dissolved in the cell culture medium, the latter serving as control group, promoted elongation of cell shapes and scaffold contraction (p < 0.05). Furthermore, scaffold-associated and dissolved TGFβ3 affected MSC musculoskeletal gene expression in a similar manner, with an upregulation of tenascin c and downregulation of other matrix molecules, most markedly decorin (p < 0.05). These results demonstrate that the bioactivity of scaffold-associated TGFβ3 is preserved, thus TGFβ3 application via absorption in decellularized tendon scaffolds is a feasible approach.

Tendon disease has been treated with multipotent mesenchymal stromal cells (MSCs) in the equine large-animal model with promising success. The aim of this study was to gain more insight into the fate and biodistribution of MSCs after local application into tendon lesions by long-term cell tracking in this large-animal model. Superficial digital flexor tendon lesions were induced in all limbs in six horses and injected with 10 × 10 6 Molday ION Rhodamine B ™ -labeled MSCs suspended in serum or serum alone. Follow-up was performed using low-field magnetic resonance imaging (MRI), flow cytometry, and histology. Cell tracking based on the hypointense artifacts induced by the superparamagnetic iron oxide (SPIO) labeling agent in MRI as well as based on Rhodamine B fluorescence was feasible. However, Prussian blue staining for assessment of histology was not entirely specific for SPIO. Labeled cells could be traced at their injection site by MRI as well as histology for the whole follow-up period of 24 weeks. Although the numbers of labeled cells within the injected tendon lesions decreased over time, part of the applied cells appeared to remain viable and integrated within the injured tissue. Furthermore, small numbers of labeled cells were identified in peripheral blood within the first 24 h after cell injection and could also be found until week 24 within the contralateral control tendon lesions that had been injected with serum. The present findings unveil details on MSC biodistribution and persistence after their local application, which are of clinical relevance with regard to MSC safety and mechanisms of action.

Cell-based therapies have been in use in veterinary medicine for years. However, the legal requirement of manufacturing, placing on the market and use of cell-based veterinary pharmaceuticals are not as well developed as the respective requirements of chemical pharmaceuticals. Cell-based veterinary pharmaceuticals are medicinal products in the sense of the pharmaceutical law of the European Union (EU). For that reason, such medicinal products principally require official approval for their manufacture and an official marketing authorisation for their placement on the market before being used by the veterinarian. The manufacture, placing on the market and use of cell-based veterinary pharmaceuticals without manufacturing approval and marketing authorisation is permitted only in certain exceptional cases determined by EU and individual Member State law. Violations of this requirement may have consequences for the respective veterinarian under criminal law and under the code of professional conduct in the respective Member State. The regular use of cell-based veterinary pharmaceuticals within the scope of a therapeutic emergency as well as the import of such veterinary pharmaceuticals from non-European countries for use in the EU are currently out of the question in the EU because of a lack of legal bases. Here, we review the general legal requirement of manufacturing, placing on the market and use of cell-based veterinary pharmaceuticals within the EU and point out different implementations of EU law within the different Member States.

Background. Mesenchymal stromal cells (MSCs) are increasingly used for clinical applications in equine patients. For MSC isolation and expansion, a laboratory step is mandatory, after which the cells are sent back to the attending veterinarian. Preserving the biological properties of MSCs during this transport is paramount. The goal of the study was to compare transport-related parameters (transport container, media, temperature, time, cell concentration) that potentially influence characteristics of culture expanded equine MSCs. Methods. The study was arranged in three parts comparing (I) five different transport containers (cryotube, two types of plastic syringes, glass syringe, CellSeal), (II) seven different transport media, four temperatures (4 °C vs. room temperature; −20 °C vs. −80 °C), four time frames (24 h vs. 48 h; 48 h vs. 72 h), and (III) three MSC concentrations (5 × 106, 10 × 106, 20 × 106 MSC/ml). Cell viability (Trypan Blue exclusion; percent and total number viable cell), proliferation and trilineage differentiation capacity were assessed for each test condition. Further, the recovered volume of the suspension was determined in part I. Each condition was evaluated using samples of six horses (n = 6) and differentiation protocols were performed in duplicates. Results. In part I of the study, no significant differences in any of the parameters were found when comparing transport containers at room temperature. The glass syringe was selected for all subsequent evaluations (highest recoverable volume of cell suspension and cell viability). In part II, media, temperatures, or time frames had also no significant influence on cell viability, likely due to the large number of comparisons and small sample size. Highest cell viability was observed using autologous bone marrow supernatant as transport medium, and “transport” at 4 °C for 24 h (70.6% vs. control group 75.3%); this was not significant. Contrary, viability was unacceptably low (

Treatment of tendon disease with multipotent mesenchymal stromal cells (MSC) is a promising option to improve tissue regeneration. To elucidate the mechanisms by which MSC support regeneration, longitudinal tracking of MSC labelled with superparamagnetic iron oxide (SPIO) by magnetic resonance imaging (MRI) could provide important insight. Nine equine patients suffering from tendon disease were treated with SPIO-labelled or nonlabelled allogeneic umbilical cord-derived MSC by local injection. Labelling of MSC was confirmed by microscopy and MRI. All animals were subjected to clinical, ultrasonographical, and low-field MRI examinations before and directly after MSC application as well as 2, 4, and 8 weeks after MSC application. Hypointense artefacts with characteristically low signal intensity were identified at the site of injection of SPIO-MSC in T1- and T2∗-weighted gradient echo MRI sequences. They were visible in all 7 cases treated with SPIO-MSC directly after injection, but not in the control cases treated with nonlabelled MSC. Furthermore, hypointense artefacts remained traceable within the damaged tendon tissue during the whole follow-up period in 5 out of 7 cases. Tendon healing could be monitored at the same time. Clinical and ultrasonographical findings as well as T2-weighted MRI series indicated a gradual improvement of tendon function and structure.

Tendon and ligament pathologies are still a therapeutic challenge, due to the difficulty in restoring the complex extracellular matrix architecture and biomechanical strength. While progress is being made in cell-based therapies and tissue engineering approaches, comprehensive understanding of the fate of progenitor cells in tendon healing is still lacking. The aim of this study was to investigate the effect of decellularized tendon matrix and moderate cyclic stretching as natural stimuli which could potentially direct tenogenic fate. Equine adipose-derived mesenchymal stromal cells (MSC) were seeded on decellularized tendon matrix scaffolds. Mechanical stimulation was applied in a custom-made cyclic strain bioreactor. Assessment was performed 4 h, 8 h, and 24 h following mechanical stimulation. Scaffold culture induced cell alignment and changes in expression of tendon-related genes, although cell viability was decreased compared to monolayer culture. Short mechanical stimulation periods enhanced most of the scaffold-induced effects. Collagen 1A2 expression levels were decreased, while collagen 3A1 and decorin levels were increased. Tenascin-C and scleraxis expression showed an initial decrease but had increased 24 h after stimulation. The results obtained suggest that decellularized tendon matrix, supported by cyclic stretching, can induce tenogenic differentiation and the synthesis of tendon components important for matrix remodeling.

Multipotent mesenchymal stromal cells (MSCs) have gained tremendous attention as potential therapeutic agents for the treatment of orthopedic diseases. Promising results have been obtained after application of MSCs for treatment of tendon and joint disease in the equine model, making it appear favorable to use these results as a basis for the translational process of the therapy. However, while the horse is considered a highly suitable model for orthopedic diseases, knowledge is lacking regarding the level of analogy of equine MSCs and their human counterparts. Therefore, the aim of this study was to assess the properties of human and equine adipose-and tendon-derived MSCs in a direct comparison. Basic properties of human and equine MSCs from both tissues were similar. The cells expressed CD29, CD44, CD90, and CD105 and lacked expression of CD73, CD14, CD34, CD45, CD79a, and MCHII/HLA-DR. No significant differences were found between proliferation potential of human and equine MSCs in early passages, but recovery of nucleated cells after tissue digestion as well as proliferation in later passages was higher in equine samples (p < 0.01). All samples showed a good migration capacity and multilineage differentiation potential. However, while osteogenic differentiation was achieved in all equine samples, it was only evident in five out of nine human tendon-derived samples. Human MSCs further showed a higher expression of collagen IIIA1 and tenascin-C, but lower expression of decorin and scleraxis (p < 0.01). Although revealing some potentially relevant differences, the study demonstrates a high level of analogy between human and equine MSCs, providing a basis for translational research in the equine model according to the guidelines issued by the authorities.

In this study, superparamagnetic iron oxide (SPIO) particle-labeled mesenchymal stromal cells (MSCs) were injected intra-articularly into osteoarthritic knee joints. Their fate and distribution were evaluated using magnetic resonance imaging (MRI) and macroscopic and histologic postmortem examination. Osteoarthritis was induced in 12 sheep by bilateral meniscectomy. After 6 weeks, one knee joint received 10 × 10 6 SPIO-labeled MSCs (Molday Ion Rhodamine B). Contralateral knees received a control injection of a) PBS, b) SPIO in PBS, c) 10 × 10 6 nonvital SPIO-labeled MSCs in PBS, or d) no injection. MR images were acquired immediately after injection and 1, 4, 8, and 12 weeks thereafter using a 0.5-T unit and a T2* sequence. Signal intensity of synovial fluid and synovial lining was assessed semiquantitatively using a scoring system. Viable SPIO-labeled MSCs produced a strong hypointense signal in the synovial fluid immediately after injection, but normal signal intensity of the synovial fluid was observed 1 week later. Synovial lining maintained its hypointensity throughout the study period. Nonvital SPIO-labeled MSCs induced hypointense signals of the synovial fluid; synovial lining appeared weak and inconsistently hypointense in the following weeks. Pure SPIO produced a strong hyperintense signal in the synovial fluid at the time of injection only. Histologically, in all knee joints receiving viable SPIO-labeled MSCs, SPIO particles were detected (Prussian blue) within the synovial lining, dorsal fat pad, and neomeniscus tissue, but not in osteochondral samples. Few SPIO particles were detected in joints injected with nonvital SPIO-labeled MSCs. Immunohistologically, no increased cell death (TUNEL) was observed in the area of detected SPIO particles, but we did observe potential chondrogenic cell differentiation (Safranin O or S100β). We conclude that viable SPIO-labeled MSCs remain detectable within the joint for 12 weeks and attach themselves to some but not all diseased joint structures.

In this study, the therapeutic effect of intra-articularly injected autologous mesenchymal stromal cells (MSCs) was evaluated in an ovine osteoarthritis (OA) model using consecutive magnetic resonance imaging (MRI), radiography, and macroscopic and histologic postmortem examination. In 12 sheep, OA was induced by bilateral, lateral meniscectomy. After 6 weeks, 20 × 10 6 bone marrow-derived MSCs (50% MSCs were superparamagnetic iron oxide particle labeled) were injected intra-articularly into one knee joint. The contralateral knee served as negative control. MR images were acquired before OA induction, immediately before and after MSC injection, and 1, 4, 8, and 12 weeks thereafter using a 0.5T unit and a T2* gradient echo sequence. Radiographs were obtained before OA induction, at MSC injection, and 12 weeks thereafter. The MRI scoring system included articular cartilage, bone, joint capsule, and synovial fluid evaluation. The radiographic scoring system included the joint space and bone. Postmortem evaluation entailed macroscopic and histologic assessment. Longitudinal MRI revealed a significant deceleration of OA progression in MSC-treated joints. However, at the conclusion of the study, there was no significant difference in the degree of OA detected by MRI, radiography, and postmortem evaluation between the treatment and control group. The degree of OA on MRI varied among the 12 animals at the time of injection, but there was no difference between the left and right limb. In conclusion, intra-articular MSCs decreased OA progression. However, no significant treatment effects were seen at the conclusion of the study at 12 weeks. This somewhat contradicts previously published results. Nevertheless, the choice of OA model, outcome measures, or lack of additional medication might explain the differences. Our results indicate that OA might benefit from intra-articular MSC injection, but further studies are needed to refine patient selection and injection parameters for a more substantially improved outcome.

Transplantation of multipotent mesenchymal progenitor cells is a valuable option for treating tendon disease. Tenogenic differentiation leading to cell replacement and subsequent matrix modulation may contribute to the regenerative effects of these cells, but it is unclear whether this occurs in the inflammatory environment of acute tendon disease. Equine adipose-derived stromal cells (ASC) were cultured as monolayers or on decellularized tendon scaffolds in static or dynamic conditions, the latter represented by cyclic stretching. The impact of different inflammatory conditions, as represented by supplementation with interleukin-1β and/or tumor necrosis factor-α or by co-culture with allogeneic peripheral blood leukocytes, on ASC functional properties was investigated. High cytokine concentrations increased ASC proliferation and osteogenic differentiation, but decreased chondrogenic differentiation and ASC viability in scaffold culture, as well as tendon scaffold repopulation, and strongly influenced musculoskeletal gene expression. Effects regarding the latter differed between the monolayer and scaffold cultures. Leukocytes rather decreased ASC proliferation, but had similar effects on viability and musculoskeletal gene expression. This included decreased expression of the tenogenic transcription factor scleraxis by an inflammatory environment throughout culture conditions. The data demonstrate that ASC tenogenic properties are compromised in an inflammatory environment, with relevance to their possible mechanisms of action in acute tendon disease.

The purpose of this study was to evaluate the use of three different superparamagnetic iron oxide (SPIO) particles for labeling of ovine and equine bone marrow (BM)-derived multipotent stromal cells (MSCs) in vitro. MSCs were obtained from five adult sheep and horses, respectively. After three passages (p3), cells were labeled with either 1) Molday ION Rhodamine B, 2) Endorem, 3) Resovist, or 4) remained unlabeled as control. Labeling efficiency, marker retention, and long-term detectability in MRI until p7 were evaluated. Further, proliferation capacity and trilineage differentiation as indicators for potential impact on stromal cell characteristics were assessed. MSCs of both species were successfully labeled with all three SPIO products. A high, exclusively intracellular, iron uptake was achieved by Molday ION Rhodamine B only. Labeling with Resovist led to prominent extracellular iron presence; labeling with Endorem was less efficient. During MRI, all labeled cells showed strong hypointense signals, contrary to unlabeled controls. Resovist induced the largest areas of hypointense signals, followed by Molday ION Rhodamine B and Endorem. MRI signal detectability decreased from p4 to p7. Proliferation, adipogenic, and osteogenic differentiation potential were not reduced by cell labeling compared to unlabeled cells. Chondrogenic differentiation capacity decreased with increasing amount of iron associated with the cells. Among the three products, Resovist and Molday were identified as promising labeling agents. While Resovist achieved superior results in most of the assessed parameters, Molday ION Rhodamine B ensured intracellular iron uptake without extracellular SPIO complexes and consistent hypointense signals on MRI.