Histo-morphological effects on equine synovium after arthroscopic synovectomy using two different motorized synovial resectors and two different intensities.

The purpose of the study was to determine the histo-morphological effects on villous synovium after synovectomy using two different motorized synovial resectors and two different intensities ex-vivo. Thirty-three (n = 33) equine metacarpophalangeal/metatarsophalangeal joints were used. Synovectomy was performed along the dorsomedial/dorsolateral synovium (n = 66) using two motorized synovial resectors (aggressive full radius resector, AFRR, used at two intensities: single treatment, n = 24 vs. triple treatment, n = 21 vs. aggressive meniscus side cutter, AMSC, n = 21). Arthroscopic images were evaluated blindly for resector type and intensity. Histological images were evaluated descriptive for synovial morphology and the extent of tissue loss using a microscopic scale. Scanning electron microscopy described the synovial morphology. The synovectomized areas were specific for each resector used and distinguishable from arthroscopic images. The AFRR demonstrated a clear demarcation between treated and non-treated areas and removed the stratum synoviale completely including parts of the underlying stratum fibrosum. In contrast, the AMSC showed less clear demarcation, villous scaffolds and no involvement of the stratum fibrosum. Triple intense treated AFFR samples resulted in significantly deeper lesions compared to single treatments (p = 0.037) but could not be distinguished on arthroscopic images. The morphological effects on villous synovium differ according to the resector type used. The extent of synovial tissue loss cannot be estimated from arthroscopic images but histologically. The type and use of motorized synovial resector determines the morphological alterations of the treated synovium. Arthroscopic control is considered unsuitable to control synovectomy depth.

Effects of mesenchymal stromal cells versus serum on tendon healing in a controlled experimental trial in an equine model.

BACKGROUND: Mesenchymal stromal cells (MSC) have shown promising results in the treatment of tendinopathy in equine medicine, making this therapeutic approach seem favorable for translation to human medicine. Having demonstrated that MSC engraft within the tendon lesions after local injection in an equine model, we hypothesized that they would improve tendon healing superior to serum injection alone. METHODS: Quadrilateral tendon lesions were induced in six horses by mechanical tissue disruption combined with collagenase application 3 weeks before treatment. Adipose-derived MSC suspended in serum or serum alone were then injected intralesionally. Clinical examinations, ultrasound and magnetic resonance imaging were performed over 24 weeks. Tendon biopsies for histological assessment were taken from the hindlimbs 3 weeks after treatment. Horses were sacrificed after 24 weeks and forelimb tendons were subjected to macroscopic and histological examination as well as analysis of musculoskeletal marker expression. RESULTS: Tendons injected with MSC showed a transient increase in inflammation and lesion size, as indicated by clinical and imaging parameters between week 3 and 6 (p < 0.05). Thereafter, symptoms decreased in both groups and, except that in MSC-treated tendons, mean lesion signal intensity as seen in T2w magnetic resonance imaging and cellularity as seen in the histology (p < 0.05) were lower, no major differences could be found at week 24. CONCLUSIONS: These data suggest that MSC have influenced the inflammatory reaction in a way not described in tendinopathy studies before. However, at the endpoint of the current study, 24 weeks after treatment, no distinct improvement was observed in MSC-treated tendons compared to the serum-injected controls. Future studies are necessary to elucidate whether and under which conditions MSC are beneficial for tendon healing before translation into human medicine.

Characterisation and intracellular labelling of mesenchymal stromal cells derived from synovial fluid of horses and sheep.

Multipotent mesenchymal stromal cells (MSCs) derived from synovial fluid (SF) are considered to be a promising cell type for therapeutic applications in joint disease. However, despite their potential relevance for clinical and experimental studies, there is insufficient knowledge about SF-derived MSCs isolated from horses and sheep. In this study, cells were recovered from healthy SF and bone marrow (BM) of sheep, and from healthy and osteoarthritic SF of horses. Ovine SF-MSCs were used to assess the efficiency of intracellular labelling with quantum dots (QDs). Colony forming units, generation times, trilineage differentiation potential and expression of CD73, CD90 and CD105 at mRNA level were assessed. QD labelling was efficient, with >98% positive cells directly after labelling at 10 nmol/L and >95% positive cells directly after labelling at 2 nmol/L. The label decreased over 7 days of culture, with more persistence at the higher labelling concentration. No significant differences in proliferation were observed. All MSCs had trilineage differentiation potential, but adipogenesis was more distinct in equine samples and chondrogenesis was most pronounced in ovine SF-MSCs. CD73, CD90 and CD105 were expressed in equine and ovine MSCs.

Systematic arthroscopic investigation of the bovine stifle joint.

The objective of the present study was to establish a protocol for arthroscopic exploration of the bovine stifle joint using craniomedial, caudolateral and caudomedial approaches. An anatomic and arthroscopic study using 26 cadaveric limbs from 13 non-lame adult dairy cows was performed. The craniomedial approach was created between the middle and medial patellar ligaments to investigate the cranial pouches of the stifle joint. The inter-condylar eminence, the proximal aspect of the medial femoral trochlear ridge and the lateral aspect of the lateral femoral condyle were used as starting points for systematic examination of the medial femorotibial, the femoropatellar and the lateral femorotibial joints, respectively. The observed structures were: the suprapatellar pouch, articular surfaces of the patella, femoral trochlear ridges, cruciate ligaments, menisci, and the meniscotibial ligaments. The arthroscopic portal for the caudomedial femorotibial pouch was about 6-8 cm caudal to the medial collateral ligament. The proximal and distal caudolateral femorotibial pouches were explored 3 cm and 1.5 cm caudal to the ipsilateral collateral ligament, respectively. The observed structures were the caudal aspect of femoral condyles, menisci, caudal cruciate ligament, popliteal tendon and the meniscofemoral ligament. Restricted joint size and risk of common peroneal nerve damage were the major limitations for exploration of the caudal femorotibial compartments. The study described the arthroscopic portals and normal intra-articular anatomy of the bovine stifle joint but further investigations are warranted to validate these techniques in clinical cases.

[Comparison of the findings of rectal examination and ultrasonographic findings in horses with colic]. / Vergleich rektaler und sonographischer Untersuchungsbefunde bei der Kolik des Pferdes.

OBJECTIVE: The examination of patients suffering from an acute abdomen routinely comprises both clinical and rectal examinations, and is ever more frequently accompanied by an ultrasonographic abdominal examination. The aim of the study was to compare the findings as defined through rectal examination with the results of the ultrasonographic examination for different forms of colic. MATERIAL AND METHODS: In a retrospective study, the patient records of the Large Animal Clinic of the University of Leipzig from 2012 and 2013 were analysed, and those of horses suffering from colic were included. Diagnoses made through rectal and ultrasonographic examination were grouped and compared with the diagnoses made during colic surgery or pathologic examination, which served as the gold standard. Horses that underwent conservative treatment had a definitive diagnosis assigned only in cases where a pathognostic rectal finding defined the diagnosis. Based on these data, sensitivity, specificity and positive and negative predictive values were calculated for both techniques. RESULTS: Ultrasonography was more sensitive than rectal examination in cases of small intestinal occlusion (97.1% vs. 50.7%), torsion of the large colon in the long axis (63.2% vs. 26.3%) and dislocation of the large colon into the nephrosplenic space (90.9% vs. 72.7%). Rectal examination was more sensitive than ultrasonographic examination in cases of other types of dislocation of the large colon (96.5% vs. 8.8%) and of constipations of the large colon (93.6% vs. 29.8%). CONCLUSION AND CLINICAL RELEVANCE: In cases of severe diseases, including small intestinal occlusions and torsions of the large colon, ultrasonography helps to better identify and more precisely diagnose conditions that in most cases require abdominal surgery than rectal examination. Therefore, under hospital conditions, it is highly advisable to include ultrasonography in the routine examination of the equine acute abdomen. However, this technique does not replace the traditional rectal examination, which is superior in the diagnosis of dislocations and constipations of the large colon as well as diseases of the caecum.

Influence of cryopreservation and mechanical stimulation on equine Autologous Conditioned Plasma (ACP®).

OBJECTIVE: To determine the influence of cryopreservation at two different temperatures on platelet concentration, growth factor (GF) levels and platelet activation parameters in equine ACP®; moreover, to determine if adding mechanical ACP® stimulation to freeze-thaw activation amplifies GF release from platelets. MATERIAL AND METHODS: Firstly, blood from five horses was used to prepare ACP®. Platelet, platelet derived growth factor BB (PDGF-BB) and transforming growth factor ß1 (TGF-ß1) concentrations as well as mean platelet volume (MPV) and mean platelet component (MPC) were determined in fresh and corresponding ACP® samples after 2 months cryopreservation at -20 °C and -80 °C, respectively. Secondly, ACP® was prepared from blood of nine horses. Half of ACP® was activated using one freeze-thaw-cycle at -20 °C, whereas the rest was first vortexed. Their PDGF-BB and TGF-ß1 concentrations were subsequently determined. RESULTS: Platelet concentration significantly decreased after -80 °C cryopreservation. PDGF-BB level augmented significantly after both storage methods, whereas TGF-ß1 concentration was not significantly altered. MPV significantly increased after -20 °C cryopreservation. Both storage regimens induced a significant MPC decrease. No significant differences in GF concentrations between the vortexed and non-vortexed samples were detected. DISCUSSION: Both cryopreservation methods induced platelet activation, but storage at -80 °C apparently harmed the platelets without generating higher GF release than -20 °C. The mechanical stimulation process could not enhance GF release in subsequently frozen-thawed ACP®. CONCLUSION AND CLINICAL RELEVANCE: Storage of ACP® at -20 °C could be useful in equine practice, but, before this procedure can be recommended, further qualitative tests are needed. The mechanical stimulation technique should be adjusted in order to increase platelet activation.

Influence of in vitro maturation of engineered cartilage on the outcome of osteochondral repair in a goat model.

This study was designed to determine if the maturation stage of engineered cartilage implanted in a goat model of cartilage injury influences the repair outcome. Goat engineered cartilage was generated from autologous chondrocytes cultured in hyaluronic acid scaffolds using 2 d, 2 weeks or 6 weeks of pre-culture and implanted above hydroxyapatite/hyaluronic acid sponges into osteochondral defects. Control defects were left untreated or treated with cell-free scaffolds. The quality of repair tissues was assessed 8 weeks or 8 months post implantation by histological staining, modified O'Driscoll scoring and biochemical analyses. Increasing pre-culture time resulted in progressive maturation of the grafts in vitro. After 8 weeks in vivo, the quality of the repair was not improved by any treatment. After 8 months, O'Driscoll histology scores indicated poor cartilage architecture for untreated (29.7 ± 1.6) and cell-free treated groups (24.3 ± 5.8). The histology score was improved when cellular grafts were implanted, with best scores observed for grafts pre-cultured for 2 weeks (16.3 ± 5.8). As compared to shorter pre-culture times, grafts cultured for 6 weeks (histology score: 22.3 ± 6.4) displayed highest type II/I collagen ratios but also inferior architecture of the surface and within the defect, as well as lower integration with native cartilage. Thus, pre-culture of engineered cartilage for 2 weeks achieved a suitable compromise between tissue maturity and structural/integrative properties of the repair tissue. The data demonstrate that the stage of development of engineered cartilage is an important parameter to be considered in designing cartilage repair strategies.

Successful treatment of equine sarcoids by topical aciclovir application.

Based on the anecdotally reported eradication of a sarcoid using aciclovir cream, the curative potential of this ointment was investigated in 22 sarcoid-affected horses referred to the Equine Clinic Tillysburg, Austria, between 2006 and 2009. Sarcoid disease was diagnosed by clinical examination and bovine papillomavirus types 1 and 2 from intact skin and tumour tissue. As nine horses had more than one lesion, a total of 47 sarcoids were treated by daily topical application of aciclovir 5 per cent cream for a period of two to six months; in four horses, surgical tumour ablation was performed before treatment. Disease parameters, including the tumour type, number, location and size, were recorded before and after aciclovir therapy. All 47 (100 per cent) of the sarcoids responded to treatment, with complete tumour regression observed for 32 (68 per cent) lesions and no recurrences reported thus far. Incomplete resolution was observed for 15 (32 per cent) lesions, probably due to their thickness. Aciclovir is proposed to be routinely used for the treatment of mild-type sarcoids and as an adjuvant therapeutic agent in combination with surgery.

Basic science and clinical application of stem cells in veterinary medicine.

Stem cells play an important role in veterinary medicine in different ways. Currently several stem cell therapies for animal patients are being developed and some, like the treatment of equine tendinopathies with mesenchymal stem cells (MSCs), have already successfully entered the market. Moreover, animal models are widely used to study the properties and potential of stem cells for possible future applications in human medicine. Therefore, in the young and emerging field of stem cell research, human and veterinary medicine are intrinsically tied to one another. Many of the pioneering innovations in the field of stem cell research are achieved by cooperating teams of human and veterinary medical scientists.Embryonic stem (ES) cell research, for instance, is mainly performed in animals. Key feature of ES cells is their potential to contribute to any tissue type of the body (Reed and Johnson, J Cell Physiol 215:329-336, 2008). ES cells are capable of self-renewal and thus have the inherent potential for exceptionally prolonged culture (up to 1-2 years). So far, ES cells have been recovered and maintained from non-human primate, mouse (Fortier, Vet Surg 34:415-423, 2005) and horse blastocysts (Guest and Allen, Stem Cells Dev 16:789-796, 2007). In addition, bovine ES cells have been grown in primary culture and there are several reports of ES cells derived from mink, rat, rabbit, chicken and pigs (Fortier, Vet Surg 34:415-423, 2005). However, clinical applications of ES cells are not possible yet, due to their in vivo teratogenic degeneration. The potential to form a teratoma consisting of tissues from all three germ lines even serves as a definitive in vivo test for ES cells.Stem cells obtained from any postnatal organism are defined as adult stem cells. Adult haematopoietic and MSCs, which can easily be recovered from extra embryonic or adult tissues, possess a more limited plasticity than their embryonic counterparts (Reed and Johnson, J Cell Physiol 215:329-336, 2008). It is believed that these stem cells serve as cell source to maintain tissue and organ mass during normal cell turnover in adult individuals. Therefore, the focus of attention in veterinary science is currently drawn to adult stem cells and their potential in regenerative medicine. Also experience gained from the treatment of animal patients provides valuable information for human medicine and serves as precursor to future stem cell use in human medicine.Compared to human medicine, haematopoietic stem cells only play a minor role in veterinary medicine because medical conditions requiring myeloablative chemotherapy followed by haematopoietic stem cell induced recovery of the immune system are relatively rare and usually not being treated for monetary as well as animal welfare reasons.In contrast, regenerative medicine utilising MSCs for the treatment of acute injuries as well as chronic disorders is gradually turning into clinical routine. Therefore, MSCs from either extra embryonic or adult tissues are in the focus of attention in veterinary medicine and research. Hence the purpose of this chapter is to offer an overview on basic science and clinical application of MSCs in veterinary medicine.

[Prevalence of hereditary diseases in three-year-old Swiss Warmblood horses]. / Erhebung der Prävalenz von Erbkrankheiten bei dreijährigen Schweizer Warmblutpferden.

The objective of this study was to investigate clinical signs indicating hereditary diseases like equine sarcoid, osteochondrosis (OC) and the idiopathic laryngeal hemiplegia (ILH), and to demonstrate relationships between environment, feeding habits and conformation ("exterieur" evaluation) of the horses. For this purpose, we analyzed veterinary examinations of 403 stallions at the approvals since 1994 examined 493 three-year-old Swiss Warmblood horses, which were shown at the Swiss-Field-Tests in 2005. With the help of the owners a questionnaire on health, environment and feeding habits of the animals was completed. At the same time, the horses were assessed and graded for their "exterieur" (type, conformation, gaits) by judges of the Swiss Sporthorse breeding association. In 11.5% of horses sarcoids were found, 8.7% showed one and 2.8% several tumors. The prevalence of sarcoids in offspring of sires with known sarcoids was not significantly higher than in descendants from stallions without a known history of sarcoids. We found distended joints as a possible symptom of OC in 11.4% of the horses, 3.9% (n = 19) in both tarsal joints. We did not find a relationship between enlarged joints in the offspring and the presence of OC in the sires. Abnormal respiratory noise at work, as a possible sign for ILH, was heard only in 1.2% (n = 6). It is important to note that while we found a high number of sarcoid affected horses compared to other studies, presence of enlarged joints was not very frequent and very few horses showed abnormal respiratory noise. Additionally, we found no correlation between "exterieur" marks and the horse's general health.

Repair of superficial osteochondral defects with an autologous scaffold-free cartilage construct in a caprine model: implantation method and short-term results.

OBJECTIVE: To compare four different implantation modalities for the repair of superficial osteochondral defects in a caprine model using autologous, scaffold-free, engineered cartilage constructs, and to describe the short-term outcome of successfully implanted constructs. METHODS: Scaffold-free, autologous cartilage constructs were implanted within superficial osteochondral defects created in the stifle joints of nine adult goats. The implants were distributed between four 6-mm-diameter superficial osteochondral defects created in the trochlea femoris and secured in the defect using a covering periosteal flap (PF) alone or in combination with adhesives (platelet-rich plasma (PRP) or fibrin), or using PRP alone. Eight weeks after implantation surgery, the animals were killed. The defect sites were excised and subjected to macroscopic and histopathologic analyses. RESULTS: At 8 weeks, implants that had been held in place exclusively with a PF were well integrated both laterally and basally. The repair tissue manifested an architecture similar to that of hyaline articular cartilage. However, most of the implants that had been glued in place in the absence of a PF were lost during the initial 4-week phase of restricted joint movement. The use of human fibrin glue (FG) led to massive cell infiltration of the subchondral bone. CONCLUSIONS: The implantation of autologous, scaffold-free, engineered cartilage constructs might best be performed beneath a PF without the use of tissue adhesives. Successfully implanted constructs showed hyaline-like characteristics in adult goats within 2 months. Long-term animal studies and pilot clinical trials are now needed to evaluate the efficacy of this treatment strategy.

Effectiveness of a worksite smoking cessation program in the military. Program evaluation.

1. A program evaluation was conducted to evaluate the effectiveness of the smoking cessation program. Substantiating data enhances the value of health promotion programs as key components of the health care delivery system. 2. Persons enrolled in the 8 week smoking cessation program and members of a comparison group were administered an evaluation tool prior to participating in the program, with a second evaluation tool upon completion of the program. Afterward, the smoking status of the members of both groups was evaluated at 3 month intervals for a period of 1 year. 3. Of the participants, 26.7% were not smoking 12 months after the program, compared to 6.9% of the comparison group. Of those participants who resumed smoking, 64% smoked less than half the amount they smoked before taking the class.

Triage of abnormal postmenopausal bleeding: a comparison of endometrial biopsy and transvaginal sonohysterography versus fractional curettage with hysteroscopy.

OBJECTIVE: We sought to compare the combined diagnostic reliability of sonohysterography and endometrial biopsy with fractional curettage with hysteroscopy in the initial evaluation of postmenopausal women with abnormal uterine bleeding. STUDY DESIGN: This year-long, prospective, controlled, clinical investigation was initiated Sept. 1, 1995. All postmenopausal women with abnormal uterine bleeding were offered inclusion, with 104 enrolled. An endometrial biopsy was performed at the time of initial evaluation. Routine transvaginal ultrasonography was then used to measure the uterus, ovaries, and endometrial stripe thickness, followed immediately by sonohysterography to evaluate the symmetry of endometrial wall thickness and delineate any intraluminal masses. Definitive histopathologic sampling was obtained by fractional curettage with hysteroscopy and statistically compared with the diagnoses arrived at by endometrial biopsy and sonohysterography. RESULTS: The combination of endometrial biopsy and transvaginal sonohysterography positively correlated with the surgical findings >95% of the time, with a sensitivity and specificity of 94% and 96%, respectively (confidence interval 91% to 99%). No patients with endometrial hyperplasia or cancer were misdiagnosed. CONCLUSIONS: Sonohysterography combined with endometrial biopsy is a reliable office tool for evaluating postmenopausal women with abnormal uterine bleeding. Medical management of those patients identified as having no endometrial abnormalities can be considered with confidence, while saving the cost and surgical risk of fractional curettage. Patients with intraluminal masses should be referred for surgical management in a timely fashion.

Economic consequences of tobacco use for the Department of Defense, 1995.

This study used Centers for Disease Control and Prevention methodology to analyze the costs of smoking-attributable mortality and morbidity within the Department of Defense (DoD) based on health care costs, characteristics of the beneficiary population, and disease characteristics. Direct health care costs attributed to smoking were estimated at $584 million. Smoking-attributable disease accounted for 16% of the deaths as well as approximately 10% of all hospital bed days and 1.5% of all active duty hospitalizations. Lost productivity among active duty personnel for hospitalization and smoke breaks was valued at $346 million. A significant portion of DoD health care resources is spent caring for smoking-attributable disease, which is preventable. This report reinforces the benefits possible from the new emphasis on wellness promotion within the DoD. Efforts are needed to prevent the initiation of smoking and encourage smoking cessation in order to reduce health care costs and increase the probability for long and healthy lives for DoD beneficiaries.

Physical and sexual abuse in female patients with fibromyalgia.

Sexual and physical abuse during childhood seem to be common. The purpose of this study was to determine the prevalence of sexual and physical abuse reported by female fibromyalgia (FM) patients in a referral-based rheumatology practice, and whether patients with FM report greater frequencies of abuse than do patients with rheumatoid arthritis (RA).Patients from two tertiary care centers, 205 with FM and 84 with RA, were mailed a self-administered questionnaire requesting information about demographics, mental health care use, and history of sexual and/ or physical abuse. There were responses from 105 FM and 44 RA patients.A history of sexual or physical abuse was reported by 54% of the sample. Any type of abuse was significantly more prevalent among patients with FM (62%) compared with those with RA (34%, p = 0.02). FM patients had a significantly increased prevalence of both sexual abuse (51% vs. 32%, p = 0.028) and physical abuse (39 vs. 16%, p = 0.006) compared with RA patients. FM patients were more likely than RA patients to report a history of multiple sexual abusers, increased duration of sexual abuse, and more violent physical abuse. Irritable bowel syndrome was more common in FM (44%) than RA patients (9%, p < 0.001), and 57% of FM patients had seen a mental health professional compared with 30% of RA patients (p = 0.002).