Systematic review and meta-analysis of positive long-term effects after intra-articular administration of orthobiologic therapeutics in horses with naturally occurring osteoarthritis.

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.

Synovial Fluid Analysis and Microscopic Assessment of Macrophage Quantities and Morphology in Equine Septic Arthritis.

OBJECTIVE: Research and provision of data on macrophages by cytological synovial fluid analysis and light microscopy in horses with septic arthritis MATERIAL AND METHODS: Records of 167 synovial fluid samples were evaluated and subdivided into different groups: (1) non-septic, (2) haematogenous septic arthritis in foals and (3) traumatic/iatrogenic septic arthritis. The effect of joint lavage on synovial fluid cytology and on the occurrence of macrophage phenotypes was investigated. RESULTS: Regardless of aetiology and age of the horse, macrophage concentrations in synovial sepsis are decreased to a median of 5-6 % (unaffected joints: 23.5 %) and further diminished by joint lavage. Microscopic assessment led to the identification of 4 phenotypes. Morphological characteristics of type 1 showed similarities to monocytes and predominated in unaffected and in septic joints after lavage. CONCLUSION AND CLINICAL RELEVANCE: Macrophages are highly versatile by altering their phenotype. A morphological assessment by light microscopy is easily applicable. Type 1 presumably contributes to joint homeostasis.

MSC in Tendon and Joint Disease: The Context-Sensitive Link Between Targets and Therapeutic Mechanisms.

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.

[Cell-based therapeutic strategies for osteoarthritis in equine patients - Basic knowledge for clinical practitioners]. / Zellbasierte Therapeutika zur Behandlung der Osteoarthritis beim Pferd.

Cell-based therapies for the treatment of osteoarthritis in equine patients experienced a real boom within the last few years. In every day medical practice, attending veterinary surgeons extract patient's blood or other autologous tissue samples and process the material for the purpose of administering the resulting product to the same patient under their own responsibility. Although being consistently classified as treatment option within the framework of regenerative medicine, the manufacturing processes, ingredients, and mechanisms of action remain highly diverse among cell-based therapies. Thus, sound knowledge about the latter ones forms the basis for therapeutic decision-making and best possible treatment regimes.

Evaluation of Villus Synovium From Unaffected Metacarpophalangeal Joints of Adult and Juvenile Horses.

Horses are a widely accepted model for osteoarthritis (OA) research. Synovial tissue sampling is commonly used in studies to evaluate and grade the progress of OA or to assess treatment effects. Synovial explants play an important role in ex-vivo studies, increasingly replacing the use of living animals. To understand histomorphological changes in the process of joint-related diseases such as OA, detailed information about histomorphometric parameters of unaffected synovial villi is necessary. The objective of the present study was to evaluate the mean width of the intimal synovial lining and its cellularity as well as the vascularization of the subintimal layer in juvenile and adult horses not affected by a joint-related disease. One hundred synovial samples from both metacarpophalangeal joints from 25 horses (one day to 24 years old) were collected to evaluate the following parameters on digitalized hematoxylin-eosin stained samples: Width of intimal synovial lining measured by the distance from the inner joint surface to the subintimal layer; density of the cells making up the intimal synovial lining by counting cell nuclei; vascularization of the subintimal layer measured by the number and size of vessels in relation to the subintimal area. The median width of the intimal lining did not differ among juvenile (22.34 µm) and adult (23.34 µm) horses. The cellularity of the intimal lining was significantly lower in juvenile (one cell/143.8 µm2) than in adult (one cell /188.7µm2), (P < .001) horses as well as the density of blood vessels per mm2 within the subintimal layer (juveniles 1/mm2 vs. adults 0.05/mm2), (P < .001). This study provides morphometric data regarding synovial intimal width, intimal cellularity, and vascularization of equine synovial villi of unaffected horses. For future studies, age-related characteristics should be taken into consideration when synovial tissue samples are used for in-vivo and in-vitro studies.

An anatomical study of the dorsal and ventral nasal conchal bullae and middle nasal conchae in normal Shetland ponies: Computed tomographic anatomical and morphometric findings.

Equine paranasal sinuses are susceptible to inflammation. Insufficient drainage through the nasal passages and meatus may lead to the accumulation of inspissated purulent discharge. Particularly in ponies, these anatomical structures are suspected to be relatively small. To date, there are no reports considering the morphology of nasal conchal bullae in small horse breeds such as Shetland ponies. The aim of the present study was to evaluate the size of the conchal bullae and the medial nasal conchae of Shetland ponies and their relation to the skull dimension using computed tomography. Reconstructed images of healthy adult heads of Shetland ponies were used. Linear skull measurements as well as two cranial indices of the head dimensions were taken. Length, width and height of the dorsal and ventral conchal bullae and the medial nasal conchae were measured in relation to the skull and compared with the data of skulls of large breed horses. The anatomical proportions of pony heads were characterized by a smaller cranial index and a greater nasal index than those of large breed horses. Shetland ponies showed a longer cranial length compared with the nasal length. Heads are consistently smaller, and the relationship of the bullae to the head length was also smaller than those measured in large breed horses. A negative correlation between the head and bullae size was found. In conclusion, this study suggests that Shetland ponies have distinguishing proportions of the head. These findings are relevant for clinical examination and surgical treatment of equine sinus disease in those breeds.

Retrospective Evaluation of Hemithyroidectomy in 14 Horses.

OBJECTIVE: To describe the presentation, presurgical diagnostic findings, treatment, and outcome of horses with histologically confirmed, unilateral thyroid neoplasia. The complications, particularly laryngeal hemiplegia, were investigated. STUDY DESIGN: Retrospective case series. ANIMALS: Client-owned horses (n=14). METHODS: Medical records of horses presenting with a unilateral thyroid mass due to neoplasia from 2003-2015 were reviewed. Horses must have undergone preoperative clinical evaluations that included ultrasound examination of the mass and upper airway endoscopy. Short-term follow-up at 2 weeks after surgery and owner questionnaire for long-term follow-up at >6 months were completed. RESULTS: Fourteen horses aged 6-21 years were included. The majority of tumors were adenocarcinomas (11/14), mainly diagnosed in mares (9/14). Intraoperative complications included hemorrhage (1/14) and postoperative complications included seroma formation (4/14). No horse developed postoperative laryngeal hemiplegia. No horses developed clinical signs of metastases or a thyroid disorder long term (mean follow-up 4.9 years). All owners reported a successful long-term outcome. CONCLUSION: The clinical findings of thyroid neoplasia in horses are not associated with the diagnosis of malignancy. Complete surgical resection of the abnormal lobe prevents local recurrence of neoplastic thyroid tissue. The modified hemithyroidectomy technique preserves the function of the recurrent laryngeal nerve.

Comparison of humoral insulin-like growth factor-1, platelet-derived growth factor-BB, transforming growth factor-ß1, and interleukin-1 receptor antagonist concentrations among equine autologous blood-derived preparations.

OBJECTIVE To compare humoral insulin-like growth factor (IGF)-1, platelet-derived growth factor (PDGF)-BB, transforming growth factor (TGF)-ß1, and interleukin-1 receptor antagonist (IL-1Ra) concentrations in plasma and 3 types of equine autologous blood-derived preparations (ABPs). SAMPLE Blood and ABP samples from 12 horses. PROCEDURES Blood samples from each horse were processed by use of commercial systems to obtain plasma, platelet concentrate, conditioned serum, and aqueous platelet lysate. Half of the platelet concentrate samples were additionally treated with a detergent to release intracellular mediators. Humoral IGF-1, PDGF-BB, TGF-ß1, and IL-1Ra concentrations were measured with ELISAs and compared statistically. RESULTS Median IGF-1 concentration was highest in conditioned serum and detergent-treated platelet concentrate, followed by platelet concentrate and plasma; IGF-1 was not detected in platelet lysate. Mean PDGF-BB concentration was highest in platelet lysate, followed by detergent-treated platelet concentrate and conditioned serum; PDGF-BB was not detected in plasma and platelet concentrate. Median TGF-ß1 concentration was highest in detergent-treated platelet concentrate, followed by conditioned serum, platelet lysate, and platelet concentrate; TGF-ß1 was not detected in most plasma samples. Median IL-1Ra concentration was highest in platelet lysate, followed by conditioned serum; IL-1Ra was not detected in almost all plasma, detergent-treated platelet concentrate, and platelet concentrate samples. CONCLUSIONS AND CLINICAL RELEVANCE Each ABP had its own cytokine profile, which was determined by the specific processing method. Coagulation and cellular lysis strongly increased humoral concentrations of cell-derived cytokines. No ABP had the highest concentrations for all cytokines. Further studies are needed to assess clinical relevance of these findings.