Corneal sequestra in cats: 175 eyes from 172 cases (2000-2016).

OBJECTIVES: To report the clinical outcomes of different surgical treatments used to manage feline corneal sequestra in a large number of cases. MATERIALS AND METHODS: Medical records of 172 cats affected by feline corneal sequestra and surgically managed by different techniques were retrospectively evaluated. Signalment, surgical technique, visual outcomes, postoperative corneal clarity and recurrence were evaluated. RESULTS: One hundred and seventy-two cats (175 eyes) of different breeds, ages and sex were included in the retrospective case series. The Persian was the most represented breed (123/172; 71.5%), followed by Exotic Shorthair breed (21/172; 12.2%), Domestic shorthair breed (21/172; 12.2%) and a smaller group of other different breeds (7/172). Surgical management included a superficial (84/175 eyes) or deep (91/175 eyes) lamellar keratectomy in association with the following covering techniques (tectonic support and/or protective support): nictitans membrane flap (n=84), conjunctival pedicle graft (n=52), BioSISt graft (n=9), corneoconjunctival transposition (n=7), corneoconjunctival transposition+nictitans membrane flap (n=6), conjunctival free island graft (n=6), BioSISt+conjunctival pedicle graft (n=6), BioSISt+nictitans membrane flap (n=4) and bridge conjunctival graft (n=1). At the end of the retrospective case series, visus was present in all patients and no or mild corneal opacity was detected in 86% (151/175 eyes) of the patients. Sequestra recurred in 20 eyes (20/175; 11.4%) within 2 years. CLINICAL SIGNIFICANCE: This paper describes the clinical outcomes of a large number of cases of feline corneal sequestra treated with varying surgical techniques and provides useful insight regarding the prognosis and outcomes of these techniques within feline ophthalmology.

Hyaluronic acid, Manuka honey and Acemannan gel: Wound-specific applications for skin lesions.

Healing of open wounds is of great medical importance. Wound healing is a complex process that aims to restore the function and structure of damaged tissue. This study was conducted to compare secondary intention healing of wounds treated daily with a topical application of commercially available hyaluronic acid (HA), Manuka honey (MH), Acemannan gel (AG), or a placebo. Bilateral wounds were surgically created on the backs of six sheep. At two and six weeks post-wound creation, biopsies were obtained to perform histological, immunohistochemical, and molecular analyses of the wound site. Daily clinical evaluations were performed and weekly photographs were taken of the wounds. HA treatment promoted a physiological progression of the healing process in all wound healing phases, while stimulating an abundant cutaneous adnexa and promoting rapid healing, representing the most compelling treatment. MH-treated wounds were slightly dry. However, the main effect of MH was to promote cell proliferation and neovascularization, with an overall pro-inflammatory effect. Results suggest that MH treatment enhances the healing process. AG treatment dehydrated the wounds and stimulated late granulation tissue and cell proliferation. Moreover, AG-treated wounds produced a mild late pro-inflammatory and neovascularization effect. Our data indicate that AG treatment can have a positive influence on moist wounds with abundant granulation tissue and exudate.

Effect of storage on long-term stability of salivary α-amylase, lysozyme, lactate dehydrogenase, calcium and phosphorus in dogs.

Research or diagnostic conditions may require the storage of salivary samples for long periods before analysis is processed. The aim of this study was to evaluate the stability of canine salivary α-amylase, lysozyme, lactate dehydrogenase (LDH), and total calcium and phosphorus after storage for 1, 3 and 6 months at -20°C. Available saliva samples were 75 immediately after collection (T0) and 46 at 1 month (T1), 31 at 3 months (T3) and 18 at 6 months (T6) of storage, according to the number of aliquots collected from each saliva sample. Compared to T0, LDH declined by 92.3% after 1 month of storage (P<0.001), whereas lysozyme concentration significantly decreased as storage time increased (-29.3%, -43.4% and -59.1% at T1, T3, and T6 respectively, P<0.001). Amylase maintained basically the same concentration for the entire experimental period, whereas total calcium and phosphorus concentration decreased over time (calcium, P<0.001). Assessing the long-term stability of canine salivary analytes stored at -20°C may have important implications in diagnosis and research.

Allogeneic mesenchymal stem cells improve the wound healing process of sheep skin.

BACKGROUND: Skin wound healing includes a system of biological processes, collectively restoring the integrity of the skin after injury. Healing by second intention refers to repair of large and deep wounds where the tissue edges cannot be approximated and substantial scarring is often observed. The objective of this study was to evaluate the effects of mesenchymal stem cells (MSCs) in second intention healing using a surgical wound model in sheep. MSCs are known to contribute to the inflammatory, proliferative, and remodeling phases of the skin regeneration process in rodent models, but data are lacking for large animal models. This study used three different approaches (clinical, histopathological, and molecular analysis) to assess the putative action of allogeneic MSCs at 15 and 42 days after lesion creation. RESULTS: At 15 days post-lesion, the wounds treated with MSCs showed a higher degree of wound closure, a higher percentage of re-epithelialization, proliferation, neovascularization and increased contraction in comparison to a control group. At 42 days, the wounds treated with MSCs had more mature and denser cutaneous adnexa compared to the control group. The MSCs-treated group showed an absence of inflammation and expression of CD3+ and CD20+. Moreover, the mRNA expression of hair-keratine (hKER) was observed in the MSCs-treated group 15 days after wound creation and had increased significantly by 42 days post-wound creation. Collagen1 gene (Col1α1) expression was also greater in the MSCs-treated group compared to the control group at both days 15 and 42. CONCLUSION: Peripheral blood-derived MSCs may improve the quality of wound healing both for superficial injuries and deep lesions. MSCs did not induce an inflammatory response and accelerated the appearance of granulation tissue, neovascularization, structural proteins, and skin adnexa.

Investigations of the corneal epithelium in Veterinary Medicine: State of the art on corneal stem cells found in different mammalian species and their putative application.

The existence of progenitor cells that can readily differentiate into a specific cell type is a common cellular strategy for physiological tissue growth and repair mechanisms. In the mammalian cornea, many aspects regarding the nature and location of these cells are still unclear. In the human limbus (peripheral area of the cornea) progenitor cells have been found and characterized but in non-human mammals, the picture is not so clear. In this review, we examine current knowledge about the morphology of limbus and the localization of corneal epithelial stem cells in all species studied so far, comparing data with humans. We have also explored different research directions in the veterinary field in order to discuss the: i) currently used protocols and ii) best range of treatments for ocular pathologies in which corneal stem cells are involved.

Morphological description of limbal epithelium: searching for stem cells crypts in the dog, cat, pig, cow, sheep and horse.

The cornea provides protection and transparency to the eye, allowing an optimal sharpness view. In some pathological conditions the cornea is able to regenerate thanks to the presence of a stem cells reservoir present at the level of the transition area between cornea and sclera (limbus). Corneal cell therapies in Veterinary Medicine are really limited due to the lacking of knowledge about the anatomy of the limbal area, the putative presence of stem cells and their identification in domestic species. The aim of this study was to provide an overview of the main distinctive structural features of the sclero-corneal junction and conjunctival-corneal junction areas in some species of veterinary importance, using optic microscope observations of histological sections. The resulting data were compared with cornea from humans adapting protocols already used to identify stem cells by means of a specific cellular marker. We tested the expression of ΔNp63α isoform in the cornea basal cells, trying to correlate the distribution profile with areas of highly proliferative turnover. The results obtained from this study represent a first step towards the identification of a corneal stem cells reservoir in different animals.

The Inferior Alveolar Nerve of the Horse: Course and Anatomical Relationship with Mandibular Cheek Teeth.

The precise location of neurovascular structures within the relatively long mandibular canal of the horse is of paramount importance in surgical procedures of the area. The inferior alveolar nerve (IAN) enters the mandibular canal on the medial (lingual) surface of the mandible and innervates all the mandibular teeth. During its course, the nerve moves laterally, crossing the roots of the inferior cheek teeth. However, the exact anatomical relationships occurring between the IAN and the roots of the equine mandibular cheek teeth have not been described in detail. In this study, the mandibles of 40 horses were examined with CT scans and then used for bilateral detailed anatomical dissection, to assess the path of the IAN and its relationship to the roots of the lower cheek teeth. The data obtained show that the equine IAN is located ventral to the apices of the molar teeth (311/411, 310/410, 309/409 according to the Triadan numerical system). At the level of PM(4) (308/408), the IAN is located on the lingual side of the roots and coronally to its apices. At the level of PM(3) (307/407), the IAN is then found on the lingual side of the roots but in proximity to the apices. In 2 of 40 horses (=5%), the IAN moves towards the lingual side between the mesial and the distal root of PM(4). Our observations are valuable for planning a surgical approach to the ventral side of the mandible in the horse and to avoid potential post-operative complications.

Influence of temperature, time and different media on mesenchymal stromal cells shipped for clinical application.

Cell-based therapies, such as the use of mesenchymal stromal cells (MSCs), are becoming popular in veterinary medicine. When MSCs are not cryopreserved, they are shipped in suspension, but no previous studies have analyzed MSC viability during delivery. Here, the impact of several experimental shipping conditions on the number of equine blood-derived (ePB-MSC) and canine adipose-derived (cA-MSC) MSCs were evaluated. Among the different parameters tested, only time and temperature influenced MSC number during the experimental shipping conditions. Cells were monitored over different time intervals for gene expression of typical MSC markers and to evaluate acquired resistance to apoptosis and beta-galactosidase activity. Overall, these results indicate that ePB-MSC and cA-MSC should be delivered in phosphate buffered saline at room temperature and within 9-12 h.

Pharmacokinetics of gallium nitrate after oral administration in adult horses--pilot study.

Gallium (Ga), a metal in group IIIA of the periodic table, has shown a remarkable activity against bone resorption and could therefore possibly prove useful in the treatment of certain diseases in sport horses, for example navicular disease. The aim of this study was to gain more information concerning the kinetics of Ga after oral administration of gallium nitrate (GaN) in adult horses. Six horses received a single dose of 10 mg/kg of GaN mixed with the food ration. Absorption was slow (T(max) = 10 ± 3 h, T(½abs) = 2 ± 0.8 h), and a C(max) of 26 ± 11 µg/L was achieved. Excretion followed a one-phase elimination model, with a long half-life (T(½el) = 52 ± 14 h). By means of a mathematical model, we estimated that the plasmatic levels should reach 93 µg/L (1.33 µm) at steady state, following the repeated daily administration of 10 mg/kg of GaN. A three times lower concentration has been demonstrated as effective in inhibiting the osteolytic activity of osteoclasts in vitro. The results of this study suggest that the administration of oral GaN at a rate of 10 mg/kg per day may be considered for future clinical studies.