Therapy of Canine Deep Pyoderma with Cephalexins and Immunomodulators

Pyodermas are among bacterial skin diseases often resisting antibiotic therapy. We therefore examined how the dogs with deep pyoderma (n = 29) respond to therapeutic effect of antibiotic cephalexins (Ceporex®, 30 mg kg-1 p.o., once a day for 9 - 11 weeks) combined with immunomodulators (Baypamune®, once a week i.m. pro toto). The dogs with the first occurrence of pyoderma (n = 11) were treated by antibiotics alone, whereas the dogs with recurrent pyoderma (n = 18) were treated by either antibiotics alone (n = 8) or antibiotics combined with Baypamune® (n = 10). Of 11 dogs with the first occurrence of disease, 8 (73%) were successfully cured. However, only 5 of them (45%) stayed recovered after a period of two months that elapsed from the completion of therapy. Of the 8 dogs with recurrent pyoderma treated by antibiotics only, 6 (75%) recovered quickly but only 3 of them (38%) stayed healthy after 2 months elapsing from the therapy termination. Of the 10 dogs treated by antibiotics combined with immunomodulators, 8 (80%) regained health within a therapeutic period and 7 of them (70%) remained completely cured after 2 months from completion of therapy. The durations of treatment in dogs with the first occurrence of pyoderma and those with recurrent pyoderma were 8.4 and 10.5 weeks, respectively, the difference begin significant. Hair lenght and percentage of the skin area affected had no effect on the therapy duration. The disappearance of pruritus preceded the successful treatment. The results suggest that the joint treatment of deep pyodermas in dogs by antibiotics and immunomodulators may be superior to the purely antibiotic therapy, because of a stronger suppressive effect on the disease relapse

Healing and angiogenic properties of collagen/chitosan scaffolds enriched with hyperstable FGF2-STAB® protein: in vitro, ex ovo and in vivo comprehensive evaluation

Wound healing is a process regulated by a complex interaction of multiple growth factors including fibroblast growth factor 2 (FGF2). Although FGF2 appears in several tissue engineered studies, its applications are limited due to its low stability both in vitro and in vivo. Here, this shortcoming is overcome by a unique nine-point mutant of the low molecular weight isoform FGF2 retaining full biological activity even after twenty days at 37 °C. Crosslinked freeze-dried 3D porous collagen/chitosan scaffolds enriched with this hyper stable recombinant human protein named FGF2-STAB® were tested for in vitro biocompatibility and cytotoxicity using murine 3T3-A31 fibroblasts, for angiogenic potential using an ex ovo chick chorioallantoic membrane assay and for wound healing in vivo with 3-month old white New Zealand rabbits. Metabolic activity assays indicated the positive effect of FGF2-STAB® already at very low concentrations (0.01 µg/mL). The angiogenic properties examined ex ovo showed enhanced vascularization of the tested scaffolds. Histological evaluation and gene expression analysis by RT-qPCR proved newly formed granulation tissue at the place of a previous skin defect without significant inflammation infiltration in vivo. This work highlights the safety and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB® protein. Moreover, these sponges could be used as scaffolds for growing cells for dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration

In Vitro Influence of Mycophenolic Acid on Selected Parameters of Stimulated Peripheral Canine Lymphocytes.

Mycophenolic acid (MPA) is an active metabolite of mycophenolate mofetil, a new immunosuppressive drug effective in the treatment of canine autoimmune diseases. The impact of MPA on immunity is ambiguous and its influence on the canine immune system is unknown. The aim of the study was to determine markers of changes in stimulated peripheral canine lymphocytes after treatment with MPA in vitro. Twenty nine healthy dogs were studied. Phenotypic and functional analysis of lymphocytes was performed on peripheral blood mononuclear cells cultured with mitogens and different MPA concentrations- 1 μM (10(-3) mol/m(3)), 10 μM or 100 μM. Apoptotic cells were detected by Annexin V and 7-aminoactinomycin D (7-AAD). The expression of antigens (CD3, CD4, CD8, CD21, CD25, forkhead box P3 [FoxP3] and proliferating cell nuclear antigen [PCNA]) was assessed with monoclonal antibodies. The proliferation indices were analyzed in carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled cells. All analyses were performed using flow cytometry. The influence of MPA on apoptosis was dependent on the mechanism of cell activation and MPA concentration. MPA caused a decrease in the expression of lymphocyte surface antigens, CD3, CD8 and CD25. Its impact on the expression of CD4 and CD21 was negligible. Its negative influence on the expression of FoxP3 was dependent on cell stimulation. MPA inhibited lymphocyte proliferation. In conclusion, MPA inhibited the activity of stimulated canine lymphocytes by blocking lymphocyte activation and proliferation. The influence of MPA on the development of immune tolerance-expansion of Treg cells and lymphocyte apoptosis-was ambiguous and was dependent on the mechanism of cellular activation. The concentration that MPA reaches in the blood may lead to inhibition of the functions of the canine immune system. The applied panel of markers can be used for evaluation of the effects of immunosuppressive compounds in the dog