Hesperidin Promotes Osteogenesis and Modulates Collagen Matrix Organization and Mineralization In Vitro and In Vivo.

This study evaluated the direct effect of a phytochemical, hesperidin, on pre-osteoblast cell function as well as osteogenesis and collagen matrix quality, as there is little known about hesperidin's influence in mineralized tissue formation and regeneration. Hesperidin was added to a culture of MC3T3-E1 cells at various concentrations. Cell proliferation, viability, osteogenic gene expression and deposited collagen matrix analyses were performed. Treatment with hesperidin showed significant upregulation of osteogenic markers, particularly with lower doses. Mature and compact collagen fibrils in hesperidin-treated cultures were observed by picrosirius red staining (PSR), although a thinner matrix layer was present for the higher dose of hesperidin compared to osteogenic media alone. Fourier-transform infrared spectroscopy indicated a better mineral-to-matrix ratio and matrix distribution in cultures exposed to hesperidin and confirmed less collagen deposited with the 100-µM dose of hesperidin. In vivo, hesperidin combined with a suboptimal dose of bone morphogenetic protein 2 (BMP2) (dose unable to promote healing of a rat mandible critical-sized bone defect) in a collagenous scaffold promoted a well-controlled (not ectopic) pattern of bone formation as compared to a large dose of BMP2 (previously defined as optimal in healing the critical-sized defect, although of ectopic nature). PSR staining of newly formed bone demonstrated that hesperidin can promote maturation of bone organic matrix. Our findings show, for the first time, that hesperidin has a modulatory role in mineralized tissue formation via not only osteoblast cell differentiation but also matrix organization and matrix-to-mineral ratio and could be a potential adjunct in regenerative bone therapies.

Retrospective evaluation of outpatient canine parvovirus treatment in a shelter-based low-cost urban clinic.

OBJECTIVE: To evaluate survival and associated risk factors when utilizing an outpatient treatment protocol for treatment of canine parvovirus (CPV) performed in a shelter-based low-cost urban clinic. DESIGN: Retrospective study. SETTING: Pennsylvania Society for the Prevention of Cruelty to Animals. ANIMALS: Ninety-five CPV positive dogs presented between June 1 and July 31, 2016. Owners elected for outpatient care when inpatient care was not financially feasible and the dog was considered medically stable for outpatient care. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of the 95 CPV positive dogs, 79 (83%) survived treatment. Logistic regression indicated that an increasing number of days with clinical signs prior to treatment and an increase in percent body weight during treatment were significantly associated with survival (odds ratio [OR], 3.15, P = 0.020; and OR, 1.29, P = 0.027, respectively). Hypothermia upon presentation (T < 37℃) was negatively associated with survival (OR, 0.002; P = 0.002). CONCLUSIONS AND CLINICAL RELEVANCE: The survival rate of this clinic suggests that an outpatient program may be a potential alternative treatment to inpatient care. Longer duration of clinical signs prior to treatment and an increase in percent body weight during treatment appear to be associated with increased survival outcomes, while hypothermia on presentation appears to be associated with decreased survival outcomes.