Dataset Information

Concentration Dependent Influence of Lipopolysaccharides on Separation of Hoof Explants and Supernatant Lactic Acid Concentration in an Ex Vivo/In Vitro Laminitis Model.

ABSTRACT: Laminitis is one of the most common diseases in horses. It is not only painful for the animal, but also has a significant financial impact on the equine industry. This multifactorial disease affects the connective tissue of the hoof. However, the pathogenesis of laminitis is still not fully understood. Endotoxins, also known as lipopolysaccharides (LPS), and bacterial exotoxins seem to play an important role during the development of laminitis. The aim of our study was to investigate the effect of increasing LPS concentrations (0, 2.5, 5, 10, and 100 μg/mL) on cell viability of isolated epidermal and dermal hoof cells as well as on the tissue integrity of hoof explants. Furthermore, glucose, acetic acid, lactic acid, and propionic acid concentrations in explant supernatants were measured to evaluate the energy metabolism in the hoof tissue. LPS did not exhibit cytotoxic effects on epidermal or dermal cells. Force required to separate LPS treated hoof explants decreased in a concentration dependent manner. Specifically, explants incubated with 10 and 100 μg/mL needed significantly less force to separate compared to control explants. Lactic acid concentrations were significantly decreased in explants incubated with 5, 10, or 100 μg/mL LPS, while glucose, acetic acid and propionic acid concentrations were unaffected by LPS treatment. Our study indicates that LPS has no cytotoxic effect on epidermal and dermal cells isolated from hoof tissue, but impairs integrity of hoof explants. In addition, LPS led to an alteration of the lactic acid production in the lamellar tissue. Since our data highlight that LPS can affect the integrity of the equine hoof tissue in vitro, endotoxins should be further explored for their contribution to facilitate the development of laminitis.

SUBMITTER: Reisinger N

PROVIDER: S-EPMC4657978 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:BackgroundSprifermin (recombinant human fibroblast growth factor 18) is in clinical development as a potential disease-modifying osteoarthritis drug (DMOAD). In vitro studies have shown that cartilage regenerative properties of sprifermin involve chondrocyte proliferation and extracellular matrix (ECM) production. To gain further insight into the process of sprifermin in the cartilage tissue, this study aimed at investigating the ECM turnover of articular cartilage explants in a longitudinal manner.MethodsBovine full-depth articular cartilage explants were stimulated with sprifermin or placebo at weekly intervals, similar to the dosing regimen used in clinical trials. Pre-culturing with oncostatin M and tumour necrosis factor-α, was also used to induce an inflammatory state before treatment. Metabolic activity was measured using AlamarBlue, and chondrocyte proliferation was visualized by immuno-histochemical detection of proliferating cell nuclear antigen. ECM turnover was quantified by biomarker ELISAs; ProC2 reflecting type II collagen formation, CS846 reflecting aggrecan formation, active MMP9, C2M and AGNx2 reflecting matrix metalloproteinase activity, and AGNx1 reflecting aggrecanase activity.ResultsSprifermin was able to reach the chondrocytes through the extracellular matrix, as it increased cell proliferation and metabolic activity of explants. ProC2 and CS846 was dose-dependently increased (P < 0.05) by sprifermin compared to placebo, while C2M and AGNx2 were unaffected, active MMP9 was slightly decreased, and AGNx1 was slightly increased. Over the course of treatment, the temporal order of ECM turnover responses was AGNx1, then ProC2, followed by CS846 and MMP9. Pro-inflammatory activation of the explants diminished the ECM turnover responses otherwise observed under non-inflammatory conditions.ConclusionsThe data suggest that sprifermin has chondrogenic effects on articular cartilage ex vivo, exerted through a sequential process of ECM turnover; aggrecan degradation seems to occur first, while type II collagen and aggrecan production increased at a later time point. In addition, it was observed that these chondrogenic effects are dependent on the inflammatory status of the cartilage prior to treatment.

Project description:Background: The objectives of this study were to, (1) investigate the impact of the Dutch five-step hoof trimming (HT) technique on time to lameness and hoof lesion prevalence in grazing (GR) and non-grazing (NGR) dairy cows, and (2) determine the association between potential benefits of HT and animal-based welfare measures during lactation. A total of 520 non-lame cows without hoof lesions from 5 dairy farms (GR = 2, NGR = 3) were enrolled at early (within 30 days in milk; DIM) and late lactation (above 200 DIM), and randomly allocated to either trimmed (HGR or HNGR) or control groups (CON-GR and CON-NGR). Locomotion scores, body condition, hock condition, leg hygiene, and hoof health were assessed at monthly intervals until the following 270 days in milk. The data were analyzed using Kaplan-Meier survival analysis, multivariable Cox, and logistic regression models. The overall incidence rate of lameness was 36.2 cases/100 cows/month, with corresponding rates of 27.4, 31.9, 48.4, and 45.8 cases/100 cows/month in HGR, HNGR, CON-GR, and CON-NGR, respectively. Time to first lameness event was significantly higher in HGR (mean ± S.E; 8.12 ± 0.15) compared to CON-GR (7.36 ± 0.26), and in HNGR (8.05 ± 0.16) compared to CON-NGR (7.39 ± 0.23). The prevalence of hoof lesions in the enrolled cows was 36.9%, with a higher occurrence in CON-GR (48.8%) than HGR (23.2%), and in CON-NGR (52.6%) compared to HNGR (32.2%). The majority of hoof lesions were non-infectious in grazing (HGR vs. CON-GR; 21.3 vs. 33.3%) and non-grazing herds (HNGR vs. CON-NGR; 25.0 vs. 40.4%). The risk of lameness was higher in underconditioned cows (Hazard ratio; HR = 3.1, 95% CI 1.2-7.4), presence of hoof lesion (HR = 33.1, 95% CI 17.6-62.5), and there was variation between farms. Aside HT, lower parity (OR = 0.4, 95% CI 0.2-0.8), normal hock condition (OR = 0.06; 95% 0.01-0.29), and absence of overgrown hoof (OR = 0.4; 95% 0.2-0.7) were protective against non-infectious hoof lesions. Functional HT is beneficial as a lameness preventive strategy during lactation; however, ensuring older cows are in good body condition and free from hock injuries are equally important.

Dataset Information

Concentration Dependent Influence of Lipopolysaccharides on Separation of Hoof Explants and Supernatant Lactic Acid Concentration in an Ex Vivo/In Vitro Laminitis Model.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets