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Background context The clinical outcome of patients with ankylosing spinal disorders (ASDs) sustaining a spinal fracture has been described to be worse compared with the general trauma population. Purpose To investigate clinical outcome (neurologic deficits, complications, and mortality) after spinal injury in patients with ankylosing spondylitis (AS) and diffuse idiopathic skeletal hyperostosis (DISH) compared with control patients. Study design Retrospective cohort study. Patient sample All patients older than 50 years and admitted with a traumatic spinal fracture to the Emergency Department of the University Medical Center Utrecht, the Netherlands, a regional level-1 trauma center and tertiary referral spine center. Outcome measures Data on comorbidity (Charlson comorbidity score), mechanism of trauma, fracture characteristics, neurologic deficit, complications, and in-hospital mortality were collected from medical records. Methods With logistic regression analysis, the association between the presence of an ASD and mortality was investigated in relation to other known risk factors for mortality. Results A total of 165 patients met the inclusion criteria; 14 patients were diagnosed with AS (8.5%), 40 patients had DISH (24.2%), and 111 patients were control patients (67.3%). Ankylosing spinal disorder patients were approximately five years older than control patients and predominantly of male gender. The Charlson comorbidity score did not significantly differ among the groups, but Type 2 diabetes mellitus and obesity were more prevalent among DISH patients. In many AS and DISH cases, fractures resulted from low-energy trauma and showed a hyperextension configuration. Patients with AS and DISH were frequently admitted with a neurologic deficit (57.1% and 30.0%, respectively) compared with controls (12.6%; p=.002), which did not improve in the majority of cases. In AS and DISH patients, complication and mortality rates were significantly higher than in controls. Logistic regression analysis showed the parameters age and presence of DISH to be independently, statistically significantly related to mortality. Conclusions Many patients with AS and DISH showed unstable (hyperextension) fracture configurations and neurologic deficits. Complication and mortality rates were higher in patients with ASD compared with control patients. Increasing age and presence of DISH are predictors of mortality after a spinal fracture.

Summary The recent development of 3D-liver stem cell cultures (hepatic organoids) opens up new avenues for gene and/or stem cell therapy to treat liver disease. To test safety and efficacy, a relevant large animal model is essential but not yet established. Because of its shared pathologies and disease pathways, the dog is considered the best model for human liver disease. Here we report the establishment of a long-term canine hepatic organoid culture allowing undifferentiated expansion of progenitor cells that can be differentiated toward functional hepatocytes. We show that cultures can be initiated from fresh and frozen liver tissues using Tru-Cut or fine-needle biopsies. The use of Wnt agonists proved important for canine organoid proliferation and inhibition of differentiation. Finally, we demonstrate that successful gene supplementation in hepatic organoids of COMMD1-deficient dogs restores function and can be an effective means to cure copper storage disease., Highlights • Canine hepatic organoids can be grown long term with chromosomal stability • Organoids can be derived from liver tissues as small as fine-needle biopsies • Differentiation and proliferation of canine organoids involve Wnt and Notch • Successful gene supplementation proves potential for stem cell-based gene therapy, Schotanus and colleagues show that the development of canine hepatic organoids ensures successful use of valuable patient material and demonstrate the use of this cell system for in vitro disease modeling and gene supplementation for gene therapy. This is a step forward for stem cell therapy in liver diseases in human, because numerous (inherited) diseases present in similar molecular biological ways in men and dogs

Canine hereditary copper-associated hepatitis is characterized by gradual hepatic copper accumulation eventually leading to liver cirrhosis. Therapy is aimed at creating a negative copper balance with metal chelators, of which d-penicillamine is the most commonly used. d-penicillamine often causes gastro-intestinal side effects and life-long continuous therapy may lead to a deficiency of copper and zinc. The aim of the current study was to investigate the effect of a low-copper, high-zinc diet as an alternative to continuous d-penicillamine treatment for the long-term management of canine copper-associated hepatitis. Sixteen affected Labrador retrievers were followed for a median time period of 19.1 months (range, 5.9–39 months) after being effectively treated with d-penicillamine. The dogs were maintained on a diet containing 1.3 ± 0.3 mg copper/1000 kcal and 64.3 ± 5.9 mg zinc/1000 kcal. Liver biopsies were taken every 6 months for histological evaluation and copper determination. Plasma alanine aminotransferase (ALT) and alkaline phosphatase, as well as serum albumin were determined. Dietary treatment alone was sufficient to maintain hepatic copper concentration below 800 mg/kg dry weight liver in 12 dogs during the study period. Four dogs needed re-treatment with d-penicillamine. ALT activity and albumin concentration were not associated with hepatic copper concentration, but showed a significant association with the stage and grade of hepatitis respectively. In conclusion, a low-copper, high-zinc diet can be a valuable alternative to continuous d-penicillamine administration for long-term management of dogs with copper-associated hepatitis. The copper re-accumulation rate of an individual dog should be considered in the design of a long-term management protocol and in determining re-biopsy intervals.

Hip dysplasia, an abnormal laxity of the hip joint, is seen in humans as well as dogs and is one of the most common skeletal disorders in dogs. Canine hip dysplasia is considered multifactorial and polygenic, and a variety of chromosomal regions have been associated with the disorder. We performed a genome-wide association study in Dutch Labrador Retrievers, comparing data of nearly 18,000 single nucleotide polymorphisms (SNPs) in 48 cases and 30 controls using two different statistical methods. An individual SNP analysis based on comparison of allele frequencies with a χ2 statistic was used, as well as a simultaneous SNP analysis based on Bayesian variable selection. Significant association with canine hip dysplasia was observed on chromosome 8, as well as suggestive association on chromosomes 1, 5, 15, 20, 25 and 32. Next-generation DNA sequencing of the exons of genes of seven regions identified multiple associated alleles on chromosome 1, 5, 8, 20, 25 and 32 (p

Additive manufacturing forms a potential route towards economically viable production of cellular constructs for tissue engineering. Hydrogels are a suitable class of materials for cell delivery and 3D culture, but are generally unsuitable as construction materials. Gelatine-methacrylamide is an example of such a hydrogel system widely used in the field of tissue engineering, e.g. for cartilage and cardiovascular applications. Here we show that by the addition of gellan gum to gelatine-methacrylamide and tailoring salt concentrations, rheological properties such as pseudo-plasticity and yield stress can be optimised towards gel dispensing for additive manufacturing processes. In the hydrogel formulation, salt is partly substituted by mannose to obtain isotonicity and prevent a reduction in cell viability. With this, the potential of this new bioink for additive tissue manufacturing purposes is demonstrated. This journal is © the Partner Organisations 2014.

Cartilage damage and osteoarthritis (OA) impose an important burden on society, leaving both young, active patients and older patients disabled and affecting quality of life. In particular, cartilage injury not only imparts acute loss of function but also predisposes to OA. The increase in knowledge of the consequences of these diseases and the exponential growth in research of regenerative medicine have given rise to different treatment types. Of these, cell-based treatments are increasingly applied because they have the potential to regenerate cartilage, treat symptoms, and ultimately prevent or delay OA. Although these approaches give promising results, they require a costly in vitro cell culture procedure. The answer may lie in single-stage procedures that, by using cell combinations, render in vitro expansion redundant. In the last two decades, cocultures of cartilage cells and a variety of (mesenchymal) stem cells have shown promising results as different studies report cartilage regeneration in vitro and in vivo. However, there is considerable debate regarding the mechanisms and cellular interactions that lead to chondrogenesis in these models. This review, which included 52 papers, provides a systematic overview of the data presented in the literature and tries to elucidate the mechanisms that lead to chondrogenesis in stem cell cocultures with cartilage cells. It could serve as a basis for research groups and clinicians aiming at designing and mplementing combined cellular technologies for single-stage cartilage repair and treatment or prevention of OA.

tHip as well as elbow dysplasia (HD, ED) are developmental disorders leading to malforma-tion of their respective joints. For a long time both disorders have been scored and targetedfor improvement using selective breeding in several Dutch dog populations. In this paperall scores for both HD and ED, given to pure bred dogs in the Netherlands from 2002 to2010, were analyzed. Heritabilities and correlations between HD and ED were calculatedfor the 4 most frequently scored breeds. Heritabilities ranged from 0.0 to 0.37 for HD relatedtraits (FCI-score, osteoarthritis, congruity, shape and laxity (Norberg angle); FCI: Fédéra-tion Cynologique Internationale) and from 0.0 to 0.39 for ED related traits (IEWG score,osteoarthritis, sclerosis and indentation; IEWG: International Elbow Working Group). HDrelated traits showed high genetic and residual correlations among each other but were onlyto a minor extent correlated with ED related traits, which also showed high correlationsamong each other. Genetic correlations were higher than residual correlations. Phenotypicand genetic trends since 2001 for the four most scored breeds were slightly positive butdecreasing over time, indicating that selection over the past decade has not been effective.

Degeneration of the intervertebral disc (IVD) is associated with a loss of notochordal cells (NCs) from the nucleus pulposus (NP) and their replacement by chondrocyte-like cells. NCs are known to maintain extracellular matrix quality and stimulate the chondrocyte-like NP cells, making NCs attractive for designing new tissue engineering approaches for IVD regeneration. However, optimal conditions, such as osmolarity and other characteristics of the culture media, for long-term culture of NCs are not known. The purpose of this study was to investigate the effects of different culture media and osmolarity on the physiology of NCs in vitro. NC clusters isolated from canine IVDs were suspended in alginate beads and cultured at 37°C under normoxic conditions for 28 days. Three different culture conditions were investigated; (1) DMEM/F12 (300 mOsm/L), (2) α-MEM (300 mOsm/L) and (3) α-MEM adjusted to 400 mOsm/L to mimic a hyperosmolar environment. NC morphology, expression of genes related to NC markers, matrix production and remodelling, and DNA- and GAG analyses were performed on 1, 7, 14, and 28 days in culture. Large, vesicle-containing cells organised in clusters, characterized as NCs, remained present during 28 days for all culture conditions. However, the proportion of the NC clusters decreased over time, whereas the proportion of spindle-shaped cells increased. Gene expression profiling at 7, 14, and 28 days in culture compared to day 1 indicated a initial loss of NC phenotype followed by some recovery of brachyury and aggrecan expression after 28 days of culture supporting a potential recovery of NC phenotype. NCs cultured in α-MEM adjusted to 400 mOsm/L showed the highest expression of brachyury, cytokeratin 18, and aggrecan, the highest GAG production, and the lowest collagen 1α1 gene expression. In conclusion, NCs cultured in alginate in native cell clusters, partially retained their characteristic morphology and recovered their phenotype in long-term culture. The type of culture medium and medium osmolarity appear to be important factors for culturing NC clusters. These findings provide additional information concerning the maintenance of NCs in vitro that may aid further mechanistic inquiry into the biology of NCs. PMID:24304309[PubMed - as supplied by publisher]

Keratin 19 marks poor differentiation and a more aggressive behaviour in canine and human hepatocellular tumours Renee GHM van Sprundel1, Ted SGAM van den Ingh2, Valeer J Desmet3, Azeam Katoonizadeh3, Louis C Penning1, Jan Rothuizen1, Tania Roskams3 and Bart Spee13* * Corresponding author: Bart Spee B.Spee@uu.nl Author Affiliations 1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary medicine, Utrecht University, Utrecht, The Netherlands 2 TCCI Consultancy BV, Utrecht, The Netherlands 3 Department of Morphology and Molecular Pathology, University Hospitals Leuven, Leuven, Belgium For all author emails, please log on. Comparative Hepatology 2010, 9:4 doi:10.1186/1476-5926-9-4 The electronic version of this article is the complete one and can be found online at: http://www.comparative-hepatology.com/content/9/1/4 Received: 23 November 2009 Accepted: 18 February 2010 Published: 18 February 2010 © 2010 van Sprundel et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.