Inflammation and Immune Response of Intra-Articular Serotype 2 Adeno-Associated Virus or Adenovirus Vectors in a Large Animal Model

Intra-articular gene therapy has potential for the treatment of osteoarthritis and rheumatoid arthritis. To quantify in vitro relative gene transduction, equine chondrocytes and synovial cells were treated with adenovirus vectors (Ad), serotype 2 adeno-associated virus vectors (rAAV2), or self-complementary (sc) AAV2 vectors carrying green fluorescent protein (GFP). Using 6 horses, bilateral metacarpophalangeal joints were injected with Ad, rAAV2, or scAAV2 vectors carrying GFP genes to assess the in vivo joint inflammation and neutralizing antibody (NAb) titer in serum and joint fluid. In vitro, the greater transduction efficiency and sustained gene expression were achieved by scAAV2 compared to rAAV2 in equine chondrocytes and synovial cells. In vivo, AAV2 demonstrated less joint inflammation than Ad, but similar NAb titer. The scAAV2 vectors can induce superior gene transduction than rAAV2 in articular cells, and both rAAV2 and scAAV2 vectors were showed to be safer for intra-articular use than Ad vectors

The Temporal Expression of Adipokines During Spinal Fusion

Background Context Adipokines are secreted by white adipose tissue and have been associated with fracture healing. Our goal was to report the temporal expression of adipokines during spinal fusion in an established rabbit model. Purpose Our goal was to report the temporal expression of adipokines during spinal fusion in an established rabbit model. Study Design The study design included a laboratory animal model. Methods New Zealand white rabbits were assigned to either sham surgery (n=2), unilateral posterior spinal fusion (n=14), or bilateral posterior spinal fusion (n=14). Rabbits were euthanized 1–6 and 10 weeks out from surgery. Fusion was evaluated by radiographs, manual palpation, and histology. Reverse transcription-polymerase chain reaction on the bone fusion mass catalogued the gene expression of leptin, adiponectin, resistin, and vascular endothelial growth factor (VEGF) at each time point. Results were normalized to the internal control gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (2^ΔCt), and control bone sites (2^ΔΔCt). Quantitative data were analyzed by two-factor analysis of variance (p\u3c.05). Results Manual palpation scores, radiograph scores, and histologic findings showed progression of boney fusion over time (p Conclusions Leptin expression is likely associated with the maturation phase of bone fusion. Adiponectin and resistin may play a role early on during the fusion process. Our results suggest that leptin expression may be upstream of VEGF expression during spinal fusion, and both appear to play an important role in bone spinal fusion

Black holes, gravitational waves and fundamental physics: a roadmap

The grand challenges of contemporary fundamental physics—dark matter, dark energy, vacuum energy, inflation and early universe cosmology, singularities and the hierarchy problem—all involve gravity as a key component. And of all gravitational phenomena, black holes stand out in their elegant simplicity, while harbouring some of the most remarkable predictions of General Relativity: event horizons, singularities and ergoregions. The hitherto invisible landscape of the gravitational Universe is being unveiled before our eyes: the historical direct detection of gravitational waves by the LIGO-Virgo collaboration marks the dawn of a new era of scientific exploration. Gravitational-wave astronomy will allow us to test models of black hole formation, growth and evolution, as well as models of gravitational-wave generation and propagation. It will provide evidence for event horizons and ergoregions, test the theory of General Relativity itself, and may reveal the existence of new fundamental fields. The synthesis of these results has the potential to radically reshape our understanding of the cosmos and of the laws of Nature. The purpose of this work is to present a concise, yet comprehensive overview of the state of the art in the relevant fields of research, summarize important open problems, and lay out a roadmap for future progress. This write-up is an initiative taken within the framework of the European Action on 'Black holes, Gravitational waves and Fundamental Physics'

Biochemical and biomechanical alterations in equine articular cartilage following an experimentally-induced synovitis

SummaryThe effects of inflammation on the biochemical and biomechanical properties of articular cartilage at two sites (dorsal and palmar) from the radial facet of the equine third carpal bone were examined in response to a synovitis induced with Escherichia coli lipopolysaccharide (LPS). Four groups were studied. In group 1 synovitis was induced at time zero and evaluated at week 6. Group 2 was the sham-treated control for group 1. In group 3 synovitis was induced at time zero and evaluated at week 2. Group 4 was the sham-treated control for group 3.There was a significant increase (P < 0.05) in newly synthesized proteoglycan PG from both sites in group 3 as compared to the sham-treated groups and group 1. No significant difference in the endogenous PG concentration between groups or sites was detected. Sepharose CL-2B revealed two peaks of newly synthesized PG in all groups; an early peak (Kav 0.11–0.13) and a late peak (Kav 0.48–0.64). Newly synthesized PG profiles from sham-treated groups and group 3 were similar, but the group 3 PG profile exhibited a more pronounced early peak. Conversely, the PG profile from group 1 demonstrated a more prominent late peak. Electrophoresis and Western blot analysis of the pooled late PG peak fractions from the sham-treated and group 1 showed a single toluidine blue stained band from both sites which reacted with monoclonal antibody (MAb) 1C6. By contrast, the late peak from the palmar site in group 3 showed an additional faster moving component on composite gels which did not react with MAb 1C6.There was a significant decrease in Poisson's ratio and a significant increase in cartilage thickness in groups 1 and 3 which had received synovitis. The increase in cartilage thickness of groups 1 and 3 was also significantly affected by site (dorsal > palmar). There was no significant difference in aggregate modulus or permeability constant among groups.Primary joint inflammation induced by LPS alters the biochemical and biomechanical properties of the articular cartilage as a function of time and site. An increase in chondrocyte PG synthesis in the early period following synovitis may be a reparative response to the inflammatory insult. Continued alterations in the qualitative PG composition in the later period following synovitis may represent a shift in chondrocyte metabolism to repopulate the existing cartilage matrix