BackgroundOsteoarthritis (OA) is a major joint disease in humans and many other animals. Consequently, medical countermeasures for OA have been investigated diligently. This study was designed to examine the regeneration of articular cartilage and subchondral bone using three-dimensional (3D) constructs of adipose tissue-derived mesenchymal stem cells (AT-MSCs).MethodsAT-MSCs were isolated and expanded until required for genetical and immunological analysis and construct creation. A construct consisting of about 760 spheroids that each contained 5.0 × 104 autologous AT-MSCs was implanted into an osteochondral defect (diameter: 4 mm; depth: 6 mm) created in the femoral trochlear groove of two adult microminipigs. After implantation, the defects were monitored by computed tomography every month for 6 months in animal no. 1 and 12 months in animal no. 2.ResultsAT-MSCs were confirmed to express the premature genes and to be positive for CD90 and CD105 and negative for CD34 and CD45. Under specific nutrient conditions, the AT-MSCs differentiated into osteogenic, chondrogenic, and adipogenic lineages, as evidenced by the expressions of related marker genes and the production of appropriate matrix molecules. A radiopaque area emerged from the boundary between the bone and the implant and increased more steadily upward and inward for the implants in both animal no. 1 and animal no. 2. The histopathology of the implants after 6 months revealed active endochondral ossification underneath the plump fibrocartilage in animal no. 1. The histopathology after 12 months in animal no. 2 showed not only that the diminishing fibrocartilage was as thick as the surrounding normal cartilage but also that massive subchondral bone was present.ConclusionsThe present results suggest that implantation of a scaffold-free 3D construct of AT-MSCs into an osteochondral defect may induce regeneration of the original structure of the cartilage and subchondral bone over the course of 1 year, although more experimental cases are needed.Electronic supplementary materialThe online version of this article (doi:10.1186/s13018-015-0173-0) contains supplementary material, which is available to authorized users.
Abstract. Atherosclerotic lesions were observed in male and ovariectomized female Microminipig (MMP) fed a high fat and cholesterol diet with sodium cholate (HFCD/SC) for 3 months. HFCD/SC induced hypercholesterolemia accompanied by an increase in serum total cholesterol (T-Cho), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and cholesterol ester (CE). Unlike the mouse or rabbit, a dominant LDL-C fraction in the intact MMP, similar to that in humans, was observed by serum lipoprotein analysis. HFCD/SC increased body weight gain. At the end of the experiment, computed tomography scans of conscious animals showed that HFCD/SC had decreased liver attenuation values (Hounsfield unit) and increased subcutaneous and abdominal fat, suggesting the induction of fatty liver and obesity. HFCD/SC induced atherosclerotic lesions in systemic arteries, including the external and internal iliac arteries, abdominal aorta, coronary artery, and cerebral arterial circle. Atherosclerosis and pathological findings induced by HFCD/SC in MMP were similar to those in humans. The MMP is a potentially suitable tool for investigating human atherosclerosis.
Uterine torsion secondary to sacculation of the uterine horns was diagnosed in two non-gravid bitches which were presented with anorexia, polydipsia and an acutely swollen abdomen. On the basis of the radiological and ultrasonographic findings, which indicated the presence of an enlarged spherical or tubular structure filled with hypoechoic material in the caudal abdomen, a tentative diagnosis of pyometra was made. Exploratory laparotomy revealed unilateral uterine horn torsion along the longitudinal axis, with bilateral fluid-filled sacculations. Ovariohysterectomy was performed in both cases. Pathological examination of the uteri demonstrated haematometra in one dog and pyometra in the other.
ABSTRACT. We hypothesized that semi-quantitative determinations of changeable blood flows in granulation and scar tissues during tendon healing could be helpful for differentiation between the acute phase rich in blood vessels and the remodeling phase with fewer vessels. Ten Thoroughbreds with injured superficial digital flexor tendon (SDFT) in a forelimb were used for evaluation of blood flows in the injured tendons of racehorses using color Doppler (CD) ultrasonography. Using longitudinal CD images, features of maximum color activities were defined. These were rhythmically blinking, tiny to small signals (grade 1), pulsatile expanded dots (grade 2), and dynamic streams (grade 3). Grade of color activity in CD ultrasonography could be useful for quantitative assessment of equine SDFT repair.
Summary
Reasons for performing study: Cartilage oligomeric matrix protein (COMP) is abundant within cartilage; its turnover and/or degradation have been investigated in various equine joint diseases and it has been suggested that COMP fragmentation might be useful for monitoring such conditions.
Objectives: To determine whether COMP metabolism is compromised in equine osteoarthritis (OA) and whether COMP degradation is a useful joint marker representing cartilage destruction.
Hypothesis: A monoclonal antibody (mAb) with a higher affinity for degraded COMP allows discrimination of diseased joints by quantifying COMP levels and fragmentation.
Methods: A mAb (clone14G4) was generated against equine cartilage COMP. The NH2‐terminal sequence of enzyme‐cut COMP fragments recognised by 14G4 was determined, as was the efficiency of binding to COMP (using a generated COMP peptide). COMP concentration and fragmentation were analysed in synovial fluid (SF) from normal horses and those with OA.
Results: The mAb 14G4 had a higher affinity for the smaller fragments of equine COMP, compared with a mAb (clone 12C4) generated against human COMP. The 14G4 epitope was identified as between C134 and F147. The COMP values in OA (mean ± s.d. 205.8 ± 90.9 μg/ml) were significantly higher than in the normal (133.1 ± 31.5 μg/ml) SF. On the immunoblots of OA sample, the proportions of intact COMP were significantly lower, while smaller fragments ranging from 75 to 290 kDa were higher compared with the normal SF.
Conclusions and potential relevance: The mAb 14G4 reliably detects COMP degradation as well as synthesis, and fragmentation analysis combined with quantification in SF could be useful to study equine OA.
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