Effects of growth factors on equine synovial fibroblasts seeded on synthetic scaffolds for avascular meniscal tissue engineering

Across species, the avascular portion of the knee meniscus cannot heal spontaneously if severely injured. The most common treatment is meniscectomy which results in osteoarthritis. The objective of this study was to assess the fibrochondrogenic potential of equine fibroblast-like synoviocytes (FLS) seeded on scaffolds under the influence of growth factors in vitro to determine the potential of developing a novel cell-based repair strategy. Cultured FLS were seeded onto synthetic scaffolds in a rotating bioreactor under the influence of three growth factor regimens: none, basic fibroblast growth factor (bFGF) alone, and bFGF plus transforming growth factor (TGF-β₁) and insulin-like growth factor (IGF-1). Constructs were analyzed for mRNA expression and production of fibrochondroid extracellular matrix constituents. Type II collagen and aggrecan mRNA were significantly higher in growth factor-treated groups (p < 0.05). Despite sub-optimal extracellular matrix production, FLS can exhibit fibrochondral characteristics and may have potential for cell-based tissue engineering for avascular meniscal regeneration.     

Evaluation of two dynamic axial fixators for large ruminants

OBJECTIVES: To evaluate healing of a radial osteotomy repaired by application of dynamic axial fixation devices (DAF) in large ruminants. STUDY DESIGN: In vivo study of bone healing after application of 2 DAF types. Model I had 2 sidebars, each with a central cylindrical cuff (internally threaded) with 2 detachable connecting rods telescoping within the cuff. Model II had 2 side bars with 2 moveable clamps with multiple holes. SAMPLE POPULATION: Bull calves (n=8; aged, 1.5-2.0 years; weighing, 175-250 kg). METHODS: A mid-diaphyseal radial osteotomy was repaired by use of a model I (n=4) or model II (n=4) DAF. Calves were monitored for weight bearing, stability of fixation, and radiographically for fracture reduction, alignment and healing at intervals for 6 months. Fixators were removed when there was radiographic evidence of healing. RESULTS: Both the fixators were well tolerated with free movement of adjacent joints. Fragment fixation was maintained until healing in all but 1 model I calf where failure occurred within 7 days. Model II DAF provided more rigid fixation as indicated by early full weight bearing and fracture healing with less callus formation. Functional recovery of repaired limbs occurred within 60 days in surviving calves. CONCLUSIONS: Both bilateral DAFs were easy to apply; however, the model II DAF provided better fixation. CLINICAL RELEVANCE: The model II DAF made of low carbon steel was economical and may be useful for treating long bone fractures in large ruminants.     

The effect of supplementing layer diets with shark cartilage or chitosan on egg components and yolk lipids

1. An experiment was designed to evaluate the effects of the addition of shark cartilage (SC) or chitosan (CH) to layer diets on egg component weights, yolk lipids and hen plasma lipids. 2. Hy-Line laying hens (80) were used during a 56 d feeding trial. Treatments were: basal diet (BD), BD + 20 g/kg SC, BD + 30 g/kg SC, BD + 20 g/kg CH and BD + 30 g/kg CH. Eggs were analysed on d 14, 28, 42 and 56. 3. Egg weight and egg component weights were not affected by these treatments throughout the experimental period. 4. After 14d of experimental feeding, cholesterol levels were higher in eggs from birds given BD + 20 g/kg CH and BD + 30 g/kg CH than in those from birds given BD. 5. Furthermore, eggs from hens given BD + 20 g/kg SC or BD + 20 g/kg CH were higher in palmitic and stearic acids and lower in oleic acid than those from birds fed on BD. After 56 d feeding, however, palmitic and stearic acid contents in eggs from hens given any of the supplemented diets were lower than in those from hens given BD, and oleic acid in eggs from hens given BD + 20 g/kg SC, BD + 30 g/kg SC and BD + 30 g/kg CH was higher than in those from birds fed on BD. 6. Plasma cholesterol and triacylglycerol levels were not significantly affected by dietary treatment. 7. Shark cartilage or chitosan at up to 30 g/kg in layer diets did not affect egg component weights (yolk, white and shell) and total lipid contents. During the period from 42 to 56d of experimental feeding, diets containing up to 30 g/kg chitosan reduced egg yolk contents of cholesterol, palmitic and stearic acids and increased the content of oleic acid.     

The comparison of equine articular cartilage progenitor cells and bone marrow-derived stromal cells as potential cell sources for cartilage repair in the horse

A chondrocyte progenitor population isolated from the surface zone of articular cartilage presents a promising cell source for cell-based cartilage repair. In this study, equine articular cartilage progenitor cells (ACPCs) and equine bone marrow-derived stromal cells (BMSCs) were compared as potential cell sources for repair. Clonally derived BMSCs and ACPCs demonstrated expression of the cell fate selector gene, Notch-1, and the putative stem cell markers STRO-1, CD90 and CD166. Chondrogenic induction revealed positive labelling for collagen type II and aggrecan. Collagen type X was not detected in ACPC pellets but was observed in all BMSC pellets. In addition, it was observed that BMSCs labelled for Runx2 and matrilin-1 antibodies, whereas ACPC labelling was significantly less or absent. For both cell types, osteogenic induction revealed positive von Kossa staining in addition to positive labelling for osteocalcin. Adipogenic induction revealed a positive result via oil red O staining in both cell types. ACPCs and BMSCs have demonstrated functional equivalence in their multipotent differentiation capacity. Chondrogenic induction of BMSCs resulted in a hypertrophic cartilage (endochondral) phenotype, which can limit cartilage repair as the tissue can undergo mineralisation. ACPCs may therefore be considered superior to BMSCs in producing cartilage capable of functional repair.     

Culture of canine synoviocytes on porcine intestinal submucosa scaffolds as a strategy for meniscal tissue engineering for treatment of meniscal injury in dogs

Meniscal injury is a common cause of canine lameness. Tissue engineered bioscaffolds may be a treatment option for dogs suffering from meniscal damage. The aim of this study was to compare in vitro meniscal-like matrix formation and biomechanical properties of porcine intestinal submucosa sheets (SIS), used in canine meniscal regenerative medicine, to synoviocyte-seeded SIS bioscaffold (SSB), cultured with fetal bovine serum (SSBfbs) or chondrogenic growth factors (SSBgf). Synoviocytes from nine dogs were seeded on SIS and cultured for 30 days with 17.7% fetal bovine serum or recombinant chondrogenic growth factors (IGF-1, TGFβ1 and bFGF). The effect on fibrochondrogenesis was determined by comparing mRNA expression of collagen types Iα and IIα, aggrecan, and Sry-type homeobox protein-9 (SOX9) as well as protein expression of collagens I and II, glycosaminoglycan (GAG), and hydroxyproline.The effect of synoviocyte seeding and culture conditions on biochemical properties was determined by measuring peak load, tensile stiffness, resilience, and toughness of bioscaffolds. Pre-culture SIS contained 13.6% collagen and 2.9% double-stranded DNA. Chondrogenic growth factor treatment significantly increased SOX9, collagens I and IIα, aggrecan gene expression (P < 0.05), and histological deposition of fibrocartilage extracellular matrix (GAG and collagen II). Culture with synoviocytes increased SIS tensile peak load at failure, resilience, and toughness of bioscaffolds (P < 0.05). In conclusion, culturing SIS with synoviocytes prior to implantation might provide biomechanical benefits, and chondrogenic growth factor treatment of cultured synoviocytes improves in vitro axial meniscal matrix formation.     

谷氨酰胺、壳聚糖及二者复合添加对断奶仔猪生长性能和血液生化指标的影响

选取192头体重相近(平均体重为5.27kg)的(21±2)日龄PIC断奶仔猪,按胎次、公母分4个处理组,每组3个重复,每个重复16头。试验分两个阶段:21～35日龄、35～49日龄,研究谷氨酰胺、壳聚糖单独和复合添加对断奶仔猪生产性能和血液生化指标的影响。结果表明:①日粮中单独添加1%谷氨酰胺或0.04%壳聚糖有提高断奶仔猪日增重、降低料重比的趋势。②谷氨酰胺与壳聚糖复合添加使用在一些指标上优于单独添加,体现出一定的协同作用。     

Evaluation of sodium bisulphate and phosphoric acid as urine acidifiers for cats.

Eighteen cats were used to compare the urine acidifying properties of sodium bisulphate to phosphoric acid. Acidifying agents were added at one of three concentrations (0.4, 0.6, or 0.8%, as-is basis). Cats were offered a commercial diet to determine basal urinary pH, and then again for a 1 week period between blocks 1 and 2. Cats were acclimated to the diets for 6 days, and urine samples were collected on day 7 at 0, 4, and 8 h postfeeding to obtain pre- and postprandial urinary pH. Intakes of diets containing sodium bisulphate tended (P < 0.07) to be lower than intakes of diets containing phosphoric acid. Cats consuming the 0.8% phosphoric acid diet had higher (P < 0.05) food intakes than cats consuming either the 0.4 or 0.6% phosphoric acid-containing diets. There was significant (P = 0.01) linear and quadratic response for food intake in cats consuming the sodium bisulphate-containing diet. Cats consuming the 0.4 and 0.8% phosphoric acid-containing diets tended (P = 0.07) to have higher water intakes than cats consuming the 0.6% phosphoric acid-containing diet. There were no differences (P > 0.05) in urine pH and specific gravity between cats fed the different acidifier types. Cats consuming the 0.6% phosphoric acid-containing diet tended (P = 0.07) to have a higher urine pH 8 h post-feeding than cats consuming the 0.4 and 0.8% phosphoric acid-containing diets. Urine pH was highest at 4 h post-feeding except for cats fed the 0.4% sodium bisulphate- and the 0.6% phosphoric acid-containing diets. No differences (P > 0.05) between acidifiers were found in faecal score or in faecal dry matter and organic matter concentrations. A quadratic response was detected in faecal score for cats consuming the phosphoric acid-containing diets. Cats consuming the 0.6% phosphoric acid diet tended (P = 0.06) to have a lower faecal score than cats consuming the 0.4 and 0.8% phosphoric acid diets. For faecal dry matter, a linear trend was detected in cats consuming the sodium bisulphate (P = 0.08) and phosphoric acid-containing (P = 0.04) diets. Sodium bisulphate and phosphoric acid generally behaved in a similar fashion when incorporated in dry cat diets.     

鲜牦牛肉氨基酸与脂肪酸组成分析评价

为分析评价鲜牦牛肉的氨基酸及脂肪酸组成及营养价值,采用氨基酸分析仪测定了氨基酸并进行氨基酸评分(AAS),脂肪酸甲酯的气相色谱分析法分析了脂肪酸。结果表明:从新鲜牦牛肉中检测出十七种氨基酸和十种脂肪酸。新鲜牦牛肉的氨基酸总量为19.19g/100g,EAA/TAA为38.83%,EAA/NEAA为63.31%;脂肪含量3.99g/100g,脂肪酸含量为51.63%。鲜牦牛肉是一种具有独特风味的优质蛋白质来源,也是具有均衡脂肪酸的高营养价值的鲜肉。     

CNCPS氨基酸模型评定奶牛氨基酸供给和需求

1　氨基酸供应量奶牛可吸收且最终能够利用的氨基酸是由瘤胃微生物合成的蛋白、日粮过瘤胃蛋白和分泌到消化道中的内源性蛋白来提供 (Richardson和Hatfield ,1978)。以Russell等 (1992 )和Sniffen等 (1992 )的模型作为基础而得出氨基酸供应模型 ,用其模型可估计由瘤胃微生物合成的蛋白和由日粮过瘤胃蛋白提供的氨基酸量。由于描述奶牛内源性分泌的氨基酸数量资料有限 ,模型不考虑由内源性分泌蛋白提供的氨基酸量 (Mantysaari等 ,1989)。因此 ,在模型中 ,依据由瘤胃微生物蛋白和日粮过瘤胃蛋白而得到的氨基酸量 ,来确定由特定日粮而获得的…     

Evaluation of canine mast cell tumors for presence of estrogen receptors

Ten tumors from 7 dogs were analyzed for estrogen receptors. Of 9 determined to be mast cell tumors, 6 were determined not to have estrogen receptors (less than 3 fmol of estradiol/mg of cytosol protein) and 3 were questionable (3 to 10 fmol of estradiol/mg). One tumor was a mixed mammary tumor and was determined to have estrogen receptors (12 fmol of estradiol/mg). Histologic grading of the mast cell tumors did not suggest a correlation with estrogen receptor values.     

温带果园适宜草种及其播量的初步筛选

为确定沈阳地区果园生草适宜草种及适宜播量, 在沈阳农业大学苹果园套种了不同播量的紫花苜蓿(Medicago sativa cv.Zhaodong)、白三叶(Trifolium repens cv. Rivendel)、红三叶(T.pretense cv.Renegade)。通过调查越冬率, 确定适宜草种, 以产草量、生长速度、粗蛋白含量为参考指标确定适宜播量。结果表明, 白三叶和红三叶的最高越冬率分别为12%和48%, 而紫花苜蓿的自然越冬率达100%, 表明紫花苜蓿更适合冷凉地区果园生草栽培。紫花苜蓿全年可刈割3次, 中播量(15 kg·hm-2)的全年干草产量、生长速度和平均粗蛋白均为最高, 分别为15 783.74 kg·hm-2、2.63 cm·d-1和19.79%;低播量(12 kg·hm-2)的平均净同化率最高, 为3.32 g·m-2·d-1。由此可知, 冷凉地区果园生草适宜草种为紫花苜蓿, 从全年干草产量、生长速度、粗蛋白含量等指标综合考虑, 适宜播量为15 kg·hm-2。     

A new cell culture protocol for enrichment and genetic modification of adult canine Schwann cells suitable for peripheral nerve tissue engineering

Easily applicable techniques are presented to obtain high numbers of enriched canine Schwann cells (cSC) in a short time-window. The potential of adult SC for tissue engineering of peripheral nerves and ex vivo gene therapy is obvious from physiological events taking place after peripheral nerve transection [Haastert, K., Grothe, C., 2007. Gene therapy in peripheral nerve reconstruction approaches. Curr. Gene Ther. 7, 221-228]. The presented techniques were modified from a protocol for cultivation and expansion of adult cSC by others [Pauls, J., Nolte, C., Forterre, F., Brunnberg, L., 2004. Cultivation and expansion of canine Schwann cells using reexplantation. Berl. Munch. Tierarztl. Wochenschr. 117, 341-352] and own experiences in rodent and human SC cultivation and transfection [Haastert, K., Mauritz, C., Chaturvedi, S., Grothe, C., 2007. Human and rat adult Schwann cell cultures: fast and efficient enrichment and highly effective non-viral transfection protocol. Nat. Protoc. 2, 99-104]. A purity of about 80% cSC achieved by immunopanning techniques and selective culture conditions is 2.5 fold higher as previously reported (Pauls et al., 2004). Additionally, highly enriched cSC populations are available in 3-4 weeks, only half the time period reported previously (Pauls et al., 2004). Furthermore, electroporation and genetic modification of cSC is reported for the first time.     

Mass screening of Irish wolfhound puppies for portosystemic shunts by the dynamic bile acid test.

Five hundred and sixty-six Irish wolfhound puppies aged six to 15 weeks were tested for congenital portosystemic shunts by the dynamic bile acid method. Plasma ammonia concentration was also measured in 165 of the puppies both fasting and postprandially. Nineteen puppies (3.4 per cent), nine males and 10 females, had portosystemic shunts. Smaller litters appeared to be more likely to contain affected puppies. The postprandial bile acid concentration was a reliable predictor of the presence of a shunt, with the highest concentration in a normal puppy being 38 mumol/litre and the lowest in an affected puppy being 43 mumol/litre. In contrast, fasting bile acid concentrations were normal in the majority of the affected puppies. There was considerable overlap in fasting plasma ammonia concentrations between normal and affected puppies (26 puppies, 15.8 per cent of those tested). Postprandial ammonia concentration appeared to give better separation between the two groups, apart from two individuals which produced bizarre results. It was concluded that the postprandial or dynamic bile acid test is an appropriate test for the mass screening of Irish wolfhound puppies for portosystemic shunts, and guidelines are proposed for the interpretation and follow-up of the test.     

Evaluation of physical and chemical properties of citric acid industrial waste

Tropical Animal Health and Production - This study aimed to evaluate physical and chemical properties and nutritive values of citric acid by-product (CABP) from cassava and to compare its...     

壳寡糖对断奶仔猪回肠养分消化、小肠形态及隐窝细胞增殖的影响

本研究旨在探讨日粮添加壳寡糖对断奶仔猪生长性能、营养物质消化率和小肠功能的影响,评估其作为断奶仔猪日粮中抗生素替代品的效果。试验选择21 d断奶的三元仔猪280头,随机分为5组,每组4个重复,每个重复14头猪,各组猪只分别饲喂在基础日粮中分别添加0、80、160、240 mg/kg壳寡糖和45 mg/kg盐霉素的日粮,试验分为1~28 d和29~56 d两个阶段,共进行8周。结果显示,日粮添加160 mg/kg壳寡糖显著提高回肠粗蛋白质、粗脂肪、灰分、钙和磷的表观消化率(P <0.05),显著提高小肠绒毛高度及绒毛高度与隐窝深度的比值(P <0.05),同时显著提高56 d十二指肠和空肠隐窝细胞增殖活力(P <0.05)。结论:日粮添加160 mg/kg壳寡糖可以通过提高回肠养分表观消化率、绒毛高度和肠上皮细胞的数量来促进消化吸收能力,进而提高断奶仔猪生长性能和健康状态,其作用与盐霉素的效果相当。