Bone marrow-derived stromal cells and cartilage tissue engineering

Articular cartilage repair is one of the most challenging issues which remains to be resolved in clinic work. Discovering of bone marrow stromal cells (BMSC) and its application in tissue engineering provide new methods for the treatment of cartilage defects. High seeding density, appropriate cytokines and three-dimensional culture play important roles in the process of inducing BMSC differentiating into chondrocytes, suitable scaffold is also essential in reconstructing cartilage in vitro by methods of tissue engineering.     

Identification of stable normalization genes for quantitative real-time PCP, in porcine articular cartilage

Background: Expression levels for genes of interest must be normalized with an appropriate reference, or housekeeping gene, to make accurate comparisons of quantitative real-time PCR results. The purpose of this study was to identify the most stable housekeeping genes in porcine articular cartilage subjected to a mechanical injury from a panel of 10 candidate genes. Results: Ten candidate housekeeping genes were evaluated in three different treatment groups of mechanically impacted porcine articular cartilage. The genes evaluated were: beta actin, beta-2-microglobulin, glyceraldehyde-3-phosphate dehydrogenase, hydroxymethylbilane synthase, hypoxanthine phosphoribosyl transferase, peptidylprolyl isomerase A (cyclophilin A), ribosomal protein L4, succinate dehydrogenase flavoprotein subunit A, TATA box binding protein, and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein-zeta polypeptide. The stability of the genes was measured using geNorm, BestKeeper, and NormFinder software. The four most stable genes measured via geNorm were (most to least stable) succinate dehydrogenase flavoprotein, subunit A, peptidylprolyl isomerase A, glyceraldehyde-3-phosphate dehydrogenase, beta actin; the four most stable genes measured via BestKeeper were glyceraldehyde-3-phosphate dehydrogenase, peptidylprolyl isomerase A, beta actin, succinate dehydrogenase flavoprotein, subunit A; and the four most stable genes measured via NormFinder were peptidylprolyl isomerase A, succinate dehydrogenase flavoprotein, subunit A, glyceraldehyde-3-phosphate dehydrogenase, beta actin. Conclusions: BestKeeper, geNorm, and NormFinder all generated similar results for the most stable genes in porcine articular cartilage. The use of these appropriate reference genes will facilitate accurate gene expression studies of porcine articular cartilage and suggest appropriate housekeeping genes for articular cartilage studies in other species.     

Identification of stable normalization genes for quantitative real-time PCR in porcine articular cartilage

Background

Expression levels for genes of interest must be normalized with an appropriate reference, or housekeeping gene, to make accurate comparisons of quantitative real-time PCR results. The purpose of this study was to identify the most stable housekeeping genes in porcine articular cartilage subjected to a mechanical injury from a panel of 10 candidate genes.

Results

Ten candidate housekeeping genes were evaluated in three different treatment groups of mechanically impacted porcine articular cartilage. The genes evaluated were: beta actin, beta-2-microglobulin, glyceraldehyde-3-phosphate dehydrogenase, hydroxymethylbilane synthase, hypoxanthine phosphoribosyl transferase, peptidylprolyl isomerase A (cyclophilin A), ribosomal protein L4, succinate dehydrogenase flavoprotein subunit A, TATA box binding protein, and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein—zeta polypeptide. The stability of the genes was measured using geNorm, BestKeeper, and NormFinder software. The four most stable genes measured via geNorm were (most to least stable) succinate dehydrogenase flavoprotein, subunit A, peptidylprolyl isomerase A, glyceraldehyde-3-phosphate dehydrogenase, beta actin; the four most stable genes measured via BestKeeper were glyceraldehyde-3-phosphate dehydrogenase, peptidylprolyl isomerase A, beta actin, succinate dehydrogenase flavoprotein, subunit A; and the four most stable genes measured via NormFinder were peptidylprolyl isomerase A, succinate dehydrogenase flavoprotein, subunit A, glyceraldehyde-3-phosphate dehydrogenase, beta actin.

Conclusions

BestKeeper, geNorm, and NormFinder all generated similar results for the most stable genes in porcine articular cartilage. The use of these appropriate reference genes will facilitate accurate gene expression studies of porcine articular cartilage and suggest appropriate housekeeping genes for articular cartilage studies in other species.     

鲟鱼硫酸软骨素的制备工艺研究

以史氏鲟鱼软骨为原料,按碱提-酶解-乙醇沉淀工艺流程进行提取硫酸软骨素的各主要工序工艺的研究.通过单因素试验及正交试验,得出最佳碱提工艺为以两倍量6%的NaOH为溶剂,于40℃的水浴中搅拌提取4 h.通过正交试验,得出最佳酶解工艺条件是先将浸提液调节pH至8.6,加入胰酶0.5%,50℃保温水解3 h;后降温至40℃,调pH至5.7,加入胃蛋白酶0.3%,保温水解2h.通过单因素试验,得出最佳的乙醇沉淀工艺条件是将滤液pH调为6.5,加入95%的乙醇至醇沉浓度为70%,静置过夜.制备的硫酸软骨素经检测符合WS1-C3-0030-2000(硫酸软骨素口服片)和WS-10001-(HD-0892)-2002(硫酸软骨素注射品)标准的要求.     

丝素组织工程支架的研究进展

随着组织工程学的兴起,丝素组织工程支架也已成为丝素生物材料研究的热点,以丝素为材料的多种形态的组织工程支架已用于各种细胞组织构建的研究。主要介绍丝素组织工程支架的制备,以及丝素组织工程支架用于骨、软骨、韧带和肌腱等细胞组织构建的研究进展,并展望丝素作为组织工程支架材料的研究发展趋势,以期为丝素组织工程支架的深入研究提供参考。     

The Effect of Glucosamine and Chondroitin on Stressed Equine Cartilage Explants

Effective prevention and treatment of osteoarthritis for horses is still needed. This research tests the ability of glucosamine and chondroitin sulfate (GLN/CS) to mitigate inflammatory and mechanical stress in vitro. In this study, GLN/CS mediate this effect by a decrease of the synthesis of nitric oxide (NO) and a decrease of proteoglycan release from the extracellular matrix in stressed cartilage explants. Explants were cultured with interleukin-1 (IL-1) + mechanical trauma with and without GLN/CS. NO and prostaglandin E2 were measured as indicators of an inflammatory response. Glycosaminoglycans were measured as an indicator of cartilage breakdown. NO levels in the stressed explants with GLN/CS treatment were lower than the IL-1 + mechanical impact treatment alone and did not differ from control group. The glycosaminoglycan release was also lower in the GLN/CS treatment than the IL-1 + mechanical impact treatment, although the prostaglandin E2 concentration was not affected. This study offers some evidence that GLN/CS treatment can partially mitigate the catabolic response to inflammatory stress and mechanical trauma in equine cartilage explants. These results provide additional support for the continued study on the benefit of GLN/CS for horses with cartilage degeneration.     

Minimally Invasive Surgical Pastern Arthrodesis: An Ex Vivo Study Comparing Three Different Drilling Techniques for Cartilage Destruction

The objective was to compare the degree of cartilage destruction obtained with three different drilling techniques for minimally invasive arthrodesis of the equine pastern joint. Three drilling techniques were randomly tested on 36 cadaver distal limbs (six front and six hind limbs in each group). The basic drilling pattern consisted of several passes in a dorsopalmar and/or plantar direction across the joint space (group 1) and was either supplemented with one additional pass in a lateromedial and/or mediolateral direction (group 2) or two additional passes in a distodorsal–proximopalmar and/or plantar direction (group 3). After drilling, the pastern joints were disarticulated, the articular surfaces of P1 and P2 were digitally photographed, and the area of removed cartilage was measured using planimetry. The mean percentage of cartilage removed in the entire pastern joint was significantly lower in group 1 (34.1 ± 4.0%) compared with groups 2 and 3 (45.0 ± 5.2% and 43.0 ± 4.0%, respectively; P < .001). There was significantly more cartilage removed in the hind (47.1 ± 4.4%) versus the forelimbs (42.0 ± 5.0%) of group 2 (P = .003), whereas in group 3, there was significantly more cartilage removed in the forelimbs (44.6 ± 3.0%) compared with the hind limbs (40.6 ± 3.0%) (P = .039). The technique of group 2 gave significantly more cartilage destruction compared with technique 1 while being practical to perform. Therefore, this technique seems to be the most promising for further evaluation in a clinical situation.