Objective To establish a method for quantitative detection of the sulfate glycosaminoglycans ( GAG) content in extracellular matrix of in vitro cultured chondrocytes so as to evaluate the biological characteristics of...Objective To establish a method for quantitative detection of the sulfate glycosaminoglycans ( GAG) content in extracellular matrix of in vitro cultured chondrocytes so as to evaluate the biological characteristics of epiphyseal, articular and rib chondrocytes. Methods Sulfate GAG content in extracellular matrix of three chondrocytes was measured by the modified dimethylmethylene blue (DMB) method. The changes of the toluidine blue (TB) stain of chondrocytes were observed by light microscope. Results Primary chondrocytes had the highest content of sulfate GAG in the extracellular matrix, ie, epiphyseal chondrocytes reached ( 70. 12 ± 7. 72 )μg/cm2, articular chondrocytes (92.00 ± 10.15) μg/cm2 and rib chondrocytes (80.61 ± 11. 40) μg/cm2, respectively. On the third pasage chondrocytes, epiphyceal chondrocytes decreased to (53.27 ± 9. 50 ) μg/cm2, articular chondrocytes to (63.88 ± 11.92) μg/cm2 and rib chondrocytes to (58.94 ±8.21) μg/cm2, respectively. The change of TB in every passage展开更多
Background:Tendinopathy alters the compositional properties of the Achilles tendon by increasing fluid and glycosaminoglycan content.It has been speculated that these changes may affect intratendinous pressure,but the...Background:Tendinopathy alters the compositional properties of the Achilles tendon by increasing fluid and glycosaminoglycan content.It has been speculated that these changes may affect intratendinous pressure,but the extent of this relationship remains unclear.Therefore,we aimed to investigate the impact of elevated fluid and glycosaminoglycan content on Achilles tendon intratendinous pressure and to determine whether hyaluronidase(HYAL) therapy can intervene in this potential relationship.Methods:Twenty paired fresh-frozen cadaveric Achilles tendons were mounted in a tensile-testing machine and loaded up to 5% strain.Intratendinous resting(at 0% strain) and dynamic pressure(at 5% strain) were assessed using the microcapillary infusion technique.First,intratendinous pressure was measured under native conditions before and after infusion of 2 mL physiological saline.Next,80 mg of glycosaminoglycans were administered bilaterally to the paired tendons.The right tendons were additionally treated with 1500 units of HYAL.Finally,both groups were retested,and the glycosaminoglycan content was analyzed.Results:It was found that both elevated fluid and glycosaminoglycan content resulted in higher intratendinous resting and dynamic pressures(p <0.001).HYAL treatment induced a 2.3-fold reduction in glycosaminoglycan content(p=0.002) and restored intratendinous pressures.Conclusion:The results of this study demonstrated that elevated fluid and glycosaminoglycan content in Achilles tendinopathy contribute to increased intratendinous re sting and dynamic pressures,which can be explained by the associated increased volume and reduced permeability of the tendon matrix,respectively.HYAL degrades glycosaminoglycans sufficiently to lower intratendinous pressures and may,therefore,serve as a promising treatment.展开更多
目的:探讨黄芪多糖对大鼠骨关节炎(OA)的影响。方法:4%木瓜蛋白酶关节腔注射建立大鼠OA模型,1周后50只大鼠分成5组:正常组、模型组、0.10,0.25,0.50 g/L的黄芪多糖治疗组,分组后即开始用不同浓度的黄芪多糖关节腔注射治疗,每3 d 1次,每...目的:探讨黄芪多糖对大鼠骨关节炎(OA)的影响。方法:4%木瓜蛋白酶关节腔注射建立大鼠OA模型,1周后50只大鼠分成5组:正常组、模型组、0.10,0.25,0.50 g/L的黄芪多糖治疗组,分组后即开始用不同浓度的黄芪多糖关节腔注射治疗,每3 d 1次,每次双膝各注射0.2 ml,对照组注射生理盐水。6周后处死动物,抽取血液、取关节滑膜检测超氧化物歧化酶(SOD)、丙二醛(MDA),切取关节,进行大体及组织学观察,用免疫组织化学方法检测基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶抑制剂-1(TIMP-1)在关节软骨中的表达。结果:黄芪多糖在高浓度可明显修复关节软骨的退变(P<0.01),降低关节滑膜、血清的MDA水平(均P<0.01),增加SOD活性(P<0.01);在低浓度时对关节软骨的修复作用并不明显(P>0.05),也不能显著降低关节滑膜、血清的MDA水平(均P>0.05)和增加SOD活性(P>0.05)。黄芪多糖可以抑制MMP-3在关节软骨中的表达(P<0.01),黄芪多糖在高浓度时可明显增加关节软骨中葡糖氨基聚糖(GAG)含量(P<0.01)。结论:黄芪多糖关节腔注射治疗大鼠OA可促进退变软骨的修复。展开更多
文摘Objective To establish a method for quantitative detection of the sulfate glycosaminoglycans ( GAG) content in extracellular matrix of in vitro cultured chondrocytes so as to evaluate the biological characteristics of epiphyseal, articular and rib chondrocytes. Methods Sulfate GAG content in extracellular matrix of three chondrocytes was measured by the modified dimethylmethylene blue (DMB) method. The changes of the toluidine blue (TB) stain of chondrocytes were observed by light microscope. Results Primary chondrocytes had the highest content of sulfate GAG in the extracellular matrix, ie, epiphyseal chondrocytes reached ( 70. 12 ± 7. 72 )μg/cm2, articular chondrocytes (92.00 ± 10.15) μg/cm2 and rib chondrocytes (80.61 ± 11. 40) μg/cm2, respectively. On the third pasage chondrocytes, epiphyceal chondrocytes decreased to (53.27 ± 9. 50 ) μg/cm2, articular chondrocytes to (63.88 ± 11.92) μg/cm2 and rib chondrocytes to (58.94 ±8.21) μg/cm2, respectively. The change of TB in every passage
基金funded by Ghent University Hospital(FIKO21/TYPE2/013)。
文摘Background:Tendinopathy alters the compositional properties of the Achilles tendon by increasing fluid and glycosaminoglycan content.It has been speculated that these changes may affect intratendinous pressure,but the extent of this relationship remains unclear.Therefore,we aimed to investigate the impact of elevated fluid and glycosaminoglycan content on Achilles tendon intratendinous pressure and to determine whether hyaluronidase(HYAL) therapy can intervene in this potential relationship.Methods:Twenty paired fresh-frozen cadaveric Achilles tendons were mounted in a tensile-testing machine and loaded up to 5% strain.Intratendinous resting(at 0% strain) and dynamic pressure(at 5% strain) were assessed using the microcapillary infusion technique.First,intratendinous pressure was measured under native conditions before and after infusion of 2 mL physiological saline.Next,80 mg of glycosaminoglycans were administered bilaterally to the paired tendons.The right tendons were additionally treated with 1500 units of HYAL.Finally,both groups were retested,and the glycosaminoglycan content was analyzed.Results:It was found that both elevated fluid and glycosaminoglycan content resulted in higher intratendinous resting and dynamic pressures(p <0.001).HYAL treatment induced a 2.3-fold reduction in glycosaminoglycan content(p=0.002) and restored intratendinous pressures.Conclusion:The results of this study demonstrated that elevated fluid and glycosaminoglycan content in Achilles tendinopathy contribute to increased intratendinous re sting and dynamic pressures,which can be explained by the associated increased volume and reduced permeability of the tendon matrix,respectively.HYAL degrades glycosaminoglycans sufficiently to lower intratendinous pressures and may,therefore,serve as a promising treatment.
文摘目的:探讨黄芪多糖对大鼠骨关节炎(OA)的影响。方法:4%木瓜蛋白酶关节腔注射建立大鼠OA模型,1周后50只大鼠分成5组:正常组、模型组、0.10,0.25,0.50 g/L的黄芪多糖治疗组,分组后即开始用不同浓度的黄芪多糖关节腔注射治疗,每3 d 1次,每次双膝各注射0.2 ml,对照组注射生理盐水。6周后处死动物,抽取血液、取关节滑膜检测超氧化物歧化酶(SOD)、丙二醛(MDA),切取关节,进行大体及组织学观察,用免疫组织化学方法检测基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶抑制剂-1(TIMP-1)在关节软骨中的表达。结果:黄芪多糖在高浓度可明显修复关节软骨的退变(P<0.01),降低关节滑膜、血清的MDA水平(均P<0.01),增加SOD活性(P<0.01);在低浓度时对关节软骨的修复作用并不明显(P>0.05),也不能显著降低关节滑膜、血清的MDA水平(均P>0.05)和增加SOD活性(P>0.05)。黄芪多糖可以抑制MMP-3在关节软骨中的表达(P<0.01),黄芪多糖在高浓度时可明显增加关节软骨中葡糖氨基聚糖(GAG)含量(P<0.01)。结论:黄芪多糖关节腔注射治疗大鼠OA可促进退变软骨的修复。