目的本研究旨在探索膝关节损伤后特定的透明软骨退化酶,如基质金属蛋白酶(MMP)、蛋白聚糖酶(Agg)的基因表达是否增加。方法试验纳入138例因关节组织如半月板、前交叉韧带(anterior cruciate ligament,ACL)、透明软骨损伤而行膝关节镜检...目的本研究旨在探索膝关节损伤后特定的透明软骨退化酶,如基质金属蛋白酶(MMP)、蛋白聚糖酶(Agg)的基因表达是否增加。方法试验纳入138例因关节组织如半月板、前交叉韧带(anterior cruciate ligament,ACL)、透明软骨损伤而行膝关节镜检查的患者,男57例,女81例,平均年龄38.8岁。检查前采集全血样本,检查时采集滑膜样本。利用RT-PCR和分光光度法处理样本。结果患者中,有56例ACL损伤,65例内侧半月板损伤,5例外侧半月板损伤,软骨损伤者根据Outerbridge标准进行分级。实验室检测显示,ACL撕裂和基因表达(MMP1、MMP2、MMP8、MMP9、MMP13、MMP14、Agg1、Agg2、IL1、TNFα和TIMP2,除外血浆TIPM1)显著相关:ACL患者滑膜中基因表达水平较非ACL患者滑膜中的对应基因表达水平显著上调:MMP1 0.920±0.068 vs 0.794±0.061,MMP2 1.075±0.053 vs 0.668±0.035,MMP8 0.951±0.047 vs0.766±0.045,MMP9 1.354±0.032 vs 0.947±0.042,MMP13 1.148±0.058 vs 0.991±0.058,MMP14 1.379±0.049vs 0.777±0.036,Agg1 1.309±0.071 vs 0.647±0.034,Agg2 1.043±0.072 vs 0.684±0.069,IL1 1.320±0.054 vs0.857±0.049,TNFα1.101±0.050 vs 0.802±0.039,TIMP1 1.197±0.060 vs 1.035±0.071,TIMP2 1.110±0.048vs 0.861±0.048(P值均<0.05)。除TIMP1外(1.092±0.053 vs 1.081±0.033,P=0.32),ACL患者血液中基因表达水平较非ACL患者血液中的对应基因表达水平显著上调:MMP1 1.163±0.030 vs 0.948±0.049,MMP20.918±0.056 vs 0.827±0.049,MMP8 1.172±0.029 vs 0.783±0.034,MMP9 1.304±0.059 vs 0.94±0.04,MMP131.155±0.060 vs 0.824±0.043,MMP14 1.095±0.058 vs 0.759±0.033,Agg1 1.270±0.077 vs 0.676±0.038,Agg21.244±0.037 vs 0.871±0.037,IL1 1.128±0.056 vs 0.912±0.045,TNFα1.018±0.076 vs 0.724±0.047,TIMP11.092±0.053 vs 1.081±0.033,TIMP2 0.906±0.056 vs 0.875±0.049(P值均<0.05)。所有基因在血浆和滑膜中的表达具有相关性(Agg1:r=0.89,MMP1:r=0.88,MMP2:r=0.87,P<0.001)。结论 ACL损伤引起蛋白酶基因表达增加,而其它损伤造成的改变尚不确切。�展开更多
The limited ability of cartilage tissue to repair itself poses a functionally impairing health problem. While many treatment methods are available, full restoration of the tissue to its original state is rare. Often, ...The limited ability of cartilage tissue to repair itself poses a functionally impairing health problem. While many treatment methods are available, full restoration of the tissue to its original state is rare. Often, complete joint replacement surgery is required to obtain long-term relief. Tissue engineering approaches, however, provide new opportunities for cartilage replacement. They seek to provide mechanisms to repair or replace lost tissue or function. A theoretical method is presented here for regenerating hyaline cartilage in vitro using a chondrocyte-seeded three-dimensional biomimetic engineered scaffold with mechanical properties similar to those occurring naturally. The scaffold composition, type II collagen, aggrecan, hyaluronan, hyaluronan binding protein (for link protein), and BMP-7, were chosen to encourage synthesis of hyaline cartilage by providing a more native environment and signaling cue for the seeded chondrocytes. The scaffold components mimic the macrofibrillar collagen network found in articular cartilage. Type II collagen provides tensile strength, and aggrecan, the predominant proteoglycan, provides compressive strength.展开更多
目的研究聚乙烯亚胺(polyethylenimine,PEI)包被的超顺磁性氧化铁粒子(superparamagnetic iron oxid,SPIO)对透明软骨细胞增殖和分泌Ⅱ型胶原蛋白功能的影响,探讨其标记软骨细胞的适宜标记浓度和适宜孵育时间。方法取1～3月龄巴马小香...目的研究聚乙烯亚胺(polyethylenimine,PEI)包被的超顺磁性氧化铁粒子(superparamagnetic iron oxid,SPIO)对透明软骨细胞增殖和分泌Ⅱ型胶原蛋白功能的影响,探讨其标记软骨细胞的适宜标记浓度和适宜孵育时间。方法取1～3月龄巴马小香猪后膝关节透明软骨体外培养的第2代细胞,以2、4、6、8、10、12、14μg/mL浓度的PEI-SPIO标记液标记的软骨细胞为实验组,没有PEI-SPIO标记的软骨细胞作为对照组,分别在37℃培养箱中培育6、12、18、24、30 h,通过普鲁士蓝染色、透射电镜、细胞内铁含量、WST-1和半定量RT-PCR法检测,筛选出PEI-SPIO的最佳标记浓度和最佳孵育时间。结果 6、8、10、12、14μg/mL组标记后的普鲁士蓝染色显示90%以上的细胞被铁粒子标记;细胞内铁含量的测定结果显示标记时间超过24 h,各组细胞内铁含量基本不再增加;WST-1检测磁标记的细胞毒性结果显示,对照组与实验组无统计学差异(P>0.05)。8μg/mL标记的软骨细胞,其Ⅱ型胶原蛋白RT-PCR条带的灰度值比值与未标记软骨细胞比较有明显降低[(13.04±0.60)%vs(18.89±1.26)%,P<0.05];6μg/mL标记的软骨细胞,其条带的灰度值比值与未标记软骨细胞比较无明显差异[(27.32±1.02)%vs(28.56±1.15)%,P>0.05]。结论 PEI-SPIO可达到安全有效标记软骨细胞的要求,PEI-SPIO标记液为6μg/mL,标记24 h是PEI-SPIO标记软骨细胞的适宜标记浓度和标记时间,既可达到标记软骨细胞的目的,又不对软骨细胞的活性、增殖和Ⅱ型胶原蛋白基因的表达造成影响。展开更多
文摘目的本研究旨在探索膝关节损伤后特定的透明软骨退化酶,如基质金属蛋白酶(MMP)、蛋白聚糖酶(Agg)的基因表达是否增加。方法试验纳入138例因关节组织如半月板、前交叉韧带(anterior cruciate ligament,ACL)、透明软骨损伤而行膝关节镜检查的患者,男57例,女81例,平均年龄38.8岁。检查前采集全血样本,检查时采集滑膜样本。利用RT-PCR和分光光度法处理样本。结果患者中,有56例ACL损伤,65例内侧半月板损伤,5例外侧半月板损伤,软骨损伤者根据Outerbridge标准进行分级。实验室检测显示,ACL撕裂和基因表达(MMP1、MMP2、MMP8、MMP9、MMP13、MMP14、Agg1、Agg2、IL1、TNFα和TIMP2,除外血浆TIPM1)显著相关:ACL患者滑膜中基因表达水平较非ACL患者滑膜中的对应基因表达水平显著上调:MMP1 0.920±0.068 vs 0.794±0.061,MMP2 1.075±0.053 vs 0.668±0.035,MMP8 0.951±0.047 vs0.766±0.045,MMP9 1.354±0.032 vs 0.947±0.042,MMP13 1.148±0.058 vs 0.991±0.058,MMP14 1.379±0.049vs 0.777±0.036,Agg1 1.309±0.071 vs 0.647±0.034,Agg2 1.043±0.072 vs 0.684±0.069,IL1 1.320±0.054 vs0.857±0.049,TNFα1.101±0.050 vs 0.802±0.039,TIMP1 1.197±0.060 vs 1.035±0.071,TIMP2 1.110±0.048vs 0.861±0.048(P值均<0.05)。除TIMP1外(1.092±0.053 vs 1.081±0.033,P=0.32),ACL患者血液中基因表达水平较非ACL患者血液中的对应基因表达水平显著上调:MMP1 1.163±0.030 vs 0.948±0.049,MMP20.918±0.056 vs 0.827±0.049,MMP8 1.172±0.029 vs 0.783±0.034,MMP9 1.304±0.059 vs 0.94±0.04,MMP131.155±0.060 vs 0.824±0.043,MMP14 1.095±0.058 vs 0.759±0.033,Agg1 1.270±0.077 vs 0.676±0.038,Agg21.244±0.037 vs 0.871±0.037,IL1 1.128±0.056 vs 0.912±0.045,TNFα1.018±0.076 vs 0.724±0.047,TIMP11.092±0.053 vs 1.081±0.033,TIMP2 0.906±0.056 vs 0.875±0.049(P值均<0.05)。所有基因在血浆和滑膜中的表达具有相关性(Agg1:r=0.89,MMP1:r=0.88,MMP2:r=0.87,P<0.001)。结论 ACL损伤引起蛋白酶基因表达增加,而其它损伤造成的改变尚不确切。�
基金supported by the National Key Research and Development Program of China(2018YFF0301100)the National Natural Science Foundation of China(82172420 and 82202723)。
文摘The limited ability of cartilage tissue to repair itself poses a functionally impairing health problem. While many treatment methods are available, full restoration of the tissue to its original state is rare. Often, complete joint replacement surgery is required to obtain long-term relief. Tissue engineering approaches, however, provide new opportunities for cartilage replacement. They seek to provide mechanisms to repair or replace lost tissue or function. A theoretical method is presented here for regenerating hyaline cartilage in vitro using a chondrocyte-seeded three-dimensional biomimetic engineered scaffold with mechanical properties similar to those occurring naturally. The scaffold composition, type II collagen, aggrecan, hyaluronan, hyaluronan binding protein (for link protein), and BMP-7, were chosen to encourage synthesis of hyaline cartilage by providing a more native environment and signaling cue for the seeded chondrocytes. The scaffold components mimic the macrofibrillar collagen network found in articular cartilage. Type II collagen provides tensile strength, and aggrecan, the predominant proteoglycan, provides compressive strength.
