碱性成纤维细胞生长因子缓释微球对兔膝骨关节炎治疗作用及其作用机制的研究
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摘要
目的:骨关节炎(osteoarthritis, OA)是一种以关节软骨损伤为特点的、原因不明的慢性退行性疾病。OA是全球分布最广、最为流行的疾病之一,严重影响了患者的日常和社会生活。OA的主要病理改变为关节软骨的损伤、退变及软骨下骨的硬化、增生、囊变,继而导致关节间隙狭窄。关节软骨是成熟的透明软骨没有血管和修复细胞,软骨细胞的分裂潜能及其修复能力非常有限,当关节软骨发生严重退行性变时进行治疗很困难。目前OA的保守治疗方法虽然很多,但是治疗效果都有限。对于严重OA的病例只能采用人工关节置换来治疗,然而关节置换术存在着诸如年龄、手术损伤大、费用高、长期预后、再次翻修等一系列问题。因此寻找有效的保守治疗骨关节炎的方法至关重要。碱性成纤维细胞生长因子(basic fibroblast growth factor, bFGF)是最有效的体外软骨细胞分裂素之一,具有加速软骨代谢,促进关节软骨再生的作用,但bFGF体内半衰期很短仅为3~5min,全身或局部应用的效果均不能满足临床上治疗需要。因此应用bFGF缓释制剂对长期维持损伤局部的有效药物浓度有重要意义。目前研究发现关节软骨组织中的基质金属蛋白酶-13(matrix metalloproteinases-13, MMP-13)、基质金属蛋白酶抑制剂-1(tissue inhibitors of metallomatrix-1, TIMP-1)可调控软骨细胞外基质的合成与降解,参与关节软骨的损伤过程。因此,本实验应用壳聚糖包被bFGF制作缓释微球以长时间维持关节内的有效药物浓度,并观察其抑制OA进展的作用及其对MMP-13、TIMP-1 mRNA表达的影响。探讨bFGF持续释放对实验性兔膝关节OA的治疗作用的部分作用机制。
方法:健康成年新西兰大白兔54只随机选取9只作为对照组,其余45只作为实验组给予实验干预。于兔双后腿膝关节腔内注射木瓜蛋白酶建立兔膝骨关节炎模型,将其随机分为5组,每组9只,分别为模型组、PBS微球组(PBS-M)、bFGF溶液组(bFGF-S)、10μg bFGF微球组(10-bFGF-M)及100μg bFGF微球组(100-bFGF-M)。用壳聚糖制成含PBS及含bFGF的缓释微球,干冻保存备用。分别于第3周及第6周两次于各治疗组动物关节腔内注射溶液或微球,于第9周处死动物后取材。肉眼观察兔软骨组织大体形态,采用印度Ink评分法比较各组动物大体软骨损伤程度;通过HE染色及PAS染色方法观察软骨组织病理改变,采用Mankin评分系统评价;应用实时定量RT-PCR检测关节软骨组织MMP-13、TIMP-1 mRNA表达变化。
结果:
1微球的外观形态及粒径分布
光学及电子显微镜观察到壳聚糖微球外观圆整,粒径分布均匀,大部分微球粒径分布在3~7μm范围内,平均粒径为4.69±0.06μm。
2各组关节软骨标本Ink评分结果:与对照组相比,模型组关节软骨损伤程度明显加重(3.38±0.65 vs. 0.15±0.37, P<0.05);PBS-M组、bFGF-S组与模型组相比,关节软骨损伤程度无明显减轻(3.08±0.64, 3.31±0.63 vs. 3.38±0.65, P值均>0.05),而10-bFGF-M组和100-bFGF-M组关节软骨损伤程度较模型组明显减轻(1.92±0.49, 1.31±0.48 vs. 3.38±0.65, P值均<0.05)。并且100-bFGF-M组与10-bFGF-M组相比关节软骨损伤程度明显减轻(1.31±0.48 vs. 1.92±0.49, P<0.05)。
3各组关节软骨标本镜下Mankin评分结果:与对照组相比,模型组关节软骨损伤程度明显加重(13.08±0.64 vs. 0.53±0.51, P<0.05);bFGF-S组、PBS-M组与模型组相比,关节软骨损伤程度无明显减轻(12.15±1.07, 12.85±0.80 vs. 13.08±0.64, P值均>0.05),而关节软骨损伤程度较模型组明显减轻(8.08±0.49, 5.85±0.69 vs. 13.08±0.64, P值均<0.05),100-bFGF-M组与10-bFGF-M组相比关节软骨损伤程度明显减轻(5.85±0.69 vs. 8.08±0.49, P<0.05)。
4关节软骨组织MMP-13、TIMP-1mRNA表达改变:与对照组相比,模型组MMP-13 mRNA表达明显上调, TIMP-1 mRNA表达明显下降(P值均<0.05)。PBS-M组、bFGF-S组MMP-13、TIMP-1mRNA表达与模型组比较无明显差异(P值均>0.05),10-bFGF-M组和100-bFGF-M组较模型组MMP-13 mRNA表达明显下降,TIMP-1 mRNA表达明显上调(P值均<0.05)。并且100-bFGF-M组较10-bFGF-M组MMP-13 mRNA表达明显下降,TIMP-1 mRNA表达明显上调(P值均<0.05)。
结论:
1关节腔内注射木瓜蛋白酶,可建立膝骨关节炎模型。用沉淀/凝聚法成功制备bFGF壳聚糖微球。
2壳聚糖包被bFGF与单纯bFGF溶液注射组相比,具有明显减轻兔膝骨关节炎软骨损伤的作用,提示本实验制备的bFGF缓释微球能维持bFGF相对稳定的关节腔内浓度,保证了软骨损伤的修复。
3兔膝骨关节炎模型组膝关节软骨组织MMP-13 mRNA表达明显增加,TIMP-1 mRNA表达明显下降。而通过bFGF缓释微球的治疗,可明显降低MMP-13 mRNA表达,增加TIMP-1 mRNA表达,推测bFGF缓释微球治疗骨关节炎的作用可能通过调节MMP-13/TIMP-1 mRNA表达实现。
Object: Osteoarthritis (OA) is a chronic, degenerative disorder of unknown cause characterized by gradual loss of articular cartilage. It is the most prevalent disease in our society, with a worldwide distribution. Since joint function is remarkably affected, daily living and social activities of patients with OA are restricted. OA process results in cystic degeneration of the bone surrounding the joint, with loss of cartilage and irregular, abnormal bone formation at the edges of the joint and narrowing of the joint space. The articular cartilage, which is the matured hyaline cartilage, has no vessels, and the cells for repair cannot be provided. There are limits to the division potential of the cartilage cells and their ability to be repaired. Therefore, it becomes difficult to treat OA when it progresses and the articular cartilage degenerates. At present, there are many conservative treatments for OA, but their clinical outcomes are limited. Advanced OA only be managed by artificial joint replacements, but there are problems concerning the degree of invasion, cost, and long-term prognosis. For these reasons, an effective conservative treatment to inhibit OA progression is needed. Basic fibroblast growth factor (bFGF) is regarded as one of the most potent mitogens for chondrocytes in vitro, which can accelerate cartilage metabolism or promote cartilage regeneration. The effect of bFGF cannot be maintained, even when it is locally injected, because its biologic half-life is short (3~5 minutes). Therefore, it is important to maintain the bFGF concentrations by using a sustained-release system of gelatin hydrogel microspheres. At present, many studies showed that the tissue inhibitors of metallomatrix-1 (TIMP-1) and tissue inhibitors of metallomatrix-1 (TIMP-1) could regulate the synthesis and degradation metabolism of extra-cellular matrix (ECM), and play important roles in the development the degradation of articular cartilage. So, in this experiment, bFGF was made into chitosan microspheres to maintain the bFGF local effective concentrations. To observe the inhibitory effects of bFGF in microspheres in treating OA in the rabbit knee and influence to express of TIMP-1 and MMP-13 mRNA. Our objective was to investigate the partial mechanism by intraarticular injections of bFGF in microspheres.
Methods: The experiment was performed on 54 adult New Zealand white rabbits. We used intra-articular injection caroid in rabbit knee to induce osteoarthritis models. These animals were divided into a model group and four therapeutic groups (PBS-M, bFGF-S, bFGF-10-M, bFGF-100-M) at random, at the same time we made a control group. Meanwhile, we used chitosan to make microsphere with PBS or bFGF impregnated. We injected the PBS-impregnated or bFGF impregnated microsphere or bFGF-liquid into rabbit knee at week 3 and 6, and killed the animals at week 9. Cartilage pathology was scored by Indian Ink system in naked eyes. And scored by Mankin system in HE or PAS staining. The expression of cartilage TIMP-1 and MMP-13 mRNA was monitored by real-time PCR.
Results:
1 Shape and size of Chitosan microspheres
It can be got that chitosan microspheres made from the optimal formula have good sphericity and narrow size distribution by optical microscopy and SEM photography. The size distribution was range from 3μm to 7μm. The average diameter was 4.69±0.06μm.
2 The result of Ink score in each group: Compared with control animals, the injury of model animals was dramatic (3.38±0.65 vs. 0.15±0.37, P<0.05), There were no differences between the model group and the therapeutic groups that used PBS-impregnated microsphere or bFGF-liquid at the injury of knee (3.08±0.64, 3.31±0.63 vs. 3.38±0.65, P>0.05). However, in bFGF-impregnated microsphere group the injury of cartilage decreased (1.92±0.49, 1.31±0.48 vs. 3.38±0.65, P<0.05). And compared with 10-bFGF-impregnated microsphere group, in 100-bFGF-impregnated microsphere group the injury of cartilage was notably decreased (1.31±0.48 vs. 1.92±0.49, P <0.05).
3 The result of Mankin score in each group: Compared with control animals, the injury of model animals was dramatic (13.08±0.64 vs. 0.53±0.51, P<0.05), There were no differences between the model group and the therapeutic groups that used PBS-impregnated microsphere or bFGF-liquid at the injury of knee (12.15±1.07, 12.85±0.80 vs. 13.08±0.64, P>0.05). However, in bFGF-impregnated microsphere group the injury of cartilage decreased (8.08±0.49, 5.85±0.69 vs. 13.08±0.64, P<0.05). And compared with 10-bFGF-impregnated micro- sphere group, in 100-bFGF-impregnated microsphere group the injury of cartilage was notably decreased (5.85±0.69 vs. 8.08±0.49, P<0.05).
4 Charges of cartilage TIMP-1、MMP-13 mRNA expression in each group: Compared with control group, the injury of model animals was dramatic, and the expression of TIMP-1 mRNA was decreased, meanwhile MMP-13 mRNA was increased (P<0.05). There were no differences between the model group and the therapeutic groups that used PBS-impregnated microsphere or bFGF-liquid at the expression of TIMP-1 and MMP-13 mRNA (P>0.05). However, in bFGF-impregnated microsphere group the expression of TIMP-1 mRNA was increased, and MMP-13 mRNA was notably decreased (P <0.05). And compared with 10-bFGF- impregnated microsphere group, in 100-bFGF-impregnated microsphere group the expression of TIMP-1 mRNA was increased, and MMP-13 mRNA was notably decreased (P <0.05).
Conclusion:
1 The intra-articular injection caroid can successfully induce osteoarthritis model in the rabbit knee. bFGF-impregnated microsphere is successfully made from Chitosan.
2 The bFGF can Sustained release to recovery the damage of cartilage of rabbit osteoarthritis only when it impregnated in microsphere.
3 Compared with control animals, the injury of model animals was dramatic, and the expression of TIMP-1 mRNA was decreased, meanwhile MMP-13 mRNA was increased. However, the injury of knee was released in the bFGF-impregnated microsphere group, the expression of TIMP-1 mRNA was increased, and MMP-13 mRNA was notably decreased. So the pathogenesis of this role may through up-regulated TIMP-1 mRNA and down-regulated MMP-13 mRNA.
方法:健康成年新西兰大白兔54只随机选取9只作为对照组,其余45只作为实验组给予实验干预。于兔双后腿膝关节腔内注射木瓜蛋白酶建立兔膝骨关节炎模型,将其随机分为5组,每组9只,分别为模型组、PBS微球组(PBS-M)、bFGF溶液组(bFGF-S)、10μg bFGF微球组(10-bFGF-M)及100μg bFGF微球组(100-bFGF-M)。用壳聚糖制成含PBS及含bFGF的缓释微球,干冻保存备用。分别于第3周及第6周两次于各治疗组动物关节腔内注射溶液或微球,于第9周处死动物后取材。肉眼观察兔软骨组织大体形态,采用印度Ink评分法比较各组动物大体软骨损伤程度;通过HE染色及PAS染色方法观察软骨组织病理改变,采用Mankin评分系统评价;应用实时定量RT-PCR检测关节软骨组织MMP-13、TIMP-1 mRNA表达变化。
结果:
1微球的外观形态及粒径分布
光学及电子显微镜观察到壳聚糖微球外观圆整,粒径分布均匀,大部分微球粒径分布在3~7μm范围内,平均粒径为4.69±0.06μm。
2各组关节软骨标本Ink评分结果:与对照组相比,模型组关节软骨损伤程度明显加重(3.38±0.65 vs. 0.15±0.37, P<0.05);PBS-M组、bFGF-S组与模型组相比,关节软骨损伤程度无明显减轻(3.08±0.64, 3.31±0.63 vs. 3.38±0.65, P值均>0.05),而10-bFGF-M组和100-bFGF-M组关节软骨损伤程度较模型组明显减轻(1.92±0.49, 1.31±0.48 vs. 3.38±0.65, P值均<0.05)。并且100-bFGF-M组与10-bFGF-M组相比关节软骨损伤程度明显减轻(1.31±0.48 vs. 1.92±0.49, P<0.05)。
3各组关节软骨标本镜下Mankin评分结果:与对照组相比,模型组关节软骨损伤程度明显加重(13.08±0.64 vs. 0.53±0.51, P<0.05);bFGF-S组、PBS-M组与模型组相比,关节软骨损伤程度无明显减轻(12.15±1.07, 12.85±0.80 vs. 13.08±0.64, P值均>0.05),而关节软骨损伤程度较模型组明显减轻(8.08±0.49, 5.85±0.69 vs. 13.08±0.64, P值均<0.05),100-bFGF-M组与10-bFGF-M组相比关节软骨损伤程度明显减轻(5.85±0.69 vs. 8.08±0.49, P<0.05)。
4关节软骨组织MMP-13、TIMP-1mRNA表达改变:与对照组相比,模型组MMP-13 mRNA表达明显上调, TIMP-1 mRNA表达明显下降(P值均<0.05)。PBS-M组、bFGF-S组MMP-13、TIMP-1mRNA表达与模型组比较无明显差异(P值均>0.05),10-bFGF-M组和100-bFGF-M组较模型组MMP-13 mRNA表达明显下降,TIMP-1 mRNA表达明显上调(P值均<0.05)。并且100-bFGF-M组较10-bFGF-M组MMP-13 mRNA表达明显下降,TIMP-1 mRNA表达明显上调(P值均<0.05)。
结论:
1关节腔内注射木瓜蛋白酶,可建立膝骨关节炎模型。用沉淀/凝聚法成功制备bFGF壳聚糖微球。
2壳聚糖包被bFGF与单纯bFGF溶液注射组相比,具有明显减轻兔膝骨关节炎软骨损伤的作用,提示本实验制备的bFGF缓释微球能维持bFGF相对稳定的关节腔内浓度,保证了软骨损伤的修复。
3兔膝骨关节炎模型组膝关节软骨组织MMP-13 mRNA表达明显增加,TIMP-1 mRNA表达明显下降。而通过bFGF缓释微球的治疗,可明显降低MMP-13 mRNA表达,增加TIMP-1 mRNA表达,推测bFGF缓释微球治疗骨关节炎的作用可能通过调节MMP-13/TIMP-1 mRNA表达实现。
Object: Osteoarthritis (OA) is a chronic, degenerative disorder of unknown cause characterized by gradual loss of articular cartilage. It is the most prevalent disease in our society, with a worldwide distribution. Since joint function is remarkably affected, daily living and social activities of patients with OA are restricted. OA process results in cystic degeneration of the bone surrounding the joint, with loss of cartilage and irregular, abnormal bone formation at the edges of the joint and narrowing of the joint space. The articular cartilage, which is the matured hyaline cartilage, has no vessels, and the cells for repair cannot be provided. There are limits to the division potential of the cartilage cells and their ability to be repaired. Therefore, it becomes difficult to treat OA when it progresses and the articular cartilage degenerates. At present, there are many conservative treatments for OA, but their clinical outcomes are limited. Advanced OA only be managed by artificial joint replacements, but there are problems concerning the degree of invasion, cost, and long-term prognosis. For these reasons, an effective conservative treatment to inhibit OA progression is needed. Basic fibroblast growth factor (bFGF) is regarded as one of the most potent mitogens for chondrocytes in vitro, which can accelerate cartilage metabolism or promote cartilage regeneration. The effect of bFGF cannot be maintained, even when it is locally injected, because its biologic half-life is short (3~5 minutes). Therefore, it is important to maintain the bFGF concentrations by using a sustained-release system of gelatin hydrogel microspheres. At present, many studies showed that the tissue inhibitors of metallomatrix-1 (TIMP-1) and tissue inhibitors of metallomatrix-1 (TIMP-1) could regulate the synthesis and degradation metabolism of extra-cellular matrix (ECM), and play important roles in the development the degradation of articular cartilage. So, in this experiment, bFGF was made into chitosan microspheres to maintain the bFGF local effective concentrations. To observe the inhibitory effects of bFGF in microspheres in treating OA in the rabbit knee and influence to express of TIMP-1 and MMP-13 mRNA. Our objective was to investigate the partial mechanism by intraarticular injections of bFGF in microspheres.
Methods: The experiment was performed on 54 adult New Zealand white rabbits. We used intra-articular injection caroid in rabbit knee to induce osteoarthritis models. These animals were divided into a model group and four therapeutic groups (PBS-M, bFGF-S, bFGF-10-M, bFGF-100-M) at random, at the same time we made a control group. Meanwhile, we used chitosan to make microsphere with PBS or bFGF impregnated. We injected the PBS-impregnated or bFGF impregnated microsphere or bFGF-liquid into rabbit knee at week 3 and 6, and killed the animals at week 9. Cartilage pathology was scored by Indian Ink system in naked eyes. And scored by Mankin system in HE or PAS staining. The expression of cartilage TIMP-1 and MMP-13 mRNA was monitored by real-time PCR.
Results:
1 Shape and size of Chitosan microspheres
It can be got that chitosan microspheres made from the optimal formula have good sphericity and narrow size distribution by optical microscopy and SEM photography. The size distribution was range from 3μm to 7μm. The average diameter was 4.69±0.06μm.
2 The result of Ink score in each group: Compared with control animals, the injury of model animals was dramatic (3.38±0.65 vs. 0.15±0.37, P<0.05), There were no differences between the model group and the therapeutic groups that used PBS-impregnated microsphere or bFGF-liquid at the injury of knee (3.08±0.64, 3.31±0.63 vs. 3.38±0.65, P>0.05). However, in bFGF-impregnated microsphere group the injury of cartilage decreased (1.92±0.49, 1.31±0.48 vs. 3.38±0.65, P<0.05). And compared with 10-bFGF-impregnated microsphere group, in 100-bFGF-impregnated microsphere group the injury of cartilage was notably decreased (1.31±0.48 vs. 1.92±0.49, P <0.05).
3 The result of Mankin score in each group: Compared with control animals, the injury of model animals was dramatic (13.08±0.64 vs. 0.53±0.51, P<0.05), There were no differences between the model group and the therapeutic groups that used PBS-impregnated microsphere or bFGF-liquid at the injury of knee (12.15±1.07, 12.85±0.80 vs. 13.08±0.64, P>0.05). However, in bFGF-impregnated microsphere group the injury of cartilage decreased (8.08±0.49, 5.85±0.69 vs. 13.08±0.64, P<0.05). And compared with 10-bFGF-impregnated micro- sphere group, in 100-bFGF-impregnated microsphere group the injury of cartilage was notably decreased (5.85±0.69 vs. 8.08±0.49, P<0.05).
4 Charges of cartilage TIMP-1、MMP-13 mRNA expression in each group: Compared with control group, the injury of model animals was dramatic, and the expression of TIMP-1 mRNA was decreased, meanwhile MMP-13 mRNA was increased (P<0.05). There were no differences between the model group and the therapeutic groups that used PBS-impregnated microsphere or bFGF-liquid at the expression of TIMP-1 and MMP-13 mRNA (P>0.05). However, in bFGF-impregnated microsphere group the expression of TIMP-1 mRNA was increased, and MMP-13 mRNA was notably decreased (P <0.05). And compared with 10-bFGF- impregnated microsphere group, in 100-bFGF-impregnated microsphere group the expression of TIMP-1 mRNA was increased, and MMP-13 mRNA was notably decreased (P <0.05).
Conclusion:
1 The intra-articular injection caroid can successfully induce osteoarthritis model in the rabbit knee. bFGF-impregnated microsphere is successfully made from Chitosan.
2 The bFGF can Sustained release to recovery the damage of cartilage of rabbit osteoarthritis only when it impregnated in microsphere.
3 Compared with control animals, the injury of model animals was dramatic, and the expression of TIMP-1 mRNA was decreased, meanwhile MMP-13 mRNA was increased. However, the injury of knee was released in the bFGF-impregnated microsphere group, the expression of TIMP-1 mRNA was increased, and MMP-13 mRNA was notably decreased. So the pathogenesis of this role may through up-regulated TIMP-1 mRNA and down-regulated MMP-13 mRNA.
引文
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9 Garcia-Molina JA, Vallespi-Miro G, Greco-Machado Y, et al. The role of fibroblast growth factor (FGF) and type beta transforming growth factor (TGF-beta 1 -beta 2 -beta 3) during rat craniofacial development. Bull Group Int Rech Sci Stomatol Odontol, 2003, 45(2-3): 66~78
10 Weisser J, Rahfoth B, Timmermann A, et al. Role of growth factors in rabbit articular cartilage repair by chondrocytes in agarose. Osteoarthritis Cartilage, 2001, 9 (Suppl A): S48~54
11 Kato Y, Iwamoto M, Koike T. Fibroblast growth factor stimulates colony formation of differentiated chondrocytes in soft agar. Cell Physiol, 1987, 133(3): 491~498
12 Yamamoto T, Wakitani S, Imoto K, et al. Fibroblast growth factor-2 promotes the repair of partial thickness defects of articular cartilage in immature rabbits but not in mature rabbits. Osteoarthritis Cartilage, 2004, 12(8): 636~641
13 Fujimoto E, Ochi M, Kato Y, Mochizuki Y, et al. Beneficial effect of basic fibroblast growth factor on the repair of full-thickness defects in rabbit articular cartilage. Arch Orthop Trauma Surg, 1999, 119(3-4): 139~145
14 Cuevas P, Burgos J, Baird A. Basic fibroblast growth factor (FGF) promotes cartilage repair in vivo. Biochem BiophysRes Commun, 1988, 156(2): 611~618
15 Sah RL, Chen AC, Grodzinsky AJ, et al. Differential effects of bFGF and IGF-I on matrix metabolism in calf and adult bovine cartilage explants. Arch Biochem Biophys 1994, 308(1): 137~147
16 Matsumura M, Kakishita H, Suzuki M, et al. Dexamethasone suppresses iNOS gene expression by inhibiting NF-kappaB in vascular smooth muscle cells. Life Sci, 2001, 69(9): 1067~1077
17 Varde NK, Pack DW. Microspheres for controlled release drug delivery. Expert Opin Biol Ther, 2004, 4(1): 35~51
18 Kato Y, Onishi H, Machida Y. Application of chitin and chitosan derivatives in the pharmaceutical field. Curr Pharm Biotechnol, 2003, 4(5): 303~309
19 Sinha VR, Singla AK, Wadhawan S, et al. Chitosan microspheres as a potential carrier for drugs. Int J Pharm, 2004, 274(1-2): 1~33
20 方华丰, 周宜开. 壳聚糖微球的研究进展. 国外医药?合成药生化药制剂分册, 1999, 20(5): 315~318
21 谢希, 高洁生. 骨关节炎动物模型研究进展. 医学综述, 2005, 11(1): 67~69
22 Kopp S, Mejersjo C, Clemenssson E. Induction of osteo- arthrosis in the guinea pig knee by papain. Oral Surg Oral Med Oral Pathol, 1983, 55 (3): 259~266
23 Marcelon G, Cros J, Guiraud R. Experimental model of osteoarthritis. Agents Actions, 1976, 6 (1~3): 191~194
24 Scheck M, Sakovich L. Degenerative joint disease of the canine hip: experimental production by multiple papain and prednisone injections. Clin Orthop, 1972, 86: 115~120
25 Chuma H, Mizuta H, Kudo S, et al. One day exposure to FGF-2 was sufficient for the regenerative repair of full- thickness defects of articular cartilage in rabbits. Osteo- arthritis Cartilage, 2004, 12(10): 834~842
26 Murphy G, Knauper V, Cowell S, et al. Evaluation of some newer matrix metalloproteinases. Ann NY Acad Sci, 1999, 878: 25~39
27 Nagase H, Woessner JF Jr. Matrix metalloproteinases. J Biol Chem, 1999, 274(31): 21491~21494
28 MurPhy G,KnauPer V,Cowell s, et al. Evaluation of some newer matrix metalloproteinases. Ann NY Aead Sei,1999; 878: 25~39
29 Kugler A. Matrix metalloproteinases and their inhibitors. Aticancer Res, 1999, 19(2c): 1589~1592
30 Marartel-Pelletier J, MeCollum R, Fujimoto N, et al. Excess of metalloproteases over tissue inhibitor of metalloprotease may contribute to cartilage degradation in osteoarthritis and rheumatoid arthritis. Lab Invest, 1994, 70(6): 807~815
31 Vincent T, Hermansson M, Bolton M, et al. Basic FGF mediates an immediate response of articular cartilage to mechanical injury. Proc Natl Acad Sci USA, 2002, 99(12): 8259~ 8264
32 Vincent TL, Hermansson MA, Hansen UN, et al. Basicfibroblast growth factor mediates transduction of mechanical signals when articular cartilage is loaded. Arthritis Rheum, 2004, 50(2): 526~533
1 Murray CJL, Lopez AD. The global burden of disease. Geneva: World Health Organisation, 1996
2 Reginster J-Y. The prevalence and burden of arthritis. Rheu- matology, 2002, 41(suppl 1): 3~6
3 Arthritis Research Campaign. Available at: http: //www.arc.org. uk/ about_arth/astats. htm
4 Felson DT, Zhang Y, Hannan MT, et al. The incidence and natural history of knee osteoarthritis in the elderly: the Framingham Osteoarthritis Study. Arthritis Rheum, 1995, 38(10): 1500~1505
5 Felson DT, Zhang Y. An update on the epidemiology of knee and hip osteoarthritis with a view to prevention. Arthritis Rheum, 1998, 41(8): 1343~1355
6 Oliveria SA, Felson DT, Reed JI, et al. Incidence of symptomatic hand, hip, and knee osteoarthritis among patients in a health maintenance organization. Arthritis Rheum, 1995, 38(8): 1134~1141
7 Cicuttini FM, spector T, Baker J. Risk factors for osteoarthritis in the tibiofemoral and patellofemoral joins of the knee. Rheum, 1997, 24(6): 1164~1167
8 McAlindon TE, Felson DT, Zhang Y, et al. Relation of dietaryintake and serum levels of vitamin D to progression of osteoarthritis of the knee among participants in the Framingham study. Ann Intern Med, 1996, 125(5): 353~359
9 Spector TD, Nandra D, Hart DJ, et al. Is hormone re- placement therapy protective for hand and knee osteoarthritis in women?: The Chingford Study. Ann Rheum Dis, 1997, 56(7): 432~434
10 Sowers MF, Lachance L. Vitamins and arthritis. The roles of vitamins A, C, D and E. Rheum Clin North Am, 1999, 25(2): 315~332
11 Brand C, Snaddon J, Bailey M, et al. Vitamin E is ineffective for symptomatic relief of knee osteoarthritis: a six month double blind, randomized, placebo controlled study. Ann Rheum Dis, 2001, 60(10): 946~949
12 Hunter DJ, March L, Sambrook PN. Knee osteoarthritis: the influence of environmental factors. Clin Exp Rheum, 2002, 20(1): 93~100
13 Loughlin J. Genome studies and linkage in primary osteo- arthritis. Rheum Dis Clin North Am, 2002, 28(1): 95~ 109
14 Goldring SR, Goldring MB. The role of cytokines in carti- lage matrix degeneration in osteoarthritis. Clinical Orthopae- dics Related Reserch. 2004, 427(Suppl): S27~36
15 Massicotte F, Fernandes JC, Martel-Pelletier J, et al. Modulation of insulin-like growth factor 1 levels in human osteoarthritic subchondral bone osteoblasts. Bone, 2006, 38 (3): 333~341
16 Kardel R, Ulfgren AK, Reinholt FP, et al. Inflammatory cell and cytokine patterns in patients with painful clicking and osteoarthritis in the temporomandibular joint. International Journal of Oral and Maxillofacial Surgery, 2003, 32(4): 390~ 396
17 ErtenliI, Kiraz S, Calguneri M. Synovial fluid cytokine levels in Behcet's disease. Clinical and Experimental Rheum, 2001, 19(5 Suppl 24): S37~41
18 Van den Berg WB. Uncoupling of inflammatory and destructive mechanisms in arthritis. Semin Arthritis Rheum, 2001, 30(5 Suppl 2): 7~16
19 Coffin JD, Florkiewicz RZ, Neumann J, et al. Abnormal bone growth and selective translational regulation in basic fibroblast growth factor (FGF-2) transgenic mice. Mol Biol Cell, 1995, 6(12): 1861~1873
20 Litinsky I, Paran D, Levartovsky D, et al. The effects of leflunomide on clinical parameters and serum levels of IL-6, IL-10, MMP-1 and MMP-3 in patients with resistant rheumatoid arthritis. Cytokine, 2006, 33(2): 106~110
21 Blanco FJ, Guitian R, Vazquez-Martul E de Toro FJ, Galdo FJ.Osteoarthritis chondrocytes die by apoptosis. A possible pathway for osteoarthritis pathology. Arthritis Rheum, 1998, 41(2): 284~289
22 Martin JA, Brown TD, Heiner AD, et al. Chondrocyte senescence, joint loading and osteoarthritis. Clin Orthop Relat Res, 2004, 427(Suppl): S96~103
23 Kuhn K, D’Lima DD, Hashimoto S, et al. Cell death in cartilage. Osteoarthritis Cartilage, 2004, 12(1): 1~16
24 Hill DJ, Logan A, De Sousa D. Stimulation of DNA and protein systhesis in epiphyseal growth plate chondrocytes by fibroblast growh factor. Interactions with other peptide growth factors. Ann NY Acad Sci, 1991, 638: 449~452
25 Avral X, Dougados M, Listrat V, et al. Arthroscopic evaluation of chondropathy in osteoarthritis of the knee. Rheum, 1996, 23(4): 698~706
26 Pelletier JP, Martel-Pelletier J, Abramson SB. Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis Rheum, 2001, 44(6): 1237~ 1247
27 Benito MJ, Veale DJ, FitzGerald O, et al. Synovial tissue inflammation in early and late osteoarthritis. Ann Rheum Dis, 2005, 64(9): 1263~1267
28 van’t Hof RJ, Ralston SH. Nitric oxide and bone. Immunology, 2001, 103(3): 255~261
29 Bettica P, Cline G, Hart DJ, et al. Evidence for increased bone resorption in patients with progressive knee osteoarthritis. Longitudinal results from the Chingford study. Arthritis Rheum, 2002, 46(12): 3178~3184
30 Thomas KS, Muir KR, Doherty M, et al. Home based exercise programme for knee pain and knee osteoarthritis: randomised controlled trial. BMJ, 2002, 325(7567): 752~ 756
31 Halbert J, Crotty M, Weller D, et al. Primary care-based physical activity programs: effectiveness in sedentary older patients with osteoarthritis symptoms. Arthritis Care and Research, 2001, 45(3): 228~234
32 Messier SP, Loeser RF, Mitchell MN, et al. Exercise and weight loss in obese older adults with knee osteoarthritis: a preliminary study. Am Geriatric Soc, 2000, 48(9): 1062~ 1072
33 Eccles M, Freemantle N, Mason J, et al. North of England evidence based guideline development project: summary guideline for non-steroidal anti-inflammatory drugs versus basic analgesia in treating the pain of degenerative arthritis. BMJ, 1998, 317(7157): 526~530
34 Smalley WE, Griffin MR. The risks and costs of upper gas- trointestinal disease attributable to NSAIDs. Gastroenterol Clin North Am, 1996, 25(2): 373~396
35 Waddell DD. Osteoarthritis: new insights. Ann Intern Med, 2002, 136(1): 86~88
36 Pelletier JP, Yaron M, Haraoui B, et al. Efficacy and safety of diacerein in osteoarthritis of the knee: a double-blind, placebo-controlled trial. The Diacerein Study Group. Arthritis Rheum, 2000, 43(10): 2339~ 22348
37 Takahashi K, Hashimoto S, Kubo T, et al. Effect of hyal- uronan on chondrocyte apoptosis and nitric oxide production in experimentally induced osteoarthritis. Rheum, 2000, 27(7): 1713~1720
38 Frisbie DD, Ghivizzani SC, Robbins PD, et al. Treatment of experimental equine osteaoarthritis by in vivo delivery of the equine interleukin-1 receptor antagonist gene. Gene Ther, 2002, 9(1): 12~20
39 Chemajovsky Y, Adams G, Podhajcer OL, et al. Inhibition of transfer of collagen-induced arthritis into SCID mice by ex vivo infection of spleen cells with retroviruses expressing soluble tumor necrosis factor receptor. Gene Trer, 1995, 2(10): 731~732
40 Ikeda T, Kubo T, Arai Y, et al. Adenovirus mediated gene delivery to the joints of guinea pigs. Rheum, 1998, 25(9): 1666~1673
41 Kang R, Marui T, Ghivizzani SC, et al. Ex vivo gene transfer to chondrocytes in full-thickness articular cartilage defects: A feasibility study. Osteoarthritis Cartilage, 1997, 5(2): 139~ 143
42 Doherty PJ, Zhang H, Manolopoulos V, et al. Adhesion of transplanted chondrocytes onto cartilage in vitro and in vivo. Rheum, 2000, 27(7): 1725~1731
43 Oligino T, Ghivzzani S, Wolfe D, et al. Intraarticular delivery of a herpes simplex virus IL-1R a gene vector transduces inflammation in a rabbit model of arthritis. Gene Ther, 1999, 6(10): 1713~1720
44 Tomita T, Hashimoto H, Tomita H, et al. In vivo direct gene transfer into articular cartilage by intraarticular injection mediated by HVJ (sendai virus) and liposomes. ArthritisRheum, 1997, 40(5): 901~906
45 Frisbie DD, Mc wraith CW. Evaluation of gene therapy as a treatment for equine traumatic arthritis and osteoarthritis. Clin Orthop, 2000, 379 (Suppl): S273~287
46 National Institute for Clinical Excellence. Technology appraisal guidance No 16. Guidance on the use of autologous cartilage transplantation for full thickness cartilage defects in knee joints. London: NICE, December 2000
47 Schmalzried TP, Callaghan JJ. Wear in total hip and knee replacements. Bone Joint Surg Am, 1999, 81(1): 115~136.
2 陆彬, 药物新剂型与新技术. 北京: 人民卫生出版社, 1998: 221
3 Reginster JY, Deroisy R, Rovati LC, et al. Long-term effects of glucosamine sulphate on osteoarthritis progression: a randomised, placebo-controlled clinical trial. Lancet, 2001, 357(9252): 251~256
4 Pavelka K, Gatterova J, Olejarova M, et al. Glucosamine sulfate use and delay of progression of knee osteoarthritis: a
3-year, randomized, placebo-controlled, double-blind study. Arch Intern Med, 2002, 162(18): 2113~2123
5 Yamashita F, Sakakida K, Suzu F, et al. The transplantation of an autogeneic osteochondral fragment for osteochondritis dissecans of the knee. Clin Orthop Relat Res, 1985, 201: 43~50
6 Brittberg M, Lindahl A, Nilsson A, et al. Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med, 1994, 331(14): 889~895
7 Nakao K, Itoh M, Tomita Y, et al. FGF-2 potently induces both proliferation and DSP expression in collagen type-1 gel cultures of adult incisor immature pulp cells. BiochemBiophys Res Commun, 2004, 325(3): 1052~1059
8 Diez del Corral R, Storey KG. Opposing FGF and retinoid pathways: a signalling switchthat controls differentiation and patterning onset in the extending vertebrate body axis. Bioessays, 2004, 26(8): 857~869
9 Garcia-Molina JA, Vallespi-Miro G, Greco-Machado Y, et al. The role of fibroblast growth factor (FGF) and type beta transforming growth factor (TGF-beta 1 -beta 2 -beta 3) during rat craniofacial development. Bull Group Int Rech Sci Stomatol Odontol, 2003, 45(2-3): 66~78
10 Weisser J, Rahfoth B, Timmermann A, et al. Role of growth factors in rabbit articular cartilage repair by chondrocytes in agarose. Osteoarthritis Cartilage, 2001, 9 (Suppl A): S48~54
11 Kato Y, Iwamoto M, Koike T. Fibroblast growth factor stimulates colony formation of differentiated chondrocytes in soft agar. Cell Physiol, 1987, 133(3): 491~498
12 Yamamoto T, Wakitani S, Imoto K, et al. Fibroblast growth factor-2 promotes the repair of partial thickness defects of articular cartilage in immature rabbits but not in mature rabbits. Osteoarthritis Cartilage, 2004, 12(8): 636~641
13 Fujimoto E, Ochi M, Kato Y, Mochizuki Y, et al. Beneficial effect of basic fibroblast growth factor on the repair of full-thickness defects in rabbit articular cartilage. Arch Orthop Trauma Surg, 1999, 119(3-4): 139~145
14 Cuevas P, Burgos J, Baird A. Basic fibroblast growth factor (FGF) promotes cartilage repair in vivo. Biochem BiophysRes Commun, 1988, 156(2): 611~618
15 Sah RL, Chen AC, Grodzinsky AJ, et al. Differential effects of bFGF and IGF-I on matrix metabolism in calf and adult bovine cartilage explants. Arch Biochem Biophys 1994, 308(1): 137~147
16 Matsumura M, Kakishita H, Suzuki M, et al. Dexamethasone suppresses iNOS gene expression by inhibiting NF-kappaB in vascular smooth muscle cells. Life Sci, 2001, 69(9): 1067~1077
17 Varde NK, Pack DW. Microspheres for controlled release drug delivery. Expert Opin Biol Ther, 2004, 4(1): 35~51
18 Kato Y, Onishi H, Machida Y. Application of chitin and chitosan derivatives in the pharmaceutical field. Curr Pharm Biotechnol, 2003, 4(5): 303~309
19 Sinha VR, Singla AK, Wadhawan S, et al. Chitosan microspheres as a potential carrier for drugs. Int J Pharm, 2004, 274(1-2): 1~33
20 方华丰, 周宜开. 壳聚糖微球的研究进展. 国外医药?合成药生化药制剂分册, 1999, 20(5): 315~318
21 谢希, 高洁生. 骨关节炎动物模型研究进展. 医学综述, 2005, 11(1): 67~69
22 Kopp S, Mejersjo C, Clemenssson E. Induction of osteo- arthrosis in the guinea pig knee by papain. Oral Surg Oral Med Oral Pathol, 1983, 55 (3): 259~266
23 Marcelon G, Cros J, Guiraud R. Experimental model of osteoarthritis. Agents Actions, 1976, 6 (1~3): 191~194
24 Scheck M, Sakovich L. Degenerative joint disease of the canine hip: experimental production by multiple papain and prednisone injections. Clin Orthop, 1972, 86: 115~120
25 Chuma H, Mizuta H, Kudo S, et al. One day exposure to FGF-2 was sufficient for the regenerative repair of full- thickness defects of articular cartilage in rabbits. Osteo- arthritis Cartilage, 2004, 12(10): 834~842
26 Murphy G, Knauper V, Cowell S, et al. Evaluation of some newer matrix metalloproteinases. Ann NY Acad Sci, 1999, 878: 25~39
27 Nagase H, Woessner JF Jr. Matrix metalloproteinases. J Biol Chem, 1999, 274(31): 21491~21494
28 MurPhy G,KnauPer V,Cowell s, et al. Evaluation of some newer matrix metalloproteinases. Ann NY Aead Sei,1999; 878: 25~39
29 Kugler A. Matrix metalloproteinases and their inhibitors. Aticancer Res, 1999, 19(2c): 1589~1592
30 Marartel-Pelletier J, MeCollum R, Fujimoto N, et al. Excess of metalloproteases over tissue inhibitor of metalloprotease may contribute to cartilage degradation in osteoarthritis and rheumatoid arthritis. Lab Invest, 1994, 70(6): 807~815
31 Vincent T, Hermansson M, Bolton M, et al. Basic FGF mediates an immediate response of articular cartilage to mechanical injury. Proc Natl Acad Sci USA, 2002, 99(12): 8259~ 8264
32 Vincent TL, Hermansson MA, Hansen UN, et al. Basicfibroblast growth factor mediates transduction of mechanical signals when articular cartilage is loaded. Arthritis Rheum, 2004, 50(2): 526~533
1 Murray CJL, Lopez AD. The global burden of disease. Geneva: World Health Organisation, 1996
2 Reginster J-Y. The prevalence and burden of arthritis. Rheu- matology, 2002, 41(suppl 1): 3~6
3 Arthritis Research Campaign. Available at: http: //www.arc.org. uk/ about_arth/astats. htm
4 Felson DT, Zhang Y, Hannan MT, et al. The incidence and natural history of knee osteoarthritis in the elderly: the Framingham Osteoarthritis Study. Arthritis Rheum, 1995, 38(10): 1500~1505
5 Felson DT, Zhang Y. An update on the epidemiology of knee and hip osteoarthritis with a view to prevention. Arthritis Rheum, 1998, 41(8): 1343~1355
6 Oliveria SA, Felson DT, Reed JI, et al. Incidence of symptomatic hand, hip, and knee osteoarthritis among patients in a health maintenance organization. Arthritis Rheum, 1995, 38(8): 1134~1141
7 Cicuttini FM, spector T, Baker J. Risk factors for osteoarthritis in the tibiofemoral and patellofemoral joins of the knee. Rheum, 1997, 24(6): 1164~1167
8 McAlindon TE, Felson DT, Zhang Y, et al. Relation of dietaryintake and serum levels of vitamin D to progression of osteoarthritis of the knee among participants in the Framingham study. Ann Intern Med, 1996, 125(5): 353~359
9 Spector TD, Nandra D, Hart DJ, et al. Is hormone re- placement therapy protective for hand and knee osteoarthritis in women?: The Chingford Study. Ann Rheum Dis, 1997, 56(7): 432~434
10 Sowers MF, Lachance L. Vitamins and arthritis. The roles of vitamins A, C, D and E. Rheum Clin North Am, 1999, 25(2): 315~332
11 Brand C, Snaddon J, Bailey M, et al. Vitamin E is ineffective for symptomatic relief of knee osteoarthritis: a six month double blind, randomized, placebo controlled study. Ann Rheum Dis, 2001, 60(10): 946~949
12 Hunter DJ, March L, Sambrook PN. Knee osteoarthritis: the influence of environmental factors. Clin Exp Rheum, 2002, 20(1): 93~100
13 Loughlin J. Genome studies and linkage in primary osteo- arthritis. Rheum Dis Clin North Am, 2002, 28(1): 95~ 109
14 Goldring SR, Goldring MB. The role of cytokines in carti- lage matrix degeneration in osteoarthritis. Clinical Orthopae- dics Related Reserch. 2004, 427(Suppl): S27~36
15 Massicotte F, Fernandes JC, Martel-Pelletier J, et al. Modulation of insulin-like growth factor 1 levels in human osteoarthritic subchondral bone osteoblasts. Bone, 2006, 38 (3): 333~341
16 Kardel R, Ulfgren AK, Reinholt FP, et al. Inflammatory cell and cytokine patterns in patients with painful clicking and osteoarthritis in the temporomandibular joint. International Journal of Oral and Maxillofacial Surgery, 2003, 32(4): 390~ 396
17 ErtenliI, Kiraz S, Calguneri M. Synovial fluid cytokine levels in Behcet's disease. Clinical and Experimental Rheum, 2001, 19(5 Suppl 24): S37~41
18 Van den Berg WB. Uncoupling of inflammatory and destructive mechanisms in arthritis. Semin Arthritis Rheum, 2001, 30(5 Suppl 2): 7~16
19 Coffin JD, Florkiewicz RZ, Neumann J, et al. Abnormal bone growth and selective translational regulation in basic fibroblast growth factor (FGF-2) transgenic mice. Mol Biol Cell, 1995, 6(12): 1861~1873
20 Litinsky I, Paran D, Levartovsky D, et al. The effects of leflunomide on clinical parameters and serum levels of IL-6, IL-10, MMP-1 and MMP-3 in patients with resistant rheumatoid arthritis. Cytokine, 2006, 33(2): 106~110
21 Blanco FJ, Guitian R, Vazquez-Martul E de Toro FJ, Galdo FJ.Osteoarthritis chondrocytes die by apoptosis. A possible pathway for osteoarthritis pathology. Arthritis Rheum, 1998, 41(2): 284~289
22 Martin JA, Brown TD, Heiner AD, et al. Chondrocyte senescence, joint loading and osteoarthritis. Clin Orthop Relat Res, 2004, 427(Suppl): S96~103
23 Kuhn K, D’Lima DD, Hashimoto S, et al. Cell death in cartilage. Osteoarthritis Cartilage, 2004, 12(1): 1~16
24 Hill DJ, Logan A, De Sousa D. Stimulation of DNA and protein systhesis in epiphyseal growth plate chondrocytes by fibroblast growh factor. Interactions with other peptide growth factors. Ann NY Acad Sci, 1991, 638: 449~452
25 Avral X, Dougados M, Listrat V, et al. Arthroscopic evaluation of chondropathy in osteoarthritis of the knee. Rheum, 1996, 23(4): 698~706
26 Pelletier JP, Martel-Pelletier J, Abramson SB. Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis Rheum, 2001, 44(6): 1237~ 1247
27 Benito MJ, Veale DJ, FitzGerald O, et al. Synovial tissue inflammation in early and late osteoarthritis. Ann Rheum Dis, 2005, 64(9): 1263~1267
28 van’t Hof RJ, Ralston SH. Nitric oxide and bone. Immunology, 2001, 103(3): 255~261
29 Bettica P, Cline G, Hart DJ, et al. Evidence for increased bone resorption in patients with progressive knee osteoarthritis. Longitudinal results from the Chingford study. Arthritis Rheum, 2002, 46(12): 3178~3184
30 Thomas KS, Muir KR, Doherty M, et al. Home based exercise programme for knee pain and knee osteoarthritis: randomised controlled trial. BMJ, 2002, 325(7567): 752~ 756
31 Halbert J, Crotty M, Weller D, et al. Primary care-based physical activity programs: effectiveness in sedentary older patients with osteoarthritis symptoms. Arthritis Care and Research, 2001, 45(3): 228~234
32 Messier SP, Loeser RF, Mitchell MN, et al. Exercise and weight loss in obese older adults with knee osteoarthritis: a preliminary study. Am Geriatric Soc, 2000, 48(9): 1062~ 1072
33 Eccles M, Freemantle N, Mason J, et al. North of England evidence based guideline development project: summary guideline for non-steroidal anti-inflammatory drugs versus basic analgesia in treating the pain of degenerative arthritis. BMJ, 1998, 317(7157): 526~530
34 Smalley WE, Griffin MR. The risks and costs of upper gas- trointestinal disease attributable to NSAIDs. Gastroenterol Clin North Am, 1996, 25(2): 373~396
35 Waddell DD. Osteoarthritis: new insights. Ann Intern Med, 2002, 136(1): 86~88
36 Pelletier JP, Yaron M, Haraoui B, et al. Efficacy and safety of diacerein in osteoarthritis of the knee: a double-blind, placebo-controlled trial. The Diacerein Study Group. Arthritis Rheum, 2000, 43(10): 2339~ 22348
37 Takahashi K, Hashimoto S, Kubo T, et al. Effect of hyal- uronan on chondrocyte apoptosis and nitric oxide production in experimentally induced osteoarthritis. Rheum, 2000, 27(7): 1713~1720
38 Frisbie DD, Ghivizzani SC, Robbins PD, et al. Treatment of experimental equine osteaoarthritis by in vivo delivery of the equine interleukin-1 receptor antagonist gene. Gene Ther, 2002, 9(1): 12~20
39 Chemajovsky Y, Adams G, Podhajcer OL, et al. Inhibition of transfer of collagen-induced arthritis into SCID mice by ex vivo infection of spleen cells with retroviruses expressing soluble tumor necrosis factor receptor. Gene Trer, 1995, 2(10): 731~732
40 Ikeda T, Kubo T, Arai Y, et al. Adenovirus mediated gene delivery to the joints of guinea pigs. Rheum, 1998, 25(9): 1666~1673
41 Kang R, Marui T, Ghivizzani SC, et al. Ex vivo gene transfer to chondrocytes in full-thickness articular cartilage defects: A feasibility study. Osteoarthritis Cartilage, 1997, 5(2): 139~ 143
42 Doherty PJ, Zhang H, Manolopoulos V, et al. Adhesion of transplanted chondrocytes onto cartilage in vitro and in vivo. Rheum, 2000, 27(7): 1725~1731
43 Oligino T, Ghivzzani S, Wolfe D, et al. Intraarticular delivery of a herpes simplex virus IL-1R a gene vector transduces inflammation in a rabbit model of arthritis. Gene Ther, 1999, 6(10): 1713~1720
44 Tomita T, Hashimoto H, Tomita H, et al. In vivo direct gene transfer into articular cartilage by intraarticular injection mediated by HVJ (sendai virus) and liposomes. ArthritisRheum, 1997, 40(5): 901~906
45 Frisbie DD, Mc wraith CW. Evaluation of gene therapy as a treatment for equine traumatic arthritis and osteoarthritis. Clin Orthop, 2000, 379 (Suppl): S273~287
46 National Institute for Clinical Excellence. Technology appraisal guidance No 16. Guidance on the use of autologous cartilage transplantation for full thickness cartilage defects in knee joints. London: NICE, December 2000
47 Schmalzried TP, Callaghan JJ. Wear in total hip and knee replacements. Bone Joint Surg Am, 1999, 81(1): 115~136.