- Email: [email protected]
Influence of sodium hyaluronate on iNOS expression in synovium and NO content in synovial fluid of rabbits with traumatic osteoarthritis
. 293 .
Chinese Journal of Traumatology 2008; 11(5):293-296
Influence of sodium hyaluronate on iNOS expression in synovium and NO content in synovial fluid of rabbits with traumatic osteoarthritis QIU Bo 邱波 *, LIU Shi-qing 刘世清 and PENG Hao 彭昊 Objective: To observe the influence of intra-articular injection of sodium hyaluronate (SH) on the expression of inducible nitric oxide synthase (iNOS) in the synovium and nitric oxide (NO) content in synovial fluid of rabbits with traumatic osteoarthritis (OA). Methods: Sixteen white rabbits underwent unilateral anterior cruciate ligament transection and were randomly divided into 2 groups 5 weeks after the operation. Rabbits in the experimental group received intra-articular injection of 0.3 ml of 1% SH, once a week for 5 weeks. Animals in the control group were treated under the same conditions using physiological saline. All the animals were sacrificed at the 10th week after surgery. The mRNA expression of iNOS in the synovium was analyzed using reverse transcriptionpolymerase chain reaction. The content of NO in the synovial fluid was assayed.
Results: The level of iNOS expression of the synovium in the experimental group was lower than that in control group (0.47 ± 0.09 vs. 0.65 ± 0.12, t =3.45, P<0.01). Compared with control group, the content of NO decreased significantly in synovial fluid of SH injection group (134.11 µmol/L ± 12.47 µmol/L vs. 152.17 µmol/L ± 15.69 µmol/L, t =2.55, P<0.05). Conclusions: SH significantly decreases the content of NO in the synovial fluid of rabbits with traumatic OA. SH may exert the effect on synovial fluid NO level as a result of the suppression of iNOS expression in the synovium. It may be one of the mechanisms of the therapeutic effect of SH on early traumatic OA. Key words: Osteoarthritis; Nitric oxide; Synovial fluid Chin J Traumatol 2008; 11(5):293-296
O
steoarthritis (OA) is one of the most common arthropathy. Traumatic OA resulted from the injury or operation of the joints is commonly seen. It is recognized that nitric oxide (NO) generated enzymatically by nitric oxide synthase (NOS) plays a vital role during the development of OA. Intra-articular injection of sodium hyaluronate (SH) has been reported to have a positive effect on the treatment of OA. 1-3 However, the mechanisms of such beneficial effect of SH on OA have not yet been fully established. To further clarify the effect of SH on OA, we investigated the mechanisms of SH on traumatic OA in this study. The expression of inducible NOS (iNOS) in the synovium and NO content in synovial fluid were assessed in experimentally induced traumatic OA rabbits with or without intra-articular injection of SH. Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan 430060, China (Qiu B, Liu SQ and Peng H) *Corresponding author: Tel: 86-27-88041911 ext 2263, E-mail: [email protected] This work was supported by the project of Science and Technology of Hubei Province (2003AA301C11).
METHODS Experimental animals Sixteen white rabbits weighing 2.4-2.8 kg were used in this study. All animals provided by the Experimental Animal Centre of Medical College of Wuhan University were anesthetized intravenously with ketamine hydrochloride (1.0 mg/kg). The animals received unilateral anterior cruciate ligament transection (ACLT). Rabbits were divided into 2 groups randomly 5 weeks after operation. Each group had 8 rabbits. Experimental group received 0.3 ml of 1% SH by intra-articular injection, once a week for 5 weeks. The control group was treated under the same condition using physiological saline. After surgery, the animals were housed individually in stainless-steel cages without any immobilization and maintained under the same environmental condition. All animals were killed 10 weeks after surgery. Reverse transcription-polymerase chain reaction (RT-PCR) assay Primers used in this study were synthesized by
. 294 .
Shanghai Sangon Biological Engineering Technology and Service Company. Primers used for rabbit glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and iNOS were as follows. GAPDH (444bp)4 sense, 5’- ATC ACT GCC ACC CAG AAG AC -3’antisense, 5’- ATG AGG TCC ACC ACC CTG TT -3’. iNOS (262bp)5 sense, 5’-CGC CCT TCC GCA GTT CT-3’; antisense, 5’-TCC AGG AGG ACA TGC AGC AC-3’. The synovium closed to the position of degenerative cartilage was harvested. The synovium tissue was powdered in liquid nitrogen by hand milling. Total RNA extraction was performed according to the instruction of Trizol Reagent (Invitrogen Co.USA). Reverse transcription and polymerase chain reaction were undertaken according to the literature described previously.6 Amplification consisted of 45 seconds at 95°C, 45 seconds at 57°C (GAPDH) and 65°C (iNOS) for annealing, 45 sec at 72°C for extension. Thirty cycles and 35 cycles of amplification were performed for GAPDH and iNOS respectively. Electrophoresis of the PCR products on a 1.5% agarose gel with 0.5 pg/ml of ethidium bromide was performed to evaluate amplification and size of generated fragments. French VL analysis system was used to scan the RT-PCR agarose gel. GAPDH was used to verify that equal amount of RNA was added to the reaction. The band intensities of gene expression were reported as the ratio of iNOS to GAPDH on the expression quantities.
Chinese Journal of Traumatology 2008; 11(5):293-296
RESULTS In the synovium, iNOS mRNA expression of the experimental group (0.47 ± 0.09) was significantly lower than that of the control group (0.65 ± 0.12) (t=3.45, P=0.004), (Figs.1-2). A significant decrease of NO level in synovial fluid was detected in SH treated group compared with control group (134.11 ± 12.47) µmol/L vs. (152.17 ± 15.69) µmol/L, t=2.55, P=0.023).
M 1 2 3 Fig.1. An electrophoregram of GAPDH. M:PCR marker, from downside to upside, the size is 1543bp,994bp, 697bp, 515bp, 377bp and 237bp, respectively. 1: the synovium of experimental group. 2: the synovium of control group. 3: negative control.
Assay of NO content in the joint fluid NO is chemically active and can be converted into nitrite (NO2) and nitrate (NO3) quickly in vivo. So the total concentration of NO2 and NO3 can exactly indicate the level of NO. The synovial fluids were aspirated from the knee joints and stored at -80°C until analysis. The synovial fluid of the knee joint was collected and then analyzed according to the instruction of the NO detection kit (Nanjing Jiancheng Bioengineering Institute, China) for NO2 and NO3 determination. The method used nitrate reductase to determine the NO level of the synovial fluids. Statistical analysis The data were expressed as x ± s. Using a commercial software SPSS10.0, statistical analysis was performed by Student’s t test and statistical significance was defined as P<0.05.
M 1 2 3 Fig.2. An electrophoregram of iNOS shows the effect of SH on iNOS mRNA expression.
DISCUSSION NO is an inorganic, gaseous free radical. NO is produced in large amounts by chondrocytes and synoviocytes. Chondrocytes and synovium cells are known to produce a large amount of NO when they are
. 295 .
Chinese Journal of Traumatology 2008; 11(5):293-296
stimulated by proinflammatory cytokines. In contrast to normal cartilage, OA cartilage sponstaneously produces NO. A high-level of NO has been detected in the synovial joint fluid and serum of the patients with OA. Increasing evidence indicates that NO may contribute to the pathophysiology of OA. It has been demonstrated that high local concentration of NO may exert detrimental effects on chondrocyte functions including the inhibition of collagen and proteoglycan synthesis, the activity of metalloproteinases, the decreased expression of interleukin-1 (IL-1) receptor antagonist, the inhibition of chondrocyte proliferation, the induction of apoptosis, and so on. NO is involved in the pathogenesis of OA. Therefore, the role of NO located in the joints on pathogenesis of OA have attracted the attention of researchers. None of the synovium samples examined from normal joints demonstrated detectable amounts of iNOS.7 A high-level of iNOS expression in cartilages of OA has been detected. Large amounts of NO can be produced by iNOS catalysis. Strong iNOS expression is observed in the synovial lining layer and subsynovium.8 NO produced by the constitutive isoform of nitric oxide synthase (cNOS) is a key regulator of homeostasis, whereas the generation of NO by the iNOS plays an important role in inflammation, host defense response, and tissue repair. Cytokines such as IL-1 and tumor necrosis factor (TNF), which induce iNOS production in chondrocytes and synovium, have been implicated in the destruction of cartilage in OA. iNOS is calcium independent and, once activated, produces large amounts of NO. In the present study, iNOS was observed to be expressed strongly in the synovium and NO in the joint fluid was detected at high levels in the control group. The results indicate that NO is one of the mechanisms of degeneration of cartilage and plays an important role in the progression of OA. These findings make us thinking that NO in synovial fluid is a potent mediator in cartilage damage in OA.
the expression of MMP-3 and IL-1 beta in the synovium. Guarda et al9 have observed that injection of SH caused significant improvement in the main clinical symptoms. Among patients who received SH injection, those who reached a good outcome showed the lowest basal levels of NO. Kobayashi et al.10 have reported that the treatment of OA with intra-articular injection of SH led to a significant reduction of NO level in the joint fluid.10 Takahashi et al.11 reported that SH exerted the inhibitory effect on NO level in the synovial fluid as a result of the suppression of NO production in the menisci and synovium in patients with OA.11 SH does not show definite effect on iNOS expression in cartilage of OA.6 Our results showed that the level of iNOS expression in the synovia among rabbits treated with SH intra-articular injection was significantly lower than that in control group and SH had inhibitory effect on NO production of the synovium. We also observed that the level of NO decreased significantly in synovial fluid of SH injection group. It might be related to the down-regulation effect of SH on iNOS of the synovium. The present study provides the evidence that intraarticular injection of SH significantly decreases the NO level in synovial joint fluid in the ACLT rabbit OA model. Exogenous SH treatment has an inhibiting effect on iNOS expression in the synivium. We conclude that low NO levels in synovial fluid are related to the suppressive effects of SH on iNOS expression of the synovium. It may be one of the mechanisms of the therapeutic effect of SH on traumatic OA.
REFERENCES 1.Turajane T, Tanavaree A, Labpiboonpong V, et al. Outcomes of intra-articular injection of sodium hyaluronate for the treatment of osteoarthritis of the knee. J Med Assoc Thai 2007; 90(9):1845-1852. 2. Petrella RJ, Petrella M. A prospective, randomized, doubleblind, placebo controlled study to evaluate the efficacy of intraarticular hyaluronic acid for osteoarthritis of the knee. J
Intra-articular injection of SH has been used in OA treatment to prevent degeneration of articular cartilage. Several studies have reported that SH has beneficial effects on cartilage during OA development. Exogenous SH has been used as a lubricative agent, but it is also known to delay the degradation of cartilage by inhibiting the release of proteoglycan from articular cartilage. SH has anti-inflammatory effects and down-regulates
Rheumatol 2006;33(5):951-956. 3.Bjornland T, Gjaerum AA, Moystad A. Osteoarthritis of the temporomandibular joint: an evaluation of the effects and complications of corticosteroid injection compared with injection with sodium hyaluronate. J Oral Rehabil 2007;34(8):583-589. 4.Bluteau G, Conrozier T, Mathieu P, et al. Matrix metalloprotease-1, -3, -13 and aggrecanase-1and -2 are differentially expressed in experimental osteoarthritis. Biochim Biophys
. 296 .
Acta 2001;1526(2):147-158.
Chinese Journal of Traumatology 2008; 11(5):293-296
Rheumatol 1997;36(6):651-655.
5. Kobayashi K, Mishima H, Hashimoto S, et al. Chondro-
9. Guarda Nardini L, Oliviero F, Ramonda R, et al. Influence
cyte apoptosis and regional differential expression of nitric oxide
of intra-articular injections of sodium hyaluronate on clinical fea-
in the medial meniscus following partial meniscectomy. J Orthop
tures and synovial fluid nitric oxide levels of temporomandibular
Res 2001;19(5):802-808.
osteoarthritis. Reumatismo 2004;56(4):272-277.
6. Hao YR, Qiu B, Liu SQ et al. The effects of sodium
10. Kobayashi K, Matsuzaka S, Yoshida Y, et al. The effects
hyaluroante on inducible nitric oxide synthase mRNA expression
of intraarticularly injected sodium hyaluronate on levels of intact
in cartilage of rabbit osteoarthritis. Chin J Rheumatol 2005;9(7):
aggrecan and nitric oxide in the joint fluid of patients with knee
405-408.
osteoarthritis. Osteoarthritis Cartilage 2004;12(7):536-542.
7. Pei M, Qu MY, Yu CL. Induced nitric oxide synthase and
11.Takahashi K, Hashimoto S, Kubo T, et al. Hyaluronan
interleukin 1 receptor antagonist protein production and effect in
suppressed nitric oxide production in the meniscus and synovium
synovial and chondral tissue from patients with osteoarthritis.
of rabbit osteoarthritis model. J Orthop Res 2001;19(3):500-503.
Chin J Sports Med 2000;19(3):246-248. 8. Grabowski PS, Wright PK, Van't Hof RJ, et al.
(Received January 28, 2008)
Immunolocalization of inducible nitric oxide synthase in synovium
Edited by SONG Shuang-ming
and cartilage in rheumatoid arthritis and osteoarthritis. Br J
Chinese Journal of Traumatology 2008; 11(5):293-296
Influence of sodium hyaluronate on iNOS expression in synovium and NO content in synovial fluid of rabbits with traumatic osteoarthritis QIU Bo 邱波 *, LIU Shi-qing 刘世清 and PENG Hao 彭昊 Objective: To observe the influence of intra-articular injection of sodium hyaluronate (SH) on the expression of inducible nitric oxide synthase (iNOS) in the synovium and nitric oxide (NO) content in synovial fluid of rabbits with traumatic osteoarthritis (OA). Methods: Sixteen white rabbits underwent unilateral anterior cruciate ligament transection and were randomly divided into 2 groups 5 weeks after the operation. Rabbits in the experimental group received intra-articular injection of 0.3 ml of 1% SH, once a week for 5 weeks. Animals in the control group were treated under the same conditions using physiological saline. All the animals were sacrificed at the 10th week after surgery. The mRNA expression of iNOS in the synovium was analyzed using reverse transcriptionpolymerase chain reaction. The content of NO in the synovial fluid was assayed.
Results: The level of iNOS expression of the synovium in the experimental group was lower than that in control group (0.47 ± 0.09 vs. 0.65 ± 0.12, t =3.45, P<0.01). Compared with control group, the content of NO decreased significantly in synovial fluid of SH injection group (134.11 µmol/L ± 12.47 µmol/L vs. 152.17 µmol/L ± 15.69 µmol/L, t =2.55, P<0.05). Conclusions: SH significantly decreases the content of NO in the synovial fluid of rabbits with traumatic OA. SH may exert the effect on synovial fluid NO level as a result of the suppression of iNOS expression in the synovium. It may be one of the mechanisms of the therapeutic effect of SH on early traumatic OA. Key words: Osteoarthritis; Nitric oxide; Synovial fluid Chin J Traumatol 2008; 11(5):293-296
O
steoarthritis (OA) is one of the most common arthropathy. Traumatic OA resulted from the injury or operation of the joints is commonly seen. It is recognized that nitric oxide (NO) generated enzymatically by nitric oxide synthase (NOS) plays a vital role during the development of OA. Intra-articular injection of sodium hyaluronate (SH) has been reported to have a positive effect on the treatment of OA. 1-3 However, the mechanisms of such beneficial effect of SH on OA have not yet been fully established. To further clarify the effect of SH on OA, we investigated the mechanisms of SH on traumatic OA in this study. The expression of inducible NOS (iNOS) in the synovium and NO content in synovial fluid were assessed in experimentally induced traumatic OA rabbits with or without intra-articular injection of SH. Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan 430060, China (Qiu B, Liu SQ and Peng H) *Corresponding author: Tel: 86-27-88041911 ext 2263, E-mail: [email protected] This work was supported by the project of Science and Technology of Hubei Province (2003AA301C11).
METHODS Experimental animals Sixteen white rabbits weighing 2.4-2.8 kg were used in this study. All animals provided by the Experimental Animal Centre of Medical College of Wuhan University were anesthetized intravenously with ketamine hydrochloride (1.0 mg/kg). The animals received unilateral anterior cruciate ligament transection (ACLT). Rabbits were divided into 2 groups randomly 5 weeks after operation. Each group had 8 rabbits. Experimental group received 0.3 ml of 1% SH by intra-articular injection, once a week for 5 weeks. The control group was treated under the same condition using physiological saline. After surgery, the animals were housed individually in stainless-steel cages without any immobilization and maintained under the same environmental condition. All animals were killed 10 weeks after surgery. Reverse transcription-polymerase chain reaction (RT-PCR) assay Primers used in this study were synthesized by
. 294 .
Shanghai Sangon Biological Engineering Technology and Service Company. Primers used for rabbit glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and iNOS were as follows. GAPDH (444bp)4 sense, 5’- ATC ACT GCC ACC CAG AAG AC -3’antisense, 5’- ATG AGG TCC ACC ACC CTG TT -3’. iNOS (262bp)5 sense, 5’-CGC CCT TCC GCA GTT CT-3’; antisense, 5’-TCC AGG AGG ACA TGC AGC AC-3’. The synovium closed to the position of degenerative cartilage was harvested. The synovium tissue was powdered in liquid nitrogen by hand milling. Total RNA extraction was performed according to the instruction of Trizol Reagent (Invitrogen Co.USA). Reverse transcription and polymerase chain reaction were undertaken according to the literature described previously.6 Amplification consisted of 45 seconds at 95°C, 45 seconds at 57°C (GAPDH) and 65°C (iNOS) for annealing, 45 sec at 72°C for extension. Thirty cycles and 35 cycles of amplification were performed for GAPDH and iNOS respectively. Electrophoresis of the PCR products on a 1.5% agarose gel with 0.5 pg/ml of ethidium bromide was performed to evaluate amplification and size of generated fragments. French VL analysis system was used to scan the RT-PCR agarose gel. GAPDH was used to verify that equal amount of RNA was added to the reaction. The band intensities of gene expression were reported as the ratio of iNOS to GAPDH on the expression quantities.
Chinese Journal of Traumatology 2008; 11(5):293-296
RESULTS In the synovium, iNOS mRNA expression of the experimental group (0.47 ± 0.09) was significantly lower than that of the control group (0.65 ± 0.12) (t=3.45, P=0.004), (Figs.1-2). A significant decrease of NO level in synovial fluid was detected in SH treated group compared with control group (134.11 ± 12.47) µmol/L vs. (152.17 ± 15.69) µmol/L, t=2.55, P=0.023).
M 1 2 3 Fig.1. An electrophoregram of GAPDH. M:PCR marker, from downside to upside, the size is 1543bp,994bp, 697bp, 515bp, 377bp and 237bp, respectively. 1: the synovium of experimental group. 2: the synovium of control group. 3: negative control.
Assay of NO content in the joint fluid NO is chemically active and can be converted into nitrite (NO2) and nitrate (NO3) quickly in vivo. So the total concentration of NO2 and NO3 can exactly indicate the level of NO. The synovial fluids were aspirated from the knee joints and stored at -80°C until analysis. The synovial fluid of the knee joint was collected and then analyzed according to the instruction of the NO detection kit (Nanjing Jiancheng Bioengineering Institute, China) for NO2 and NO3 determination. The method used nitrate reductase to determine the NO level of the synovial fluids. Statistical analysis The data were expressed as x ± s. Using a commercial software SPSS10.0, statistical analysis was performed by Student’s t test and statistical significance was defined as P<0.05.
M 1 2 3 Fig.2. An electrophoregram of iNOS shows the effect of SH on iNOS mRNA expression.
DISCUSSION NO is an inorganic, gaseous free radical. NO is produced in large amounts by chondrocytes and synoviocytes. Chondrocytes and synovium cells are known to produce a large amount of NO when they are
. 295 .
Chinese Journal of Traumatology 2008; 11(5):293-296
stimulated by proinflammatory cytokines. In contrast to normal cartilage, OA cartilage sponstaneously produces NO. A high-level of NO has been detected in the synovial joint fluid and serum of the patients with OA. Increasing evidence indicates that NO may contribute to the pathophysiology of OA. It has been demonstrated that high local concentration of NO may exert detrimental effects on chondrocyte functions including the inhibition of collagen and proteoglycan synthesis, the activity of metalloproteinases, the decreased expression of interleukin-1 (IL-1) receptor antagonist, the inhibition of chondrocyte proliferation, the induction of apoptosis, and so on. NO is involved in the pathogenesis of OA. Therefore, the role of NO located in the joints on pathogenesis of OA have attracted the attention of researchers. None of the synovium samples examined from normal joints demonstrated detectable amounts of iNOS.7 A high-level of iNOS expression in cartilages of OA has been detected. Large amounts of NO can be produced by iNOS catalysis. Strong iNOS expression is observed in the synovial lining layer and subsynovium.8 NO produced by the constitutive isoform of nitric oxide synthase (cNOS) is a key regulator of homeostasis, whereas the generation of NO by the iNOS plays an important role in inflammation, host defense response, and tissue repair. Cytokines such as IL-1 and tumor necrosis factor (TNF), which induce iNOS production in chondrocytes and synovium, have been implicated in the destruction of cartilage in OA. iNOS is calcium independent and, once activated, produces large amounts of NO. In the present study, iNOS was observed to be expressed strongly in the synovium and NO in the joint fluid was detected at high levels in the control group. The results indicate that NO is one of the mechanisms of degeneration of cartilage and plays an important role in the progression of OA. These findings make us thinking that NO in synovial fluid is a potent mediator in cartilage damage in OA.
the expression of MMP-3 and IL-1 beta in the synovium. Guarda et al9 have observed that injection of SH caused significant improvement in the main clinical symptoms. Among patients who received SH injection, those who reached a good outcome showed the lowest basal levels of NO. Kobayashi et al.10 have reported that the treatment of OA with intra-articular injection of SH led to a significant reduction of NO level in the joint fluid.10 Takahashi et al.11 reported that SH exerted the inhibitory effect on NO level in the synovial fluid as a result of the suppression of NO production in the menisci and synovium in patients with OA.11 SH does not show definite effect on iNOS expression in cartilage of OA.6 Our results showed that the level of iNOS expression in the synovia among rabbits treated with SH intra-articular injection was significantly lower than that in control group and SH had inhibitory effect on NO production of the synovium. We also observed that the level of NO decreased significantly in synovial fluid of SH injection group. It might be related to the down-regulation effect of SH on iNOS of the synovium. The present study provides the evidence that intraarticular injection of SH significantly decreases the NO level in synovial joint fluid in the ACLT rabbit OA model. Exogenous SH treatment has an inhibiting effect on iNOS expression in the synivium. We conclude that low NO levels in synovial fluid are related to the suppressive effects of SH on iNOS expression of the synovium. It may be one of the mechanisms of the therapeutic effect of SH on traumatic OA.
REFERENCES 1.Turajane T, Tanavaree A, Labpiboonpong V, et al. Outcomes of intra-articular injection of sodium hyaluronate for the treatment of osteoarthritis of the knee. J Med Assoc Thai 2007; 90(9):1845-1852. 2. Petrella RJ, Petrella M. A prospective, randomized, doubleblind, placebo controlled study to evaluate the efficacy of intraarticular hyaluronic acid for osteoarthritis of the knee. J
Intra-articular injection of SH has been used in OA treatment to prevent degeneration of articular cartilage. Several studies have reported that SH has beneficial effects on cartilage during OA development. Exogenous SH has been used as a lubricative agent, but it is also known to delay the degradation of cartilage by inhibiting the release of proteoglycan from articular cartilage. SH has anti-inflammatory effects and down-regulates
Rheumatol 2006;33(5):951-956. 3.Bjornland T, Gjaerum AA, Moystad A. Osteoarthritis of the temporomandibular joint: an evaluation of the effects and complications of corticosteroid injection compared with injection with sodium hyaluronate. J Oral Rehabil 2007;34(8):583-589. 4.Bluteau G, Conrozier T, Mathieu P, et al. Matrix metalloprotease-1, -3, -13 and aggrecanase-1and -2 are differentially expressed in experimental osteoarthritis. Biochim Biophys
. 296 .
Acta 2001;1526(2):147-158.
Chinese Journal of Traumatology 2008; 11(5):293-296
Rheumatol 1997;36(6):651-655.
5. Kobayashi K, Mishima H, Hashimoto S, et al. Chondro-
9. Guarda Nardini L, Oliviero F, Ramonda R, et al. Influence
cyte apoptosis and regional differential expression of nitric oxide
of intra-articular injections of sodium hyaluronate on clinical fea-
in the medial meniscus following partial meniscectomy. J Orthop
tures and synovial fluid nitric oxide levels of temporomandibular
Res 2001;19(5):802-808.
osteoarthritis. Reumatismo 2004;56(4):272-277.
6. Hao YR, Qiu B, Liu SQ et al. The effects of sodium
10. Kobayashi K, Matsuzaka S, Yoshida Y, et al. The effects
hyaluroante on inducible nitric oxide synthase mRNA expression
of intraarticularly injected sodium hyaluronate on levels of intact
in cartilage of rabbit osteoarthritis. Chin J Rheumatol 2005;9(7):
aggrecan and nitric oxide in the joint fluid of patients with knee
405-408.
osteoarthritis. Osteoarthritis Cartilage 2004;12(7):536-542.
7. Pei M, Qu MY, Yu CL. Induced nitric oxide synthase and
11.Takahashi K, Hashimoto S, Kubo T, et al. Hyaluronan
interleukin 1 receptor antagonist protein production and effect in
suppressed nitric oxide production in the meniscus and synovium
synovial and chondral tissue from patients with osteoarthritis.
of rabbit osteoarthritis model. J Orthop Res 2001;19(3):500-503.
Chin J Sports Med 2000;19(3):246-248. 8. Grabowski PS, Wright PK, Van't Hof RJ, et al.
(Received January 28, 2008)
Immunolocalization of inducible nitric oxide synthase in synovium
Edited by SONG Shuang-ming
and cartilage in rheumatoid arthritis and osteoarthritis. Br J