OBJECTIVE: To investigate the efficacy and safety of oral bovine type II collagen (C II) in the treatment of rheumatoid arthritis (RA). METHODS: Forty-five patients with active RA were enrolled and randomized to receive placebo or oral C II for 3 months. Efficacy parameters were assessed monthly. Cumulative response rates (percentages of patients meeting the criteria for response at anytime during the study) were analyzed utilizing 3 set of composite criteria : Paulus criteria, ACR criteria for improvement in RA, and a requirement for > or = 30% reduction in both swollen and tender joint counts. RESULTS: The C II-treated group (n=25) showed significant higher response rate by the Paulus criteria compared to placebo group (n=20, p=0.04), and MHAQ scores between baseline and 3 months of treatment were also significantly decreased in the C II-treated group (p<0.05). However, there were no significant differences in tender and swollen joint count, and physician and patient global scores between C II-treated and placebo groups. Only one patient treated with C II had a urticaria 1 week after administration, but no serious side effects were found in the two groups. Patients treated with C II (n=15) showed the decreased levels of circulating IgG antibodies to bovine C II 3 months after treatment (p=0.02), whereas significant changes of IgG antibodies to C II were not found in placebo group (n=12). CONCLUSION: Oral administration of C II was safe and effective for the treatment of rheumatoid arthritis. The finding that serum IgG antibodies to bovine C II was decreased in patients who treated with C II suggest that autoimmune response to C II could be decreased by repetitive administration of C II.
Administration, Oral ; Antibodies ; Arthritis, Rheumatoid* ; Autoimmunity ; Collagen Type II* ; Humans ; Immunoglobulin G ; Joints ; Urticaria

OBJECTIVETo observe the therapeutic effect of acupoint application with Leima type II plaster on collagen-induced arthritis (CIA) in rats and probe its mechanism.

METHODSBovine type II collagen was injected intradermally into the middle line of the back to induce CIA model with 48 Wistar rats. Then the rats were randomly divided into a model control group (group A), a matrix control group (group B), acupoint application group with plaster of low concentration (group C) and high concentration plaster group (group D), 12 rats in each group. Group C and group D were treated with low and high concentration of Leima type II plaster, and "Shenzhu" (GV 12), "Zhiyang" (GV 9) and "Mingmen" (GV 4) were selected, each application for about 15 hours, once each day for 30 days. Group B was used the same method of acupoint application except using non-drug matrix plaster, and group A was not given any treatment. The morphous and the histopathological changes of affection joint were observed.

RESULTSThe paw edema volume after 30 days treatment in group C was significantly lower than that in group B (P < 0.01), and the anti-type II collagen antibody level after 15 days treatment in group C was significantly lower than that in group A (P < 0.05), and the synoviocytes proliferation of the knee joint in group C was significantly lower than that in group A and group B (both P < 0.01). The paw edema volume after 25 days treatment, arthritic index after 20 days treatment, pathological change of the paw and the synoviocytes proliferation of the knee joint in group D were significantly lower than those in group A and group B (P < 0.01, P < 0.05), and the anti-type II collagen antibody level after 15 days treatment in group D was significantly lower than that in group A (P < 0.05), and the paw edema volume and the arthritic index after 25 days treatment in group D were significantly lower than those in group C (P < 0.05, P < 0.01).

CONCLUSIONAcupoint application with Leima type II plaster has a good therapeutic effect on CIA rats and the protective mechanism is related to the reduction of anti-type II collagen antibody level so as to carry out anti-inflammatory effect and immunosuppression.

Acupuncture Points ; Animals ; Arthritis, Experimental ; therapy ; Collagen Type II ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; Male ; Rats ; Rats, Wistar

Oral administration of antigen has long been used in the induction of immune tolerance in various animal models of autoimmune diseases including rheumatoid arthritis (RA). Alleveation of arthritogenic symptoms has been reported from RA patients who received oral administration of type II collagen (CII) without side effects, however its rather inconsistent therapeutic efficacy and variation among patients calls for more detailed investigation on the mechanism of oral tolerance to be settled as regular treatment for RA. In an attempt to understand the immunogenic processes underpinning tolerance induction by orally administered CII, we analyzed changes in the expression of costimulatory molecules and STAT/SOCS signaling messengers in the mouse model of collagen induced arthritis (CIA). We found thatin the spleen of CIA mice, that has been undergone repeated oral feeding of CII prior to the induction of arthritis, showed increased promortion of CTLA4 expressing lymphocytes than in the spleen of PBS fed control. On the other hand, cells expressing CD28 or ICOS were decreased in the spleen of tolerized mice. Tolerance induction by oral CII administration also enhanced the expression of STAT6 in both RNA and protein level, while not affecting the expression of STAT3. The expression of SOCS3, which hasbeen known to transmit STAT-mediated signals from Th2 type cytokines, remained unchanged in the spleen of tolerized mice. Interestingly transcript of SOCS1, which has been associated with Th1 related pathways, was only visible in the spleen of tolerized but not of control mice, suggesting that as in the case of IL-6 signaling, it may exert a feed back inhibition toward the Th1 type stimulation.
Administration, Oral* ; Animals ; Arthritis* ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Collagen ; Collagen Type II* ; Cytokines ; Hand ; Humans ; Immune Tolerance ; Interleukin-6 ; Lymphocytes ; Mice* ; Models, Animal ; RNA ; Spleen

collagen contents of the articular cartilage using Masson trichrome stain. Safranin O–Fast Green revealed low proteoglycan contents. The collagen type II was also declined. The manikin score was 7.8. Risedronate improved this manifestation slightly more than glucosamine, but combination of booth drugs caused significant improvement of the damaged articular cartilage caused by immobilization. Oral administration of glucosamine and risedronate improved the degenerative changes of rat knee articular cartilage that follow immobilization. This improvement was more remarkable when both drugs were used in combination.]]>
Administration, Oral ; Adult ; Animals ; Cartilage, Articular ; Chondrocytes ; Collagen ; Collagen Type II ; Glucosamine ; Humans ; Immobilization ; Knee ; Male ; Manikins ; Models, Theoretical ; Osteoarthritis ; Proteoglycans ; Rats ; Risedronate Sodium

To detect the cellular immunity state of New Zealand white rabbit immunized by pig type II collagen. The New Zealand white rabbit was immunized by type II collagen for sixty days. The plasma was collected at a regular interval and the anti-type II collagen antibodies were examined. At the sixtieth day, the peripheral circular lymphocytes and the lymphocytes separated from spleen cells of rabbit and lymph nodes were collected and were stimulated by type II collagen in vivo again. The regulation of reactive cellular proliferation caused by the stimulation was detected. The experiment samples were divided into two groups. The first group was the positive control group by adding different concentrations of PHA and the non-specific immunity was assayed. The different concentrations of type II collagen were added to the second group and the specific immunity was assayed. The lymphocytes of normal rabbits showed proliferation by PHA stimulation but no proliferation by the first stimulation of type II collagen. Obvious proliferation due to the stimulation of both PHA and type II collagen in the immunized rabbit were observed. It shows that certain concentration of heterogeneous collagen may cause an increase of anti-type II collagen antibody in immunized rabbit and may cause a proliferation of lymphocytes in rabbit spleen and peripheral blood. The heterogeneous type II collagen causes cellular immunity in vivo.
Animals ; Cell Division ; drug effects ; Collagen Type II ; administration & dosage ; immunology ; Cytotoxicity, Immunologic ; Female ; Histocompatibility Antigens ; Lymphocytes ; cytology ; immunology ; Male ; Rabbits ; Spleen ; cytology ; Swine ; Transplantation, Heterologous

OBJECTIVETo repair cartilage defects at non-weight-bearing area of the femoral condyle in rabbits with invitro amplified cartilage cell using calcium alginate column scaffold combined with calcium alginate gel injection, and to study repair effects of combination with different form of the same material.

METHODSTwenty-four New Zealand rabbits were divided into 4 groups randomly. The wounds of rabbits in the Group 1 were repaired with injection of calcium alginate gel; the wounds of rabbits in the Group 2 were repaired with in planting of calcium alginate column scaffold; the wounds of rabbits in the Group 3 were repaired with in planting of calcium alginate column supporter firstly, and then injection of calcium alginate gel at the surrounding; and Group 4 is control group, the rabbits in the group were repair and without any support. The repair effects were demonstrated with Xij, and the effects of all animals were studied with statistical analysis.

RESULTSThe Xij scores of each rabbits were calculated, and the scores in four groups were compared. The statistical results showed that combination therapy was better than other methods (F = 69.0, P < 0.05).

CONCLUSIONThe calcium alginate with column shape has better shaping effects and certain mechanical strength. The calcium alginate gel has better stick nature and can be used to integrate artifical material with normal structure. They can be used together, which meeting the desire of repair and integration in cartilaginous tissue engineering.

Alginates ; administration & dosage ; Animals ; Cartilage, Articular ; pathology ; surgery ; Collagen Type II ; analysis ; Female ; Gels ; Glucuronic Acid ; administration & dosage ; Hexuronic Acids ; administration & dosage ; Immunohistochemistry ; Knee Joint ; pathology ; surgery ; Male ; Rabbits ; Tissue Engineering

Oral administration of antigen has long been considered as a promising alternative for the treatment of chronic autoimmune diseases including rheumatoid arthritis (RA), and oral application of type II collagen (CII) has been proven to improve pathogenic symptoms in RA patients without problematic side effects. To further current understandings about the immune suppression mechanisms mediated by orally administered antigens, we examined the changes in IgG subtypes, T-cell proliferative response, and proportion of interleukin (IL)-10 producing Th subsets in a time course study of collagen induced arthritis (CIA) animal models. We found that joint inflammation in CIA mouse peaked at 5 weeks after first immunization with CII, which was significantly subdued in mice pre-treated by repeated oral administration of CII. Orally tolerized mice also showed increase in their serum level of IgG1, while the level of IgG2a was decreased. T-cell proliferation upon CII stimulation was also suppressed in lymph nodes of mice given oral administration of CII compared to non-tolerized controls. When cultured in vitro in the presence of CII, T-cells isolated from orally tolerized mice presented higher proportion of CD4+ IL-10+ subsets compared to non-tolerized controls. Interestingly, such increase in IL-10 producing cells were obvious first in Peyer's patch, then by 5 weeks after immunization, in mesenteric lymph node and spleen instead. This result indicates that a particular subset of T-cells with immune suppressive functions might have migrated from the original contact site with CII to inflamed joints via peripheral blood after 5 weeks post immunization.
Administration, Oral ; Animals* ; Arthritis* ; Arthritis, Rheumatoid ; Autoimmune Diseases ; Collagen ; Collagen Type II ; Humans ; Immunity, Cellular* ; Immunization ; Immunoglobulin G ; Inflammation ; Interleukin-10 ; Interleukins ; Joints ; Lymph Nodes ; Mice ; Models, Animal* ; Spleen ; T-Lymphocytes

BACKGROUND/OBJECTIVES: The inhibitory effect of Hijikia fusiforme (HF) extracts on degenerative osteoarthritis was examined in primary cultured rat cartilage cells and a monosodium iodoacetate (MIA)-induced osteoarthritis rat model. MATERIALS/METHODS: In vitro, cell survival and the expression of matrix metalloproteinases (MMPs), collagen type I, collagen type II, aggrecan, and tissue inhibitor of metalloproteinases (TIMPs) was measured after H2O2 (800 µM, 2 hr) treatment in primary chondrocytes. In vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats, and then RH500, HFE250 and HFE500 were administered orally once a day for 28 days. To determine the anti-inflammatory effects of HFE, nitric oxide (NO), prostaglandin E2 (PGE2) expression were measured. In addition, real-time PCR was performed to measure the genetic expression of MMPs, collagen type I, collagen type II, aggrecan, and TIMPs. RESULTS: In the in vitro assay, cell survival after H2O2 treatment was increased by HFE extract (20% EtOH). In addition, anabolic factors (genetic expression of collagen type I, II, and aggrecan) were increased by HFE extract (20% EtOH). However, the genetic expression of MMP-3 and 7, known as catabolic factors were significantly inhibited by treatment with HFE extract (20% EtOH). In the in vivo assay, anabolic factors (genetic expression of collagen type I, II, aggrecan, and TIMPs) were increased by oral administration of HFE extract. However, the genetic expression of MMP-3 and 7, known as catabolic factors, and production of NO and PGE2 were significantly inhibited by treatment with oral administration of HFE extract. CONCLUSIONS: HFE extract inhibited articular cartilage degeneration through preventing extracellular matrix degradation and chondrocyte injury.
Administration, Oral ; Aggrecans ; Animals ; Cartilage ; Cartilage, Articular ; Cell Survival ; Chondrocytes ; Collagen ; Collagen Type I ; Collagen Type II ; Dinoprostone ; Extracellular Matrix ; In Vitro Techniques* ; Injections, Intra-Articular ; Knee Joint ; Matrix Metalloproteinases ; Models, Animal ; Nitric Oxide ; Osteoarthritis* ; Rats ; Real-Time Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinases

BACKGROUND/OBJECTIVES: The inhibitory effect of Hijikia fusiforme (HF) extracts on degenerative osteoarthritis was examined in primary cultured rat cartilage cells and a monosodium iodoacetate (MIA)-induced osteoarthritis rat model. MATERIALS/METHODS: In vitro, cell survival and the expression of matrix metalloproteinases (MMPs), collagen type I, collagen type II, aggrecan, and tissue inhibitor of metalloproteinases (TIMPs) was measured after H2O2 (800 µM, 2 hr) treatment in primary chondrocytes. In vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats, and then RH500, HFE250 and HFE500 were administered orally once a day for 28 days. To determine the anti-inflammatory effects of HFE, nitric oxide (NO), prostaglandin E2 (PGE2) expression were measured. In addition, real-time PCR was performed to measure the genetic expression of MMPs, collagen type I, collagen type II, aggrecan, and TIMPs. RESULTS: In the in vitro assay, cell survival after H2O2 treatment was increased by HFE extract (20% EtOH). In addition, anabolic factors (genetic expression of collagen type I, II, and aggrecan) were increased by HFE extract (20% EtOH). However, the genetic expression of MMP-3 and 7, known as catabolic factors were significantly inhibited by treatment with HFE extract (20% EtOH). In the in vivo assay, anabolic factors (genetic expression of collagen type I, II, aggrecan, and TIMPs) were increased by oral administration of HFE extract. However, the genetic expression of MMP-3 and 7, known as catabolic factors, and production of NO and PGE2 were significantly inhibited by treatment with oral administration of HFE extract. CONCLUSIONS: HFE extract inhibited articular cartilage degeneration through preventing extracellular matrix degradation and chondrocyte injury.
Administration, Oral ; Aggrecans ; Animals ; Cartilage ; Cartilage, Articular ; Cell Survival ; Chondrocytes ; Collagen ; Collagen Type I ; Collagen Type II ; Dinoprostone ; Extracellular Matrix ; In Vitro Techniques* ; Injections, Intra-Articular ; Knee Joint ; Matrix Metalloproteinases ; Models, Animal ; Nitric Oxide ; Osteoarthritis* ; Rats ; Real-Time Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinases

This study aims to investigate the effect of total flavones of Fructus Chorspondiatis (TFFC) on the mRNA and protein expression of collagen type I and III of rat cardiac fibroblasts (CFs) induced by angiotensin II (Ang II), and explore its anti-myocardial fibrosis molecular mechanism. Neonatal rat CFs were prepared from Sprague-Dawley rats (1-3 d after birth). The expression of collagen type I and III mRNA and protein were measured by RT-PCR and Western blotting, respectively. The study showed that stimulation of neonatal rat CFs with 100 nmol.L-1 of Ang II for 72 h resulted in a significant increase of the expression of collagen type I and III mRNA and protein. The changes on the expression level were blocked by TFFC. The results demonstrated that TFFC can inhibit myocardial fibrosis induced by Ang II in rats, which is probably associated with the collagen type I and III mRNA and protein levels up-regulated by Ang II, and TFFC was shown to decrease the expression levels of collagen type I and III mRNA and protein.
Anacardiaceae ; chemistry ; Angiotensin II ; pharmacology ; Animals ; Animals, Newborn ; Cells, Cultured ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Flavones ; administration & dosage ; isolation & purification ; pharmacology ; Fruit ; chemistry ; Myocardium ; cytology ; metabolism ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley