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OBJECTIVES@#This study aims to investigate the effects and mechanisms of chondroitin sulfate (CS), dermatan sulfate (DS), and heparin (HEP) on chondrogenesis of murine chondrogenic cell line (ATDC5) cells and the maintenance of murine articular cartilage in vitro.@*METHODS@#ATDC5 and articular cartilage tissue explant were cultured in the medium containing different sulfated glycosaminoglycans. Cell proliferation, differentiation, cartilage formation, and mechanism were observed using cell proliferation assay, Alcian blue staining, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blot, respectively.@*RESULTS@#Results showed that HEP and DS primarily activated the bone morphogenetic protein (BMP) signal pathway, while CS primarily activated the protein kinase B (AKT) signal pathway, further promoted ATDC5 cell proliferation and matrix production, and increased Sox9, Col2a1, and Aggrecan expression.@*CONCLUSIONS@#This study investigated the differences and mechanisms of different sulfated glycosaminoglycans in chondrogenesis and cartilage homeostasis maintenance. HEP promotes cartilage formation and maintains the normal state of cartilage tissue in vitro, while CS plays a more effective role in the regeneration of damaged cartilage tissue.
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Animals , Mice , Cartilage/metabolism , Cell Differentiation , Cells, Cultured , Chondrocytes/metabolism , Chondrogenesis/physiology , Glycosaminoglycans/pharmacologyABSTRACT
Objective:To investigate the adhesion of polydopamine-modified collagen membrane composites to cartilage tissues and the effect on chondrocyte proliferation, and further explore the possibility of their application in autologous chondrocyte transplantation.Methods:Porous collagen membranes were prepared, and the polydopamine-modified collagen membrane composites were constructed by the adsorption method. The physical and chemical properties and structural characteristics of the membranes, such as thermal stability, thermal properties, porous structure, and surface element composition, were characterized by infrared spectroscopy, thermogravimetric analysis, differential thermal analysis, scanning electron microscopy, and X-ray photoelectron spectroscopy, respectively. The adhesion between the polydopamine-modified collagen membrane and fresh cartilage tissue was tested by a mechanical testing machine. The effects of the membranes on the adhesion and proliferation of rabbit chondrocytes were investigated by in vitro cell culture.Results:The structure and surface element composition of the membranes altered with the increase in the adsorption time of polydopamine, and the capacity of polydopamine increased with the increase in the adsorption time. The thermal stability and thermal properties of collagen membrane materials were not significantly affected by the adsorption of polydopamine. The adhesion of the membrane to cartilage tissue increased with the increase in the amount of absorbed polydopamine. The membranes showed a time-dependent promoting effect on the proliferation of the chondrocytes.Conclusions:The polydopamine-modified collagen membrane has potential application in articular cartilage repair, but more research is required to optimize the membrane before it is used in articular cartilage repair.
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Objective:To observe any effect of exosomes derived from umbilical cord mesenchymal stem cells on pain, cartilage repair and the expression of transcriptional activator 3 (ATF-3) and growth related protein 43 (GAP-43) in the dorsal root ganglia (DRG), as well as to explore the mechanism of their relieving pain.Methods:Fifty-four male Sprague-Dawley rats were randomly divided into a sham-operation group, a monoiodoacetate group and an exosome group, each of 18. The knee cavities of the left hind limbs of all of the rats except those in the sham-operation group were injected with 50μl of monoiodoacetate to establish an arthritis pain model. The sham-operation group received only 50μl of saline solution as controls. Two weeks after the modelling, the knee joint cavities of the exosome group were injected with 50μl of exosomes, while the other two groups were injected with 50μl of normal saline. The rats′ mechanical and thermal pain thresholds were measured 1 day before the modeling, 7 and 14 days after the monoiodoacetate injection, as well as 7, 14 and 28 days after the exosome injection. Western blotting was used to detect the expression of ATF-3 and GAP-43 in the rats′ DRG, while hematoxylin and eosin staining was used to detect any cartilage repair.Results:Compared with the monoiodoacetate group, the latency of the mechanical and thermal pain thresholds had increased significantly in the exosome group 7 days after the exosome injection. The difference remained significant until the 28th day after the injection. The expression of ATF-3 protein decreased significantly and that of the GAP-43 protein increased significantly. Significant differences were observed in the average Osteoarthritis Research Society International (OARSI) knee cartilage score.Conclusions:Exosomes can alleviate the pain induced by monoiodoacetate adjuvant. The analgesic mechanism may be related to reducing nerve injury and promoting nerve and cartilage repair, with the nerve repair earlier than cartilage repair.
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Articular cartilage injury is common in orthopedics. Improper exercises and physical trauma can lead to the injury of cartilage. Since articular cartilage lacks blood supply, once damaged, it is difficult for the cartilage to repair itself. If not treated effectively, cartilage injuries will develop into severe osteoarthritis affecting the whole joint. Arthroscopic microfracture technique can achieve better therapeutic effects than regular joint debridement, with simple procedures, minimal invasion, and low cost. However, the microfracture technique is limited by the patients′ age (under 45 years old) and the size of the cartilage defect area (less than 4 cm 2) Additionally, postoperative patients need to conduct strict and long-term rehabilitation trainings. Generally speaking, the short-term prognosis of microfracture is satisfactory. However, the repair tissue is mainly composed of fibrocartilage, which is inferior to hyaline cartilage because of its poor mechanical properties and anti-wear abilities. Therefore, the long-term effect is controversial. To conclude, arthroscopic microfracture is a recommended method for young patients with small cartilage defect areas, but its exact long-term clinical effects still need to be verified by further research. This paper reviews the operation protocol, clinical efficacy, and the mechanism of arthroscopic microfracture surgery, and aims to provides theoretical basis for its application in clinical treatment.
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@#Introduction: Autologous matrix-induced chondrogenesis (AMIC) is a one-step surgical cartilage repair procedure involving the insertion of a scaffold into the chondral defect after microfracture. BST-CarGel [Smith and Nephew, Watford, England] is an injectable chitosan-based scaffold which can more easily fill defects with irregular shapes and be used to treat vertical or roof chondral lesions. The study aims to evaluate the clinical outcomes of knee cartilage repair with microfracture surgery and BST-CarGel using the AMIC technique for a minimum of two years. Materials and methods: A prospective study of patients undergoing cartilage repair with microfracture surgery and BST-CarGel at our institution from 2016 to 2019 was performed. Clinical outcomes were determined using the Lysholm Knee Scoring System and Knee Injury and Osteoarthritis Outcome Score (KOOS). These questionnaires were administered before the surgery and at a minimum of two years after surgery. Results: A total of 21 patients were identified and recruited into the study. 31 cartilage defects were seen and treated in 21 knees. These included horizontal lesions (e.g., trochlear, lateral tibial plateau), vertical lesions (e.g., medial femoral condyle, lateral femoral condyle) and inverted lesions (e.g., patella). No complications or reoperations were seen in our study population. For the average duration of follow-up of 42.5±8.55 months, there was an average improvement in Lysholm score of 25.8±18.6 and an average improvement in KOOS score of 22.5±15.0. Conclusion: BST-CarGel with microfracture surgery using the AMIC technique is a safe and effective treatment for cartilage defects in the short to medium term.
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@#Osteoarthritis (OA) is a common chronic joint disease,whose main pathological changes are the degeneration of articular cartilage and secondary bone hyperplasia.The limitation of current treatment methods including pain relief and joint replacement surgery is that they cannot fundamentally improve the damage of articular cartilage.The emergence of disease-modifying osteoarthritis drugs (DMOAD) may break the above limitations.They fundamentally inhibit the structural degeneration of articular cartilage by participating in the regulation of cartilage metabolic balance, regulation of subchondral bone remodeling,and control of local inflammation.Thereby,OA patients will get symptom improvement including pain relief and joint function restoration,delay the artificial joint replacement surgery, and improve the quality of life. There are still no DMOAD drugs widely available on the market worldwide.This paper reviews the background of R&D,the classification of mechanisms of action and research progress of representative drugs under different inechanisms so as to provide reference for future research.
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Traumatic arthritis (TA) is one of the common diseases of bone and joint caused by trauma. The main pathological changes are degeneration of articular cartilage and secondary hyperplasia and ossification of cartilage. The main clinical manifestations are joint pain and dysfunction of movement. Its pathogenesis is still unclear. At present, the treatment of TA is based mainly on symptoms rather than etiology, including physical therapy, drug treatment, surgical treatment, etc. Conservative treatment (physical therapy, drug treatment) can only alleviate short-term pain, and the long-term effect is not satisfactory. Thus, patients with middle and late TA tend to choose surgical treatment. At present, the surgical treatment of TA includes arthroscopic debridement, arthrodesis, cartilage repair, osteotomy, artificial joint replacement, 3D printing technology, etc. There are differences in the postoperative efficacy. This article reviews the current situation of surgical treatment for TA.
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BACKGROUND: Graphene-related materials have good biocompatibility and can improve cartilage repair. At the same time, their excellent mechanical strength and electrical conductivity make them promising as cartilage replacement materials, which have been widely used in tissue engineering. OBJECTIVE: To review the general properties, biocompatibility and application of graphene in cartilage tissue engineering and cartilage repair. METHODS: A computer-based online search of CNKI and PubMed databases was performed using the search terms “graphene, tissue engineering, biocompatibility, cartilage” in Chinese and English to search related literatures published between January 2000 and January 2019. Preliminary screening was conducted by reading the titles and abstracts to exclude the literature irrelevant to the theme of the paper. According to inclusion and exclusion criteria, 67 literatures were included in the final analysis. RESULTS AND CONCLUSION: Graphene has good biocompatibility, and has low cytotoxicity to prokaryotic cells and eukaryotic cells, but the cytotoxicity can be further reduced by chemical modification or surface modification, so as not to affect the growth of cells. Graphene and its derivatives can promote the growth and chondrogenic differentiation of human bone marrow mesenchymal stem cells, as well as the proliferation and maturation of chondrocytes, and accelerate the repair of cartilage defects. Due to its mechanical strength and electrical conductivity, graphene can compound biomimetic cartilage material, which is suitable for cartilage tissue engineering. Graphene has several unresolved problems and challenges, but the application potential of graphene-related materials may pave the way for future breakthroughs in tissue engineering research.
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BACKGROUND: Our previous studies have found that silk fibroin-chitosan scaffold carrying bone marrow mesenchymal stem cells can repair cartilage defect in rabbits, but further exploration on the biocompatibility of tissue engineered cartilage is yet to be done. OBJECTIVE: To explore the biocompatibility of tissue engineered cartilage that is constructed in vitro by silk fibroin-chitosan scaffold with bone marrow mesenchymal stem cells. METHODS: Three-dimensional silk fibroin-chitosan scaffolds were prepared in a ratio of 1:1. Rabbit bone marrow mesenchymal stem cells were extracted, induced and seeded onto the silk fibroin-chitosan scaffold to construct the cell-scaffold composite. The composite was then implanted into a rabbit joint defect model for cartilage repair. There were three groups in the present study: Experiment group with implantation of induced bone marrow mesenchymal stem cells+silk fibroin-chitosan scaffold into the cartilage defect model, control group with implantation of silk fibroin-chitosan scaffold into the cartilage defect model, and blank group without implantation. RESULTS AND CONCLUSION: The three-dimensional silk fibroin-chitosan scaffolds were successfully prepared and combined with bone marrow mesenchymal stem cells (BMSCs) to construct the tissue engineered cartilage for repair cartilage defects in rabbits. Blood routine parameters, procalcitonin levels, erythrocyte sedimentation rates and C-reactive protein levels detected at 2, 4, 8, and 12 weeks post-implantation indicated no obvious signs of systemic infection, and there was no damage to liver and kidney functions in the three groups. There were also no significant differences between the three groups in terms of blood routines and liver and kidney functions (P > 0.05). As shown by gross observation, hematoxylin-eosin staining and scanning electron microscope, in the experimental group, cartilage defects were repaired, with scaffold degradation, no presence of inflammatory cells, and good integration with surrounding tissues. Therefore, tissue engineered cartilage constructed in vitro by silk fibroin-chitosan scaffolds carrying bone marrow mesenchymal stem cells has good biocompatibility, which provides an experimental basis for tissue engineering approaches to cartilage repair.
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Objective: To compare the effectiveness of arthroscopic osteochondral autologous transplantation (OAT) in the treatment of young and middle-aged patients with the articular cartilage injury. Methods: A clinical data of 43 patients (43 knees) with articular cartilage injury, who underwent OAT between January 2008 and August 2016, was retrospectively analyzed. There were 23 patients aged 20-40 years (young group) and 20 patients aged 40-60 years (middle-aged group). The difference in age between the two groups was significant ( t=14.120, P=0.001). There was no significant difference in gender, body mass index, complications, affected side, lesion site, lesion area, and the International Cartilage Repair Society (ICRS) grade of cartilage injury between the two groups ( P>0.05). The function of knee joint was evaluated by Lysholm score and International Knee Documentation Committee (IKDC) score during the follow-up. MRI examination was performed to observe the repair of both receiving and the donor sites. Results: All the incisions in the two groups were healed by first intention. All patients in the two groups were followed up with an average of 3.6 years (range, 2-8 years). At 2 years after operation, the Lysholm and IKDC scores were significantly improved in the two groups when compared with the preoperative scores ( P0.05). The MRI examination at 2 years after operation showed that both receiving and the donor sites healed well in the two groups. Conclusion: According to the texture, thickness, elasticity, and lesion area of the cartilage, arthroscopic OAT might be the first choice for the articular cartilage injury in middle-aged patients and can obtain the satisfactory short-term effectiveness.
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After the articular cartilage injury, the metabolic level is increased during the progressive degeneration, the chondrocytes secrete a variety of inflammatory factors, and the original cell phenotype is gradually changed. For a long time, a large number of researchers have done a lot of researches to promote anabolism of chondrocytes and to maintain the stability of chondrocyte phenotype. There are many molecular signaling pathways involved in the process of promoting cartilage repair. This review focuses on the key signaling molecules in articular cartilage repair, such as transforming growth factor-beta and bone morphogenetic protein, and reveals their roles in the process of cartilage injury and repair, so that researchers in related fields can understand the molecular mechanism of cartilage injury and repair widely and deeply. Based on this, they may find promising targets and biological methods for the treatment of cartilage injury.
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Humans , Bone Morphogenetic Proteins , Physiology , Cartilage, Articular , Wounds and Injuries , Chondrocytes , Physiology , Regeneration , Signal Transduction , Transforming Growth Factor beta , PhysiologyABSTRACT
Cartilage is a tissue simple in composition, complex in structure and difficult to repair after injury. The development of cartilage tissue engineering has been prompted by the limitations of current treatment methods, but is now faced with difficulties in further clinical transformation. With an introduction to the present situation in the treatment of cartilage injury, we analyzed the challenges opportunities in the clinical transformation of cartilage tissue engineering, hoping to provide some ideas and methods to promote its development andclinical transformation.
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BACKGROUND: Articular cartilage lesions occur frequently but unfortunately damaged cartilage has a very limited intrinsic repair capacity. Therefore, there is a high need to develop technology that makes cartilage repair possible. Since joint damage will lead to (sterile) inflammation, development of this technology has to take into account the effects of inflammation on cartilage repair. METHODS: A literature search has been performed including combinations of the following keywords; cartilage repair, fracture repair, chondrogenesis, (sterile) inflammation, inflammatory factors, macrophage, innate immunity, and a number of individual cytokines. Papers were selected that described how inflammation or inflammatory factors affect chondrogenesis and tissue repair. A narrative review is written based on these papers focusing on the role of inflammation in cartilage repair and what we can learn from findings in other organs, especially fracture repair. RESULTS: The relationship between inflammation and tissue repair is not straightforward. Acute, local inflammation stimulates fracture repair but appears to be deleterious for chondrogenesis and cartilage repair. Systemic inflammation has a negative effect on all sorts of tissue repair. CONCLUSION: Findings on the role of inflammation in fracture repair and cartilage repair are not in line. The currently widely used models of chondrogenesis, using high differentiation factor concentrations and corticosteroid levels, are not optimal. To make it possible to draw more valid conclusions about the role of inflammation and inflammatory factors on cartilage repair, model systems must be developed that better mimic the real conditions in a joint with damaged cartilage.
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Cartilage , Cartilage, Articular , Chondrogenesis , Cytokines , Immunity, Innate , Inflammation , Joints , MacrophagesABSTRACT
OBJECTIVES: Articular cartilage is vulnerable to injuries and undergoes an irreversible degenerative process. The use of amniotic fluid mesenchymal stromal stem cells for the reconstruction of articular cartilage is a promising therapeutic alternative. The aim of this study was to investigate the chondrogenic potential of amniotic fluid mesenchymal stromal stem cells from human amniotic fluid from second trimester pregnant women in a micromass system (high-density cell culture) with TGF-β3 for 21 days. METHODS: Micromass was performed using amniotic fluid mesenchymal stromal stem cells previously cultured in a monolayer. Chondrocytes from adult human normal cartilage were used as controls. After 21 days, chondrogenic potential was determined by measuring the expression of genes, such as SOX-9, type II collagen and aggrecan, in newly differentiated cells by real-time PCR (qRT-PCR). The production of type II collagen protein was observed by western blotting. Immunohistochemistry analysis was also performed to detect collagen type II and aggrecan. This study was approved by the local ethics committee. RESULTS: SOX-9, aggrecan and type II collagen were expressed in newly differentiated chondrocytes. The expression of SOX-9 was significantly higher in newly differentiated chondrocytes than in adult cartilage. Collagen type II protein was also detected. CONCLUSION: We demonstrate that stem cells from human amniotic fluid are a suitable source for chondrogenesis when cultured in a micromass system. amniotic fluid mesenchymal stromal stem cells are an extremely viable source for clinical applications, and our results suggest the possibility of using human amniotic fluid as a source of mesenchymal stem cells.
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Humans , Pregnancy , Cell Culture Techniques/methods , Chondrocytes/cytology , Chondrogenesis , Mesenchymal Stem Cells/cytology , Gene Expression , Cell Differentiation , Collagen Type II/analysis , Aggrecans/metabolism , Transforming Growth Factor beta3/metabolism , SOX9 Transcription Factor/metabolism , Amniotic FluidABSTRACT
Objective: To summarize the research progress of rehabilitation after autologous chondrocyte implantation (ACI). Methods: The literature related to basic science and clinical practice about rehabilitation after ACI in recent years was searched, selected, and analyzed. Results: Based on the included literature, the progress of the graft maturation consists of proliferation phase (0-6 weeks), transition phase (6-12 weeks), remodeling phase (12-26 weeks), and maturation phase (26 weeks-2 years). To achieve early protection, stimulate the maturation, and promote the graft-bone integrity, rehabilitation protocol ought to be based on the biomechanical properties at different phases. Weight-bearing program, range of motion (ROM), and options or facilities of exercise are importance when considering a rehabilitation program. Conclusion: It has been proved that the patients need a program with an increasingly progressive weight-bearing and ROM in principles of rehabilitation after ACI. Specific facilities can be taken at a certain phase. Evidences extracted in the present work are rather low and the high-quality and controlled trials still need to improve the rehabilitation protocol.
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Objective: To review the application and research progress of in-situ tissue engineering technology in bone and cartilage repair. Methods: The original articles about in-situ tissue engineering technology in bone and cartilage repair were extensively reviewed and analyzed. Results: In-situ tissue engineering have been shown to be effective in repairing bone defects and cartilage defects, but biological mechanisms are inadequate. At present, most of researches are mainly focused on animal experiments, and the effect of clinical repair need to be further studied. Conclusion: In-situ tissue engineering technology has wide application prospects in bone and cartilage tissue engineering. However, further study on the mechanism of related cytokines need to be conducted.
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Absrtact: Osteoarthritis (OA) is a most common form of degenerative joint disease, primarily characterized by the degradation of articular cartilage, subchondral sclerosis and inflammation of the synovial membrane. Mesenchymal stem cells (MSCs), a multipotent adult stem cell population, can be isolated from many connective tissue lineages, including those of the diarthrodial joint. Joint-resident MSCs or MSC-like progenitor cells contribute to the maintenance of healthy microenvironment or to the response to trauma. The onset of degenerative changes in the joint related to abnormal condition or depletion of these endogenous MSCs and native host hyaline cartilage cells, leading to limited self-repair potential of the joint and advance of the degradation. To date, no acknowledged medical treatment strategies, including non-operative and classical surgical techniques, are efficient in restoring normal anatomy and function of hyaline cartilage in OA. This highlights an urgent need for better celled-based therapeutic strategies that supplement these functional cells exogenously to recover the tissue homeostasis and repair in joint cavity via chondrogenic and anti- in fl ammatory functions. In this review we focus on the role of native MSCs in healthy or OA joint and recent progress in cell-based researches utilizing culture-expanded chondrocytes, pluripotent stem cells, or MSCs from different sources for treating OA.
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Treatment options for partial thickness cartilage defects are limited. The purpose of this study was to evaluate the efficacy of the chondrocyte-seeded cartilage extracellular matrix membrane in repairing partial thickness cartilage defects. First, the potential of the membrane as an effective cell carrier was investigated. Secondly, we have applied the chondrocyte-seeded membrane in an ex vivo, partial thickness defect model to analyze its repair potential. After culture of chondrocytes on the membrane in vitro, cell viability assay, cell seeding yield calculation and cell transfer assay were done. Cell carrying ability of the membrane was also tested by seeding different densities of cells. Partial defects were created on human cartilage tissue explants. Cell-seeded membranes were applied using a modified autologous chondrocyte implantation technique on the defects and implanted subcutaneously in nude mice for 2 and 4 weeks. In vitro data showed cell viability and seeding yield comparable to standard culture dishes. Time dependent cell transfer from the membrane was observed. Membranes supported various densities of cells. Ex vivo data showed hyaline-like cartilage tissue repair, integrated on the defect by 4 weeks. Overall, chondrocyte-seeded cartilage extracellular membranes may be an effective and feasible treatment strategy for the repair of partial thickness cartilage defects.
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Animals , Humans , Mice , Cartilage , Cell Survival , Chondrocytes , Extracellular Matrix , In Vitro Techniques , Lifting , Membranes , Mice, NudeABSTRACT
As a degenerative disease caused by multiple factors, osteoarthritis is characterized by articular cartilage degeneration and reactive hyperplasia of joint edge and the subchondral bone. Recently, kartogenin (KGN) was identified to promote chondrocyte differentiation. KGN can block interleukin 1β(IL-1β)-caused loss of extracellular matrix and proteoglycan. Transforming growth factor β1(TGF-β1), bone morphogenetic protein 7(BMP-7), and KGN together can synergistically promote the expression of lubricin in chondrocytes. KGN can also induce cartilage-like tissue formation in tendon-bone junction. In addition, chitosan (CHI)-KGN nanoparticles and CHI-KGN microspheres can more effectively induce chondrogenic differentiation than unconjugated KGN. Here in this paper we summarized the roles of KGN in regulating the cartilage regeneration.
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OBJECTIVE: To discuss the effect of glucosamine-hydrochloride (Glu/Ch) in protecting and repairing the cartilage in blood-induced joint damage (BJD) in vivo. METHODS: Thirty-two adult New Zealand rabbits were randomly divided into 4 groups (n=8):high-dose Glu/Ch treated group (group A), low-dose Glu/Ch treated group (group B), positive control group (group C), and negative control group (group D). A joint bleeding model was established by blood injection into articular cavity in groups A, B, and C. Glu/Ch was given by gavage in groups A (250 mg/kg) and B (21.5 mg/kg) once a day for 8 weeks, and the same dosage of saline was given in groups C and D. The serum cartilage oligomeric matrix protein (COMP), serum chondroitin sulfate 846(CS846), and urinary C-terminal telopepide of type II collagen (CTX-II) were measured at 3 days, 7 days, 2 weeks, and 8 weeks after modeling. The expressions of cytokines such as interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) in synovial fluid were analyzed by ELISA at 8 weeks after modeling. The expression of matrix metalloproteinase 13(MMP-13) was detected by immunohistochemistry. Alcian blue staining and Safranin-O staining were performed to calculate the percentage of the positive staining areas. The proteoglycan content was detected by semi-quantitative analysis in the articular cartilage. RESULTS: The COMP concentration was significantly higher in groups A, B, and C than group D, and in groups B and C than group A at 3 days after modeling (P0.05), and it was significantly lower in groups A, B, and D than group C (P0.05). Difference in CS846 concentration had no significance among 4 groups at each time point (P>0.05). The CTX-II concentration of groups A, B, and C was significantly higher than that of group D at each time point (P0.05). The IL-1β concentration was significantly higher in group C than the other groups (P0.05). The MMP-13 expression was significantly higher in group C than groups A, B, and D (P<0.05), in groups A and B than group D (P<0.05). A significant decrease in the area stained with Alcian blue and Safranin-O was observed in group C. There were significant differences in the percentage of the positive stained areas of Alcian blue and Safranin-O among 4 groups (P<0.05). The relative quantities of proteoglycan from small to large in order was groups C, B, A, and D, respectively, showing significant differences (P<0.05). CONCLUSIONS: The metabolism disorder of cartilage matrix and synovium inflammatory reaction can be observed in rat joint bleeding model. Glu/Ch has certain protective effect on the cartilage after BJD by down-regulating IL-1β, TNF-α, and MMP-13, as well as increasing proteoglycan content in the cartilage.