Isolation of RNA from small human articular cartilage specimens allows quantification of mRNA expression levels in local articular cartilage defects

Polyvinyl alcohol hydrogel has compatibility and biomechanical properties of human articular cartilage similar and good biological. The implantation in the human body can replace part of articular cartilage, which plays the role of bearing and alleviate the impact force. It has the prospect of clinical application. This paper introduces the research progress of polyvinyl alcohol hydro-gel materials. And compared with the characteristics of articular cartilage, clarify the possibility of repair of articular cartilage defects of the materials.

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Histological and Biochemical Evaluation of Perichondrial Transplants in Human Articular Cartilage Defects

The Journal of Bone and Joint Surgery (American) ◽

10.2106/00004623-200004000-00031 ◽

2000 ◽

Vol 82 (4) ◽

pp. 48

Author(s):

Phianne Bouwmeester ◽

Roel Kuijer ◽

Els Terwindt-Rouwenhorst ◽

Ton van der Linden ◽

Sjoerd Bulstra

Keyword(s):

Articular Cartilage ◽

Cartilage Defects ◽

Human Articular Cartilage ◽

Biochemical Evaluation ◽

Articular Cartilage Defects

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The molecular mechanism study of COMP involved in the articular cartilage damage of Kashin-Beck disease

Bone and Joint Research ◽

10.1302/2046-3758.99.bjr-2019-0247.r1 ◽

2020 ◽

Vol 9 (9) ◽

pp. 578-586

Author(s):

Mei Ma ◽

Xiao Liang ◽

Xi Wang ◽

Lu Zhang ◽

Shiqiang Cheng ◽

...

Keyword(s):

Articular Cartilage ◽

Mrna Expression ◽

Cartilage Damage ◽

Control Group ◽

Control Groups ◽

Expression Levels ◽

Mrna And Protein Expression ◽

Functional Relevance ◽

Beck Disease ◽

Mrna Expression Levels

Aims Kashin-Beck disease (KBD) is a kind of chronic osteochondropathy, thought to be caused by environmental risk factors such as T-2 toxin. However, the exact aetiology of KBD remains unclear. In this study, we explored the functional relevance and biological mechanism of cartilage oligosaccharide matrix protein (COMP) in the articular cartilage damage of KBD. Methods The articular cartilage specimens were collected from five KBD patients and five control subjects for cell culture. The messenger RNA (mRNA) and protein expression levels were detected by quantitative reverse transcription PCR (qRT-PCR) and western blot. The survival rate of C28/I2 chondrocyte cell line was detected by MTT assay after T-2 toxin intervention. The cell viability and mRNA expression levels of apoptosis related genes between COMP-overexpression groups and control groups were examined after cell transfection. Results The mRNA and protein expression levels of COMP were significantly lower in KBD chondrocytes than control chondrocytes. After the T-2 toxin intervention, the COMP mRNA expression of C28/I2 chondrocyte reduced and the protein level of COMP in three intervention groups was significantly lower than in the control group. MTT assay showed that the survival rate of COMP overexpression KBD chondrocytes were notably higher than in the blank control group. The mRNA expression levels of Survivin, SOX9, Caspase-3, and type II collagen were also significantly different among COMP overexpression, negative control, and blank control groups. Conclusion Our study results confirmed the functional relevance of COMP with KBD. COMP may play an important role in the excessive chondrocytes apoptosis of KBD patients. Cite this article: Bone Joint Res 2020;9(9):578–586.

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Perlecan in the Natural and Cell Therapy Repair of Human Adult Articular Cartilage: Can Modifications in This Proteoglycan Be a Novel Therapeutic Approach?

Biomolecules ◽

10.3390/biom11010092 ◽

2021 ◽

Vol 11 (1) ◽

pp. 92

Author(s):

John Garcia ◽

Helen S. McCarthy ◽

Jan Herman Kuiper ◽

James Melrose ◽

Sally Roberts

Keyword(s):

Articular Cartilage ◽

Cell Therapy ◽

Heparan Sulphate ◽

Cartilage Defects ◽

Human Articular Cartilage ◽

Normal Cartilage ◽

Embryonic Cartilage ◽

Autologous Cell Therapy ◽

Articular Cartilage Defects

Articular cartilage is considered to have limited regenerative capacity, which has led to the search for therapies to limit or halt the progression of its destruction. Perlecan, a multifunctional heparan sulphate (HS) proteoglycan, promotes embryonic cartilage development and stabilises the mature tissue. We investigated the immunolocalisation of perlecan and collagen between donor-matched biopsies of human articular cartilage defects (n = 10 × 2) that were repaired either naturally or using autologous cell therapy, and with age-matched normal cartilage. We explored how the removal of HS from perlecan affects human chondrocytes in vitro. Immunohistochemistry showed both a pericellular and diffuse matrix staining pattern for perlecan in both natural and cell therapy repaired cartilage, which related to whether the morphology of the newly formed tissue was hyaline cartilage or fibrocartilage. Immunostaining for perlecan was significantly greater in both these repair tissues compared to normal age-matched controls. The immunolocalisation of collagens type III and VI was also dependent on tissue morphology. Heparanase treatment of chondrocytes in vitro resulted in significantly increased proliferation, while the expression of key chondrogenic surface and genetic markers was unaffected. Perlecan was more prominent in chondrocyte clusters than in individual cells after heparanase treatment. Heparanase treatment could be a means of increasing chondrocyte responsiveness to cartilage injury and perhaps to improve repair of defects.

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Vitrification of particulated articular cartilage via calculated protocols

npj Regenerative Medicine ◽

10.1038/s41536-021-00123-5 ◽

2021 ◽

Vol 6 (1) ◽

Author(s):

Kezhou Wu ◽

Nadia Shardt ◽

Leila Laouar ◽

Janet A. W. Elliott ◽

Nadr M. Jomha

Keyword(s):

Articular Cartilage ◽

Cartilage Defects ◽

Human Articular Cartilage ◽

Significant Deterioration ◽

Chondrocyte Viability ◽

Long Term Storage ◽

Graft Tissue ◽

Articular Cartilage Defects ◽

Term Storage

AbstractPreserving viable articular cartilage is a promising approach to address the shortage of graft tissue and enable the clinical repair of articular cartilage defects in articulating joints, such as the knee, ankle, and hip. In this study, we developed two 2-step, dual-temperature, multicryoprotectant loading protocols to cryopreserve particulated articular cartilage (cubes ~1 mm3 in size) using a mathematical approach, and we experimentally measured chondrocyte viability, metabolic activity, cell migration, and matrix productivity after implementing the designed loading protocols, vitrification, and warming. We demonstrated that porcine and human articular cartilage cubes can be successfully vitrified and rewarmed, maintaining high cell viability and excellent cellular function. The vitrified particulated articular cartilage was stored for a period of 6 months with no significant deterioration in chondrocyte viability and functionality. Our approach enables high-quality long-term storage of viable articular cartilage that can alleviate the shortage of grafts for use in clinically repairing articular cartilage defects.

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