scholarly journals Elucidating mechano-pathology of osteoarthritis: transcriptome-wide differences in mechanically stressed aged human cartilage explants

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Evelyn Houtman ◽  
Margo Tuerlings ◽  
Janne Riechelman ◽  
Eka H. E. D. Suchiman ◽  
Robert J. P. van der Wal ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Mechanical Stress ◽  
Cellular Senescence ◽  
Protein Interactions ◽  
Differential Expression Analysis ◽  
Cartilage Explants ◽  
Human Articular Cartilage ◽  
Protein Protein Interactions ◽  
Human Cartilage ◽  
Age Related

Abstract Background Failing of intrinsic chondrocyte repair after mechanical stress is known as one of the most important initiators of osteoarthritis. Nonetheless, insight into these early mechano-pathophysiological processes in age-related human articular cartilage is still lacking. Such insights are needed to advance clinical development. To highlight important molecular processes of osteoarthritis mechano-pathology, the transcriptome-wide changes following injurious mechanical stress on human aged osteochondral explants were characterized. Methods Following mechanical stress at a strain of 65% (65%MS) on human osteochondral explants (n65%MS = 14 versus ncontrol = 14), RNA sequencing was performed. Differential expression analysis between control and 65%MS was performed to determine mechanical stress-specific changes. Enrichment for pathways and protein-protein interactions was analyzed with Enrichr and STRING. Results We identified 156 genes significantly differentially expressed between control and 65%MS human osteochondral explants. Of note, IGFBP5 (FC = 6.01; FDR = 7.81 × 10−3) and MMP13 (FC = 5.19; FDR = 4.84 × 10−2) were the highest upregulated genes, while IGFBP6 (FC = 0.19; FDR = 3.07 × 10−4) was the most downregulated gene. Protein-protein interactions were significantly higher than expected by chance (P = 1.44 × 10−15 with connections between 116 out of 156 genes). Pathway analysis showed, among others, enrichment for cellular senescence, insulin-like growth factor (IGF) I and II binding, and focal adhesion. Conclusions Our results faithfully represent transcriptomic wide consequences of mechanical stress in human aged articular cartilage with MMP13, IGF binding proteins, and cellular senescence as the most notable results. Acquired knowledge on the as such identified initial, osteoarthritis-related, detrimental responses of chondrocytes may eventually contribute to the development of effective disease-modifying osteoarthritis treatments.

scholarly journals Age-related changes in the composition and structure of human articular-cartilage proteoglycans

1978 ◽  
Vol 176 (3) ◽  
pp. 683-693 ◽  
Author(s):  
M T Bayliss ◽  
S Y Ali
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Age Groups ◽  
Human Articular Cartilage ◽  
Human Cartilage ◽  
Keratan Sulphate ◽  
Age Related ◽  
Composition And Structure ◽  
Normal Human ◽  
Cartilage Proteoglycans

1. Analysis of the purified proteoglycans extracted from normal human articular cartilage with 4M-guanidinium chloride showed that there was an age-related increase in their content of protein and keratan sulphate. 2. The hydrodynamic size of the dissociated proteoglycans also decreased with advancing age, but there was little change in the proportion that could aggregate. 3. Results suggested that some extracts of aged-human cartilage had an increased content of hyaluronic acid compared with specimens from younger patients. 4. Dissociated proteoglycans, from cartilage of all age groups, bind to hyaluronic acid and form aggregates in direct proportion to the hyaluronic acid concentration. 5. Electrophoretic heterogeneity of the dissociated proteoglycans was demonstrated on polyacrylamide/agarose gels. The number of proteoglycan species observed was also dependent on the age of the patient.

scholarly journals Age-related changes in the content of the C-terminal region of aggrecan in human articular cartilage

1996 ◽  
Vol 313 (3) ◽  
pp. 933-940 ◽  
Author(s):  
Jayesh DUDHIA ◽  
Catherine M. DAVIDSON ◽  
Terri M. WELLS ◽  
Demis H. VYNIOS ◽  
Timothy E. HARDINGHAM ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Terminal Region ◽  
Human Articular Cartilage ◽  
Amino Acid Residues ◽  
Turnover Rates ◽  
Human Cartilage ◽  
Age Related ◽  
Sds Page ◽  
High Concentrations ◽  
Age Related Changes

The content of the C-terminal region of aggrecan was investigated in samples of articular cartilage from individuals ranging in age from newborn to 65 years. This region contains the globular G3 domain which is known to be removed from aggrecan in mature cartilage, probably by proteolytic cleavage, but the age-related changes in its abundance in human cartilage have not been described previously. The analysis was performed by immunosorbant assay using an antiserum (JD5) against recombinant protein expressed from a cDNA clone encoding the terminal 598 amino acid residues of human aggrecan, on crude extracts of cartilage without further purification of aggrecan. The results showed that the content of the C-terminal region decreased with age relative to the G1 domain content (correlation coefficient = 0.463). This represented a 92% fall in the content of this region of the molecule from newborn to 65 years of age. Furthermore, when the G1 content of the cartilage extracts was corrected to only include the G1 attached to aggrecan and to exclude the G1 fragments which accumulate as a by-product of normal aggrecan turnover (free G1), the age-related decrease in the C-terminal region remained very pronounced. Analysis by composite agarose/PAGE showed that the number of subpopulations of aggrecan resolved increased from one in newborn to three in adult cartilage. All of these reacted with an antiserum to the human G1 domain, but only the slowest migrating species reacted with the C-terminal region antiserum (JD5). Similar analysis by SDS/PAGE confirmed the presence of high-molecular-mass (200 kDa) proteins reactive with JD5, but no reactive fragments of lower electrophoretic mobility were detected. In contrast, when probed with the antiserum to the human G1 domain, the immunoblots showed protein species corresponding to the free G1 and G1-G2 fragments, which were present at high concentrations in adult cartilage. The results suggest that the loss of the C-terminal region is not directly part of the process of aggrecan turnover, but it is a slow independent matrix process that occurs more extensively with aging as turnover rates become slower. Young cartilage with the fastest turnover contains least molecules lacking the C-terminal region, whereas in old tissue with slow turnover few molecules retain this region. An increase in the cleavage of this region with age may also contribute to this change. The content of the C-terminal region may thus give a measure of the abundance of newly synthesized aggrecan.

scholarly journals Small Non-Coding RNAome of Ageing Chondrocytes

2020 ◽  
Vol 21 (16) ◽  
pp. 5675
Author(s):  
Panagiotis Balaskas ◽  
Jonathan A. Green ◽  
Tariq M. Haqqi ◽  
Philip Dyer ◽  
Yalda A. Kharaz ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Differential Expression Analysis ◽  
Cartilage Tissue ◽  
Pcr Analysis ◽  
Small Rna Sequencing ◽  
High Grade ◽  
Human Cartilage ◽  
Fragment Analysis ◽  
Qrt Pcr ◽  
Age Related

Ageing is a leading risk factor predisposing cartilage to osteoarthritis. However, little research has been conducted on the effect of ageing on the expression of small non-coding RNAs (sncRNAs). RNA from young and old chondrocytes from macroscopically normal equine metacarpophalangeal joints was extracted and subjected to small RNA sequencing (RNA-seq). Differential expression analysis was performed in R using package DESeq2. For transfer RNA (tRNA) fragment analysis, tRNA reads were aligned to horse tRNA sequences using Bowtie2 version 2.2.5. Selected microRNA (miRNAs or miRs) and small nucleolar RNA (snoRNA) findings were validated using real-time quantitative Polymerase Chain Reaction (qRT-PCR) in an extended cohort of equine chondrocytes. tRNA fragments were further investigated in low- and high-grade OA human cartilage tissue. In total, 83 sncRNAs were differentially expressed between young and old equine chondrocytes, including miRNAs, snoRNAs, small nuclear RNAs (snRNAs), and tRNAs. qRT-PCR analysis confirmed findings. tRNA fragment analysis revealed that tRNA halves (tiRNAs), tiRNA-5035-GluCTC and tiRNA-5031-GluCTC-1 were reduced in both high grade OA human cartilage and old equine chondrocytes. For the first time, we have measured the effect of ageing on the expression of sncRNAs in equine chondrocytes. Changes were detected in a number of different sncRNA species. This study supports a role for sncRNAs in ageing cartilage and their potential involvement in age-related cartilage diseases.

scholarly journals Age-related accumulation of Maillard reaction products in human articular cartilage collagen

2000 ◽  
Vol 350 (2) ◽  
pp. 381 ◽  
Author(s):  
Nicole VERZIJL ◽  
Jeroen DEGROOT ◽  
Esther OLDEHINKEL ◽  
Ruud A. BANK ◽  
Suzanne R. THORPE ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Maillard Reaction ◽  
Maillard Reaction Products ◽  
Human Articular Cartilage ◽  
Reaction Products ◽  
Age Related

Effects of Radiofrequency Energy on Human Articular Cartilage

2005 ◽  
Vol 33 (7) ◽  
pp. 1035-1039 ◽  
Author(s):  
Sean Caffey ◽  
Edward McPherson ◽  
Brian Moore ◽  
Thomas Hedman ◽  
C. Thomas Vangsness
Keyword(s):  
Cell Death ◽  
Articular Cartilage ◽  
Dead Cell ◽  
Radiofrequency Energy ◽  
Human Articular Cartilage ◽  
Human Cartilage ◽  
Cellular Death ◽  
Significant Difference ◽  
Grade Ii ◽  
Grade Iii

Background Previous radiofrequency work has not rigidly controlled energy application to the articular cartilage, giving uncertain results published to date. Hypothesis At minimal settings, radiofrequency probes cause cell death in measurable areas when applied to human articular cartilage. Study Design Controlled laboratory study. Methods Simulating operating room conditions, 5 commercially available radiofrequency probes were attached to a customized jig to standardize a minimal contact pressure of each probe tip to 2.0 g. Keeping all variables the same, probes were placed on specific points of fresh grade II human cartilage with treatment times of 1 and 3 seconds at the manufacturer's recommended settings. Grade III cartilage was also tested with a treatment time of 3 seconds, and grade II cartilage was studied with the probe held 1 mm off the cartilage surface. Cartilage was blindly analyzed by confocal microscopy using a live/dead cell viability assay to determine the extent of cell death. Results Radiofrequency probes produced significant cellular death in the form of a half-circle into the cartilage to variable depths. For treatment times of 1 and 3 seconds, cell death measurements ranged from 404 to 539 μm and 1034 to 1283 μm, respectively. One probe failed to show any effect, with minimal evidence of cell death or cartilage smoothing. When probes were kept a 1.0-mm distance above the cartilage, no cell death or cartilage smoothing was noted. Radiofrequency treatment of grade III cartilage penetrated to the subchondral bone. There was no statistically significant difference between the damage caused by monopolar and bipolar probes when tested under these rigidly controlled conditions. Conclusion These results showed significant cellular death at these minimal conditions to the underlying chondrocytes with radiofrequency probes. Surgeons using this technology need to be aware of the power and dangerous potential these probes can have on articular cartilage.

scholarly journals Collagen Growth Pattern in Human Articular Cartilage of the Knee

Cartilage ◽  
2020 ◽  
pp. 194760352097101
Author(s):  
Adam E.M. Jørgensen ◽  
Peter Schjerling ◽  
Michael R. Krogsgaard ◽  
Michael M. Petersen ◽  
Jesper Olsen ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Growth Pattern ◽  
Age Determination ◽  
Joint Surface ◽  
Lateral Condyle ◽  
Human Articular Cartilage ◽  
Human Cartilage ◽  
Medial Condyle ◽  
Cartilage Formation ◽  
Tibia Plateau

Objective During skeletal growth, the articular cartilage expands to maintain its cover of bones in joints, however, it is unclear when and how cartilage grows. We aim to determine the expanding growth pattern and timing across the tibia plateau in human knees. Design Six human tibia plateaus (2 healthy, 2 with osteoarthritis, and 2 with posttraumatic osteoarthritis) were used for full-depth cartilage sampling systematically across the joint surface at 12 medial and 4 lateral sites. Methodologically, we took advantage of the performed nuclear bomb tests in the years 1955 to 1963, which increased the atmospheric 14C that was incorporated into human tissues. Cartilage was treated enzymatically to extract collagen, analyzed for 14C content, and year at formation was determined from historical atmospheric 14C concentrations. Results By age-determination, each tibia condyle had central points of formation surrounded by later-formed cartilage toward the periphery. Furthermore, the tibia plateaus contained collagen with 14C levels corresponding to mean donor age of 11.7 years (±3.8 SD). Finally, the medial condyle had lower 14C levels corresponding to formation 1 year later than the lateral condyle ( P = 0.009). Conclusions Human cartilage on the tibia plateau contains collagen that has experienced little if any turnover since school-age. The cartilage formation develops from 2 condyle centers and radially outward with the medial condyle finishing slightly later than the lateral condyle. This suggests a childhood programmed cartilage formation with a very limited adulthood collagen turnover.

scholarly journals Age-related changes in protein-related epitopes of human articular-cartilage proteoglycans

1986 ◽  
Vol 236 (1) ◽  
pp. 71-75 ◽  
Author(s):  
T T Glant ◽  
K Mikecz ◽  
P J Roughley ◽  
E Buzás ◽  
A R Poole
Keyword(s):  
Core Protein ◽  
Buoyant Density ◽  
Human Articular Cartilage ◽  
Human Cartilage ◽  
Keratan Sulphate ◽  
Cartilage Proteoglycan ◽  
Age Related ◽  
Age Dependent ◽  
Old Adult ◽  
Cartilage Proteoglycans

Monoclonal antibodies were prepared that recognize different age-related epitopes on proteoglycan subunits of high buoyant density isolated from human epiphysial and articular cartilages. Antibody EFG-4 (IgG1) recognizes a proteinase-sensitive segment associated with the core protein. Antibody BCD-4 (IgG1) reacts with keratan sulphate bound to core protein. Both epitopes are minimally expressed in foetal cartilage and increase with age after birth to become maximally expressed in adult cartilage by about 30 years of age. In contrast, monoclonal antibody alpha HFPG-846 (IgM) recognizes a core-protein-related epitope that is maximally expressed in young foetal cartilage, declines up to birth and thereafter and is almost absent after about 30 years of age. Antibody alpha HFPG-846 was used to isolate by immuno-affinity chromatography two subpopulations of proteoglycan subunits from a 16-year-old-human cartilage proteoglycan subunit preparation. Only the antibody-unbound population showed a significant reaction with antibodies EGF-4 and BCD-4. The amino acid and carbohydrate compositions of these proteoglycan fractions were different, and one (antibody-bound) resembled those of foetal and the other (antibody-unbound) resembled those of adult proteoglycans isolated from 24-27-week-old-foetal and 52-56-year-old-adult cartilage respectively. These observations demonstrate that human cartilages contain at least two chemically and immunochemically distinct populations of proteoglycans, the proportions and content of which are age-dependent. It is likely that these populations represent the products of different genes, though their heterogeneity may be compounded by the result of different post-translation modifications.

scholarly journals Studies on cathepsin B in human articular cartilage

1978 ◽  
Vol 171 (1) ◽  
pp. 149-154 ◽  
Author(s):  
M T Bayliss ◽  
S Y Ali
Keyword(s):  
Articular Cartilage ◽  
Cathepsin B ◽  
Cathepsin D ◽  
Human Articular Cartilage ◽  
Normal Cartilage ◽  
Elderly Individuals ◽  
Age Related ◽  
Hydrolysis Of ◽  
Increased Activity ◽  
Thiol Proteinase

The thiol proteinase cathepsin B (EC 3.4.22.1), previously called cathepsin B1, was assayed in human articular cartilage by its hydrolysis of the synthetic substrate alpha-N-benzoyl-DL-arginine 2-naphthylamide. The enzyme was activated by cysteine and EDTA and completely inhibited by iodoacetamide and HgCl2. It was also partially inhibited by whole human serum. Human osteoarthrotic cartilage had increased activity when compared with normal cartilage. Cathepsin B activity of normal cartilage was age-related, being high in juveniles and declining to low values in adult and elderly individuals. Cathepsin D and cathepsin B both exhibited a zonal variation through the cartilage depth; the surface cells appeared to contain more activity than those close to the subchondral bone.

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