Biomarkers Affected by Impact Severity during Osteochondral Injury.

Osteochondral injury elevates the risk for developing posttraumatic osteoarthritis (PTOA). Therefore, our objective was to evaluate the relationship between impact severity during injury to cell viability and biomarkers possibly involved in PTOA. Osteochondral explants (6 mm, n = 72) were harvested from cadaveric femoral condyles (N = 6). Using a test machine, each explant (except for No Impact) was subjected to mechanical impact at a velocity of 100 mm/s to 0.25, 0.5, 0.75, 1.0, or 1.25 mm maximum compression corresponding to Low, Low-Moderate, Moderate, Moderate-High, or High impact groups. Cartilage cell viability, collagen content, and proteoglycan content were assessed at either day 0 or after 12 days of culture. Culture media were assessed for prostaglandin E2 (PGE2); nitric oxide; granulocyte macrophage colony-stimulating factor (GM-CSF); interferon gamma (IFNγ); interleukin (IL)-2, -4, -6, -7, -8, -10, -15, -18; interferon gamma-induced protein 10 (IP-10); keratinocyte-derived chemoattractant (KC); monocyte chemoattractant protein-1 (MCP-1); tumor necrosis factor alpha (TNFα); and matrix metalloproteinase-2, -3, -8, -9, -13. There was increased impact energy absorbed for the High group compared with the Moderate-High group, Moderate group, and Low-Moderate group (p = 0.011, 0.048, 0.008, respectively). At day 0, there was decreased area cell viability for the High group compared with the Low-Moderate group (p = 0.035). At day 1, PGE2 was increased for the High group compared with the Moderate, Low-Moderate, Low, and No Impact groups (p ≤ 0.01). Cumulative PGE2 was increased for the Moderate-High and High groups compared with the Moderate, Low-Moderate, Low, and No Impact groups (p ≤ 0.036). At day 1, MCP-1 was increased for the Moderate-High and High groups compared with the Low and No Impact groups (p ≤ 0.032). Impact to osteochondral explants resulted in multiple levels of severity. PGE2 was sensitive to impact severity which may justify its use as a clinically measurable biomarker after joint injury for monitoring early PTOA.

Biomarkers affected by impact velocity and maximum strain of cartilage during injury.

Osteoarthritis is one of the most common, debilitating, musculoskeletal diseases; 12% associated with traumatic injury resulting in post-traumatic osteoarthritis (PTOA). Our objective was to develop a single impact model with cartilage "injury level" defined in terms of controlled combinations of strain rate to a maximum strain (both independent of cartilage load resistance) to study their sensitivity to articular cartilage cell viability and potential PTOA biomarkers. A servo-hydraulic test machine was used to measure canine humeral head cartilage explant thickness under repeatable pressure, then subject it (except sham and controls) to a single impact having controlled constant velocity V=1 or 100mm/s (strain rate 1.82 or 182/s) to maximum strain ε=10%, 30%, or 50%. Thereafter, explants were cultured in media for twelve days, with media changed at day 1, 2, 3, 6, 9, 12. Explant thickness was measured at day 0 (pre-injury), 6 and 12 (post-injury). Cell viability, and tissue collagen and glycosaminoglycan (GAG) were analyzed immediately post-injury and day 12. Culture media were tested for biomarkers: GAG, collagen II, chondroitin sulfate-846, nitric oxide, and prostaglandin E2 (PGE2). Detrimental effects on cell viability, and release of GAG and PGE2 to the media were primarily strain-dependent, (PGE2 being more prolonged and sensitive at lower strains). The cartilage injury model appears to be useful (possibly superior) for investigating the relationship between impact severity of injury and the onset of PTOA, specifically for discovery of biomarkers to evaluate the risk of developing clinical PTOA, and to compare effective treatments for arthritis prevention.