Chondrocyte necrosis and apoptosis in impact damaged articular cartilage

Abstract

A decrease in chondrocyte numbers is one characteristic of osteoarthritic cartilage. This decrease may be the result of apoptosis or other forms of cell death induced by mechanical damage. Furthermore, cell death may contribute to the structural and metabolic changes found in osteoarthritic cartilage. Therefore, we investigated cell viability and the mode of cell death in cartilage subjected to an increasing severity of impact loads expected to cause compositional damage and osteoarthritic-like metabolic alterations. Canine cartilage explants were subjected to cyclic indentation impacts of 5 megapascals at 0.3 Hz for 0,2,20, and 120 min and then kept in culture for 2, 4, 48, and 144 h. Cell death was assessed by the TUNEL assay and by uptake of propidium iodide. Viable cells were detected by the ability to metabolize fluorescein diacetate. Nuclear morphology and ultrastructure of the cell were examined using Hoechst 33342 fluorescent staining and transmission electron microscopy (TEM). As controls for necrosis and apoptosis, cartilage was, respectively, frozen and thawed or incubated with mitomycin-C, an apoptosis inducer. In cartilage that had been loaded for 2 h, 32% of the chondrocytes in the loaded core took up propidium iodide within 2 h after loading. Most of these were in the middle to superficial zones and reflected leaky cell membranes usually characteristic of necrosis. Less than 1% of these chondrocytes were positive in the TUNEL assay after 4 h. After additional culture for 2 days, however, the proportion of chondrocytes which were positive in the TUNEL assay reached 73%. A dose dependent response to duration of loading was detected with the TUNEL assay at this time. The TUNEL assay was not specific for apoptosis since 92% of chondrocytes in freezekhawed cartilage were TUNEL positive. However, some cells with apoptotic bodies and chromatin condensation characteristic of apoptosis were found in the transition zone between necrotic and normal chondrocytes, but not in the superficial and upper zones, in impact damaged cartilage. We concluded that in this study, necrosis occurred first, followed by apoptosis. 0

Acute repair of chondrocytes in the rabbit tibiofemoral joint following blunt impact using P188 surfactant and a preliminary investigation of its long-term efficacy

“…As a result, some cells undergo necrotic death in the short term, whereas the remaining traumatized cells may enter an apoptotic pathway later. 34 Additionally, a previous study by Natoli and Athanasiou 5 suggests that less necrosis may lead to decreased apoptosis by preventing the release of apoptotic initiators from necrotic cells, thereby preventing further death at later time points. Apoptotic initiators and other intracellular signals may be involved in the spreading of cell death through the cartilage matrix.…”

Mechanical injury of cartilage explants causes specific time‐dependent changes in chondrocyte gene expression

“…Mechanical loads applied in vitro range from single compressions of up to 50% strain (12)(13)(14)(15)(16)(17) to large-amplitude cyclic compression at varying frequencies (ϳ0.05-0.3 Hz) for up to 2 hours (18)(19)(20)(21). Injurious mechanical compression of cartilage in vitro can damage the extracellular matrix (ECM), leading to increased water content (12,16,18,22), decreased stiffness (12,22), increased hydraulic permeability (20), loss of glycosaminoglycan (GAG) to the culture medium (12)(13)(14)(15)17,20,22,23), loss of collagen to the medium (20), and temporary denaturation of collagen in the tissue (16,18,20,21).…”

“…Injurious mechanical compression of cartilage in vitro can damage the extracellular matrix (ECM), leading to increased water content (12,16,18,22), decreased stiffness (12,22), increased hydraulic permeability (20), loss of glycosaminoglycan (GAG) to the culture medium (12)(13)(14)(15)17,20,22,23), loss of collagen to the medium (20), and temporary denaturation of collagen in the tissue (16,18,20,21). In addition, injurious mechanical compression can lead to cell death by both apoptosis and necrosis (14,16,19,(21)(22)(23), as well as decreased matrix biosynthesis rates in the remaining viable cells after injury (12).…”

The limitation of acute necrosis in retro-patellar cartilage after a severe blunt impact to the in vivo rabbit patello-femoral joint

“…Necrosis was observed 2 h after cessation of loading, whereas apoptosis (TUNEL-positive cells) was not significant until 48 or more hours after loading stopped. Apoptosis was verified in some cells using transmission electron microscopy [3]. A study that induced osteochondral wounding of a joint also found significant percentages of necrotic and apoptotic cells in the tissue [29].…”