The extent and distribution of cell death and matrix damage in impacted chondral explants varies with the presence of underlying bone.

Excessive mechanical loading can lead to matrix damage and chondrocyte death in articular cartilage. Previous studies on chondral and osteochondral explants have not clearly distinguished to what extent the degree and the distribution of cell death are dependent on the presence of an underlying layer of bone. The current study hypothesized that the presence of underlying bone would decrease the amount of matrix damage and cell death. Chondral and osteochondral explants were loaded to 30 MPa at a high rate of loading (approximately 600 MPa/s) or at a low rate of loading (30 MPa/s). After 24 hours in culture, matrix damage was assessed by the total length and average depth of surface fissures. The explants were also sectioned and stained for cell viability in the various layers of the cartilage. More matrix damage was documented in chondral than osteochondral explants for each rate of loading experiment. The total amount of cell death was also less in osteochondral explants than chondral explants. The presence of underlying bone significantly reduced the extent of cell death in all zones in low rate of loading tests. The percentage of cell death was also reduced in the intermediate zone and deep zones of the explant by the presence of the underlying bone for a high rate of loading. This study indicated that the presence of underlying bone significantly limited the degree of matrix damage and cell death, and also affected the distribution of dead cells through the explant thickness. These data may have relevance to the applicability of experimental data from chondral explants to the in situ condition.

Rate of blunt impact loading affects changes in retropatellar cartilage and underlying bone in the rabbit patella.

Our laboratory has developed a small animal model using Giant Flemish rabbits to examine chronic degradative changes in joint tissues following a blunt impact. Historically, we observe surface fissuring and decreases in the elastic modulus of retropatellar cartilage along with thickening of the underlying subchondral bone. Previous studies resulted in load insults that peaked in approximately 5ms, while loads that occur during automotive accidents or heavy exercise can produce longer rise times. The objective of the current study was to examine the influence of blunt impact loading rate using our established model. We hypothesized that the extent of fissuring and softening of retropatellar cartilage following impact would not be significantly different for a high (5ms to peak) versus low (50ms to peak) rate of loading experiment. Eight animals were impacted with a high rate of loading blunt impact, while ten animals were subjected to the same impact load at a low rate of loading. An additional eight animals served as a control population. All animals were sacrificed 12 months post-impact. The study yielded unexpected results for the first hypothesis. The high rate of loading experiments generated more surface fissuring of the retropatellar cartilage than the low rate of loading experiments. However, the degree of softening was similar for the two rates, which supported the second hypothesis. Furthermore, the study documented more thickening of bone underlying retropatellar cartilage following the high versus the low rate of loading experiments. The current study suggested that chronic injury mechanisms may be highly dependent on the rate of impact loading. These data could become extremely relevant in the development of high-velocity "safety" devices, such as knee air bags, that are needed to help position an unbelted occupant in an automobile crash.

The extent of matrix damage and chondrocyte death in mechanically traumatized articular cartilage explants depends on rate of loading.

Mechanical loads can lead to matrix damage and chondrocyte death in articular cartilage. This damage has been implicated in the pathogenesis of secondary osteoarthritis. Studies on cartilage explants with the attachment of underlying bone at high rates of loading have documented cell death adjacent to surface lesions. On the other hand, studies involving explants removed from bone at low rates of loading suggest no clear spatial association between cell death and matrix damage. The current study hypothesized that the observed differences in the distribution of cell death in these studies are attributed to the rate of loading. Ninety bovine cartilage explants were cultured for two days. Sixty explants were loaded in unconfined compression to 40 MPa in either a fast rate of loading experiment (approximately 900 MPa/s) or a low rate of loading experiment (40 MPa/s). The remaining 30 explants served as a control population. All explants were cultured for four days after loading. Matrix damage was assessed by measuring the total length and average depth of surface lesions and the release of glycosaminoglycans to the culture media. Explants were sectioned and stained with calcein and ethidium bromide homodimer to document the number of live and dead cells. Greater matrix damage was documented in explants subjected to a high rate of loading, compared to explants exposed to a low rate of loading. The high rate of loading experiments resulted in cell death adjacent to fissures, whereas more dead cells were observed in the low rate of loading experiments and a more diffuse distribution of dead cells was observed away from the fissures. In conclusion, this study indicated that the rate of loading can significantly affect the degree of matrix damage, the distribution of dead cells, and the amount of cell death in unconfined compression experiments on explants of articular cartilage.

Blunt injuries to the patellofemoral joint resulting from transarticular loading are influenced by impactor energy and mass.

Various impact models have been used to study the injury mechanics of blunt trauma to diarthrodial joints. The current study was designed to study the relationship between impactor energy and mass on impact biomechanics and injury modalities for a specific test condition and protocol. A total of 48 isolated canine knees were impacted once with one of three free flight inertial masses (0.7, 1.5, or 4.8 kg) at one of three energy levels (2, 11, 22 J). Joint impact biomechanics (peak load, loading rate, contact area) generally increased with increasing energy. Injuries were typically more frequent and more severe with the larger mass at each energy level. Histological analyses of the patellae revealed cartilage injuries at low energy with deep injuries in underlying bone at higher energies.

Tissue-engineered rotator cuff tendon using porcine small intestine submucosa. Histologic and mechanical evaluation in dogs.

To determine its efficacy in stimulating the regeneration of a rotator cuff tendon, an implant of 10-ply porcine small intestinal submucosa was used to replace a completely resected infraspinatus tendon in 21 adult mongrel dogs. The contralateral infraspinatus tendon was elevated and then reattached to the greater tubercle with sutures to mimic conventional repair (sham operation). Mechanical evaluations were performed at 0, 3, and 6 months (five specimens at each time period). Histologic comparisons were made at 3 and 6 months (three specimens). At both times, the gross appearance, histologic continuity, and failure mode of the constructs mimicked those of sham-operated and native infraspinatus tendons, thus suggesting host tissue ingrowth and implant remodeling with solid integration of the regenerated tissue to muscular and bony interfaces. Tissue ingrowth occurred without histologic evidence of foreign body or immune-mediated reactions or adhesions to peripheral tissues. Sham operations simulated tendon mobilization and reimplantation procedures routinely performed to treat chronic rotator cuff tendon injuries. Although the ultimate strength of small intestinal submucosa-regenerated tendons was significantly less than that of native infraspinatus tendons (P < 0.001), it was similar to that of reimplanted tendons at 3 (P > 0.05) and 6 months (P > 0.05).

Polysulphated glycosaminoglycan treatments can mitigate decreases in stiffness of articular cartilage in a traumatized animal joint.

A single, blunt impact to the rabbit patellofemoral joint has been shown to decrease the stiffness of retropatellar cartilage and increase the thickness of the underlying bone. Polysulphated glycosaminoglycan treatments, on the other hand, have been shown to inhibit the degradation of articular cartilage and possibly increase synthesis of collagen and glycosaminoglycans in experimental studies on diseased joints. The aim of the current study was to examine the effect of early treatments with polysulphated glycosaminoglycans on cartilage using an in vivo post-trauma animal model. The study used 24 Flemish Giant rabbits in three groups: control, impacted, and impacted with treatment. Treatment consisted of intramuscular injections the day of insult and every 4 days thereafter for 6 weeks. At 30 weeks after trauma, mechanical tests were performed on the retropatellar cartilage to determine its mechanical stiffness. The patellae were also grossly evaluated for surface lesions on the retropatellar cartilage and histologically processed to measure the thickness of the subchondral bone. The rabbits that received no treatment had a statistically significant decrease in stiffness (modulus) for the cartilage of the impacted patellae compared with that of the contralateral, unimpacted patellae and compared with the cartilage of rabbits in the control group. The degradation in mechanical stiffness, however, was not observed in patellae of rabbits in the group receiving treatment. There was also a significant increase in the underlying thickness of the subchondral plate on the impacted patellae compared with that on the contralateral, unimpacted sides for rabbits in both the treated and nontreated groups. In conclusion, the polysulphated glycosaminoglycan treatments minimized a decrease in mechanical stiffness (modulus) of retropatellar articular cartilage 30 weeks after trauma. The mechanism by which the mechanical stiffness of the cartilage was preserved is unknown.

Chronic softening of cartilage without thickening of underlying bone in a joint trauma model.

We have recently developed a trauma model to study degradation of the rabbit patello-femoral joint. Our current working hypothesis is that alterations in retropatellar cartilage and underlying bone in our model are initiated independently by acute overstresses developed in each tissue during blunt insult to the joint, and that the processes of chronic degradation in each tissue are not related in a mechanical sense. The current study was conducted in an attempt to help validate our hypothesis by impacting the patello-femoral joint with a padded interface. Based upon earlier human cadaver experiments, we believe this would reduce the acute overstresses in patellar bone while the stresses developed in the overlying retropatellar cartilage would be sufficient enough to initiate a chronic softening of the tissue. Twenty-four animals received an impact to the patello-femoral joint and were sacrificed at either 0, 4.5, or 12 months post-insult. Three acute animals were impacted to develop a simplified computational model to estimate the stresses in joint tissues. The study showed there was a significant softening of the retropatellar cartilage at 4.5 and 12 months post-trauma, compared to unimpacted controls. However, no thickening of the underlying subchondral bone was documented at any timepoint. This was consistent with a reduction of stress in the bone compared to earlier studies, which document thickened subchondral bone post-insult at the same applied impact load. In conclusion, this study helped validate our hypothesis by documenting chronic softening of cartilage without remodeling of the underlying subchondral bone. Furthermore, this study, along with our earlier studies, suggest that impact load alone, which is currently used by the automobile industry to certify new automobiles, is not a good predictor of chronic injuries to a diarthrodial joint, and that simply the addition of padding to impact interfaces may not be adequate to protect occupants from chronic injuries.

The effect of loading rate on the degree of acute injury and chronic conditions in the knee after blunt impact.

accidents are often overlooked, but can have a profound societal cost. Knee injuries, for example, account for approximately 10% of the total injuries. Fracture of the knee is not only an acute issue but may also have chronic, or long term, consequences. The criterion currently used for evaluation of knee injuries in new automobiles, however, is based on experimental impact data from the 70's using seated human cadavers. These studies involved various padded and rigid impact interfaces that slightly alter the duration of contact. Based on these data and a simple mathematical model of the femur, it appears fracture tolerance increases as contact duration shortens. In contrast, more recent studies have shown mitigation of gross fractures of the knee itself using padded interfaces. The use of padded interfaces, however, result in coincidental changes in contact duration and knee contact area. Therefore, it is difficult to extract the direct effect of loading rate on fracture tolerance of the knee. The object of the current study was to isolate the effect of loading rate alone on fracture tolerance of the human knee joint. Paired experiments were conducted on eight pairs of isolated cadaver knees impacted with a rigid interface to approximately 5 kN at a high (5 ms to peak) or low (50 ms to peak) rate of loading. Gross fracture and occult microfractures of the knee joint were documented. A second part of the study examined some chronic effects of loading rate on "subfracture" injuries in an animal. Thirty-four rabbits were subjected to a "subfracture" knee load at the same rates as used in the human studies. Alterations in the mechanical properties of retropatellar cartilage and thickening of subchondral bone were documented out to one year post "subfracture" trauma to the joint. The current study documented an opposite effect than that expected based on 70's experiments with seated cadavers. There was an increase in the number of gross fractures and occult microfractures in high versus low rate of loading experiments. A similar effect was also seen in the "subfracture" chronic animal experiments, which showed relatively more degradative change in the mechanical properties of cartilage following high versus low rate of loading experiments. There was also a significant increase in subchondral bone thickening underlying cartilage and increased fissuring of cartilage in high versus low rate of loading experiments. The current study suggests a relative decrease in tolerance of the knee at high versus low rates of loading in acute experiments with human cadavers and in the chronic setting with animals. Therefore, it would appear that rate of knee loading may be an important issue in establishing a future injury criterion for the knee itself.

The tensile and stress relaxation responses of human patellar tendon varies with specimen cross-sectional area.

In order to provide insight into the mechanical response of the collagen fascicle structures in tendon, a series of constant strain rate and constant displacement, stress relaxation mechanical tests were performed on sequentially sectioned human patellar tendon specimens (protocol 1) and specimens with both small (approximately 1 mm2) and large (approximately 20 mm2) cross-sectional areas (protocol 2). These data described the stress relaxation and constant strain rate tensile responses as a function of cross-sectional area and water content. The experimental data suggested that small portions of tendon exhibit a higher tensile modulus, a slower rate of relaxation and a lower amount of relaxation in comparison to larger specimens from the same location in the same tendon. The decrease in relaxation response and the increase in tensile modulus with decreasing cross-sectional area was nonlinear. These data suggest that there may be structures other than the subfascicle, such as the epitenon and other connective tissue components, which influence the tensile and stress relaxation responses in tendon.