High-Temperature Requirement A1 Protease as a Rate-Limiting Factor in the Development of Osteoarthritis.

Preserving the mature articular cartilage of joints is a critical focus in the prevention and treatment of osteoarthritis. We determined whether the genetic inactivation of high-temperature requirement A1 (HtrA1) can significantly attenuate the degradation of articular or condylar cartilage. Two types of mouse models of osteoarthritis were used, a spontaneous mutant mouse model [type XI collagen-haploinsufficient (Col11a1+/-) mice] and two post-traumatic mouse models [destabilization of the medial meniscus (DMM) on the knee and a partial discectomy (PDE) on the temporomandibular joint]. Three different groups of mice were generated: i) HtrA1 was genetically deleted from Col11a1+/- mice (HtrA1-/-;Col11a1+/-), ii) HtrA1-deficient mice (HtrA1-/-) were subjected to DMM, and iii) HtrA1-/- mice were subjected to PDE. Knee and temporomandibular joints from the mice were characterized for evidence of cartilage degeneration. The degradation of articular or condylar cartilage was significantly delayed in HtrA1-/-;Col11a1+/- mice and HtrA1-/- mice after DMM or PDE. The amount of collagen type VI was significantly higher in the articular cartilage in HtrA1-/-;Col11a1+/- mice, compared with that in Col11a1+/- mice. The genetic removal of HtrA1 may delay the degradation of articular or condylar cartilage in mice.

MMP-3 and MMP-8 in rat mandibular condylar cartilage associated with dietary loading, estrogen level, and aging.

OBJECTIVES: The structure of the mandibular condylar cartilage (MCC) is regulated by dynamic and multifactorial processes. The aim of this study was to examine the effects of altered dietary loading, estrogen level, and aging on the structure of the condylar cartilage and the expressions of matrix metalloproteinase (MMP) -3 and MMP-8 of rat MCC. METHODS: In this study, Crl:CD (SD) female rats were randomly divided into 3 groups according to dietary hardness: hard diet (diet board), normal diet (pellet), and soft diet (powder). In each group, the rats were further divided into 2 subgroups by ovariectomy at the age of 7 weeks. The rats were sacrificed at 5- and 14-month-old. Histomorphometric analysis of the MCC thickness was performed after toluidine blue staining. Immunochemical staining was done for MMP-3 and MMP-8. A linear mixed model was used to assess the effects of dietary loading, estrogen level, and aging. RESULTS: Increased dietary loading was the main factor to increase the MMP-3 expression and the anterior and central thickness of the MCC. Lack of estrogen was the main factor associated with decreased MMP-8. Aging was associated with the thickness changes of the whole condylar cartilage and the reduced expression of MMP-8. CONCLUSION: The condylar cartilage structure and metabolism of the female rats are sensitive to dietary loading changes, estrogen level as well as aging. The proper balance of these factors seems to be essential for the maintenance of the condylar cartilage.

Immunohistochemical expression of matrix metalloprotease-2 and matrix metalloprotease-9 in the disks of patients with temporomandibular joint dysfunction.

PURPOSE: Matrix metalloproteases (MMPs) are tissue-remodeling enzymes that function during the remodeling process, such as in immune-inflammatory diseases. Metalloprotease-2 (MMP-2) and metalloprotease-9 (MMP-9) are gelatinases that degrade several types of extracellular matrix collagen. It is hypothesized that in temporomandibular joint (TMJ) dysfunction, MMP-2 and MMP-9 expression levels may be elevated. Therefore, the objective of this study is to determine the association of MMP-2 and MMP-9 expression with temporomandibular joint dysfunction using an immunohistochemical approach to evaluate the joint disk. MATERIAL AND METHODS: A total of 45 human temporomandibular joint samples were collected, with 36 samples in the test group (patients with anterior disk displacement with reduction (n = 29) and without reduction (n = 7)) and nine samples in the control group. The immunostaining of the TMJ disks was statistically compared between the groups (P < 0.05). RESULTS: There was a statistically significant difference for the area of MMP-2 immunostaining between the control group and the displacement disks with reduction group (ADDwR) (P = 0.048) and between the groups with disk displacement and without reduction (ADDwoR) (P = 0.029). The expression of MMP-2 was significantly elevated in the ADDwoR group. CONCLUSION: No statistically significant difference was found between the variable area of MMP-9 expression in the disk with and without disk displacement, as determined by immunohistochemical analysis. However, there was an elevation of MMP-2 expression in the disks of patients with displacement and without reduction (more severe alteration).

Effect of psychological stress on the structure of the temporomandibular joint and the expression of MMP-3 and TIMP-3 in the cartilage in rats.

Our aim was to observe the effects of psychological stress on the structure of the temporomandibular joint (TMJ), and to evaluate the expression of matrix metallopeptidase-3 (MMP-3) and tissue inhibitor of metalloproteinase-3 (TIMP-3) in condylar chondrocytes in rats. The rats were divided into 3 groups of 12 according to the duration of psychological stress: 3 weeks or 6 weeks, and 6 weeks of recovery. A fourth group of 12 rats was used as controls. Each rat was evaluated by the open-field test and the weight measured. The results confirmed psychological stress in 24 of the 36 rats (67%). The tissues of the TMJ were stained with haematoxylin and eosin and pathological changes were studied under a light microscope. MMP-3 and TIMP-3 expression was investigated using the SP kit. The experimental groups showed thinning of articular cartilage, shedding of collagen fibres, cracks in the articular discs, and other structural changes that were aggravated with time, from three weeks to six weeks. The 6-week recovery group showed an improvement in these changes, which indicated the initiation of joint repair. The MMP-3 expression rate correlated with the degree of joint lesion, while the TIMP-3 rate showed an opposite trend and was highest in the 6-week recovery group. Our findings clearly indicate that psychological stress may play an important part in the development of TMJ diseases in rats; further studies should be made to extrapolate the results to other models before clinical use.

Effects of low-intensity pulsed ultrasound on the expression of cyclooxygenase-2 in mandibular condylar chondrocytes.

AIMS: To determine the effect of low-intensity pulsed ultrasound (LIPUS) on cyclooxygenase-2 (COX-2) expression and related mechanisms by using cultured articular chondrocytes derived from porcine mandibular condyles after treatment with interleukin-1 beta (IL-1ß). METHODS: Chondrocytes were derived from porcine mandibular condylar cartilage and cultured. The cells were treated with or without 10 ng/mL IL-1ß. At the same time, the cells were exposed to LIPUS for 20 minutes. After LIPUS exposure, the conditioned medium was changed to a fresh one without IL-1ß, and the cells were incubated for 0 to 24 hours. The effects of LIPUS on IL-1ß-treated chondrocytes were examined in terms of the expression of p-integrin ß1, COX-2, and phosphorylated extracellular signal-related kinase (p-ERK) 1/2 by real-time polymerase chain reaction (PCR) and Western blot analyses. Differences in the means among multiple groups were examined by one-way analysis of variance (ANOVA) for all groups at each time point, followed by a Scheffé multiple comparison test as a post-hoc test; Student t test was also used. RESULTS: COX-2 mRNA level was upregulated by the treatment with IL-1ß and was suppressed significantly (P < .01) by LIPUS exposure. Furthermore, LIPUS enhanced gene expression and phosphorylation of integrin ß, and it inhibited the expression of p-ERK 1/2. CONCLUSION: LIPUS exposure inhibited IL-1ß-induced COX-2 expression through the integrin ß1 receptor followed by the phosphorylation of ERK 1/2. Despite the restricted duration of its effect, LIPUS is suggested to be a potential candidate as a preventive and auxiliary treatment to suppress the degradation of articular chondrocytes in temporomandibular joint osteoarthritis.

Influence of sleep deprivation on expression of MKK4 and c-fos in the mandibular condylar cartilage of rats.

The aim of this study was to investigate the changes in expression of mitogen-activated protein kinase kinase 4 (MKK4) and c-fos in the mandibular condylar cartilage of rats that had been subjected to sleep deprivation. One hundred and twenty female Wistar rats were randomly divided into 6 groups with 20 in each: sleep deprivation for 2 days, 4 days, 6 days, and 8 days, large-platform controls, and cage controls. After sleep deprivation by the modified multiple platform method the sleep-deprived rats were killed. The large-platform and cage control rats were killed at the same time as the rats deprived of sleep for 8 days. Haematoxylin and eosin were used to record the morphological changes in cartilage, and immunohistochemistry and real-time quantitative polymerase chain reaction (PCR) were used to detect the expression of MKK4 and c-fos. Pathological alterations were apparent after 6 and 8 days of sleep deprivation. Compared with control groups, the expression of MKK4 in the sleep-deprived groups was lower, while that of c-fos was higher. As the duration of sleep deprivation increased, the expression of MKK4 decreased. These results indicate that the variation in expression of MKK4 and c-fos may be correlated with pathological changes induced by sleep deprivation in mandibular condylar cartilage in rats.

Double-stranded RNA-dependent protein kinase regulates insulin-stimulated chondrogenesis in mouse clonal chondrogenic cells, ATDC-5.

Double-stranded RNA-dependent protein kinase (PKR) is an interferon-induced protein that has been identified and characterized as a translational inhibitor in an interferon-regulated antiviral pathway. PKR is also reported to play important roles in the regulation of cell growth and differentiation. We have previously demonstrated that PKR inactivation suppresses osteoblast calcification and osteoclast formation. However, reports concerning the roles of PKR in chondrogenesis are limited. In this study, we have demonstrated that PKR is required for the in vitro differentiation of the mouse clonal chondrogenic cell line ATDC-5. ATDC-5 cells treated with insulin differentiated into chondrocytes and produced an alcian-blue-positive cartilage matrix. The protein expression of signal transducers and activators of transcription (STAT) peaked at day 7 of differentiation, whereas the expression of SRY-box-containing gene 9 (Sox-9), which is a transcription factor for chondrocyte differentiation, increased gradually. When the cells were treated with a PKR inhibitor (2-aminopurine), the cartilage matrix formation decreased. The protein expression of STAT1 continued to increase up to day 21, whereas the expression of Sox-9 was low and did not increase. We also demonstrated that PKR was localized to a marginal region of the mandibular condyle cartilage in mouse embryos. Our findings suggest that PKR has important functions in the differentiation of chondrocytes through the modulation of STAT1 and Sox-9 expression.

Changes in the expression of aromatase, estrogen receptor α and ß in mandibular condylar cartilage of rats induced by disordered occlusion.

BACKGROUND: Estrogens play an important role in modulating the morphology and function of temporomandibular joints (TMJs), which is suggested to act via estrogen receptors (ERs). The present study was to investigate the expression of aggrecan, collagen type II (Col II), Col X, aromatase, ERα and ERß in degenerative changes of mandibular condylar cartilage. METHODS: Forty male and 40 female 8-week-old rats were enrolled in this study. In experimental groups, the disordered occlusion was created by moving the first molars mesially and the third ones distally. Immunohistochemistry and real-time PCR were performed at the end of the second or fourth week. RESULTS: Degenerative changes, characterized by interrupted continuity of hypertrophic layer, pyknotic and eosinophilic lesion with few nuclei, areas filled with eosinophilic nuclei, were observed in more joints from female experimental groups than male ones. However, thickening changes in hypertrophic layer were only found in male experimental groups. The gene expression of Col II, Col X and aggrecan increased in 4-wk male experimental subgroup (both P < 0.01), but decreased in 2-wk and 4-wk female subgroups (P < 0.05). The gene expression of ERα decreased in 2-wk male and female experimental subgroups (both P < 0.01), however, that of ERß increased except the 2-wk female experimental subgroup (all P < 0.01). The expression of aromatase decreased in both male and female experimental subgroups (all P<0.01). CONCLUSIONS: Mandibular condylar cartilage responses differently to the disordered occlusion in male and female rats. The levels of locally synthesized estrogen, ERα and ERß may have limited attribution, if any, to the sex-specific cartilage response.

Dietary loading and aggrecanase-1/TIMP-3 expression in rat mandibular condylar cartilage.

AIMS: To examine the expression of aggrecanase-1 and a tissue inhibitor of metalloproteinases (TIMP-3) in the condylar cartilage of young rats and to determine their relationship during altered dietary loading at different time points after weaning. METHODS: One hundred Sprague-Dawley rats were randomly assigned to 1 of 2 groups: the soft-diet group, which served as the control group (n=50), or the hard-diet group, which served as the experimental group (n=50). Ten soft- and 10 hard-diet rats were killed at 6 hours, 12 hours, 24 hours, 48 hours, and 9 days after weaning (i.e., after initiation of diet change for hard-diet rats). The right-side temporomandibular joints (TMJs) were prepared for immunohistochemical staining. The cartilage from the left-side mandibular condyles of all 10 animals in each group was combined for Western blot analysis. RESULTS: Immunohistochemical analysis revealed strong staining for aggrecanase-1 localized mainly in the chondrocytes of proliferative and upper hypertrophic cartilage zones at all time points in both groups. The immunohistological expression of aggrecanase-1 was significantly higher in the hard-diet group at 12 and 24 hours than in the soft-diet group. Strong staining for TIMP-3 was mainly localized in the chondrocytes of proliferative and upper hypertrophic zones at all time points in both groups. The expression of TIMP-3 in the hard-diet group was at a significantly lower level compared to the soft-diet group at 6 hours. Western blot analysis also showed time-related differences in aggrecanase-1 and TIMP-3, but there was no significant difference between the 2 groups. CONCLUSION: The temporary change in aggrecanase-1 and TIMP-3 expression reflects the complex interaction of these enzymes in the physiologic range and cartilage response to altered dietary loading.

Induction of MMP-3 by hyaluronan oligosaccharides in temporomandibular joint chondrocytes.

Low-molecular-weight hyaluronan (LMW-HA) is often increased in osteoarthritic joints; however, its biological function in cartilage has not been clarified. We hypothesize that LMW-HA causes the catabolic activation of chondrocytes through its interaction with CD44. Cartilage explants and chondrocytes, derived from bovine temporomandibular joints (TMJ), were examined for matrix loss and the expression of matrix metalloproteinase-3 (MMP-3) following treatment with hyaluronan oligosaccharides (HA(oligos)). Hyaluronan and CD44 were uniformly distributed throughout the fibrous and cartilaginous zones of the TMJ condyle. Treatment of cartilage explants with HA(oligos) resulted in cartilage matrix loss with increased secreted caseinolytic activity. HA(oligos) treatment of TMJ chondrocytes resulted in enhanced MMP-3 expression, whereas wash-out of the HA(oligos) in the middle of the experimental period reduced this induction. These results suggest that HA(oligos) activate chondrocytes, resulting in a substantial enhancement of proteinase expression, and the removal of HA(oligos) by wash-out reverses this catabolic activation.

Age-related changes in the expression of gelatinase and tissue inhibitor of metalloproteinase genes in mandibular condylar, growth plate, and articular cartilage in rats.

Mandibular condylar cartilage acts as both articular and growth plate cartilage during growth, and then becomes articular cartilage after growth is complete. Cartilaginous extracellular matrix is remodeled continuously via a combination of production, degradation by matrix metalloproteinases (MMPs), and inhibition of MMP activity by tissue inhibitors of metalloproteinases (TIMPs). This study attempted to clarify the age-related changes in the mRNA expression patterns of MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 in mandibular condylar cartilage in comparison to tibial growth plate and articular cartilage using an in situ hybridization method in growing and adult rats. MMP-2 and MMP-9 were expressed in a wide range of condylar cartilage cells during growth, and their expression domains became limited to mature chondrocytes in adults. The patterns of TIMP-1 and TIMP-2 expression were similar to those of MMP-2 and MMP-9 during growth, and were maintained until adulthood. TIMP-3 was localized to hypertrophic chondrocytes throughout the growth stage. Therefore, we concluded that TIMP-1 and TIMP-2 were general inhibitors of MMP-2 and MMP-9 in condylar cartilage, while TIMP-3 regulates the collagenolytic degradation of the hypertrophic cartilage matrix.

Immunohistochemical localization of matrix metalloproteinase 13 (MMP-13) in mouse mandibular condylar cartilage.

MMP-13 appears to be one of the most important MMPs in cartilage remodeling and mineralization, because it exhibits a substrate preference for the cartilage-specific type II collagen. The condylar process is constructed by rapid accumulation of hypertrophic chondrocytes during development, but its mechanism is still unclear. To investigate the role of MMP-13 in developing condylar cartilage, we immunohistochemically examined the localization of MMP-13 in the endochondral ossification of the mandibular condyle and tibiae of newborn mice. In the tibiae, the MMP-13 expression was detected only in the deepest layer of the terminal hypertrophic chondrocytes through every examined stage (day 1 to day 10 after birth). On the other hand, in the condylar cartilage at days 1 and 5, MMP-13 was expressed throughout the proliferating and the hypertrophic chondrocytes, and at day 10, MMP-13 was mainly localized in the deepest edge of the hypertrophic layer. A zymographical study showed that the activity of MMP-13 in the condyle was observed at day 1, earlier than in the tibia, and increased until day 7. The time-dependent and cell-specific expression of MMP-13 and its enzymatic property suggest that in the mandibular condylar cartilage, MMP-13 plays a role in making the space for cell enlargement by degradation of the cartilage matrix and in onset of mineralization during the early stage of development.

Distribution of plasma-membrane Ca2+ pump in mandibular condyles from growing and adult rabbits.

Chondrocytes may control the mineralization of the extracellular matrix of condylar cartilage by several mechanisms including the release of microvesicles involved in the initial nucleation, the creation or modification of the local matrix to help propagate or restrict mineralization, and the regulation of the ionic environment at the calcifying foci within the matrix. The plasma membrane Ca2+-Mg2+ ATPase (Ca2+ pump) is known to play a part in the vectorial efflux of calcium in a variety of cells including chondrocytes. The purpose here was to study the distribution of Ca2+-pump protein in mandibular condyles from growing and adult rabbits, and compare the expression of that protein in progressively differentiating chondrocytes whose final stage is associated with a mineralized extracellular matrix. Ca2+-pump antigen was identified immunohistochemically in six growing and six adult rabbit mandibular condyles with a Ca2+ pump-specific monoclonal antibody. The presence of Ca2+-pump antigen was established in hypertrophic chondrocytes, and in osteoblasts and osteoclasts of subchondral bone. Slot-blot analysis of nitrocellulose-immobilized chondrocyte homogenates showed that the amount of Ca2+ pump in growing cartilage was more than twice that in adult cartilage (p < 0.05). The demonstration of Ca2+-pump antigen in the hypertrophic chondrocytes of growing rabbit condyles is consistent with a role for the plasma-membrane Ca2+ pump in the calcification of mandibular condylar cartilage.

Influences of ovariectomy and orchiectomy on the remodeling of mandibular condyle in mice.

Ovariectomy (OVX) and orchiectomy (ORX) are well known to increase bone turnover; however, the influences on condylar remodeling are not fully understood. The purpose of this study was to examine histological changes of the mandibular condyles in OVX and ORX mice in comparison to the femora. Eighty 8-week-old mice were used in this study. In each of the experimental and control groups, five mice were sacrificed 2, 4, 8, and 12 weeks after surgery for OVX and ORX. Histological changes of the mandibular condyle were studied in comparison to the femur, which is liable to undergo the influences of OVX and ORX. The number of TRAP-positive cells and the thickness of articular cartilage layer were also quantified. TRAP-positive cells in the condylar head of OVX and ORX mice exhibited the most prominent increase 4 weeks after the surgery and approached the control levels at the later experimental stages. Trabecular bone volume in the condyle of the OVX and ORX mice also decreased, although the rate was less than in the femora. It is shown that influences of OVX on the remodeling of femora appear earlier than those of ORX, whereas OVX and ORX affect the remodeling of condyle during the same growth period. It is also demonstrated that the influences of OVX and ORX are less in the condyle than in the femora. These findings emphasize that the influences of OVX and ORX on bone remodeling are varied by sexual difference and the type of bones.

Immunohistochemistry of collagen types II and X, and enzyme-histochemistry of alkaline phosphatase in the developing condylar cartilage of the fetal mouse mandible.

We investigated the immunohistochemical localisation of types II and X collagen as well as the cytochemical localisation of alkaline phosphatase in the developing condylar cartilage of the fetal mouse mandible on d 14-16 of pregnancy. On d 14 of pregnancy, although no immunostaining for types II and X collagen was observed, alkaline phosphatase activity was detected in all cells in the anlage of the future condylar process. On d 15 of pregnancy, immunostaining for both collagen types was simultaneously detected in the primarily formed condylar cartilage. Alkaline phosphatase activity was also detected in chondrocytes at this stage. By d 16 of pregnancy, the hypertrophic cell zone rapidly increased in size. These findings strongly support a periosteal origin for the condylar cartilage of the fetal mouse mandible, and show that progenitor cells for condylar cartilage rapidly or directly differentiate into hypertrophic chondrocytes.

Stimulation of alkaline phosphatase activity by ascorbic acid and suppression by 1,25-dihydroxycholecalciferol in rabbit craniofacial chondrocytes in culture.

To clarify the role of vitamins D and C in chondrocyte hypertrophy of craniofacial cartilage, we have studied cultured chondrocytes from rabbit mandibular condylar cartilage (MCC), sphenooccipital synchondrosis (SOS) and nasal septal cartilage (NSC) under conditions in which these cells mature into hypertrophic chondrocytes. In cultures of MCC- and SOS-chondrocytes, alkaline phosphatase (ALPase) activity started to increase on day 9 at confluence and the cessation of cell division, and reached a maximum on day 18. The degree of the increase of ALPase activity on day 18 was higher in MCC-chondrocytes than in SOS-chondrocytes. ALPase activity was very low level in NSC- and CGC-chondrocytes. Ascorbic acid induced a marked increase in ALPase activity in MCC-, SOS-, NSC- and CGC-chondrocytes. The ALPase activities in MCC- and SOS-chondrocytes with 50 micrograms/ml ascorbic acid were 2.5-times those in its absence. Those in NSC- and CGC-chondrocytes were 10 times and 20 times, respectively. When chondrocytes were cultured with 10% charcoal-treated serum, ALPase activity decreased less than that in cultures with 10% normal serum in MCC-, SOS-, NSC- and CGC-chondrocytes. Treatment of 1,25-(OH)2D3 for 9 days from days 4 to 13 in MCC-chondrocytes and for 14 days from days 4 to 18 in SOS-, NSC- and CGC-chondrocytes inhibited ALPase activity dose-dependently at the concentrations of 10(-12) M to 10(-8) M in MCC- chondrocytes, 10(-10) M to 10(-8) M in SOS- and CGC-chondrocytes, 10(-11) M to 10(-9) M in NSC-chondrocytes. These findings suggest that 1,25-(OH)2D3 and ascorbic acid may be involved in the control of cartilage growth and terminal differentiation.

Internal derangement of the temporomandibular joint: a histochemical study.

The purpose of this study was to correlate histologic findings in temporomandibular joint (TMJ) condyles and discs with their macroscopic appearance at surgery. The 24 patients with internal derangement of the joint included 20 women and 4 men (mean age, 37 years; range, 18 to 61 years). The tissue lesions varied in degree from mild soft-tissue fraying and bone remodeling to extensive resorption and new cartilage and bone formation with high phosphatase enzyme activities, and even to loss of articular soft tissue and breakdown of cortical bone. Reactions may arise in the hard tissues before they occur in the articular surface layers.

The activity of 2,5-oligoadenylate synthetase, an interferon-induced enzyme, is coupled to the differentiation state of mouse condylar cartilage.

The enzyme 2,5-oligoadenylate synthetase (2-5A synthetase) is associated with the interferon system, with special reference to the differentiation process of various cell types. The present study investigated whether 2-5A synthetase is also involved in the differentiation of neocartilage in perinatal mice. The cartilage of the mandibular condyle, a secondary type of cartilage, develops relatively late in prenatal life; and consequently it was possible to obtain a relatively embryonic cartilage at a developmental stage that could be manipulated enzymatically, in order to separate and thereby obtain its undifferentiated, proliferative portion along with its more mature fraction. Immunohistochemical studies using antibodies against type I and type II collagen and cartilage-specific proteoglycans could have determined the differentiation status of various portions of the developing condyle. However, the above methodology lacks the necessary precision and accuracy to indicate subtle changes in cellular differentiation. It became evident that the activity of 2-5A synthetase was indeed different in cellular compartments that were at different stages of differentiation. In the neonatal condyle the highest level of activity was encountered in proliferating and as yet undifferentiated prechondrocytes, whereas fully differentiated chondrocytes showed a marked decrease in the activity of this enzyme.

Collagenolytic and phosphatase activity in the rat mandible after functional protrusion.

The effect of chronic mandibular protrusion on the collagenolytic and phosphatase activity of several mandibular bone sites and the condylar cartilage was evaluated. Ninety-three male Sprague-Dawley rats were equally divided into two experimental and one control group. One experimental group wore a protrusive appliance for 2 weeks, the other for 4 weeks. All animals were killed at 59 days of age. Collagenolytic, alkaline and cid phosphatase activities were determined in the condylar cartilage, the subchondral bone and condylar neck, and in the gonial angle and coronoid process. In the cartilage and subchondral bone, the protrusive appliance caused a reduction in collagenolytic and alkaline phosphatase activity. In the condylar neck, it caused a large increase in collagenolytic activity and a decrease in alkaline phosphatase activity in both experimental groups. In the gonial angle and coronoid process, the appliance increased the collagenolytic activity only in the 2-week group. In the 4-week group, the alkaline phosphatase and collagenolytic activities were not different from the activities in those tissues in the control animals. Thus a protrusive appliance induced quantitative changes in enzyme activities in condylar cartilage and mandibular bone. The increase in collagenolytic activity (representing increased bone resorption) occurred typically in areas of muscle attachment and might have been the result of the neuromuscular changes induced by the protrusive appliance. The recovery to normal values of collagenolytic activity in the coronoid process and gonial angle of the 4-week group suggests that at these sites the muscles (and subperiosteal bone) might have adapted to their new biomechanical environment after the longer period of appliance wear.

Effect of condylotomy on matrix synthesis and mineralization in the rat mandibular condylar cartilage.

Little information is available about the influence of biomechanical factors on the rate of matrix synthesis and mineralization in the condylar cartilage. In an attempt to temporarily disrupt articular forces at the temporomandibular joint, bilateral condylotomies were performed on 29 day-old male Sprague-Dawley rats. Animals which underwent a similar surgical procedure except for actual condylotomy comprised a sham-operated control group; a non-operated third group served as further controls. Incorporation of [35S]-sulphate did not differ among the groups at 18 h and 2 days after condylotomy, but was significantly reduced in condylotomy animals at 7 and 14 days. Total alkaline phosphatase activity was reduced at 18 h and increased at 14 days by condylotomy, with no differences at the intermediate time intervals; similarly 45Ca-uptake was increased at the 14-day interval. Thus both matrix synthesis and mineralization of the condylar cartilage were altered in the 2 weeks after condylotomy. The changes are in general similar to those noted in other studies after a lessening or removal of articular forces from the condyle.