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Objective: Fibroblast growth factors (FGFs) are a family of 22 proteins and 4 FGF receptors (FGFRs) that are crucial elements for normal development. The contribution of different FGFs and FGFRs for the homeostasis or disease of the cartilage from the mandibular condyle is unknown. Therefore, our goal was to characterize age-related alterations in the protein expression of FGF ligands and FGFRs in the mandibular condyle of mice., Method: Mandibular condyles of 1-, 6-, 12-, 18-, and 24-month-old C57BL/6J male mice (5 per group) were collected and histologically sectioned. Immunofluorescence for FGFs that have been reported to be relevant for chondrogenesis (FGF2, FGF8, FGF9, FGF18) as well as the activated/phosphorylated FGFRs (pFGFR1, pFGFR3) was carried out., Results: FGF2 and FGF8 were strongly expressed in the cartilage and subchondral bone of 1-month-old mice, but the expression shifted mainly to the subchondral bone as mice aged. FGF18 and pFGFR3 expression was limited to the cartilage of 1-month-old mice only. Meanwhile, pFGFR1 and FGF9 were mostly limited to the cartilage with a significant increase in expression as mice aged., Conclusions: Our results indicate FGF2 and FGF8 are important growth factors for mandibular condylar cartilage growth in young mice but with limited role in the cartilage of older mice. In addition, the increased expression of pFGFR1 and FGF9 and the decreased expression of pFGFR3 and FGF18 as mice aged suggest the association of these factors with aging and osteoarthritis of the cartilage of the mandibular condyle., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

For decades, it has been widely accepted that hypertrophic chondrocytes undergo apoptosis prior to endochondral bone formation. However, very recent studies in long bone suggest that chondrocytes can directly transform into bone cells. Our initial in vivo characterization of condylar hypertrophic chondrocytes revealed modest numbers of apoptotic cells but high levels of antiapoptotic Bcl-2 expression, some dividing cells, and clear alkaline phosphatase activity (early bone marker). Ex vivo culture of newborn condylar cartilage on a chick chorioallantoic membrane showed that after 5 d the cells on the periphery of the explants had begun to express Col1 (bone marker). The cartilage-specific cell lineage-tracing approach in triple mice containing Rosa 26(tdTomato) (tracing marker), 2.3 Col1(GFP) (bone cell marker), and aggrecan Cre(ERT2) (onetime tamoxifen induced) or Col10-Cre (activated from E14.5 throughout adult stage) demonstrated the direct transformation of chondrocytes into bone cells in vivo. This transformation was initiated at the inferior portion of the condylar cartilage, in contrast to the initial ossification site in long bone, which is in the center. Quantitative data from the Col10-Cre compound mice showed that hypertrophic chondrocytes contributed to ~80% of bone cells in subchondral bone, ~70% in a somewhat more inferior region, and ~40% in the most inferior part of the condylar neck (n = 4, P < 0.01 for differences among regions). This multipronged approach clearly demonstrates that a majority of chondrocytes in the fibrocartilaginous condylar cartilage, similar to hyaline cartilage in long bones, directly transform into bone cells during endochondral bone formation. Moreover, ossification is initiated from the inferior portion of mandibular condylar cartilage with expansion in one direction., (© International & American Associations for Dental Research 2015.)

Osterix (Osx) is a transcription factor essential for osteoblast differentiation and bone mineralization. Although there are indications that Osx also plays a regulatory role in cartilage, this has not been well-studied. The goal of this study was to define the function of Osx in the post-natal growth of the secondary cartilage at the mandibular condyle. Conditional Osx knockout (cKO) mice that were missing Osx only in cartilage were generated by crossing Osx-loxP mice to Aggrecan-Cre mice. Cre activity was induced by tamoxifen injection twice a week from day 12 to 1 mo of age, and specimens were collected at 1 and 5 mo of age. At 1 mo of age, the condylar hypertrophic chondrocyte zone in the cKO-mice was > three-fold thicker than that in the age-matched control, with little sign of endochondral bone formation. Immunohistochemistry and analysis of histological data revealed a defect in the coupling of chondrogenesis and osteogenesis in the cKO mice. In five-month-old mice examined to address whether late-stage removal of the Cre-deletion event would alleviate the phenotype, the hypertrophic chondrocyte zone in the cKO condyles was considerably larger than in wild-type mice. There were large discrete areas of calcified cartilage in the hypertrophic zone, few signs of endochondral bone formation, and large regions of disorganized intramembranous bone. Analysis of these data further strengthens the notion that Osterix is essential for the coupling of terminal cartilage differentiation and endochondral ossification in mandibular condylar cartilage., (© International & American Associations for Dental Research.)

Condylar regeneration with the use of functional appliances after condylectomy has been validated. However, the process during treatment remains unclear. In this study, we evaluated the condylar regeneration process and then examined mandibular growth and masticatory muscle activity after regeneration in growing rats. Seventy-five male Wistar rats aged 4 weeks were equally divided into 3 groups: unilateral condylectomy group, unilateral condylectomy + appliance group, or control group. The use of a functional appliance following condylectomy promoted mandibular growth and regeneration of the condyle 1 week after condylectomy. Condyle regeneration showing normal morphology was finally achieved 8 weeks after condylectomy. Asymmetrical masticatory muscle activity was observed after condylectomy. However, the use of a functional appliance produced symmetrical masticatory muscle activity. These results indicate a favorable regeneration process in the condylectomized area due to the use of a functional appliance. In addition, due to condylar regeneration, symmetrical masticatory muscle activity was achieved.

Primary cilia regulate limb and axial skeletal formation and hedgehog signaling, but their roles in temporomandibular joint (TMJ) development are unknown. Thus, we created conditional mouse mutants deficient in ciliary transport protein Kif3a in cartilage. In post-natal wild-type mice, primary cilia were occasionally observed on the superior, inferior, or lateral side of condylar cells. Cilia were barely detectable in mutant chondrocytes but were evident in surrounding tissues, attesting to the specificity of chondrocyte Kif3a ablation. Mutant condyles from 3-month-old mice were narrow and flat along their antero-posterior and medio-lateral axes, were often fused with the articular disc, and displayed an irregular bony surface. The polymorphic layer in P15 mutants contained fewer Sox9-expressing chondroprogenitor cells because of reduced mitotic activity, and newly differentiated chondrocytes underwent precocious hypertrophic enlargement accompanied by early activation of Indian hedgehog (Ihh). Interestingly, there was excessive intramembranous ossification along the perichondrium, accompanied by local expression of the hedgehog receptor Patched-1 and up-regulation of Osterix and Collagen I. In summary, Kif3a and primary cilia are required for coordination of chondrocyte maturation, intramembranous bone formation, and chondrogenic condylar growth. Defects in these processes in Kif3a condylar cartilage are likely to reflect abnormal hedgehog signaling topography and dysfunction.

Indian hedgehog (Ihh) is essential for embryonic mandibular condylar growth and disc primordium formation. To determine whether it regulates those processes during post-natal life, we ablated Ihh in cartilage of neonatal mice and assessed the consequences on temporomandibular joint (TMJ) growth and organization over age. Ihh deficiency caused condylar disorganization and growth retardation and reduced polymorphic cell layer proliferation. Expression of Sox9, Runx2, and Osterix was low, as was that of collagen II, collagen I, and aggrecan, thus altering the fibrocartilaginous nature of the condyle. Though a disc formed, it exhibited morphological defects, partial fusion with the glenoid bone surface, reduced synovial cavity space, and, unexpectedly, higher lubricin expression. Analysis of the data shows, for the first time, that continuous Ihh action is required for completion of post-natal TMJ growth and organization. Lubricin overexpression in mutants may represent a compensatory response to sustain TMJ movement and function.

Previous studies have indicated that an injured condyle during adolescence is a causative factor for reduced mandibular growth and resulting asymmetry of the mandible. The aim of this study was to examine the nature of mandibular growth after unilateral condylectomy and to elucidate the effects of mandibular advancement. Sixty growing mice were subjected to unilateral condylectomy, and then one-half of them underwent treatment with a functional appliance. After 4 wks, a unilateral condylectomy produced reduced growth of the mandible and a subsequent lateral shift to the affected side. However, reduced growth and a lateral shift of the mandible were eliminated by a functional appliance, and prominent regeneration of the condyle was also demonstrated. It was shown that mandibular advancement provides for the regeneration of cartilaginous tissues on injured condyles and recovery of reduced mandibular growth, leading to correction of the lateral shift of the mandible.

Orthodontic tooth movement is dependent upon the remodeling of the periodontal ligament and alveolar bone by mechanical means. Facial sutures are also fibrous articulations, and by remodeling these joints, one can alter the positional relationships of the bones of the facial skeleton. As might be expected from the structure and mobility of the temporomandibular joint (TMJ), this articulation is more resistant to mechanical deformation, and whether functional mandibular displacement can alter the growth of the condyle remains controversial. Clinical investigations of the effects of the Andresen activator and its variants on dentofacial growth suggest that the changes are essentially dento-alveolar. However, with the popularity of active functional appliances, such as the Herbst and twin-block based on 'jumping the bite', attention has focused on how they achieve dentofacial change. Animal experimentation enables informed decisions to be made regarding the effects of orthodontic treatment on the facial skeleton at the tissue, cellular, and molecular levels. Both rat and monkey models have been widely used, and the following conclusions can be drawn from such experimentation: (1) Facial sutures readily respond to changes in their mechanical environment; (2) anterior mandibular displacement in rat models does not increase the mitotic activity of cells within the condyle to be of clinical significance, and (3) mandibular displacement in non-human primates initiates remodeling activity within the TMJ and can alter condylar growth direction. This last conclusion may have clinical utility, particularly in an actively growing child.

The impact of mandibular distraction on condyles is poorly understood. To examine how condylar mineralization is affected, we performed distraction in 128 one-month-old rapidly and 126 three-month-old slowly growing rats. The rate of distraction was 0.0 mm (sham), 0.2 mm (slow), 0.4 mm (moderate), or 0.6 mm (rapid). From 7 to 9 rats from each rate (n = 29-32) were killed at 4 time periods (D6, D10, D24, and D38) following osteotomy. Calcein and alizarin were injected 6 and 3 days, respectively, prior to death. Methacrylate-embedded sagittal condylar sections were examined under epifluorescence, and mineral apposition rates were measured. Results indicated that: (1) rapidly growing rats showed higher mineral apposition rates (p < 0.01-0.001) than did slowly growing rats; (2) mineral apposition rates were lower in distracted sides at all times in rapidly growing rats (p < 0.05-0.01), while this side-dependency was seen only at D24 in slowly growing rats (p < 0.05); and (3) distraction rates had little effect on mineral apposition rates. Thus, mandibular distraction decreases condylar mineral apposition rates, but only in rapidly growing rats, which is related to surgery and its functional consequences, not to the distraction rate.

Indian hedgehog (Ihh) is a critical mediator transducing mechanical signals to stimulate chondrocyte proliferation. To clarify the cellular signal transduction pathway that senses and converts mechanical signals into tissue growth in mandibular condyle, we evaluated Ihh expression and its relation to the kinetics of replicating mesenchymal cells in condylar cartilage during natural growth and mandibular advancement. Thirty-five-day-old Sprague-Dawley rats were fitted with functional appliances. Experimental animals with matched controls were doubly labeled with iododeoxyuridine and bromodeoxyuridine so that we could evaluate the cycles of the proliferative mesenchymal cells. Mandibular advancement triggered Ihh expression in condylar cartilage. A higher level of Ihh expression coincided with the increase of the replicating mesenchymal cells' population and the shortening of the turnover time. These findings suggested that Ihh acts as a mediator of mechanotransduction that converts mechanical signals resulting from anterior mandibular displacement to stimulate cellular proliferation in condylar cartilage.

Little is known about the mechanisms of mandibular condylar growth. In this study, gene expression in the mandibular condylar cartilage of young post-natal mice was monitored by means of a cDNA microarray, real-time PCR, and laser microdissection before and after the initiation of mastication (newborn, 7 days, 21 days, initiation of mastication, and 35 days). Insulin-like growth factor-1 (IGF-I), transforming-growth-factor-beta-2 (TGFbeta2), and aggrecan mRNAs were clearly expressed at 21 days, while the expression of osteopontin mRNAs was most clear at 35 days. Parathyroid-hormone-related protein (PTHrP), Indian-hedgehog (Ihh), and insulin-like growth factor-2 (IGF-2) mRNAs were clearly expressed during lactation (newborn and 7 days). Heat-shock-protein 84 (HSP-84) and heat-shock-protein 86 (HSP-86) were clearly expressed at 35 days. These results revealed that gene expression changed during mandibular condylar cartilage growth, and that, interestingly, these changes coincided with the initiation of mastication.

Asymmetric growth occurs frequently in the mandibulofacial region, but little attention has been given to asymmetry in the temporomandibular joint. The purpose of this study was to clarify the feature of asymmetry in the condylar long axis and its relation to upper first molar rotation. Records of 148 pre-orthodontic patients were used. The angle of the condylar long axis and that of the molar rotation were both larger on the left side than on the right side. Positive correlations were found between the corresponding bilateral measurements of condylar long axes and also between those of molar rotations, whereas no correlation was found between the condylar long axis and molar rotation. These findings were found in most subgroups classified by dental age, skeletal pattern, bite force balance, or gender. These results suggest that consistent left-right differences in the condylar long axis and first molar rotation are common.

The temporomandibular joint is susceptible to diseases and trauma that may ultimately lead to structural degeneration. Current approaches for replacing degenerated mandibular condyles suffer from deficiencies such as donor site morbidity, immunorejection, implant wear and tear, and pathogen transmission. The hypothesis of this study was that a human-shaped mandibular condyle can be tissue-engineered from rat mesenchymal stem cells (MSCs) encapsulated in a biocompatible polymer. Rat bone marrow MSCs were isolated and induced to differentiate into chondrogenic and osteogenic cells in vitro, and encapsulated in poly(ethylene glycol)-based hydrogel in two stratified layers molded into the shape of a cadaver human mandibular condyle. Eight weeks following in vivo implantation of the bilayered osteochondral constructs in the dorsum of immunodeficient mice, mandibular condyles formed de novo. Microscopic evaluation of the tissue-engineered mandibular condyle demonstrated two stratified layers of histogenesis of cartilaginous and osseous phenotypes. The current approach is being refined for ultimate therapeutic applications.

PTHrP is a key factor regulating the pace of endochondral ossification during skeletal development. Mandibular advancement solicits a cascade of molecular responses in condylar cartilage. However, the pace of cellular maturation and its effects on condylar growth are still unknown. The purpose of this study was to evaluate the pattern of expression of PTHrP and correlate it to cellular dynamics of chondrocytes in condylar cartilage during natural growth and mandibular advancement. We fitted 35-day-old Sprague-Dawley rats with functional appliances. Experimental animals with matched controls were labeled with bromodeoxyuridine 3 days before their death, so that mesenchymal cell differentiation could be traced. Mandibular advancement increased the number of differentiated chondroblasts and subsequently increased the cartilage volume. Higher levels of PTHrP expression in experimental animals coincided with the slowing of chondrocyte hypertrophy. Thus, mandibular advancement promoted mesenchymal cell differentiation and triggered PTHrP expression, which retarded their further maturation to allow for more growth.

Bone and cartilage metabolism is known to be more active during rest than during periods of activity. The purpose of this study was to examine the hypothesis that mandibular retractive force could be more effective when applied to rats during rest. Mandibular retractive force caused a considerable reduction in the condylar length in experimental groups, and the magnitude of this reduction was greater in the Light-period (08:00-20:00) group than in the Dark-period (20:00-08:00) group. The differentiation and proliferation of chondrocytes were inhibited in animals in the Light-period group, compared with those in the Dark-period group. These results suggest that the orthopedic effects of mandibular retractive force vary depending on the time of day the force is applied, and that such force may be more effective while animals are resting than while they are active.

Juvenile rheumatoid arthritis (JRA) is a chronic systemic disease of childhood that affects synovial joints including the temporomandibular joint (TMJ). Individuals with JRA of the TMJ frequently show aberrations in mandibulofacial development. Since the basis for these developmental perturbations is poorly understood, they remain a perplexing clinical problem to manage. To begin dissecting the mechanisms for altered craniofacial development in JRA of the TMJ, we characterized the gross morphologic adaptations in the facial skeleton in a juvenile animal model of TMJ arthritis. Arthritis was induced in ten 87-day-old male rabbits by intra-articular challenge with ovalbumin. Eight sham-challenged and 4 unchallenged rabbits were used as controls. Serial lateral head cephalograms, taken at 73 (T1), 87 (T2), 108 (T3), 129 (T4), and 150 (T5) days of age, were evaluated by linear measures of maxillary, mandibular, and posterior dental height dimensions. Differences in the absolute dimensions and relative percent incremental changes were compared by ANOVA and Fisher's test. The body weights, as well as the absolute measures and incremental changes in maxillary and posterior dental height dimensions, were not significantly different between the antigen-challenged and control groups. In contrast, absolute measures of posterior mandibular height, condylar neck height, and total mandibular length were significantly smaller (P < 0.05) in antigen-challenged rabbits than in both control groups at T5. Furthermore, the antigen-challenged rabbits demonstrated significantly smaller (P < 0.05) relative increases in all measures of mandibular length, and in total posterior mandibular and condylar neck heights. Cephalometric superimpositions on the cranial base and tantalum implants confirmed these quantitative observations. This investigation demonstrates mandibulofacial developmental aberrations in experimental JRA-like disease of the TMJ that are similar to those observed in humans with this disease.

The sequelae of sustained, in vivo delivery of two important neurotransmitter substances, glutamate and thyrotropin-releasing hormone (TRH), upon craniofacial growth and development have previously not been investigated. Our purpose was to document and compare the relative effects of glutamate and TRH microspheres stereotactically placed in proximity to trigeminal motoneurons within the trigeminal motor nucleus. The following null hypotheses were tested: (1) TRH microspheres in proximity to trigeminal motoneurons have no significant effect upon the craniofacial skeleton, and (2) there are no significant differences between the relative effects of chronic, long-term delivery of glutamate and TRH upon the neuromusculoskeletal system of growing rats. Forty male Sprague-Dawley rats were divided into 4 experimental groups (glutamate microspheres, TRH microspheres, blank microspheres, sham surgeries) and underwent stereotactic neurosurgery at 35 days; 5 rats of each group were killed at 14 and 21 days for data collection. Histology revealed that implants were clustered in the pontine reticular formation, close to the ventrolateral tegmental nucleus. Both glutamate and TRH rats had implant-side deviation of their facial skeleton and snout regions; 4 x 2 ANOVA and post hoc t-tests revealed significant (P < or = 0.05, 0.01) differences between groups and sides for motoneuron count, muscle weight, and osteometric data. TRH rats also demonstrated larger implant-side TMJ discs and mandibular fossae in comparison with the other groups. The stated null hypotheses were therefore rejected.

The effects of mechanical factors on the growth of the mandibular condyle were studied by monitoring the maturation of the mesenchymal cells in 55 rats. Thirty-five animals were fed normal pellet food, and 20 were fed a soft diet and their incisors were cut regularly. 3H-thymidine was injected intraperitoneally three days before death at 18, 23, or 33 days. Histologic sections showed the most advanced 3H-thymidine-labeled cells to occur deep in the cartilage, in the lower hypertrophic cell layer in anterior and posterior regions of the condyle, and in the upper hypertrophic cell layer in the superior region at the age of 18 days. A distinct difference in the maturation state of the labeled cells could also be observed between these regions. In animals fed a soft diet, maturation was slower in the superior region of the condyle and faster in the posterior region than in the normal rats. The rate at which cells stepped out of the proliferating cell pool was measured by use of monoclonal antibodies against proliferating cell nuclear antigen. The ratio between labeled cells in the proliferating cell layer and the number of labeled cells beneath it was greater in control animals than in the soft-diet animals. The rate of differentiation and maturation of mesenchymal cells into chondrocytes seems to be controlled by mechanical factors.

Attempts to increase mandibular growth by the stimulation of condylar proliferative activity, either experimentally or with functional appliances in humans, have led to controversial results. The aim of this study was to measure changes in proliferative activity in the mandibular condyle after steady experimental posterior relocation of the glenoid fossa in the rabbit without actively interfering with normal masticatory action. The method differed from most previous experimental procedures, which force the mandible anteriorly to stimulate functional appliance therapy. Twelve 5-day-old rabbits underwent gluing of the interparietal, temporoparietal, and lambdoidal sutures. Three experimental and three control rabbits were injected with tritiated thymidine at 10, 15, 20, and 30 days and were killed after 2 h for histological and autoradiographic examination. The total number of labeled cells in the prechondroblastic layer was higher in the experimental group, the difference being greatest in the 20-day-old rabbits. The highest proliferative activity in the experimental group was found in the area immediately posterior to the articular contact surface. There was a tendency for the cartilage layers to be thicker in the experimental group, especially in the extreme anterior segments of the condyle.

In the present study, the role of the periosteum in mandibular growth was investigated. The orientation of the superficial bony spicules of rabbit mandibles was determined on dry skulls after perfusion of the animals with an India ink solution. The spicular orientation in the ramus area appeared to be toward the condyle, rostrally toward the incisors, and caudally toward the angular region. The behavior, during growth, of the periosteum in the caudal mandibular half was studied by implantation of metal periosteal and bone markers. A series of cephalograms revealed the migration pattern of the periosteal markers, and by that the migration pattern of the periosteum. It can be concluded that both the pattern of the superficial bony spicules and the periosteal migration pattern suggest a possible influence of the periosteum on mandibular growth.

The present study describes the behavior of mandibular condylar cartilage (MCC) cells as a function of time in primary culture, since it is not yet clear whether these cells maintain their phenotype in culture. MCC cells from New Zealand white rabbits were seeded at high density and cultured in DMEM containing 50 micrograms/mL ascorbic acid and 10% fetal bovine serum. These cells appeared as a heterogeneous population and changed their shape, size, and refractivity as cultures aged. Cartilage-like cells, which always dominated the culture, were infiltrated with a minority of fibroblast-like cells. Cell number increased progressively, and cultures reached confluence at nine days. Antibody activity for cartilage-specific glycosaminoglycan was determined by ELISA assay. This reaction reached a maximum at six days and decreased thereafter. Cultures stained with Alcian blue (pH 1.0) supported these results. Cytoplasmic mRNA analysis indicated that the transcription of type II collagen gene was present at all time points. Type I collagen and alkaline phosphatase mRNA levels showed progressive increases from 12 h to nine days, with significantly higher values in cells cultured for six, nine, and 12 days than in cells collected from earlier time points. These results suggest that in our present culture system, MCC cells undergo phenotypic changes that resemble their maturation processes in vivo.

It has been shown that a release of periosteal tension may augment the growth of a long bone; however, similar experiments in the mandible have produced equivocal results. For further definition of the role of periosteal tension in the growth of the mandibular condyle, 80 young female Sprague-Dawley rats were randomly assigned to four equal groups: (1) sham, (2) condylotomy, (3) narrow periostomy, and (4) wide periostomy. All procedures were performed unilaterally. Lateral cephalograms, transcranial condylar radiographs, and metallic implants made possible the measurement of the actual increment of condylar growth over three post-operative periods: 0 to 14 days, 14 to 28 days, and 0 to 28 days. Analysis of covariance was performed (weight gain as the covariate) and, where overall significance was found, Turkey's HSD test was used for determination of individual group differences. Conservative, circumferential periostomy of the condylar neck as well as condylotomy failed to alter the condylar growth rate, whereas removal of a wide band of periosteum led to a small decrease. These findings suggest that, at least in the rat, periosteal tension plays only a minor role-if any-in the control of condylar/mandibular growth. Furthermore, a subsequent increase or decrease in condylar growth, in experiments that use a condylotomy model, cannot be attributed solely to a disturbance in periosteal integrity caused by the condylotomy.

The objectives of the present study were to characterize the loading of the immature TMJ, and to develop a theoretical model to explain the relationship between joint loading and development of the eminence of the human TMJ. The osteological remains of forty individuals, ages ranging from birth to twenty years, were used to provide metric coordinates of the three-dimensional relationships of the anatomy of the biting apparatus. The data were used, in a numerical model of TMJ loading (Smith et al., 1986), to calculate the magnitudes and directions of condylar loading. The following conclusions were drawn: (i) static equilibrium cannot be satisfied unless the immature TMJ is loaded; (ii) in the neonate, the direction of condylar loading is approximately vertical but, as the child matures, the angle of condylar loading becomes more oblique; and (iii) evidence is given in support of the hypothesis that early development of the eminence is consequent upon the stimulation of bone growth by the appropriate position and timing of loading of the immature condyle on the temporal component of the joint.

Interrelationships of growth hormone and thyroxine effects on endochondral osteogenesis have been investigated. Evidence is offered that thyroxine augments the effect of growth hormone on the mandibular condyle just as at other osteogenetic centers, for example proximal tibial epiphysis in hypophysectomized rats. This contrasts with previous reports of a unique thyroxine antagonism to the effect of growth hormone on condylar development. In a further study on tibial epiphysis, special attention was given to assure absence of the endogenous thyroid hormone by adding thyroidectomy to hypophysectomy and reducing the possibility of thyroteopin residues in growth hormone. By histochemical tests, the responses of both calcified and noncalcified regions of the epiphyseal cartilage plate were measured. Graded growth hormone doses stimulated graded response in a noncalcified region. Thyroxine augmented this response eightfold. The width of the calcified region of the plate did not vary with treatment.