Effects of Twin Inclined Plane Device on Adaptation and Ultrastructure Variations in Condyle of Growing Rats.

OBJECTIVE: This study investigates the effects of using a twin inclined plane device (TIPD) on the remolding and ultrastructure variation of mandibular condyle in growing rats. MATERIALS AND METHODS: Forty-eight male Wistar rats (six weeks old, body weight of approximately 190-210 g) were divided into experimental group (wearing appliance, n = 32) and control group (no appliance, n = 16). Samples were collected on days 3, 14, 30, and 60. The immunohistochemical analysis for vascular endothelial growth factor (VEGF) and type II collagen was carried out. Tartrate-resistant acid phosphatase (TRAP) reaction was performed to evaluate the osteoclastic activity. Three-dimensional morphometric images were reconstructed for morphometric analysis by microcomputed tomography (micro-CT). The ultrastructure of the condylar surface was observed by scanning electron microscopy (SEM). RESULTS: The expression of VEGF significantly increased, while the expression of type II collagen decreased in the experimental group at days 30 and 60. Furthermore, the enhanced osteoclast activity was observed under the subchondral bone, which was highest at day 30, and decreased to the lowest at day 60 in the experimental group. In addition, adaptive subchondral bone remolding in the posterior part of the condyle was observed at day 60 in the experimental group, and the SEM revealed the ultrastructure variations after installation of the TIPD. However, these changes began to reverse after 30 days. CONCLUSION: Condylar tissue changes point to the osteoclastic activity in the posterior region of the condyle. These adaptive changes point to bone resorption in the posterior condyle. Type II collagen and VEGF contribute to the MCC remolding induced by the TIPD. The ultrastructural changes in the posterior condylar area in response to mechanical stresses are recoverable at the initial stage.

Early molecular response and microanatomical changes in the masseter muscle and mandibular head after botulinum toxin intervention in adult mice.

BACKGROUND: Masseter muscle paralysis induced by botulinum toxin type A (BoNTA) evokes subchondral bone loss in mandibular heads of adult rats and growing mice after 4 weeks. However, the primary cellular and molecular events leading to altered bone remodeling remain unexplored. Thus, the aim of the current work has been to assess the molecular response that precedes the early microanatomical changes in the masseter muscle and subchondral bone of the mandibular head in adult mice after BoNTA intervention. METHODS: A pre-clinical in vivo study was performed by a single intramuscular injection of 0.2 U BoNTA in the right masseter (experimental) of adult BALB/c mice. The contralateral masseter was injected with vehicle (control). Changes in mRNA levels of molecular markers of bone loss or muscle atrophy/regeneration were addressed by qPCR at day 2 or 7, respectively. mRNA levels of receptor activator of nuclear factor-κB ligand (RANKL) was assessed in mandibular heads, whilst mRNA levels of Atrogin-1/MAFbx, MuRF-1 and Myogenin were addressed in masseter muscles. In order to identify the early microanatomical changes at day 14, fiber diameters in transversal sections of masseter muscles were quantified, and histomorphometric analysis was used to determine the bone per tissue area and the trabecular thickness of subchondral bone of the mandibular heads. RESULTS: An increase of up to 4-fold in RANKL mRNA levels were detected in mandibular heads of the BoNTA-injected sides as early as 2 days after intervention. Moreover, a 4-6 fold increase in Atrogin-1/MAFbx and MuRF-1 and an up to 25 fold increase in Myogenin mRNA level were detected in masseter muscles 7 days after BoNTA injections. Masseter muscle mass, as well as individual muscle fiber diameter, were significantly reduced in BoNTA-injected side after 14 days post-intervention. At the same time, in the mandibular heads from the treated side, the subchondral bone loss was evinced by a significant reduction in bone per tissue area (-40%) and trabecular thickness (-55%). CONCLUSIONS: Our results show that masseter muscle paralysis induced by BoNTA leads to significant microanatomical changes by day 14, preceded by molecular changes as early as 2 days in bone, and 7 days in muscle. Therefore, masseter muscle atrophy and subchondral bone loss detected at 14 days are preceded by molecular responses that occur during the first week after BoNTA intervention.

Disease and functional loading effect on the structural conformation and mechanical properties of the mandibular condyle in a transgenic rheumatoid arthritis murine model: an experimental study.

AIM: The aim of the present study was to investigate the effect of rheumatoid arthritis (RA) and functional loading through diet modification on the structural conformation and the mechanical properties of the mandibular condyle in a transgenic mouse model and compare to healthy littermates. MATERIALS AND METHODS: Four-week-old hybrid male mice from mixed background CBAxC57BL/6 were used. Four groups of animals were formed consisting of five animals each, either presenting RA (transgenic line hTNF 197), or wild-type (control), half receiving ordinary (hard) diet and half receiving soft diet within each category. Following sacrifice, resin-embedded and metallographically polished condylar specimens were evaluated employing scanning electron microscopy/ Energy dispersive x-ray spectroscopy and also tested for mechanical properties, through Vickers microhardness (HV100) measurements. RESULTS: The multivariable analysis revealed significantly lower HV100 values for the RA groups after adjusting for diet (ß = -10; 95% confidence interval: -16, -4; P = 0.001), while functional loading through diet modification did not appear as a significant predictor of the outcome. CONCLUSIONS: There was evidence of compromised mechanical properties of the mandibular condylar bone for the diseased animals, whereas no association between functional loading and mechanical properties of the condyle could be established.

The microstructural and biomechanical development of the condylar bone: a review.

BACKGROUND: Bone constantly strives for optimal architecture. Mandibular condyle, which is subjected to various mechanical loads forcing it to be highly adaptive, has a unique structure and a relatively high remodelling rate. Despite the eminent clinical relevance of mandibular condyle, literature on its structural and biomechanical development and on the mechanical role of its mineralized and non-mineralized bone components is scarce. OBJECTIVE: The aim of the present review is to provide a brief introduction to basic bone mechanics and a synopsis of the growth and development of human mandibular condyle. Subsequently, the current ideas on the relationship between the structural and biomechanical properties of bone in general and of mandibular condyle in particular are reviewed. Finally, up-to-date knowledge from fundamental bone research will be blended with the current knowledge relevant to clinical dentistry, above all orthodontics. METHODS: A comprehensive literature study was performed with an emphasis on recent and innovative work focusing on the interaction between microarchitectural and micromechanical properties of bone. CONCLUSIONS: Mandibular condyle is a bone structure with a high bone turnover rate. Mechanical properties of mandibular condyle improve during adolescence and are optimal during adulthood. Local mineralization degree might not be a decisive determinant of the local bone tissue stiffness as was believed hitherto. Bone collagen and its cross links play a role in toughness and tensile strength of bone but not in its compressive properties. Clinical procedures might affect mandibular condyle, which is highly reactive to changes in its mechanical environment.

Psychological stress alters the ultrastructure and increases IL-1β and TNF-α in mandibular condylar cartilage

Psychological factors can be correlated with temporomandibular disorders (TMDs), but the mechanisms are unknown. In the present study, we examined the microstructural changes and expression of proinflammatory cytokines in mandibular condylar cartilage of the temporomandibular joint (TMJ) in a psychological stress animal model. Male Sprague-Dawley rats (8 weeks old, 210 ± 10 g) were randomly divided into 3 groups: psychological stress (PS, N = 48), foot shock (FS, N = 24), and control (N = 48). After inducing psychological stress using a communication box with the FS rats for 1, 3, or 5 weeks, PS rats were sacrificed and compared to their matched control littermates, which received no stress and were killed at the same times as the PS rats. Body and adrenal gland weight were measured and corticosterone and adrenocorticotropic hormone levels were determined by radioimmunoassay. After hematoxylin-eosin staining for histological observation, the ultrastructure of the TMJ was examined by scanning electron microscopy. Transcription and protein levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were evaluated by ELISA and semi-quantitative RT-PCR. The PS group showed a significantly higher adrenal gland weight after 3 weeks of stress and higher hormone levels at weeks 1, 3, and 5. Histopathological changes and thinning cartilage were apparent at weeks 3 and 5. In the PS group, TNF-α increased at 1, 3, and 5 weeks and IL-1β increased significantly after 1 and 3 weeks of stress, and then decreased to normal levels by 5 weeks. Psychological stress increased plasma hormone levels and RT-PCR indicated increased IL-1β and TNF-α expression in the TMJ in a time-dependent manner. These results suggest that cytokine up-regulation was accompanied by stress-induced cartilage degeneration in the mandibular condyle. The proinflammatory cytokines play a potential role in initiating the cartilage destruction that eventually leads to the TMDs.

Application of alternative fixatives to formalin in diagnostic pathology.

Fixation is a critical step in the preparation of tissues for histopathology. The objective of this study was to investigate the effects of different fixatives vs formalin on proteins and DNA, and to evaluate alternative fixation for morphological diagnosis and nucleic acid preservation for molecular methods. Forty tissues were fixed for 24 h with six different fixatives: the gold standard fixative formalin, the historical fixatives Bouin and Hollande, and the alternative fixatives Greenfix, UPM and CyMol. Tissues were stained (Haematoxylin-Eosin, Periodic Acid Schiff, Trichromic, Alcian-blue, High Iron Diamine), and their antigenicity was determined by immunohistochemistry (performed with PAN-CK, CD31, Ki-67, S100, CD68, AML antibodies). DNA extraction, KRAS sequencing, FISH for CEP-17, and flow cytometry analysis of nuclear DNA content were applied. For cell morphology the alternative fixatives (Greenfix, UPM, CyMol) were equivalent to formalin. As expected, Hollande proved the best fixative for morphology. The morphology obtained with Bouin was comparable to that with formalin. Hollande was the best fixative for histochemistry. Bouin proved equivalent to formalin. The alternative fixatives were equivalent to formalin, although with greater variability in haematoxylin-eosin staining. It proved possible to obtain immunohistochemical staining largely equivalent to that following formalin-fixation with the following fixatives: Greenfix, Hollande, UPM and CyMol. The tissues fixed in Bouin did not provide results comparable to those obtained with formalin. The DNA extracted from samples fixed with alternative fixatives was found to be suitable for molecular analysis.

Psychological stress alters the ultrastructure and increases IL-1ß and TNF-α in mandibular condylar cartilage.

Psychological factors can be correlated with temporomandibular disorders (TMDs), but the mechanisms are unknown. In the present study, we examined the microstructural changes and expression of proinflammatory cytokines in mandibular condylar cartilage of the temporomandibular joint (TMJ) in a psychological stress animal model. Male Sprague-Dawley rats (8 weeks old, 210 ± 10 g) were randomly divided into 3 groups: psychological stress (PS, N = 48), foot shock (FS, N = 24), and control (N = 48). After inducing psychological stress using a communication box with the FS rats for 1, 3, or 5 weeks, PS rats were sacrificed and compared to their matched control littermates, which received no stress and were killed at the same times as the PS rats. Body and adrenal gland weight were measured and corticosterone and adrenocorticotropic hormone levels were determined by radioimmunoassay. After hematoxylin-eosin staining for histological observation, the ultrastructure of the TMJ was examined by scanning electron microscopy. Transcription and protein levels of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) were evaluated by ELISA and semi-quantitative RT-PCR. The PS group showed a significantly higher adrenal gland weight after 3 weeks of stress and higher hormone levels at weeks 1, 3, and 5. Histopathological changes and thinning cartilage were apparent at weeks 3 and 5. In the PS group, TNF-α increased at 1, 3, and 5 weeks and IL-1ß increased significantly after 1 and 3 weeks of stress, and then decreased to normal levels by 5 weeks. Psychological stress increased plasma hormone levels and RT-PCR indicated increased IL-1ß and TNF-α expression in the TMJ in a time-dependent manner. These results suggest that cytokine up-regulation was accompanied by stress-induced cartilage degeneration in the mandibular condyle. The proinflammatory cytokines play a potential role in initiating the cartilage destruction that eventually leads to the TMDs.

Psychological stress induces alterations in temporomandibular joint ultrastructure in a rat model of temporomandibular disorder.

OBJECTIVE: The objective of this study was to investigate the effects of psychological stress on temporomandibular disorder (TMD). STUDY DESIGN: A communication box was used to induce psychological stress (PS) in rats. Then, the ultrastructure of temporomandibular was observed using scanning electron microscopy. Interleukin-1 (IL-1) and IL-6 were measured with reverse transcription polymerase chain reaction. RESULTS: The PS group showed evidence of ultrastructural changes in the condyle and articular disk after stimulation, i.e., incomplete gelatinlike material was observed on the condyle after 1 week of PS, wider waves on the articular disk and exposed condylar collagen were observed after 3 weeks of PS, and cracks were apparent on the surface of the condyle. The expression of IL-1 and IL-6 in the condyle cartilage significantly increased after exposure to psychological stress. CONCLUSIONS: These results indicate that psychological stress induces ultrastructure alterations in the temporomandibular joint and plays an important role in TMD.

Reliability of two techniques for measuring condylar asymmetry with X-rays / Confiabilidad de dos técnicas de medición de asimetría condilar con método radiográfico

Among structural alterations that can be a risk factor for temporomandibular joint disorder (TMD) is condylar asymmetry. In order to measure the condylar asymmetry index in panoramic x-rays quantitatively, two methods have been proposed: those of Habets and Kjellberg. The aim of this study was to determine whether the x-ray method of measuring condylar asymmetries in orthopantomographies presents a minor tendency to error due to slight displacements of the head in the horizontal plane. 30 patients between 18 and 25 years of age were assessed. Each of them underwent three panoramic x-rays in three different positions: orthoradial, and at 5 and 10 horizontal angles. Then the Habets and Kjellberg measurements were taken. Habets' technique did not show any statistically significant differences in the x-rays at 5° and 10° horizontal angles compared to the 0 angle. However, Kjellberg's technique showed statistically significant differences only at the 10° angle compared to the 0 angle. The 10° changes produced linear and ratio variations, but the indices did not vary. It was concluded that both methods provide acceptable clinical information within the limitations of these techniques to obtain data on condylar symmetries or asymmetries of the mandibular body or ramus.

Dentro de las alteraciones estructurales que pueden ser un factor de riesgo de desarrollo de un trastorno temporomandibular (TTM) se menciona a la asimetría condilar. Para realizar la medición cuantitativa del índice de asimetría condilar en radiografías panorámicas se han propuesto dos métodos, Habets y Kjellberg. El objetivo de este estudio fue determinar si el método radiográfico de medición de asimetrías condilares en ortopantomografías que presenta menor tendencia al error por leves desplazamientos de la cabeza en el plano horizontal. Se evaluaron 30 pacientes entre 18 y 25 años de edad. Cada uno de ellos se sometió a tres radiografías panorámicas en tres posiciones distintas: posición ortoradia, 5 y 10 de angulación horizontal. Posteriormente, se realizaron las mediciones de Habets y Kjellberg. La técnica de Habets no mostró diferencias estadísticamente significativas en las radiografías con 5° y 10° de angulación horizontal con respecto al ángulo de 0. Sin embargo, la técnica de Kjellberg mostró diferencias estadísticamente significativas sólo al ángulo de 10° con respecto al ángulo de 0. Las alteraciones de 10° produjeron variaciones lineales y de razones, sin embargo no variaron los índices. Se concluye que ambos métodos entregan información clínica aceptable con las limitaciones que estas técnicas tienen para obtener información sobre simetrías o asimetrías condilares de cuerpo o de rama.

Study of the tidemark in human mandibular condylar cartilage.

OBJECTIVES: The tidemark is the junction of mature calcified and uncalcified articular cartilage. Metabolic activity in the tidemark area makes this structure important in articular remodelling and osteoarthrosis. However, detailed study of the tidemark on the mandibular condyle has been limited. This study investigated the functional significance of the tidemark in the mandibular condyle with respect to load-bearing. DESIGN: The study sample included 87 healthy human mandibular condyles from subjects aged 14-76 years. The morphology, number, and thickness of tidemarks were observed and measured in the anterior, superior, and posterior regions. Five additional mandibular condyles were evaluated by scanning electron microscopy. RESULTS: The tidemark appeared in 80 specimens as a dense wavy line at the interface between the calcified and hypertrophic layers of the condylar cartilage. Scanning electron microscopy showed a highly wavy tidemark surface in the load-bearing areas and a relatively flat and smooth surface in the non-load-bearing areas. The thickness of the tidemark in the load-bearing areas was significantly higher than that in the non-load-bearing areas (P=0.003). CONCLUSION: The results of this study demonstrated that the thickness of the tidemark was related to load-bearing on the articular surface.

Effect of mechanical pressure on the thickness and collagen synthesis of mandibular cartilage and the contributions of G proteins.

To investigate the role of mechanical pressure on cartilage thickness and type II collagen synthesis, and the role of G protein in that process, in vitro organ culture of mandibular cartilage was adopted in this study. A hydraulic pressure-controlled cellular strain unit was used to apply hydrostatic pressurization to explant cultures. The explants were compressed by different pressure values (0 kPa, 100 kPa, and 300 kPa) after pretreatment with or without a selective and direct antagonist (NF023) for the G proteins. After 4, 8 and 12 h of cell culture under each pressure condition, histological sections of the explants were stained with hematoxylin-eosin to investigate the thickness of the cartilage. Immunohistochemical staining was used to observe type II collagen expressions. The results showed that a hydrostatic pressure of 100 kPa significantly reduced the thickness of the proliferative layer in condylar cartilage without affecting the thickness of the transitional layer. Hydrostatic pressures of 100 kPa and 300 kPa significantly enhanced the synthesis of type II collagen. G proteins are involved not only in the proliferation and differentiation of condylar cartilage regulated by prolonged pressure, but also in the process of collagen production in condylar cartilage stimulated by pressure.

Age-related changes in collagen properties and mineralization in cancellous and cortical bone in the porcine mandibular condyle.

Collagen is an important constituent of bone, and it has been suggested that changes in collagen and mineral properties of bone are interrelated during growth. The aim of this study was to quantify age-related changes in collagen properties and the degree of mineralization of bone (DMB). The DMB in cancellous and cortical bone samples from the mandibular condyle of 35 female pigs aged 0-100 weeks was determined using micro-computed tomography. Subsequently, the amount of collagen and the number of pentosidine (Pen), hydroxylysylpyridinoline (HP), and lysylpyridinoline (LP) cross-links were quantified by means of high-performance liquid chromatography. The amount of collagen increased with age in cancellous bone but remained unchanged in cortical bone. The number of Pen and LP cross-links decreased in both bone types. In contrast, the number of HP cross-links decreased only in cancellous bone. The sum of the number of HP and LP cross-links decreased with age in cancellous bone only. The DMB increased in cancellous and cortical bone. It was concluded that the largest changes in the number of mature collagen cross-links and the mineralization in porcine cancellous and cortical bone take place before the age of 40 weeks. The low number of mature cross-links after this age suggests that the bone turnover rate continues to be high and thereby prevents the development of mature cross-links.

Morphological changes in the mandible of male mice associated with aging and biomechanical stimulus.

Degenerative changes in the temporomandibular joint (TMJ) associated with aging can affect mandibular shape and reduce growth potential when stimulated by functional appliance therapy. This study was designed to evaluate the morphological changes in the mandibles of male mice associated with aging and biomechanical stimulus. Every 3 days over the course of 1 month, the lower incisors were trimmed by 1 mm to induce mandibular advancement (MA) when the animal was feeding. The left mandibles of the 23 experimental and 27 control animals were subsequently dissected, and digital images were obtained to analyze nine linear/angular measurements. Because mandibular morphology depends on the maintenance of condylar cartilage, the surfaces of the condylar cartilage and the ascending ramus of the mandible were also analyzed by scanning electron microscopy (SEM). The linear measurements of the mandible showed changes according to age in the control group and a growth response in the mandibular condyle in 7- and 15-month-old mice after MA. Moreover, SEM analysis revealed depressions in the anterior region of the condylar cartilage and inclined vascular grooves in the ascending ramus in the 7- and 15-month-old experimental mice. Although the growth potential is reduced in mice after 6 months of age, the results showed that continuous growth of the mandible occurs after maturation, except in the condyle, and that biomechanical stimulus of the TMJ of male mice leads to condylar growth. These results suggest that mature and old individuals can favorably respond to maxillary functional orthopedic therapy.

Ultrastructural and histomorphometric alterations of rat jaw bones after experimental induction of lathyrism.

OBJECTIVE: The aim of this study was to investigate the effect of beta-aminopropionitrile (beta-APN), present in Lathyrus sativus (L. sativus) seeds consumed in drought prone areas, on the maxillary and mandibular bones of rats. DESIGN: Eighteen rats were used; they were divided into experimental group (12 rats) and a control group (6 rats). In the experimental group, lathyrism was induced by a once daily subcutaneous administration of beta-APN at a dose of 5mg in 0.4ml distilled water per 100g of body weight for 40 days. The specimens were examined histologically, histomorphometrically and ultrastructurally. RESULTS: Histopathological examination of lathyritic rats showed that both mandible and maxillae were significantly affected. Irregular bone trabeculae with wide medullary cavities and abundant marrow spaces were observed. Focal resorptive areas lined with osteoclasts were also present. Histomorphometric analysis showed a decrease in the area percentage of bone in lathyritic rats as compared to the control. This decrease was statistically significant only in the mandible (p=0.018). Moreover, a significant increase in the number of osteoclasts (p=0.014, 0.012) and resorptive bays (p=0.013, 0.002) was detected in the maxillae and mandible of the experimental group, respectively. Ultrastructurally, there was a significant difference in the microarchitecture of the collagen fibres in lathyritic rats compared to the control. The collagen fibres in the lathyritic group appeared disorganized, fewer in number and more spaced. CONCLUSIONS: This study provided evidence for lathyritic disturbances in bone architecture. These disturbances are likely to affect mineralization and bone strength as well.

Biomechanical properties of the condyle created by osteodistraction.

A new condyle can be reconstructed by osteodistraction, but the biomechanical properties of the neocondyle remain unknown. This study examined the hypothesis that the biomechanical properties of neocondylar cancellous bone could reach control levels 24 weeks after its creation by osteodistraction. The right mandibular condyles were removed and reconstructed by osteo-distraction in 16 adult goats. Their contralateral condyles served as controls. Microstructural and mechanical properties were examined by microcomputed tomography and mechanical testing. At 24 weeks after distraction, the neocondyle grew larger in size, but the shape and histological features were similar to those of the controls. The cancellous bone of the neocondyle even appeared to be more dense and stiffer in comparison with the control condyle. The results of this study suggest that the neocondyle created by osteodistraction develops nearly normal biomechanical properties for functional loading by 24 weeks after creation.

Matrix metalloproteinase and its inhibitor in temporomandibular joint osteoarthrosis after indirect trauma in young goats.

Our aim was to examine the change in expression of matrix metalloproteinases (MMP-13), matrix metalloproteinases-3 (MMP-3), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in the articular cartilage of goats with experimentally-induced osteoarthrosis of the temporomandibular joint (TMJ) at various times. Osteoarthrosis was induced in 20 goats in the bilateral TMJ and 5 goats acted as controls. There were 5 goats in each group, and a group was killed at 7 days, and 1, 3, and 6 months postoperatively. The samples were collected, and the joints evaluated histologically. Immunofluorescence was used to detect the presence of MMPs and TIMP-1 in the articular disc and condylar cartilage. The ultrastructure of the articular disc and condylar surface at 1 month was examined with scanning electron microscopy (SEM). Osteoarthrosis of the TMJ progressed gradually over time. MMP-13, MMP-3, and TIMP-1 were expressed strongly in the TMJ soon after injury; MMP-13 became gradually weakened, and MMP-3 strengthened later. None of these were expressed in the normal condyle. After a month the surface of the arthrotic condyle was uneven, and the underlying collagen fibrils were exposed in irregular fissures on the surface. The secretion of TIMP-1 was related closely to the changes of MMPs during osteoarthrosis of the TMJ. The unbalanced ratio between them caused degradation of the matrix of the cartilage and might be the cause of osteoarthrosis of the TMJ.

[Fabrication of collagen/sodium hyaluronate scaffold and its biological characteristics for cartilage tissue engineering].

OBJECTIVE: To develop a scaffold material containing collagen 1 and sodium hyaluronate for the cartilage tissue engineering and to evaluate its biocompatibility by using the rabbit chondrocytes derived from a mandibular condylar process. METHODS: The porous matrices containing collagen 1 and sodium hyaluronate were fabricated by the freeze-drying technique and were crosslinked by using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC). The microstructure of the scaffold was observed under the scanning electron microscope (SEM), and the enzymatic degradation test was performed to compare the ability of the scaffold resistance to collagenase before and after the crosslinking. The chondrocytes from the rabbits' condylar process were isolated and cultured before they were seeded into the scaffold, and cell attachment and proliferation were measured by the cell count 1, 3, 5, 7 and 10 days after the cell being seeded; then, the biocompatibility of the scaffold was evaluated by the light microscopic examination, histological examination, and the SEM exmination. RESULTS: The porous structure of the scaffold facilitated the penetration and attachment of the seeded cells. The porosity was 83.7% and the pore size was 100-120 microm. The cell number increased from 3.7 x 10(4) per scaffold 1 day after the cell being seeded to 8.2 x 10(4) per scaffold 10 days after the cell being seeded. The crosslinking treatment could significantly enhance the scaffold resistance to the collagenase activity. The examinations under the light microscope and SEM indicated that the chondrocyte adhered and spread well on the scaffold, and the extracellular matrices were also observed around the chondrocytes. CONCLUSION: The porous scaffold composed of collagen I and hyaluronan has an appropriate structure and a good biocompatibility for the attachment and proliferation of the chondrocytes, which can facilitate it to become a useful scaffold in the cartilage tissue engineering.

Age-related changes in microarchitecture and mineralization of cancellous bone in the porcine mandibular condyle.

The mandibular condyle is considered a good model for developing cancellous bone because of its rapid growth and high rate of remodeling. The aim of the present study was to analyze the simultaneous changes in microarchitecture and mineralization of cancellous bone during development in a three-dimensional fashion. Eight mandibular condyles of pigs aged 8 weeks prepartum to 108 weeks postpartum were scanned using microCT with an isotropic spatial resolution of 10 microm. The number of trabeculae decreased during development, whereas both the trabecular thickness and the distance between the trabeculae increased. The bone surface to volume ratio decreased during development, possibly limiting the amount of (re)modeling. Both the mean degree of mineralization and intratrabecular differences in mineralization between the surfaces and cores of trabecular elements increased during development. The trabecular surfaces were more highly mineralized in the older condyles compared to the younger ones. Together with the observed decrease in the relative size of trabecular surface, this finding suggests a decrease in (re)modeling activity during development. In accordance with the general growth and development of the pig, it was concluded that most developmental changes in cancellous bone occur until the age of 40 weeks postpartum.

Study on the effects of mechanical pressure to the ultrastructure and secretion ability of mandibular condylar chondrocytes.

During mandibular movement, condyle is subjected to repetitive compression and the mandibular condylar chondrocytes (MCCs) can detect and respond to this biomechanical environment by altering their metabolism. The present study was undertaken to investigate the effects of pressure to the ultrastructure, aggrecan synthesis, nitric oxide (NO) and prostaglandin F(1)alpha(PGF(1)alpha) secretion in MCCs. In vitro cultured rabbit MCCs were incubated and pressed under continuous pressure of 90kPa for 60min and 360min by hydraulic pressure controlled cellular strain unit. The ultrastructure, aggrecan mRNA expression, activity of nitric oxide synthase (NOS) and PGF(1)alpha secretion were investigated. Besides, nitric oxide inhibitor was used together with pressure to investigate the role of NO in mechanical effects. The appearance of MCC on TEM showed that after been pressed under 90kPa for 60min, the cellular processes became elongated and voluminous, together with aggrecan mRNA increasing. Under 90kPa for 360min, some of the cells showed distinct sign of apotosis and the aggrecan mRNA decreased. Pressure of 90kPa could cause increase of NOS activity and decrease of PGF(1)alpha composition. Inhibitor experiments indicated that pressure-induced upregulation of aggrecan mRNA and inhibition of PGF(1)alpha synthesis was partly mediated by NO. Continuous pressure could cause changes on the ultrastructure and function of MCC, as well as up-regulation of aggrecan synthesis, increase of NO secretion and decrease of PGF(1)alpha composition. NO was the upstream molecule, which mediated the response of aggrecan and PGF(1)alpha to mechanical pressure.

Mandibular advancement and morphological changes in the mandibles of female mice of different ages.

Mandibular advancement (MA) by means of functional orthopedic techniques is currently used in young patients to stimulate mandibular growth. The aim of this study was to evaluate the morphological changes in the mandibles of 2-, 7-, and 16-month-old female mice after MA. Every 3 days during 1 month, the lower incisors were trimmed by 1mm to induce protrusion when the animal was feeding. The left mandibles of the 30 experimental and 28 control individuals were subsequently dissected and digital images were obtained to analyze nine linear/angular measurements. The condylar microstructure was also analyzed by scanning electron microscopy (SEM). The linear/angular measurements showed a growth response in different mandibular regions in 2- and 16-month-old individuals. SEM showed that, in the 7-month-old mice, the condylar cartilage had regenerated in the treated individuals but not in the controls. The results suggest that MA produces mandibular growth in 2- and 16-month-old female mice. Although there was no mandibular growth in 7-month-old mice, regeneration of the condylar cartilage was detected, thus demonstrating that different responses to the MA stimulus occur in female mice of different ages.