Effect of nickel titanium file design on the root surface strain and apical microcracks.

The aim of this study was to determine the effect of nickel titanium file design on the root surface strain generated and apical microcracks caused during canal shaping. Thirty-three mandibular incisors were distributed into LightSpeed X, FlexMaster and a control group. A strain gauge was fixed apically on the proximal root surface to determine the maximum strain during canal shaping. Except for the control group, all root canals were enlarged to size 50. Images were taken after removing the apical 1 and 2 mm of the root end. Mean maximum strain values and presence of microcracks were statistically compared using the t-test and chi-square test, respectively. During canal shaping, the strain increased cumulatively with mean maximum strains of 808.2 ± 228.8 and 525.1 ± 168.9 microstrain in LightSpeed X and FlexMaster, respectively (P = 0.004). Both systems caused comparable microcracks. Although LightSpeed X produced higher maximum strain, no difference in microcrack development was found between both systems.

Correlation between cyclic fatigue and the bending properties of nickel titanium endodontic instruments.

The effects of cyclic fatigue on bending properties of NiTi endodontic instruments were investigated. Sixteen Profiles(®) were divided into two groups (A, and B). The sequence of cantilever bending test and cyclic fatigue test was alternated repeatedly until file separation occurred. In the cyclic fatigue test, the instrument curvature was 19° in group A and 38° in group B. Fractographic examination was performed to determine fracture patterns. In group A, there were significant differences between the bending load values measured before the cyclic fatigue test and the last cantilever bending test before instrument fracture at each deflection (p<0.05). Fractographic examination showed the specific patterns of cyclic fatigue fracture. The stress required to induce martensitic transformation might be reduced due to the softening behavior caused by the cyclic fatigue under the relaxation condition of the superelasticity range (group A). The SEM images were able to display specific patterns indicating cyclic fatigue fracture.

Down-regulation of inflammatory mediator synthesis and infiltration of inflammatory cells by MMP-3 in experimentally induced rat pulpitis.

INTRODUCTION: Matrix metalloproteinase (MMP)-3 is a member of the MMP family that degrades the extracellular matrix. Application of MMP-3 to injured pulp tissue induces angiogenesis and wound healing, but its anti-inflammatory effects are still unclear. Here, we evaluated the anti-inflammatory functions of MMP-3 in vitro and in vivo. METHODS: Nitric oxide and inflammatory mediator synthesis in macrophages activated by lipopolysaccharide (LPS) was measured in the presence or absence of MMP-3. The mouse Mmp3 (mMmp3) expression vector containing full length cDNA sequence of mMmp3 or cDNA sequence of mMmp3 missing the signal peptide and pro-peptide regions was transfected to RAW264, a mouse macrophage cell line, and NO synthesis and inflammatory mediator expression were evaluated. Pulpal inflammation was histologically and immunohistochemically evaluated in a rat model of incisor pulpitis induced by the application of LPS for 9 hours in the presence or absence of MMP-3. RESULTS: NO and pro-inflammatory mediator synthesis promoted by LPS was significantly down-regulated by MMP-3 in vitro. The full length of mMmp3 down-regulated the LPS-induced NO synthesis and chemical mediator mRNA expression, however the mMmp3 missing the signal peptide failed to block the NO synthesis induced by LPS. The numbers of major histocompatibility complex class II+ and CD68+ cells, which infiltrated into the rat incisor pulp tissues in response to the topical application of LPS, were significantly decreased by the application of MMP-3 in vivo. CONCLUSIONS: These results indicate that MMP-3 possesses anti-inflammatory functions, suggesting its potential utility as an anti-inflammatory agent for pulpal inflammation.

Application of optical coherence tomography to identify pulp exposure during access cavity preparation using an Er:YAG laser.

OBJECTIVE: The study aimed to evaluate the ability of optical coherence tomography (OCT) to guide and identify pulp exposure using an erbium: yttrium-aluminum-garnet (Er:YAG) laser. BACKGROUND DATA: The Er:YAG laser has been proven to be effective in ablating dental hard tissue and offers advantages, as there is none of the vibration and noise you get with conventional methods, but it has limitations in relation to the tactile feedback that would aid in identification of entry into the pulp chamber. Based on depth-resolved optical reflectivity, OCT technology has been developed to provide high-resolution, cross-sectional images of the internal structure of biological tissues. MATERIALS AND METHODS: The pulp chambers of 20 human mandibular incisors were examined, and the average thickness of hard tissue covering the pulp chamber was assessed using micro-computed tomography (micro-CT) images. An Er:YAG laser was used to gradually penetrate the hard tissue over the pulp chamber under microscopic guidance. The preparation was constantly imaged using a swept-source OCT at 10 sec intervals until a pulp chamber exposure was identified using the technology. The pulp exposure was re-examined under the microscope and compared with micro-CT images for verification. RESULTS: The pulp exposures of 20 incisors were all verified microscopically and with micro-CT images. The thickness of hard tissue penetrated by the laser ranged from 0.44 to 1.69 mm. CONCLUSIONS: Swept-source OCT is a useful tool for identifying pulp exposure during access opening with the Er: YAG laser.

Joint haemorrhage partly accelerated immobilization-induced synovial adhesions and capsular shortening in rats.

PURPOSE: To elucidate the effects of intra-articular haemorrhage on the joint capsule of immobilized knees in rats. METHODS: The unilateral knee joints were immobilized using a plastic plate and screws. Sham operated rats had only screws inserted. A single injection of fresh autologous blood was given postoperatively into the knee joints of the immobilized blood injection (Im-B) and the Sham blood injection (Sm-B) groups. Normal saline was administered for the immobilized-saline injection (Im-S) group. Sagittal sections were prepared from the medial midcondylar region of the knee and assessed with histological, histomorphometric, and immunohistochemical methods. The range of motion (ROM) was measured, and the mechanical property of the capsule was assessed by scanning acoustic microscope. RESULTS: Absorption of the injected blood was delayed and made severe adhesions in the Im-B group. The length of the synovial membrane in the Im-B group was significantly shorter than that of the other groups. The ROM was significantly restricted in the Im-B group compared with the other groups. The elasticity of the posterior capsule in the Im-B group was significantly lower than that in the Sm-B group. Iron deposition was observed in the Im-B and Sm-B groups. Strong immunoreactivities of CD68, TGF-ß1, and α-SMA were observed in the adhesion area of the Im-B group. Joint immobilization with blood injection caused severe capsular adhesion and limited range of motion. Immunostaining related to fibrosis increased with joint haemorrhage. CONCLUSION: Intra-articular haemorrhage with joint immobilization might be an accelerated risk factor for joint contracture. It is likely that leaving a haematoma inside an immobilized joint should be avoided.

Kinetics of Th17-related cytokine expression in experimentally induced rat periapical lesions.

Th17-related cytokines are essential factors in various pathological states, including inflammatory bone destruction. This study investigated the contribution of Th17-related cytokines to the progress of experimentally induced rat periapical lesions. Periapical pathoses were induced by unsealed exposure of the pulp chamber of the lower first molars. A variety of immunocompetent cells, including CD68(+) macrophages, Ia antigen(+) cells and TCRαß(+) T cells, were observed in the lesions. The expression levels of Th17-related cytokines, IL-17 and IL-23, and of pro-inflammatory cytokines, IL-1ß and IL-6, were significantly increased at 14 days (expansion stage) compared with normal periapical tissues. The expression levels of Foxp3, a regulatory T cell (Treg)-related gene, and of IL-10, an anti-inflammatory cytokine, were higher at 28 days (chronic stage) than at 14 days. These findings suggest that Th17-related cytokines may be primary contributors to the initiation of periapical bone destruction, and that lesion expansion may be regulated by anti-inflammatory mediators.

Wnt11 expression in rat dental pulp and promotional effects of Wnt signaling on odontoblast differentiation.

Congenital anomalies of wingless-type mouse mammary tumor virus (MMTV) integration site family (Wnt) are frequently accompanied with tooth and dentin abnormality. The aim of this study was to investigate the effects of Wnt signaling on odontoblast differentiation of mouse dental papilla cells (MDPs). Mouse dental papilla cells were cultured in α-modified minimum essential medium containing 10% fetal bovine serum and antibiotics. Odontoblast differentiation was induced by bone morphogenic protein 2 (BMP2), and the expression of odontoblast-specific markers and Wnt-related signaling molecules was analyzed by real-time reverse transcription-polymerase chain reaction and immunohistochemistry. Odontoblast differentiation was evaluated by dentin sialophosphoprotein (Dspp) and dentin matrix protein (DMP) 1 expression. Localization of ß-catenin in MDPs was detected by immunocytochemistry using an anti-ß-catenin antibody. Dspp expression in MDPs was upregulated in the presence of BMP2. Wnt5a, Wnt11, Lef1 and Tcf4 expression was upregulated in BMP2-treated MDPs. Wnt11 expression was detected in rat dental pulp in vivo, and particularly strong expression of Wnt11 was detected in odontoblasts. Enhanced Dspp and DMP1 expression and alkaline phosphatase activity induced by BMP2 were completely negated by the Wnt antagonist: IWR-1-endo treatment. Nuclear translocation of ß-catenin observed in BMP2-treated MDPs was also negated by IWR-1-endo treatment. These results indicate that Wnt signaling upregulates odontoblast marker expression in MDPs, suggesting a promoting effect of Wnt signaling on odontoblast differentiation.

Joint immobilization induced hypoxic and inflammatory conditions in rat knee joints.

The purpose of this study was to examine the hypoxic and inflammatory conditions after immobilization in the joint capsule of rat knees. The unilateral knee joints of adult male rats were immobilized with an internal fixator (Im group) for 1 day, 3 days, and 1, 2, 4, 8, and 16 weeks. Sham-operated animals had holes drilled in the femur and tibia and screws inserted without a plate (control group). The number of cells and blood vessels in the capsule were histologically examined. The hypoxic condition in the capsule was histologically examined with a Hypoxyprobe™-1. The gene expressions related to the hypoxic (hypoxia inducible factor-1α, vascular endothelial growth factor, and fibroblast growth factor 2) and inflammatory conditions [interleukin-6 (IL-6), IL-1α, IL-1ß, tumor necrosis factor-α, and tumor necrosis factor-ß] were evaluated by quantitative reverse transcription polymerase chain reaction. The number of cells was unchanged at 1 day in the two groups; however, the number significantly increased at 3 days in the Im group. The number of blood vessels in the Im group gradually decreased. Strong immunostaining of Hypoxyprobe™-1 around the blood vessels was observed in the Im group. The gene expressions of hypoxia inducible factor-1α and fibroblast growth factor 2 were significantly higher in the Im group compared with those in the control group. The gene expressions of IL-6, IL-1α, IL-1ß, and tumor necrosis factor-ß were significantly higher in the Im group compared with those in the control group. These data indicated that joint immobilization induced hypoxic and inflammatory conditions in the joint capsule, which might be an initiating factor for joint contracture.

Detection of root surface fractures with swept-source optical coherence tomography (SS-OCT).

OBJECTIVE: The purpose of this study was to compare optical coherence tomography (OCT) with the existing technologies, to assess its accuracy and utility in detecting vertical root fractures of extracted human teeth. BACKGROUND DATA: The detection of root fractures in teeth that have undergone root canal treatment is challenging because of the great difficulty in differentiating these fractures from morphologic or radiographic anomalies. OCT methods are based on depth-resolved optical reflectivity and have been developed to reduce the invasiveness and radiation exposure inherent to other techniques. METHODS: Twelve extracted human mandibular teeth (totaling 25 roots) that were free of caries, calculus, and root treatment were used, and assessed by microfocus computed tomography, the current gold standard for fracture detection. The ability of appropriately trained observers to detect root fractures using visual, microscopic, and swept-source OCT (SS-OCT) techniques were compared. micro-CT and SS-OCT produce three-dimensional images of the tooth from which to diagnose fractures, but CT scanning involves radiation exposure that is not required in SS-OCT. RESULTS: Seventeen of the 25 roots were found to have fractures by microfocus CT. These findings were replicated by SS-OCT, which revealed fractures exhibiting identical origin, size, and angulation within the root. We found that SS-OCT gave results compatible to the gold standard technique, and that SS-OCT and microscopy were more effective for identifying root fractures than was visual observation alone. CONCLUSIONS: SS-OCT may represent a novel, noninvasive, noncontact and nonexposure alternative to the conventional methods used for assessing root fractures in teeth.

Promotional effects of vasoactive intestinal peptide on the development of rodent Hertwig's epithelial root sheath.

Hertwig's epithelial root sheath (HERS), a bilayered epithelial cell sheath located at the cervical loop of the enamel organ in a developing tooth, is at the forefront of root formation. However, little is known about the exact mechanisms that regulate the development of HERS. The neuropeptide vasoactive intestinal peptide (VIP) is involved in the development of various tissues and cells. In this study, we investigated the roles of VIP in HERS development. VIP-immunoreactive nerve fibers were found in the dental pulp and around the root apex of the tooth, while the expression of VIP receptor 1 (VPAC1) was observed in HERS. The expression level of VPAC1 correlated with the development of HERS and was elevated at postnatal days 14 and 21. Using ex vivo cultures of neonatal tooth germs, VIP enhanced the elongation and proliferation of HERS. In vitro, VIP also promoted the proliferation of cells from the HERS-derived cell line, HERS01a cells, and upregulated the mRNA expression of cytokeratin 14 and vimentin (typical molecular markers of HERS) in these cells. These results suggest that VIP may be an essential factor for HERS development.

Shaping of the root canal using Er:YAG laser irradiation.

OBJECTIVE: The purpose of the present study was to investigate the degree of Er:YAG laser irradiation at the apical area in vitro. BACKGROUND DATA: Since the laser was developed, advancement of laser treatment has been seen in various fields. However, few reports exist on shaping of the root canal using Er:YAG laser irradiation. METHODS: Six single-rooted human teeth were used. The working length of root canals was set at 6.5 mm, and they were enlarged to apical file size #25. An Er:YAG laser and cone-shaped irradiation tips (R135T and R200T) were used. Laser irradiation conditions were 30 m J, 20 pps, and water flow of 5 mL/min. Samples were irradiated three times for 10 sec each using each tip. To evaluate the cutting degree of horizontal area of the root canal, the laser-irradiated surfaces were observed using microfocus X-ray computed tomographic photography before and after every irradiation. The samples were observed under a scanning electron microscope. Measurement of pixels in an area was performed by image-editing software (Adobe Photoshop 7.0). Statistical analysis was performed using StatView (version 5.0). One-way ANOVA and the Tukey-Kramer tests were used; p<0.05 indicated statistical significance. RESULTS: When root canals were irradiated with R200T for 10 sec (p<0.05), a large amount of evaporation (0.12 ± 1.07 mm(2)) was acquired in their cut area compared with the other irradiation conditions. In scanning electron microscopic observation, there was no smear layer and the dentinal tubules were open. CONCLUSIONS: When the distance between the tip and root dentin was adjacent, the shaping of root dentin by Er:YAG laser irradiation was definitely observed.

Deflecting load of nickel titanium rotary instruments during cyclic fatigue.

The dispersion of the lifetime of NiTi instruments, and their deflecting load (DL) changes during cyclic fatigue were investigated. A total of 120 ProFile NiTi rotary instruments were tested using a specially designed cyclic fatigue testing apparatus with three pins. Using these pins, the instrument was bent and rotated at 300 rpm to fracture. DL was recorded using a load cell attached to the central pin. For each sample, the working time was converted to number of cycles to fracture (NCF) and the mean DL (DL(m)) was calculated. The averages of NCF and DL(m) of 120 samples were 584.3±180.5 cycles and 6.44±0.91 N, respectively. All samples showed a sequential decrease in DL during rotation. Based on the present study, it is impossible to estimate the lifetime of a NiTi instrument from NCF. Thus, the change in DL could be an alternative criterion to determine the remaining lifetime.

Remobilization does not restore immobilization-induced adhesion of capsule and restricted joint motion in rat knee joints.

Joint immobilization, which is used in orthopaedic treatments and observed in bedridden people, usually causes restricted joint motion. Decreased joint motion diminishes activities of daily living and increases burden of nursing-care. The purpose of this study was to clarify the reversibility of immobilization-induced capsular changes and restricted joint motion in rat knee joints. The unilateral knee joints of adult male rats were immobilized with an internal fixator for 1, 2, 4, 8, and 16 weeks as a model of immobilization after surgery or disuse of the joint. After the fixation devices were removed, the rats were allowed to move freely for 16 weeks. Sham-operated rats were used as controls. Sagittal sections at medial midcondylar regions were made and assessed with histological, histomorphometric, and immunohistochemical methods. Joint motion was measured using a custom-made device under x-ray control after removal of the periarticular muscles. In the 1/16-week and 2/16-week immobilization-remobilization (Im-Rm) groups, cord-like structures connecting the superior and inferior portions of the posterior capsule (partial adhesion) were observed without restricted joint motion. In the 4/16-, 8/16-, and 16/16-week Im-Rm groups, global adhesion of the posterior capsule and restricted joint motion were observed. The restricted joint motion was not completely restored after incision of the posterior capsule. These data indicate that immobilization alone causes irreversible capsular changes and arthrogenic restricted joint motion. Besides the joint capsule, other arthrogenic factors such as ligaments might influence the restricted joint motion. Prolonged immobilization over 4 weeks should be avoided to prevent irreversible joint contracture.

Effects of dentin surface modifications treated with Er:YAG and Nd:YAG laser irradiation on fibroblast cell adhesion.

OBJECTIVE: The purpose of this in vitro study was to evaluate the effect of surface modifications induced by erbium (Er):YAG and neodymium (Nd):YAG laser irradiation on cell adhesion by comparing it to that of conventional methods for surface preparation after root-end resection. BACKGROUND DATA: Many studies have been seeking a favorable method to produce a resected root end with optimal conditions for cell response. However, little improvement has been achieved. This study evaluated the biocompatibilities of resected root surfaces after Er:YAG or Nd:YAG laser irradiation on initial cell adhesion. MATERIALS AND METHODS: Dentin disks were divided into three groups. Group A was left untreated, Group B was treated with Er:YAG laser irradiation (60 mJ/pulse, 10 pps, 60 sec), and Group C with Nd:YAG laser irradiation (60 mJ/pulse, 10 pps, 60 sec). After laser irradiation, the dentin disks were incubated with NIH/3T3 fibroblasts cultured in Dulbecco's modified Eagle's medium. A morphological analysis of the dentin surface and cell adhesion was observed under a scanning electron microscope. Surface roughness was measured using a confocal laser scanning microscope. The statistical analysis was undertaken using ANOVA at a level of significance of 5% (p<0.05). RESULTS: Morphological analysis and roughness measurement showed that dentin surfaces treated with Er:YAG laser irradiation were rougher than those in Groups A and C. Group B (Er:YAG) exhibited the greatest number of attached cells among all groups after 12 and 24 h. CONCLUSIONS: Morphological alteration induced by Er:YAG laser irradiation showed a favorable effect on the attachment of fibroblasts to dentin surfaces.

Gene expression analysis of resident macrophages in lipopolysaccharide-stimulated rat molar pulps.

INTRODUCTION: In normal dental pulp, a considerable number of resident macrophages are distributed. This study was designed to analyze the expression levels of genes associated with differentiation and function of resident macrophages in rat molar pulps stimulated with lipopolysaccharide (LPS). METHODS: Mandibular first molars of 7-week-old male Wistar rats were used. After transcardiac perfusion with a culture medium to preserve tissue integrity, pulpotomy and LPS application were carried out on the experimental teeth, and then dissected mandibles were subjected to whole-tooth culture for 3 days. Normal teeth and pulpotomized teeth without LPS served as controls. The specimens were then immunostained for ED1 (CD68, a general macrophage marker) and ED2 (CD163, a resident macrophage marker). Real-time polymerase chain reaction for Toll-like receptor 4 (TLR4), CD14, chemokine receptors (CCR2 and CX3CR1), and colony-stimulating factor-1 (CSF1) mRNAs was carried out after laser capture microdissection of ED1+ and ED2+ cells. RESULTS: LPS-treated pulps showed significant increases in (1) density of ED1+ and ED2+ cells beneath the amputation site and (2) expression levels of TLR4, CD14, CSF1, and CX3CR1 mRNAs, as compared with non-LPS-treated groups. CCR2 mRNA showed no significant difference between each group. CONCLUSIONS: LPS treatment of cultured rat molars caused the accumulation of resident macrophages and enhanced the expression of TLR4, CD14, CSF1, and CX3CR1 mRNAs in these cells. Up-regulation of these molecules might be involved in the differentiation and subsequent migration of resident macrophages of the pulp.

Laser-capture microdissection for Factor VIII-expressing endothelial cells in cancer tissues.

The immunostaining based Laser-capture microdissection (LCM) method called immune-LCM allows us to quantify the mRNA. Immune-LCM has recently been introduced to enhance identification of cells carrying a particular protein from frozen tissue samples. We have recently performed the immune-LCM of formaldehyde-fixated, paraffin-embedded tissues immunostained with a monoclonal antibody Factor VIII. This method could be useful for quantitative gene expression analysis in blood vessels from tumors of patients that have been treated with antiangiogenic drugs, allowing for validation of the effect of drug on the expected targets. Such capability might be exceedingly useful for the evaluation of the bioactivity of new drugs. This method is also useful to compare gene expression patterns in tumor cells versus endothelial cells during tumor progression or tumor angiogenesis.

Crack initiation on the apical root surface caused by three different nickel-titanium rotary files at different working lengths.

INTRODUCTION: The aim of this study was to compare the effects of three brands of nickel-titanium (NiTi) rotary files with different designs on the initiation of apical root cracks when working short, at, and beyond the apical foramen. METHODS: One-hundred eight teeth with straight single canals were selected and mounted on resin blocks with simulated periodontal ligaments, and the apex was exposed. The teeth were divided into 9 groups of 12 teeth according to the NiTi rotary file type used (Profile [Dentsply Maillefer, Ballaigues, Switzerland], K3 [SybronEndo, West Collins, CA], and EndoWave [FKG Dentaire, La-Chaux-de-Fonds, Switzerland]) and working length (at CL, 1 mm short of [CL - 1 mm], and 1 mm beyond [CL + 1 mm] the apical foramen). Digital images of the apical surface of every tooth were taken during the apical enlargement sequence at each file change. These images were compared with the baseline image, and the presence of a crack was noted. RESULTS: Significantly less cracks were observed in the CL - 1 mm group than in the CL and CL + 1 mm groups. No significant difference was found between the file types used. CONCLUSIONS: Working 1 mm short of the apical foramen caused less cracks on the apical surface. In addition, more cracks were observed when using larger file sizes. Instrumentation with NiTi rotary files could potentially cause cracks on the apical root surface.

Osteogenic differentiation of mouse mesenchymal progenitor cell, Kusa-A1 is promoted by mammalian transcriptional repressor Rbpj.

Pluripotent mesenchymal stem cells possess the ability to differentiate into many cell types, but the precise mechanisms of differentiation are still unclear. Here, we provide evidence that Rbpj (recombination signal-binding protein for immunoglobulin kappa j region) protein, the primary nuclear mediator of Notch, is involved in osteogenesis. Overexpression of Rbpj promoted osteogenic differentiation of mouse Kusa-A1 cells in vitro and in vivo. Transient transfection of an Rbpj expression vector into Kusa-A1 cells upregulated promoter activities of Runx2 and Ose2. Enhanced osteogenic potentials including high alkaline phosphatase activity, rapid calcium deposition, and increased calcified nodule formation, were observed in established stable Rbpj-overexpressing Kusa-A1 (Kusa-A1/Rbpj) cell line. In vivo mineralization by Kusa-A1/Rbpj was promoted compared to that by Kusa-A1 host cells. Histological findings revealed that expression of Rbpj was primarily observed in osteoblasts. These results suggest that Rbpj may play essential roles in osteoblast differentiation.

Distribution of type A and B synoviocytes in the adhesive and shortened synovial membrane during immobilization of the knee joint in rats.

Joint immobilization is commonly used for the treatment of joint injuries and diseases, but it also causes unfavorable outcomes such as joint contracture. The purpose of this study was to examine the morphological changes of the synovial membrane that is suspected as a cause of joint contracture, and localization of type A (macrophage-like) and type B (fibroblast-like) synoviocytes in the capsule after joint immobilization. Male Sprague-Dawley rats were used in this study. Unilateral knee joints were rigidly immobilized at 150 degrees of flexion with internal fixators for 3 days, 1, 2, 4, 8, and 16 weeks (7 rats/each immobilized group), while 42 rats were sham-operated. Sagittal sections of 5 mum were prepared from the medial midcondylar region of the knee joints and assessed with histological, histomorphometric, and immunohistochemical methods. Adhesions were observed both in the anterior and posterior synovial membranes in the immobilized group after 2 weeks. In the adhesion area, the cells were mainly composed of type A synoviocytes that were positive for CD68 and type B synoviocytes positive for prolyl 4-hydroxylase subunit beta. The length of synovial membrane in the immobilized group was significantly shorter than that in the control group after 2 and 4 weeks. After 8 weeks, the adhesion area in the immobilized group became fibrous and hypocellular. The staining intensity of hyaluronic acid-binding protein was increased after 16 weeks. Adhesion and shortening of the synovial membrane and the structural changes of the adhesion area may contribute to the development of joint contracture.

Expression of collagen types I and II on articular cartilage in a rat knee contracture model.

The purpose of our study was to clarify the expression patterns of collagen types I and II on articular cartilage after immobilization in a rat knee contracture model in 3 specific areas (noncontact area, transitional area, contact area). The unilateral knee joints of adult male rats were rigidly immobilized at 150 degrees of flexion using screws and a rigid plastic plate. Sham-operated animals had holes drilled in the femur and the tibia and screws inserted but were not plated. The expression patterns of collagen types I and II in each area were evaluated by in situ hybridization (ISH), immunohistochemistry (IHC), and quantitative real-time polymerase chain reaction (qPCR). The expression of collagen type II in the noncontact area was decreased by ISH but appeared unchanged when examined by IHC. In the transitional and contact areas, the expression of collagen type II was initially shown to have decreased and then increased at the hypertrophic chondrocytes by ISH but appeared decreased by IHC. Quantitative PCR revealed the decreased expression of type II collagen in the contact area. Immunostaining of collagen type I was increased at the noncontact area and transitional areas. Alterations of collagen types I and II expression may also affect the degeneration of articular cartilage after immobilization and the changes were different in the three areas.