Does Condylar Morphology Affect Maxillary Repositioning in Bimaxillary Orthognathic Surgery?

This study aimed to measure the association between condylar morphology and a measure of the maxillary centroid following bimaxillary surgery using mandibular-dependent splints. The study included skeletal Class III and Class II malocclusion patients, excluding those with facial asymmetry. Based on computed tomography imaging patients were characterized into normal or abnormal temporomandibular joint (TMJ) groups. A computer-aided design/computer-aided manufacturing splints were fabricated to reposition the maxilla in Le Fort I osteotomy. The primary outcome measure was the absolute differences between the maxillary centroid's the planned and actual postoperative positions calculated by superimposing computed tomography scans. The secondary outcome was the measure of other variations in linear and angular maxilla discrepancies. The demographic covariates included the age and sex of the patients. The operative covariates consisted of the dentofacial deformity and the planned movement of the maxilla. Seventy patients with skeletal maxillofacial deformities were included for analysis: 44 patients in the normal and 26 in the abnormal TMJ group. The average maxillary misalignment was 1.04±0.48 mm in the normal and 1.53±0.63 mm in the abnormal TMJ group (P<0.001). A statistically significant relationship existed between the discrepancies of the maxillary centroid and dentofacial deformity (η=0.656, P<0.001). These findings suggest an increased propensity for maxillary malposition in skeletal Class II patients. Furthermore, condylar morphology is a significant prognostic factor influencing maxillary repositioning errors in bimaxillary surgery with mandibular-dependent splints.

Retrospective analysis of odontogenic myxoma and odontogenic myxofibroma in the oral and maxillofacial region: a fibrous tissue-related differentiation.

The clinical differences between odontogenic myxoma (OM) and odontogenic myxofibroma (OMF), and the clinical significance of their classifications, remain unclear. This study reviewed the clinicopathological characteristics of patients with OM or OMF and evaluated the fibrous component of the specimens. Medical records of 21 patients with OM or OMF who underwent tumour resection were reviewed. The percentage of fibrous tissue on the representative sections was evaluated using haematoxylin and eosin- and Masson's trichrome-stained specimens. Histopathological diagnoses included 11 OMs and 10 OMFs with no tumour recurrence except for two cases in which the dredging method was applied. More cortical bone perforation was observed in OM than in OMF cases, without significant differences. Location-locularity and apparent diffusion coefficient value (ADC)-cortical bone perforation were significantly correlated in all OM and OMF cases. The percentage of fibrous tissue in specimens showed bimodal distribution bordered by 45%. There was a significant association between diagnosis based on 45% fibrous tissue criterion and the final pathological diagnosis. Our study showed a tendency for cortical bone perforation in OM compared to OMF and correlation between ADC and cortical bone perforation. According to the histopathological analyses, the fibrous component of each case was bimodal with 45%, which may be a criterion to distinguish between OM and OMF. Accumulating knowledge, such as significant differences in prognosis, may allow for minimal surgical treatment options based on the diagnosis according to this novel histopathological criterion.

NF1 with 47,XYY mosaicism diagnosed by mandibular neurofibromas.

Neurofibromatosis type 1 (NF1) is an autosomal dominant nevus disease characterized by multiple manifestations, primarily café-au-lait macules and neurofibromas. Here, we present the case of an NF1 patient with 47,XYY mosaicism whose diagnosis was prompted by café-au-lait macules on the skin and mandibular neurofibromas. Targeted next-generation sequencing of the patient's blood sample revealed a novel frameshift mutation in NF1 (NM_000267.3:c.6832dupA:p.Thr2278Asnfs*8) that is considered a pathogenic variant.

Study of Anatomical Changes of the Inferior Nasal Passage After Le Fort I Osteotomy With Superior Repositioning.

The relationship between postoperative morphological changes in the inferior nasal cavity and inferior turbinate after Le Fort I osteotomy remains unclear. This study aimed to investigate how the bone volume of the inferior turbinate affects contact with the inferior nasal cavity of patients who underwent superior repositioning. We evaluated the 3-dimensional relationship between the anatomical changes in the inferior nasal passage before and after surgery in 51 patients who underwent Le Fort I osteotomy with an elevation of >4.0 mm in the first molar. The soft tissue and bone volumes of the inferior turbinate and airway volume of the inferior nasal passage were calculated using Proplan CMF 3.0 and compared according to the size of the bone volume of the inferior turbinate. In addition, we reclassified the maxillary movements in the pitch direction and compared the results. The contact rates of the postoperative inferior nasal airway and the inferior turbinate in the large-bone group was 72.3% and that in the small-bone group was 40.0% in the χ2 test. The reduction in the inferior nasal passage volume was significantly greater in the large-bone group (pitch+) than in the small-bone group (pitch+). For patients with well-developed bony tissue of the inferior turbinate, caution is advised if the maxillary elevation is ≥4.0 mm, because the possibility of postoperative obstruction of the inferior nasal passages exist, which may lead to deterioration of nasal ventilation.

Development of bioinspired damage-tolerant calcium phosphate bulk materials.

Improving the damage tolerance and reliability of ceramic artificial bone materials, such as sintered bodies of hydroxyapatite (HAp), that remain in vivo for long periods of time is of utmost importance. However, the intrinsic brittleness and low damage tolerance of ceramics make this challenging. This paper reports the synthesis of highly damage tolerant calcium phosphate-based materials with a bioinspired design for novel artificial bones. The heat treatment of isophthalate ion-containing octacalcium phosphate compacts in a nitrogen atmosphere at 1000°C for 24 h produced an HAp/ß-tricalcium phosphate/pyrolytic carbon composite with a brick-and-mortar structure (similar to that of the nacreous layer). This composite exhibited excellent damage tolerance, with no brittle fracture upon nailing, likely attributable to the specific mechanical properties derived from its unique microstructure. Its maximum bending stress, maximum bending strain, Young's modulus, and Vickers hardness were 11.7 MPa, 2.8 × 10‒2, 5.3 GPa, and 11.7 kgf/mm2, respectively. The material exhibited a lower Young's modulus and higher fracture strain than that of HAp-sintered bodies and sintered-body samples prepared from pure octacalcium phosphate compacts. Additionally, the apatite-forming ability of the obtained material was confirmed in vitro, using a simulated body fluid. The proposed bioinspired material design could enable the fabrication of highly damage tolerant artificial bones that remain in vivo for long durations of time.

Genetic Landscape of Masticatory Muscle Tendon-Aponeurosis Hyperplasia.

Limited mouth opening is a characteristic of masticatory muscle tendon-aponeurosis hyperplasia (MMTAH). Although genetic involvement is suspected where familial onset is frequently observed, the genetic background of MMTAH is yet to be elucidated. In this study, we conducted whole genome sequencing of 10 patients with MMTAH and their family members when available. We also conducted RNA sequencing of normal temporal tendon (as disease region) and Achilles tendon (as control region) from commercially available pig samples. We identified 51 genes that had rare variants in patients with MMTAH and were highly expressed in the temporal tendons of pigs. Among the 51 genes, 37 genes have not been reported to be causative for human genetic diseases so far. As an implication of genetic involvement in the pathogenesis of MMTAH, 21 of these 37 genes were identified in two independent families. In particular, PCDH1 and BAIAP3 were identified in one affected individual in a family and consistently segregated in unrelated family, indicating they could be candidate causative genes of MMTAH. Our findings will help elucidate the genetic landscape of MMTAH and provide insights into future possibilities for tendon regeneration treatment.

Postoperative stability of bioresorbable plates made of 85:15 poly (L-lactide-co-glycolide) in Le Fort I osteotomy.

OBJECTIVE: Recently, RapidSorb plates (DePuy Synthes) made of 85.15 poly (L-lactide-co-glycolide) have been used for orthognathic surgery; however, reports regarding their effectiveness are limited. We aimed to compare the postoperative stability of RapidSorb plates, RapidSorb combined with titanium (MOJ plates), and MOJ plates in patients who underwent Le Fort I osteotomy at Tokyo Medical and Dental University Hospital. STUDY DESIGN: The use of RapidSorb in the maxilla is a load-sharing application and therefore constitutes an approved indication. Discrepancies in the maxillary positions were measured using postoperative computed tomography data at 1 week and 1 year using the centroid method 3-dimensionally. Treatment with RapidSorb alone showed a more vertical discrepancy in the maxilla treatment with MOJ and RapidSorb+MOJ. The RapidSorb4 group was subdivided into 2 groups (under and over 1.0-mm) based on the change in the maxillary centroid. RESULTS: The bone gap at the lateral border of the piriform aperture was significantly larger in the over-1.0-mm group than in the 1.0-mm group. CONCLUSIONS: The fixation of RapidSorb alone is not appropriate in load-bearing and unstable applications but is not contraindicated for load-sharing indications. Fixation with RapidSorb combined with MOJ was clinically effective, with results similar to titanium plate-only fixation regarding postoperative stability.

Vitamin D3 Promotes Not Motor Coordination but Motor Skill Learning without Influence on Muscle Function.

Although motor coordination or motor skill learning are improved by taking vitamin D in the animal experiment, muscle function have not been estimated. Here we examined the effect of vitamin D3 administration on motor coordination and motor skill learning, muscle strength, and muscle volume in mice fed a vitamin D deficient diet. In mice fed a vitamin D deficient diet, serum calcium and 25(OH)D3 concentrations were measured. We then conducted Rotarod test, beam walking assay, micro-CT analysis, and forelimb grip strength test. Administration of vitamin D3 elongated the retention time in the Rotarod test in a time dependent manner. In contrast, the time to reach a beam goal box in beam walking assay was not changed in mice administered with vitamin D3, compared to the control. Oral administration of vitamin D3 did not affect muscle strength nor muscle volume. Oral administration of vitamin D3 promotes not motor coordination but motor skill learning and does not affect muscle function.

Risk factors for postoperative facial swelling at 1 week after bimaxillary orthognathic surgery in Class III patients.

OBJECTIVE: To quantify facial swelling at 1 week after Le Fort I osteotomy and bilateral sagittal splitting ramus osteotomy in Class III patients and to identify factors contributing to the swelling based on clinical, morphologic, and surgical variables. STUDY DESIGN: Data from 63 patients were examined in this single-center, retrospective study. Facial swelling was quantitatively measured by superimposing computed tomography data taken in the supine position at 1 week and 1 year postoperatively and extracting the area of maximum intersurface distance. Age, sex, body mass index, thickness of subcutaneous tissue, and of masseter muscle, maxillary length (A-VRP), mandibular length (B-VRP), and posterior maxillary height (U6-HRP), surgical movement (ΔA-VRP, ΔB-VRP, ΔU6-HRP), drainage method, and usage of facial bandages were examined. Multiple regression analysis was performed using the above factors. RESULTS: The median swelling at 1 week postoperatively was 8.35 IQR (5.99-11.47) mm. Multiple regression analysis revealed three factors that were significantly associated with facial swelling: Use of postoperative facial bandages (P=0.03), masseter muscle thickness (P=0.03), and ΔB-VRP (P=0.04). CONCLUSION: Absence of a facial bandage, thin masseter muscle, and large horizontal mandibular movement are risk factors for facial swelling at 1 week postoperatively.

Molecular Mobility of Polyrotaxane Surfaces Alleviates Oxidative Stress-Induced Senescence in Mesenchymal Stem Cells.

Polyrotaxane is a supramolecular assembly consisting of multiple cyclic molecules threaded by a linear polymer. One of the unique properties of polyrotaxane is molecular mobility, cyclic molecules moving along the linear polymer. Molecular mobility of polyrotaxane surfaces affects cell spreading, differentiation, and other cell-related aspects through changing subcellular localization of yes-associated proteins (YAPs). Subcellular YAP localization is also related to cell senescence derived from oxidative stress, which is known to cause cancer, diabetes, and heart disease. Herein, the effects of polyrotaxane surface molecular mobility on subcellular YAP localization and cell senescence following H2 O2 -induced oxidative stress are evaluated in human mesenchymal stem cells (HMSCs) cultured on polyrotaxane surfaces with different molecular mobilities. Oxidative stress promotes cytoplasmic YAP localization in HMSCs on high-mobility polyrotaxane surfaces; however, low-mobility polyrotaxane surfaces more effectively maintain nuclear YAP localization, exhibiting lower senescence-associated ß-galactosidase activity and senescence-related gene expression and DNA damage than that seen with the high-mobility surfaces. These results suggest that the molecular mobility of polyrotaxane surfaces regulates subcellular YAP localization, thereby protecting HMSCs from oxidative stress-induced cell senescence. Applying the molecular mobility of polyrotaxane surfaces to implantable scaffolds can provide insights into the prevention and treatment of diseases caused by oxidative stress.

Hyaluronic acid hydrogels support to generate integrated bone formation through endochondral ossification in vivo using mesenchymal stem cells.

Engineered cartilage tissue from differentiated mesenchymal stem cells (MSCs) can generate bone in vivo through endochondral ossification (ECO). This ECO-mediated approach has the potential to circumvent the severe problems associated with conventional MSC-based bone tissue engineering techniques that lack mechanisms to induce angiogenesis. Hyaluronic acid (HA) is a key component in the cartilage extracellular matrix. However, the ECO-supporting properties of HA remain largely unclear. This study aimed to compare the ability of HA and collagen hydrogels to support in vitro differentiation of MSC-based hypertrophic cartilage tissues and to promote endochondral bone formation in vivo. Following the chondrogenic and hypertrophic differentiation in vitro, both HA and collagen constructs accumulated sulfated glycosaminoglycan (sGAG) and type 1, type II, and type X collagen. However, HA hydrogels exhibited a more uniform distribution of sGAG, type 1 collagen, type X collagen, and osteocalcin proteins; in addition, the cells embedded in the hydrogels had more rounded cell morphologies than those in the collagen constructs. At week 5 of in vitro culture, two to three constructs were implanted into a subcutaneous pocket in nude mice and harvested after 4 and 8 weeks. Both HA and collagen constructs promoted endochondral bone formation with vascularization and bone marrow development; however, the HA constructs fused to form integrated bone tissues and the bone marrow developed along the space between the two adhered grafts in all implanted pockets (n = 5). In the collagen constructs, the integration was observed in 40% of the pockets (n = 5). Microcomputer CT analysis revealed that the bone volume of HA constructs was larger than that of collagen constructs. In conclusion, compared to collagen hydrogels, HA hydrogels had superior potential to generate integrated bone with vascularization and bone marrow development. This study provides valuable insights for applying ECO-mediated bone tissue engineering approaches for the repair of critical-sized bone defects.

Gene expression profiling of the masticatory muscle tendons and Achilles tendons under tensile strain in the Japanese macaque Macaca fuscata.

Both Achilles and masticatory muscle tendons are large load-bearing structures, and excessive mechanical loading leads to hypertrophic changes in these tendons. In the maxillofacial region, hyperplasia of the masticatory muscle tendons and aponeurosis affect muscle extensibility resulting in limited mouth opening. Although gene expression profiles of Achilles and patellar tendons under mechanical strain are well investigated in rodents, the gene expression profile of the masticatory muscle tendons remains unexplored. Herein, we examined the gene expression pattern of masticatory muscle tendons and compared it with that of Achilles tendons under tensile strain conditions in the Japanese macaque Macaca fuscata. Primary tenocytes isolated from the masticatory muscle tendons (temporal tendon and masseter aponeurosis) and Achilles tendons were mechanically loaded using the tensile force and gene expression was analyzed using the next-generation sequencing. In tendons exposed to tensile strain, we identified 1076 differentially expressed genes with a false discovery rate (FDR) < 10-10. To identify genes that are differentially expressed in temporal tendon and masseter aponeurosis, an FDR of < 10-10 was used, whereas the FDR for Achilles tendons was set at > 0.05. Results showed that 147 genes are differentially expressed between temporal tendons and masseter aponeurosis, out of which, 125 human orthologs were identified using the Ensemble database. Eight of these orthologs were related to tendons and among them the expression of the glycoprotein nmb and sphingosine kinase 1 was increased in temporal tendons and masseter aponeurosis following exposure to tensile strain. Moreover, the expression of tubulin beta 3 class III, which promotes cell cycle progression, and septin 9, which promotes cytoskeletal rearrangements, were decreased in stretched Achilles tendon cells and their expression was increased in stretched masseter aponeurosis and temporal tendon cells. In conclusion, cyclic strain differentially affects gene expression in Achilles tendons and tendons of the masticatory muscles.

Stiffness as measured with strain elastography is a prognostic factor for pT1/T2 tongue squamous cell carcinoma with muscle-layer invasion.

OBJECTIVE: The objective was to evaluate stiffness as a prognostic factor for tongue squamous cell carcinoma (TSCC). STUDY DESIGN: This retrospective study included 55 patients with pathologic stage pT1 or T2 TSCC with muscle-layer invasion who underwent preoperative strain elastography of the tongue, followed by surgery, as the primary treatment modality at our cancer center. The stiffness of TSCC was semi-quantified as the ratio of the strain value of a non-tumor site to the strain value of the tumor site (strain ratio [SR]) using ultrasound strain elastography findings. RESULTS: SR cutoff values that maximized the significance of the difference for prognosis of delayed cervical lymph node metastasis (DCLNM) and overall survival (OS) were 7.10 and 7.49, respectively. In univariate analysis, SR, age, depth of invasion, pT stage, and perineural invasion were significant risk factors for DCLNM, whereas SR, sex, and DCLNM were identified as having an association with OS. In multivariate analysis, SR was a significant risk factor for DCLNM (hazard ratio [HR] = 3.102; P = .021) and a non-significant but relevant risk factor for OS (HR = 8.774; P = .073). Age also had an association with OS (HR = 0.382; 95% CI 0.127-1.152; P = .088). CONCLUSION: Tongue stiffness is a prognostic factor in patients with pT1/T2 TSCC with muscle-layer invasion. SR values >7.10 indicate a poor prognosis, thereby warranting a strict follow-up regimen in these cases.

Suppressed Migration and Enhanced Cisplatin Chemosensitivity in Human Cancer Cell Lines by Tuning the Molecular Mobility of Supramolecular Biomaterials.

Cancer cells recognize physical cues transmitted from the surrounding microenvironment, and accordingly alter the migration and chemosensitivity. Cell adhesive biomaterials with tunable physical properties can contribute to the understanding of cancer cell responses, and development of new cancer therapies. Previously, it was reported that polyrotaxane-based surfaces with molecular mobility effectively modulate cellular functions via the yes-associated protein (YAP)-related signaling pathway. In the present study, the impact of molecular mobility of polyrotaxane surfaces on the migration and chemosensitivity of lung (A549), pancreatic (BxPC-3), and breast cancer (MDA-MB-231) cell lines is investigated, and it is found that the cellular spreading of adherent A549 and BxPC-3 cells and nuclear YAP translocation are promoted on low-mobility surfaces, suggesting that cancer cells alter their subcellular YAP localization in response to molecular mobility. Furthermore, low-mobility surfaces suppress cellular migration more than high-mobility surfaces. Additionally, low-mobility surfaces promote the cisplatin chemosensitivity of each cancer cell line to a greater extent than high-mobility surfaces. These results suggest that the molecular mobility of polyrotaxane surfaces suppresses cellular migration and enhances chemosensitivity via the subcellular translocation of YAP in cancer cells. Biointerfaces based on polyrotaxanes can thus be a new platform for elucidating cancer cell migration and chemoresistance mechanisms.

Comparison of magnetic resonance imaging findings in patients with intermittent closed lock and acute closed lock of the temporomandibular joint: a cross-sectional retrospective study.

OBJECTIVE: This study aimed to estimate the disc status in intermittent closed lock (ICL) and permanent closed lock (CL) temporomandibular disorders (TMDs) to test the hypothesis that the disc morphology and degree of anterior disc displacement affect the outcomes of these disorders. MATERIALS AND METHODS: All patients were clinically examined according to the Diagnostic Criteria for Temporomandibular Disorders Axis I protocol, and magnetic resonance imaging (MRI) confirmed ICL and CL. Fifty-six joints of 56 patients with ICL and 110 joints of 110 patients with acute CL with a locking period of less than 3 months were included. Patients with acute CL were further classified into two groups: those with CL that could be successfully manipulated (CLs group) and those with acute CL without the possibility of unlocking (CLu group). MRI was used to assess the degree of anterior displacement, lateral displacement of the disc, disc deformity, and joint effusion. MRI findings were compared among the joints in the ICL, CLs, and CLu groups. RESULTS: The degree of anterior displacement and disc deformity prevalence significantly differed among the ICL, CLs, and CLu groups. No significant intergroup differences were observed in terms of lateral displacement or joint effusion. CONCLUSIONS: These results suggest that anteriorly displaced discs and deformation of discs associated with TMD progression affect disc reducibility.

Risk factors of neurosensory disturbances at 1 year postoperatively after bilateral sagittal split osteotomy.

OBJECTIVE: To investigate morphologic and surgical risk factors causing neurosensory disturbances (NSDs) after bilateral sagittal split osteotomy (BSSO). STUDY DESIGN: A total of 237 patients (with 474 sides) who underwent BSSO were followed up for 1 year. Parameters examined included age, sex, asymmetry, mandibular movement direction, mandible cutting devices, split type, intraoperative exposure of the inferior alveolar nerve (IAN), contact between the IAN and screw, distance between mandibular canal and inner surface of the cortical bone (distance A), distance from lateral osteotomy to mental foramen (distance B), and NSD at 1 year postoperatively. RESULTS: NSD was observed in 62 (13.1%) sides of 51 patients. Exploratory factor analysis determined 4 factors (factor 1: distance A; factor 2: direction of mandibular movement; factor 3: distance B and cutting devices; factor 4: IAN exposure). Logistic regression analysis was performed using the above factors and age, sex, and asymmetry, making a total of 7 variables. Age, factor 1, and factor 4 were significant predictors of NSD. CONCLUSIONS: Advanced age, close distance between mandibular canal and inner surface of the cortical bone, and IAN intraoperative exposure are risk factors for NSD 1 year postoperatively. Cases at high risk for NSD must be treated with great care.

Supramolecular Surface Coatings with Acetylated Polyrotaxane-Based Triblock Copolymers for Thermal Regulation of Cell Adhesion and Fabrication of Cell Sheets.

Polyrotaxanes (PRXs) containing acetylated α-cyclodextrins exhibit a temperature-dependent phase transition in aqueous solutions across their lower critical solution temperature (LCST) of approximately 26.6 °C. To gain insights into the interactions of acetylated PRXs (Ac-PRXs) with biological components, thermoresponsive supramolecular surfaces were prepared by coating tissue culture polystyrene (TCPS) surfaces with Ac-PRX triblock copolymers, and their surface properties across the LCST were evaluated. The wettability and protein adsorption of Ac-PRX-coated surfaces changed significantly between 10 and 37 °C, whereas the uncoated TCPS and unmodified PRX-coated surfaces did not alter the wettability and protein adsorption at 10 and 37 °C. The adhesion, proliferation, morphology, and adhesion strength of NIH/3T3 cells on Ac-PRX-coated surfaces were found to be similar to those of the uncoated and unmodified PRX-coated surfaces. However, the adhesion strength of NIH/3T3 cells on Ac-PRX-coated surfaces decreased drastically at 10 °C. Consequently, the cells spontaneously detached from the Ac-PRX-coated surfaces without enzymatic treatment. Additionally, when incubating confluent cells at 10 °C, the cells detached from Ac-PRX-coated surfaces as cell sheets while retaining extracellular matrix proteins. The findings of this study provide new directions for the design of thermoresponsive supramolecular biointerfaces for applications in bioseparation and cell manipulation.

Independent Roles of Molecular Mobility and Zeta Potential on Supramolecular Surfaces in the Sequence of RAW264.7 Macrophage Responses.

Surface properties of biomaterials affect the morphologies and inflammatory responses of macrophages. Recently, biomaterial design utilizing these properties has been explored to build a scaffold for balancing the immune system in vivo. In the present study, polyrotaxane surfaces with different functional groups including methyl, amino, and sulfo groups are utilized to clarify the effect of molecular mobility and zeta potential of these surfaces on RAW264.7 macrophage responses. At 24 h post-seeding, the majority of the cells adhere onto each surface, and the initial spreading is suppressed by more negatively-charged polyrotaxane surfaces. From 24 to 48 h of incubation, the spreading areas on the unmodified and methylated surfaces significantly increase, whereas those on the aminated and sulfonated surfaces remain unchanged. These results suggest that the initially cellular spreading process depends on the zeta potential, while the subsequent spreading process is governed by the molecular mobility. After lipopolysaccharide stimulation, the less mobile surfaces induce higher expression of inflammation-related genes than highly mobile surfaces, suggesting that molecular mobility is the main factor modulating the inflammatory activity in macrophages. These findings indicate that the zeta potential and molecular mobility of polyrotaxane surfaces may play independent roles in the sequence of macrophage responses.

Mitigating RANKL-induced cholesterol overload in macrophages with ß-cyclodextrin-threaded polyrotaxanes suppresses osteoclastogenesis.

Free cholesterol acts as an endogenous agonist for estrogen-related receptor α (ERRα), a nuclear receptor that regulates osteoclastogenesis. Because stimulation of macrophages with receptor activator of nuclear factor κB ligand (RANKL) induces an overload of free cholesterol and activates ERRα, we hypothesized that direct removal of cellular cholesterol would suppress osteoclastogenesis. In this study, the effect of 2-hydroxypropyl ß-cyclodextrin (HP-ß-CD), a highly water-soluble cyclic glucopyranose, and ß-CD-threaded polyrotaxanes (PRXs), supramolecular polymers designed to release threaded ß-CDs in acidic lysosomes, on RANKL-induced cholesterol overload and osteoclast differentiation of murine macrophage-like RAW264.7 cells were investigated. PRXs suppressed RANKL-induced cholesterol overload. Additionally, RANKL-induced osteoclast differentiation of RAW264.7 cells was inhibited by PRXs. In contrast, HP-ß-CD did not reduce cholesterol levels or inhibit osteoclast differentiation in RAW264.7 cells. Gene expression analysis of osteoclast markers suggested that PRXs suppress only the early stage of osteoclast differentiation, as PRXs cannot be internalized into multinucleated osteoclasts. However, modification of PRXs with cell-penetrating peptides facilitated their cellular uptake into multinucleated osteoclasts and inhibited osteoclast maturation. Thus, PRXs are promising candidates for inhibiting osteoclast differentiation by suppressing cholesterol overload and may be useful for treating osteoporosis or other bone defects caused by the overactivity of osteoclasts.