Modified five times simulated body fluid for efficient biomimetic mineralization.

Simulated body fluid (SBF) is widely utilized in preclinical research for estimating the mineralization efficacy of biomaterials. Therefore, it is of great significance to construct an efficient and stable SBF mineralization system. The conventional SBF solutions cannot maintain a stable pH value and are prone to precipitate homogeneous calcium salts at the early stages of the biomimetic process because of the release of gaseous CO2. In this study, a simple but efficient five times SBF buffered by 5 % CO2 was developed and demonstrated to achieve excellent mineralized microstructure on a type of polymer-aligned nanofibrous scaffolds, which is strikingly similar to the natural human bone tissue. Scanning electron microscopy and energy-dispersive X-ray examinations indicated the growth of heterogeneous apatite with a high-calcium-to-phosphate ratio on the aligned nanofibers under 5 times SBF buffered by 5 % CO2. Moreover, X-ray diffraction spectroscopy and Fourier transform infrared analyses yielded peaks associated with carbonated hydroxyapatite with less prominent crystallization. In addition, the biomineralized aligned polycaprolactone nanofibers demonstrated excellent cell attachment, alignment, and proliferation characteristics in vitro. Overall, the results of this study showed that 5 × SBFs buffered by 5 % CO2 partial pressure are attractive alternatives for the efficient biomineralization of scaffolds in bone tissue engineering, and could be used as a model for the prediction of the bone-bonding bioactivity of biomaterials.

TSF/FHA induces osteogenic differentiation of Mc3t3 cells via Pygo2 dependent Wnt/ß-catenin signaling pathway.

OBJECTIVES: This study aimed to investigate whether tussah silk fibroin (TSF)/fluoridated hydroxyapatite (FHA) can promote osteogenic differentiation of Mc3t3 cells and explore the role of Wnt/ß-catenin signaling in this process. METHODS: TSF/FHA was gained via freeze drying technique and cyclic phosphate immersion method. The relative expression levels of bone-related genes and proteins of Mc3t3 cells seeded on different materials were examined by RT-qPCR and Western blotting. Knockdown or overexpression of Pygo2 in Mc3t3 cells was achieved using lentiviral transfection. Cell proliferation, the expression of bone-related genes and proteins were subsequently examined. Animal experiment was also performed to observe the osteogenesis effect. RESULTS: Different ratios of fluorine of TSF/FHA accelerated the osteogenic differentiation of Mc3t3 cells and increased the Pygo2 expression. The Wnt/ß-catenin signaling pathway was activated after TSF/FHA induction, accompanied by the increased expression of related genes. In SD rats with skull defect, the newly formed bone increased significantly and the Pygo2 overexpressing Mc3t3 cells promoted osteogenesis. However, Pygo2 knockdown markedly compromised the osteogenesis of Mc3t3 cells after TSF/FHA induction. CONCLUSION: TSF/FHA facilitates osteogenic differentiation of Mc3t3 cells via upregulating Pygo2 and activating Wnt/ß-catenin signaling pathway.

Blocking circ-SCMH1 (hsa_circ_0011946) suppresses acquired DDP resistance of oral squamous cell carcinoma (OSCC) cells both in vitro and in vivo by sponging miR-338-3p and regulating LIN28B.

BACKGROUND: Circular RNAs (circRNAs) could participate in cis-dichlorodiammineplatinum (DDP) resistance of human cancers. However, circRNAs role in DDP resistance of oral squamous cell carcinoma (OSCC) progression remains largely undeveloped. Here, we attempted to explore the role of circ-SCMH1 (ID hsa_circ_0011946) in acquired DDP resistance. METHODS: Expression of circ-SCMH1, microRNA (miR)-338-3p and Lin-28 homolog B (LIN28B) was detected by real-time quantitative PCR and western blotting, and their interactions were confirmed by dual-luciferase reporter assay, RNA immunoprecipitation and RNA pull-down assay. DDP resistance was assessed by MTT assay, colony formation assay, flow cytometry, transwell assays, western blotting, and xenograft experiment. Transmission electron microscopic analysis, nanoparticle tracking analysis and western blotting confirmed the characterizations of extracellular vesicles (EVs). RESULTS: Circ-SCMH1 was upregulated in DDP-resistant OSCC tissues and cells (SCC-15/DDP and CAL-27/DDP). Circ-SCMH1 knockdown suppressed the half-maximal inhibitory concentration of DDP, colony formation, and migration/invasion in SCC-15/DDP and CAL-27/DDP cells, but promoted apoptosis rate and apoptotic proteins (Bax and cleaved-caspase-3) expression. However, silencing miR-338-3p abrogated above effects, and overexpressing miR-338-3p mimicked that. Similarly, miR-338-3p overexpression role could be counteracted by restoring LIN28B. Moreover, interfering circ-SCMH1 retarded tumor growth of SCC-15/DDP cells in vivo with DDP treatment or not. Mechanistically, circ-SCMH1 directly sponged miR-338-3p in regulating LIN28B, a target gene for miR-338-3p. Notably, circ-SCMH1 was an EVs cargo, and DDP-resistant OSCC cells-derived EVs could provoke circ-SCMH1 upregulation in parental cells. CONCLUSION: Circ-SCMH1 contributes to chemoresistance of DDP-resistant OSCC cells partially via EVs secretion and circ-SCMH1/miR-338-3p/LIN28B axis.

A multicenter randomized phase II trial of hyperthermia combined with TPF induction chemotherapy compared with TPF induction chemotherapy in locally advanced resectable oral squamous cell carcinoma.

BACKGROUND: Hyperthermia has been reported to cause cancer stage regression, thus providing surgical opportunities in patients with unresectable tumors and improving the quality of life of patients by preserving certain organs. METHODS: A prospective open-label phase II trial was conducted to evaluate the efficacy of hyperthermia combined with induction chemotherapy in patients with locally advanced resectable oral squamous cell carcinoma (OSCC). Patients received hyperthermia combined with two cycles of 5-fluorouracil, cisplatin, and docetaxel (TPF) induction chemotherapy regimens or TPF induction chemotherapy alone, followed by radical surgery with postoperative radiotherapy. The primary endpoint was the clinical response rate of the induction chemotherapy. The secondary endpoints were overall survival (OS), disease-free survival (DFS), and toxicity. RESULTS: A total of 120 patients were enrolled, and 115 patients were included in the clinical response analysis. The clinical response rate was significantly higher in the experimental arm than in the control arm (65.45% vs. 40.00%, p = 0.0088). There were no unexpected toxicities, and hyperthermia and induction chemotherapy did not increase the perioperative morbidity rate. Moreover, there was a significant improvement in DFS, but no significant difference in OS between the two arms. In the subgroup analysis, increased OS and DFS rates were associated with patients with favorable clinical response after induction chemotherapy in the total population, experimental arm, and control arm. CONCLUSIONS: Our study demonstrates that hyperthermia combined with induction chemotherapy is associated with a high response rate and provides a new treatment option for patients with resectable stage III or IVA OSCC.

Determining the optimal magnetic resonance imaging sequences for the efficient diagnosis of temporomandibular joint disorders.

BACKGROUND: To compare and analyze nine MRI sequences of the TMJ and determine the optimum sequence for the rapid diagnosis of TMDs so as to develop new clinical guidelines. METHODS: Twenty young volunteers (a total of 40 joints) aged 22-26 years were recruited. Three basic sequences, T1-weighted imaging (T1WI), T2-weighted imaging (T2WI), and proton density-weighted imaging (PDWI), together with three positions, oblique sagittal (OSag) with closed mouth, oblique coronal (OCor) with closed mouth, and OSag with opened mouth, were selected in combination for testing. In the OCor position, four regions of interest (ROIs), the condyle (C), the disc (D), the disc outside (DO), and fat (F), were analyzed. For the OSag with closed mouth position and the OSag with opened mouth position sequences, the four ROIs were the condyle (C), the disc (D), the disc ahead (DA), and the disc rear (DR). The signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and signal intensity ratio (SIR) were calculated and analyzed using independent sample t-tests and one-way analysis of variance. Two senior radiologists scored the images of the nine MRI sequences subjectively and selected three optimal sequences. Using the three selected sequences, 1479 patients with anterior disc displacement with reduction (ADDwR) or anterior disk displacement without reduction (ADDwoR) were evaluated by comparing the preoperative TMJ MRI with the outcomes of the maxillofacial arthroscopy or open surgery. RESULTS: The T1WI sequence showed the highest SNR while the T2WI group had the lowest SNR. The ROIs of the T2WI group had the highest CNR and SIR values in the OCor and OSag sequences. In the OCor sequence, the value for the SIR F/DO group was higher than the SIR C/D and SIR C/DO values. Using subjective analysis to evaluate the quality of the scans, the highest total scores were obtained for the OSag T2WI with opened mouth and OSag PDWI with closed mouth sequences. From the objective and subjective analysis, the three optimal sequences selected were OSag PDWI, OCor T2WI with closed mouth, and OSag T2WI with opened mouth. In patients with anterior disc displacement, the comparisons of the surgery and the selected MRI sequences indicated that the total diagnostic accuracy of the MRI was 96.3% (1,425/1,479 cases). For patients with ADDwoR, the diagnostic accuracy was 98.5% (1,372/1,393 cases), and for those with ADDwR it was 61.6% (53/86 cases). There were significant differences between the ADDwoR and ADDwR groups (χ2=312.92, P<0.01). CONCLUSIONS: The three optimal MRI sequences for the rapid and efficient diagnosis of TMD were determined to be OSag PDWI, OCor T2WI with closed mouth, and OSag T2WI with opened mouth.

The magnetic resonance imaging evaluation of condylar new bone remodeling after Yang's TMJ arthroscopic surgery.

To evaluate the post-operative condylar bone remodeling after the treatment of Yang's arthroscopic surgery. Consecutive cases from Jan 2017 to May 2018 that received Yang's arthroscopic surgery were included in this study, the TMJ MRI examinations were performed preoperatively and postoperatively (follow up for 1 year or more), and condylar bone remodeling was estimated. A total of 229 patients (29 male and 200 female) were included in the study, 161 patients had new bone formation, and the average age was 17.5 ± 2.1a. There was no new bone formation in 68 patients with an average age of 24.5 ± 0.7a. The percentage of new bone formation patients in 10-15 years of age was 94.33% and decreases as the age increases. In the position of new bone formation, the posterior slope of condyle was the most (129 joints), the second was the top of condyle (54 joints), the third was around condyle (33 joints), only 25 joints had new bone on the anterior slope of condyle. After TMJ arthroscopic surgery, the condyle has the ability to form new bone, and the younger the age, the stronger the ability of new bone formation. The formation of new bone was most in posterior slope and least in anterior slope of condyle.

Tumor Suppressor LINC02487 Inhibits Oral Squamous Cell Carcinoma Cell Migration and Invasion Through the USP17-SNAI1 Axis.

PURPOSE: The aim of this study was to explore the functions and associated mechanisms of long noncoding RNA LINC02487 in oral squamous cell carcinoma (OSCC). METHODS: The relative expression levels of LINC02487 in OSCC cell lines and tissue samples were examined by RT-qPCR. Intracellular localization was determined using RNA fluorescence in situ hybridization. LINC02487 was cloned into the pCMV-puro vector and then introduced into OSCC cells using lentiviral transfection. Cell processes, such as proliferation, apoptosis, migration, and invasion, were subsequently examined. LINC02487-binding proteins were identified by ChIRP-MS and confirmed by RNA immunoprecipitation. Protein expression was determined by western blotting assay. RESULTS: LINC02487 has been reported to be downregulated in OSCC. Here, we confirmed that the expression of LINC02487 was reduced in 6 OSCC cell lines compared with that in immortalized normal oral epithelial cells and in 50 OSCC samples compared with paired adjacent normal tissue in a Chinese population and that LINC02487 expression levels were associated with cancer metastasis. We further identified that LINC02487 was localized to the cytoplasm, aggregated around the nuclear membrane. Functional studies demonstrate that overexpression of LINC02487 significantly suppresses cell migration and invasion and also inhibits cell proliferation. For the mechanism, we reveal that LINC02487 directly binds to USP17, a deubiquitinating enzyme, and regulates cell migration and invasion through the USP17-SNAI1 axis in a process that involves epithelial-mesenchymal transition (EMT). CONCLUSION: Our results confirm that long noncoding RNA LINC02487 is downregulated in OSCC tissue samples and cell lines. We also find that LINC02487 acts as a tumor suppressor through the USP17-SNAI1 axis.

Myositis ossificans of the masticatory muscle monitored over three generations: A case report and review of the literature.

Myositis ossificans (MO) is a rare disease and its major feature is the formation of heterotropic bone involving muscle or any other type of soft tissue (tendons, ligament, fascia and connective tissue). In the present study, a case report of a patient diagnosed with MO is presented. The diagnosis was established by evaluation of the medical history of the patient and the patient's family, as well as clinical data, radiology and post-operative pathology. The patient underwent excision surgery of the calcified lesion. In addition, genomic DNA was examined from blood samples of the patient and the patient's father with their consent. A mutation in the non-coding region was detected but any direct causative effect remains elusive. The present case report provided significant information with regard to the incidence of MO in four members of the same family assessed over three generations. The disease exhibited a unique localization in the maxillofacial region.

Nomogram model to predict postoperative infection after mandibular osteoradionecrosis surgery.

Osteoradionecrosis of the mandible (ORNM) is one of the most dreaded complications of radiotherapy. The poor healing capacity of soft tissue after radiation may lead to surgical failure. The current study was designed to identify prognostic factors for postoperative infection (PPI) and propose corresponding prophylaxis and intervention protocols. A retrospective study was conducted concerning ORNM patients from 2000 to 2015. A risk-stratification score and nomogram model were established to predict the risk of PPI. A total of 257 patients were analyzed, and the total incidence of PPI was 23.3% (60/257). In multiple logistic regression analysis, radiation dose [Formula: see text]80 Gy (versus <80 Gy, OR = 2.044, P = 0.035, 95% CI: 1.05-3.979), bilateral ORNM (versus unilateral, OR = 4.120, P = 0.006, 95% CI: 1.501-11.307), skin fistula (versus none, OR = 3.078, P = 0.040, 95% CI: 1.05-9.023), and implant utilization (versus none, OR = 2.115, P = 0.020, 95% CI: 1.125-3.976) were significantly associated with PPI. The susceptibility to PPI in patients with risk-stratification scores of 14-22 was 2.833 times that of patients with scores of 7-13, and 7.585 times that of cases defined as scores of 0-6. The discrimination capability of the nomogram model was estimated using a ROC curve with an AUC of 0.708, revealing potentially useful predictive abilities. In conclusion, current risk-stratification scores and nomogram models effectively predicted the risk of PPI in ORNM patients.

Reconstruction of Maxillary and Orbital Floor Defect With Free Fibula Flap and Whole Individualized Titanium Mesh Assisted by Computer Techniques.

PURPOSE: We sought to investigate the clinical application of free fibula flap and individualized titanium mesh through the use of a virtual planning and guiding template to assist the reconstruction of maxilla and orbital floor defects. PATIENTS AND METHODS: Between 2015 and 2016, a total of 6 adult patients with maxillary and orbital floor defects were enrolled in this study. Preoperative virtual planning, including virtual maxillary resection and fibular reconstruction, was performed in all cases according to 3-dimensional radiographic and clinical findings. A 3-dimensionally printed resin model and prebent templates were used to guide the harvesting and positioning of the fibula flap during surgery. Then, an individualized titanium mesh was used to support the orbital floor and restore the maxillary contour. The results were confirmed by postoperative computed tomography scans and clinical follow-up. RESULTS: Preoperative virtual planning and prebent templates can be used to guide the harvesting and positioning of the fibula flap, as well as the forming and positioning of the individualized titanium mesh, with satisfactory results. All flaps survived, and symmetrical facial contours were achieved with normal lower jaw movement and proper vertical distance for dental implants in all patients. CONCLUSIONS: Computer-aided techniques such as virtual planning, 3-dimensionally printed models, and prebent guide templates can be used to harvest and position a free fibula flap, form personalized titanium mesh, and ultimately improve the clinical efficacy of maxillary and orbital floor reconstruction.

Role of fine needle aspiration cytology in the differential diagnosis of vascular anomalies from other lesions in the head and neck region.

Objective To summarise the role of fine needle aspiration cytology in the differential diagnosis of vascular anomalies from other lesions in the head and neck region by presenting five interesting cases reported from our hospital data. Method Five patients ranging in age from 3 months to 25 years old were diagnosed with vascular anomalies based on their histories, clinical examinations, imaging examinations and ultrasonic testing. The previous diagnosis of these patients were corrected by fine needle aspiration cytology in our hospital and the proper treatment was administered. Result Two cases were wrongly treated in a local hospital. All five of the patients underwent operations after excluding vascular anomalies by fine needle aspiration cytology. Conclusion Fine needle aspiration cytology should be used in the following situations to prevent misdiagnoses and the mistreatment as vascular anomalies: when imaging examination and ultrasonic testing lead to a diagnosis of vascular malformation, but the history or clinical examination does not confirm this diagnosis; when lesions are treated as "vascular malformations" by sclerotherapy, and the hydatid fluid is not like blood or lymph fluid; and when propranolol is used to treat the proliferation stage of "haemangiomas" with without any effect.

Regulation of tumorigenesis in oral epithelial cells by defined reprogramming factors Oct4 and Sox2.

Oct4 and Sox2 are pluripotent stem cell factors but the interplay between them in tumorigenesis is unclear. The aim of the present study was to investigate the roles of Oct4 and Sox2 in the reprogramming of oral cancer stem cells. One or both Oct4 and Sox2 were overexpressed in immortalized oral epithelial (hTERT+-OME) cells by lentivirus transduction. In addition, Oct4 and Sox2 proteins in two oral squamous cell carcinoma cell (OSCC) lines (Cal27 and primary cultured OSCC from a T2N2M0 patient) were individually or combinedly knocked down by shRNA. The results showed that the doubly transduced (Oct4+Sox2+) cells could trigger neoplasms in immunodeficient mice after lentivirus transduction, but single transduced (Oct4+ or Sox2+) cells had no tumor formation ability. The knockdown Sox2low and knockdown Oct4lowSox2low cells resulted in decreased tumor size in the immunodeficient mice but the single knockdown Oct4low cancer cells acquired more aggressive xenografts. Our findings suggest that Oct4+Sox2+ cells may be reprogrammed cancer stem cells inducing oral carcinogenesis.

Can a Novel Surgical Approach to the Temporomandibular Joint Improve Access and Reduce Complications?

PURPOSE: This clinical study investigated whether the vascular-guided multilayer preauricular approach (VMPA) to the temporomandibular joint (TMJ) could improve access and decrease complications. PATIENTS AND METHODS: This retrospective evaluation consisted of a consecutive series of patients who underwent TMJ surgeries through the VMPA from January through December 2013. Patients with a history of TMJ surgery were excluded. Clinical data, including operating times, subjective complaints of incision scars, functional conditions of the auriculotemporal nerve and facial nerve, and other complications, were recorded and analyzed. All patients in this study were followed for at least 6 months. RESULTS: All patients (606 joints) had successful TMJ surgeries through the VMPA. All incisions healed favorably with an uneventful recovery. No patient developed permanent weakness of the facial nerve or other severe complications. CONCLUSION: The VMPA can provide direct access and favorable visibility to the TMJ region and yield good esthetic and functional results. The VMPA can be considered the approach of choice for common TMJ surgeries.

Superficial circumflex iliac artery perforator flap's imaging, anatomy and clinical applications in oral maxillofacial reconstruction.

PURPOSE: To assess the feasibility of superficial circumflex iliac artery perforator (SCIP) flap for oral maxillofacial reconstruction and evaluate computed tomography angiography (CTA) and color Doppler sonography (CDS) in mapping superficial circumflex iliac artery (SCIA) and its perforators. MATERIAL AND METHODS: Fifteen patients were carried out surgery. Perforator identification and the relationship between SCIA, deep circumflex iliac artery (DCIA) and superficial inferior epigastric artery (SIEA) were performed preoperatively and intra-operatively. RESULTS: The relationship between SCIA, DCIA and SIEA was depicted and subdivided into type 1 (8/15), type 2 (2/15), type 3 (2/15), type 4 (2/15) and type 5 (1/15). Surgical procedures and SCIP flap anatomy were described. 14/15 of the SCIP flaps survived and one with necrosis. CONCLUSIONS: The SCIP flap is a reliable, thin and pliable flap with hidden donor site morbidity for oral maxillofacial reconstruction. CTA and CDS are valuable methods for preoperative assessment of the perforator's location and type.

Primary intra- and juxta-articular vascular malformations of the temporomandibular joint: a clinical analysis of 8 consecutive patients.

OBJECTIVE: To analyze primary intra- and juxta-articular vascular malformations of the temporomandibular joint. PATIENTS AND METHODS: This study retrospectively reviewed eight patients (seven venous malformations and one lymphatico-venous malformation) who were treated for intra- or juxta-articular vascular malformations of the temporomandibular joint from November 2005 to January 2011. All patients underwent magnetic resonance imaging (MRI) preoperatively. RESULTS: According to MRI findings, vascular malformations involving TMJ could be divided into 3 types; homogenous, lacunar and mixed types. All patients underwent surgical resection, and the final clinical diagnoses were confirmed by postoperative histopathology and immunohistochemical examinations. All treated patients had no clinical or radiographic signs of recurrence. CONCLUSION: Owing to the lower incidence and nonspecific clinical presentations, preoperative diagnosis of vascular malformations involving the TMJ region is very difficult. The classification based on MRI manifestations is proposed first, then it may greatly help in the initial diagnosis. Surgical resection is considered the first option for these TMJ lesions with excellent results.

Congenital huge submandibular and neck teratoid cyst in newborn.

Head and neck teratoid cysts are the least common congenital cysts in the head and neck region, accounting for approximately 1.8% of all dermoid cysts. Teratoid cysts exhibiting mesodermal elements may be lined by gastric, intestinal, respiratory, squamous, or cilitated epithelium. We present a case of huge submandibular and neck teratoid cyst in newborn with airway obstruction and feeding difficulty. Surgical extirpation is the treatment of choice. However, before operation, some other cystic diseases in the head and neck region needs to be excluded. Fine-needle aspiration biopsy was necessary in differential diagnosis of lesions before treatment, especially that the principle of treatment of those diseases is different.

Surgical management of osteoradionecrosis of the jaws.

OBJECTIVE: The objective of this study is to evaluate our 10-year clinical experience in surgical management of patients with osteoradionecrosis (ORN) unresolved with conservative nonoperative treatment. PATIENTS AND METHODS: The medical records of 120 patients who had been surgically treated for ORN during a 10-year period (January 2003 to January 2013) were retrospectively reviewed. RESULTS: The most predilection ORN site was mandible (82.5%), followed by the maxilla (11.7%). ORN developed within initial 12 months in 39.2% and within the first 3 years in 68.3%. The median radiation dose was 68.1 Gy (range, 35-148 Gy), but 51 patients (42.5%) experienced ORN even though radiation doses were controlled under 60 Gy. Surgical trauma, as we believed, was the most important factor leading to this result. Among all the patients, 12 (10.0%) patients were found ineligible for operative treatment due to comorbid systematic diseases whereas none healed or improved. In terms of surgical management of the rest of the 108 patients, 90 (75.0%) patients underwent radical resection (4 patients unhealed), and 18 (15.0%) patients underwent mild surgical procedures such as sequestrectomy or debridement (1 patient unhealed). Of the 90 radical resection patients, 58 patients underwent radical resections and immediate microvascular flap reconstruction (19 bone flaps and 39 soft flaps), and 32 patients only experience radical resection (5 patients received second-stage reconstruction). According to follow-up information, 55 patients were free of disease. CONCLUSION: Though priority should be given to surgical treatment for the patient whose ORN does not respond to conservative nonoperative treatment, we may as well take into account more individualized regimens based on ORN severity. A hard lesson learned from our article is that the oral maxillofacial surgeon should minimize the trauma for jaws as possible as he can, especially to patients who need to receive postoperative radiotherapy.

Post-infectious myositis ossificans in medial, lateral pterygoid muscles: A case report and review of the literature.

Myositis ossificans (MO) is a disease where heterotropic bone forms within a muscle or other type of soft tissue. MO is classified into two groups, MO progressiva and post-traumatic MO. It rarely occurs in the masticatory muscles and thus, only 20 cases involving the masticatory muscles have been reported since 2001. The majority of the reported cases occurred due to trauma, repeated injury or surgical manipulation. However, in a small number of cases, no specific traumatic event was identified as the cause of MO. To the best of our knowledge, this is the first case of post-infectious MO to be reported in the medial and lateral pterygoid muscles.