Increased salivary microvesicles are associated with the prognosis of patients with oral squamous cell carcinoma.

Microvesicles (MVs), which are cell-derived membrane vesicles present in body fluids, are closely associated with the development of malignant tumours. Saliva, one of the most versatile body fluids, is an important source of MVs. However, the association between salivary MVs (SMVs) and oral squamous cell carcinoma (OSCC), which is directly immersed in the salivary milieu, remains unclear. SMVs from 65 patients with OSCC, 21 patients with oral ulcer (OU), and 42 healthy donors were purified, quantified and analysed for their correlations with the clinicopathologic features and prognosis of OSCC patients. The results showed that the level of SMVs was significantly elevated in patients with OSCC compared to healthy donors and OU patients. Meanwhile, the level of SMVs showed close correlations with the lymph node status, and the clinical stage of OSCC patients. Additionally, the ratio of apoptotic to non-apoptotic SMVs was significantly decreased in OSCC patients with higher pathological grade. Consistently, poorer overall survival was observed in patients with lower ratio of apoptotic to non-apoptotic SMVs. In conclusion, the elevated level of SMVs is associated with clinicopathologic features and decreased survival in patients with OSCC, suggesting that SMVs are a potential biomarker and/or regulator of the malignant progression of OSCC.

Osteoradionecrosis of the Zygoma.

Osteoradionecrosis occurs in 4.74% to 37.5% of patients following radiation therapy for head and neck cancer. Osteoradionecrosis mostly happens in the mandible but seldom occurs in other maxillofacial bones. Here, the authors reported a rare case of zygomatic osteoradionecrosis which occurred after maxillectomy and then radiotherapy because of maxillary myoepithelial carcinoma. After resection of zygoma sequestrum, the defect was repaired with forehead flap and healed uneventfully.

Down-regulation of connexin43 and connexin32 in keratocystic odontogenic tumours: potential association with clinical features.

AIMS: The objective of this study was to explore the potential involvement of connexin43 (Cx43) and connexin32 (Cx32), two vital members of the connexin families, in the pathogenesis of keratocystic odontogenic tumours (KCOT). METHODS AND RESULTS: The expression levels of Cx43 and Cx32 in human KCOT and normal oral mucosa (OM) tissues were measured using immunohistochemistry and real-time quantitative polymerase chain reaction (qPCR). The relationship between Cx43 and Cx32 expression and markers of proliferation [proliferating cell nuclear antigen (PCNA), cyclin D1], anti-apoptosis [B cell lymphoma 2 (Bcl-2)] and autophagy [light chain 3 (LC3), Sequestosome 1 p62 (p62)] was then investigated in the KCOT samples. The results showed that Cx43 and Cx32 expression was down-regulated significantly in KCOT samples relative to OM samples. Meanwhile, the expression levels of Cx43 and Cx32 were correlated negatively with the expression levels of PCNA, cyclin D1, Bcl-2, LC3 and p62, as confirmed further by double-labelling immunofluorescence analyses. CONCLUSIONS: This study reveals for the first time that Cx43 and Cx32 are down-regulated in KCOT and suggests an association with growth regulation, anti-apoptosis and autophagy in KCOT.

Evaluation of the clinical photographs in the Journal of Oral and Maxillofacial Surgery: from readers' perspectives.

PURPOSE: This study was designed to evaluate clinical photographs published in the Journal of Oral and Maxillofacial Surgery (JOMS) and understand the current status of oral and maxillofacial surgery. MATERIALS AND METHODS: A total of 1,317 photographs from the JOMS Volume 69 were assessed. These photographs were scored from 1 to 10 for the following parameters: sharpness; depth of field; exposure; composition; color or grayscale; background; position; distortion; label consistency; and white balance. Then, the distributions of scores were analyzed. Each score was compared with the average score. The effects of different subjects; emergency or nonemergency situations; and intraoperative, preoperative, or postoperative conditions on the quality of photographs were analyzed by conducting a nonparametric Kruskal-Wallis test. RESULTS: The total score of each photograph showed a left-skewed distribution, varying from 3 to 10, with an average score of 6.82. Four parameters, including sharpness, depth of field, exposure, and white balance, scored less than the average score. Photographs with an intraoral subject yielded the lowest score, with a significant difference (P < .05). The score of photographs taken during a nonemergency situation was significantly higher than that during an emergency situation (6.84 vs 6.03; P < .001). Photographs of an intraoperative condition yielded a score significantly lower than those of pre- and postoperative conditions (6.53 vs 7.11 and 6.75, respectively; P < .001). Approximately 45.5% of photographs (148 of 325) displayed uncovered eyes and 57.1% of specimens (40 of 70) did not appear with a plotting scale. CONCLUSIONS: Sharpness, depth of field, exposure, and white balance should be considered to a greater extent than the other parameters when oral and maxillofacial photographs are taken, particularly for intraoral conditions, emergency situations, and intraoperative conditions. Enhanced parameters and protection of a patient's identity may significantly improve the average level of photographic quality.

A retrospective study of the malignant change of odontogenic keratocyst.

The odontogenic keratocyst (OKC) is a common cystic lesion of the maxilla and mandible. Squamous cell carcinoma (SCC) arising from OKC or dysplasia occurring in OKC is rare. This study aimed to explore the incidence and clinical features of the dysplasia and malignant transformation of OKC. In this study, 544 patients diagnosed with OKC were collected. Among them, 3 patients were diagnosed as SCC arising from OKC, and 12 patients were diagnosed as OKC with dysplasia. The incidence was calculated. Clinical features were analyzed by chi-square test. In addition, a representative case reconstructing mandible with vascularized fibula flap under general anesthesia was reported. And cases reported before were reviewed. The incidence of the dysplasia and malignant transformation of OKC, which are highly associated with the clinical features of swelling and chronic inflammation, is about 2.76%. But the relevance between the dysplasia and malignant transformation and age, gender together with pain is not statistically high. All in all, the clinical features of swelling and chronic inflammation can be considered as characteristics of the dysplasia and malignant transformation of OKC. Although the pain isn't statistically relevant, it may be a dangerous clew. Also, combined with earlier literatures, the dysplasia and malignant transformation of OKC shows unique features of radiographs and histopathology.

Mandibular Reconstruction With the Contralateral Vascularized Iliac Flap Using Individual Design: Iliac Crest Used to Reconstruct the Ramus and the Anterior Border of the Iliac Wing Used to Reconstruct the Inferior Border: A Case Report.

Mandible defects resulting from resection of benign or malignant lesions, trauma, or radionecrosis are commonly encountered in the oral and maxillofacial department. Vascularized bone flaps, in general, provide the best functional and aesthetic outcome. The iliac crest provides a large piece of curved cortico-cancellous bone, measuring 6-16 cm in length. It has a natural curvature that complements the curve of the lateral and sometimes anterior mandible and can be placed accordingly to fill defects. In the paper, we report a mandibular reconstruction with a vascularized iliac flap using individual virtual preoperative planning and 3D printing technology. We want to offer a new design idea for mandibular defect reconstruction.

Myofascial iliac crest flap for reconstruction of combined oral mucosa-mandibular defects: A single-centre experience.

BACKGROUND: Deep circumflex iliac artery (DCIA) myofascial iliac crest flap has been used for combined oral mucosa-mandibular defects reconstruction. The bone component of this composite flap can reconstruct the mandible with superior contour match, and the muscle fascia which used for repairing the oral mucosa defect will transform into an oral mucosa-like appearance. To explore its scope of clinical application and how the fascia transformed into oral mucosa will give surgeons flexibility to reconstruct the combined oral mucosa-mandibular defects. METHODS: A retrospective review of 18 patients who received combined oral mucosa-mandibular defects reconstruction with DCIA myofascial iliac crest flaps from Dec 2016 to Dec 2020 was performed. The characteristics of the mandibular defects and the flaps were recorded. The postoperative dynamic changes of one graft's fascia were observed from serial photographs. RESULTS: All myofascial iliac crest flaps survived successfully. The bone grafts were from 4.0 to 9.5 cm (mean 7.6 ± 1.5 cm) in length and from 2.0 to 3.5 cm (mean 2.7 ± 0.4 cm) in height. The sizes of fascia were from 13.5 to 48.0 cm2 (mean 27.2 ± 9.4 cm2). The grafted fascia firstly changed into a yellow pseudomembrane-like appearance, and then experienced muscle oedema before finally transformed into an oral mucosa-like appearance at about 60 days after operation. CONCLUSION: Myofascial iliac crest flap is a good option for reconstruction of combined oral mucosa-mandibular defects because of its excellent bone and oral mucosa matches.

A deep learning algorithm for detection of oral cavity squamous cell carcinoma from photographic images: A retrospective study.

BACKGROUND: The overall prognosis of oral cancer remains poor because over half of patients are diagnosed at advanced-stages. Previously reported screening and earlier detection methods for oral cancer still largely rely on health workers' clinical experience and as yet there is no established method. We aimed to develop a rapid, non-invasive, cost-effective, and easy-to-use deep learning approach for identifying oral cavity squamous cell carcinoma (OCSCC) patients using photographic images. METHODS: We developed an automated deep learning algorithm using cascaded convolutional neural networks to detect OCSCC from photographic images. We included all biopsy-proven OCSCC photographs and normal controls of 44,409 clinical images collected from 11 hospitals around China between April 12, 2006, and Nov 25, 2019. We trained the algorithm on a randomly selected part of this dataset (development dataset) and used the rest for testing (internal validation dataset). Additionally, we curated an external validation dataset comprising clinical photographs from six representative journals in the field of dentistry and oral surgery. We also compared the performance of the algorithm with that of seven oral cancer specialists on a clinical validation dataset. We used the pathological reports as gold standard for OCSCC identification. We evaluated the algorithm performance on the internal, external, and clinical validation datasets by calculating the area under the receiver operating characteristic curves (AUCs), accuracy, sensitivity, and specificity with two-sided 95% CIs. FINDINGS: 1469 intraoral photographic images were used to validate our approach. The deep learning algorithm achieved an AUC of 0·983 (95% CI 0·973-0·991), sensitivity of 94·9% (0·915-0·978), and specificity of 88·7% (0·845-0·926) on the internal validation dataset (n = 401), and an AUC of 0·935 (0·910-0·957), sensitivity of 89·6% (0·847-0·942) and specificity of 80·6% (0·757-0·853) on the external validation dataset (n = 402). For a secondary analysis on the internal validation dataset, the algorithm presented an AUC of 0·995 (0·988-0·999), sensitivity of 97·4% (0·932-1·000) and specificity of 93·5% (0·882-0·979) in detecting early-stage OCSCC. On the clinical validation dataset (n = 666), our algorithm achieved comparable performance to that of the average oral cancer expert in terms of accuracy (92·3% [0·902-0·943] vs 92.4% [0·912-0·936]), sensitivity (91·0% [0·879-0·941] vs 91·7% [0·898-0·934]), and specificity (93·5% [0·909-0·960] vs 93·1% [0·914-0·948]). The algorithm also achieved significantly better performance than that of the average medical student (accuracy of 87·0% [0·855-0·885], sensitivity of 83·1% [0·807-0·854], and specificity of 90·7% [0·889-0·924]) and the average non-medical student (accuracy of 77·2% [0·757-0·787], sensitivity of 76·6% [0·743-0·788], and specificity of 77·9% [0·759-0·797]). INTERPRETATION: Automated detection of OCSCC by deep-learning-powered algorithm is a rapid, non-invasive, low-cost, and convenient method, which yielded comparable performance to that of human specialists and has the potential to be used as a clinical tool for fast screening, earlier detection, and therapeutic efficacy assessment of the cancer.

M2-polarized macrophages in keratocystic odontogenic tumor: relation to tumor angiogenesis.

The purpose of this study was to evaluate the presence of M2-polarized macrophages and their relationships to angiogenesis in keratocystic odontogenic tumor (KCOT). M2-polarized macrophages were detected in KCOT samples by immunohistochemistry and immunofluorescence. Meanwhile, microvessel density measured with antibody against CD31 was closely correlated with the presence of M2-polarized macrophages. In addition, macrophage colony-stimulating factor (M-CSF) significantly contributed to the activation of M2-polarized macrophages. Moreover, the results of in vitro wound healing, cell migration and tube formation assays further revealed the pro-angiogenic function of M2-polarized macrophage-like cells. This function might be associated with secretion of angiogenic cytokines, such as vascular endothelial growth factor (VEGF), transforming growth factor-ß (TGF-ß) and matrix metalloprotein-9 (MMP-9). This study demonstrates for the first time that M2-polarized macrophages are prevalent in KCOT, and their presence is dependent on M-CSF expression. More importantly, these tumor-supportive cells can also promote tumor angiogenesis by secreting angiogenic cytokines.

Cryopreserved lip mucosa tissue derived keratinocytes can fabricate tissue engineered palatal mucosa equivalent.

Clinical application of tissue engineered palatal mucosa is hampered by unavailability of suitable oral keratinocytes as seeding cells. The aim of this study is to fabricate a tissue engineered palatal mucosa equivalent from the oral keratinocytes which cultured from cryopreserved lip mucosa tissues. Abundant lip mucosa tissues during cheilorrhaphy were firstly cryopreserved in liquid nitrogen for four to six months, and then recovered to culture oral keratinocytes for the fabrication of oral mucosa equivalent. In the control groups, oral keratinocytes cultured from fresh lip mucosa, fresh palate mucosa, and cryopreserved palate mucosa were used to fabricate oral mucosa equivalents. Attachment rate of the oral keratinocytes derived from cryopreserved lip mucosa was lower than that of the keratinocytes from fresh lip mucosa samples, however, the cell cycle distribution of oral keratinocytes cultured from all four groups of mucosa samples were similar. Histologically, the fabricated mucosa equivalents from these four groups had four- to six epithelial layers, the basal cells were cubic and the outmost cells were flatten with narrow nuclei which paralleled to the surface of the dermal matrix. Additionally, Ki-67 positive stained cells were mainly located in the basal layer of the epithelium of these equivalents. These characteristics disclosed that the oral mucosa equivalent cultured from the cryopreserved lip mucosa tissue was not different with the equivalents from other groups and similar to the native palate mucosa tissue. It suggested that the cryopreserved lip mucosa tissues could be used for the construction of palatal mucosal equivalent for clinical application.

In vitro engineering of a palatal mucosa equivalent with acellular porcine dermal matrix.

The objective of this study was to develop a palatal mucosa equivalent composed of multilayered oral keratinocytes grown on the acellular porcine dermal matrix. Acellular porcine dermal matrix was prepared through a series of procedures and assessed by histological, immunohistochemical, and scanning electron microscopy examination. The palatal mucosa equivalent was fabricated by seeding oral keratinocytes, which cultured from human palate mucosa, onto the acellular dermal matrix. After 4 days submerged in medium, this composite was raised to the air-liquid interface for another 7 or 14 days of cultivation. The results demonstrated the processed porcine dermal matrix was totally cell-free. The resultant palatal mucosa equivalent showed a multilayered oral epithelium that had been formed, and the number of cell layers was correlated with the culture period at the air-liquid interface. Oral keratinocytes infiltrated into the empty hair follicles of the acellular porcine dermal matrix and formed an anchor-like structure, which exhibited resemblance to the rete ridges of the native palate mucosa. Immunohistochemical staining for CK10/13, CK19, Ki-67 nuclear antigen, and Heparan sulphate indicated the cultured palatal mucosa equivalent shared the same characteristics with that of the native palate mucosa. In conclusion, our fabricated palatal mucosa equivalent exhibited the characteristics of the native counterpart, and this equivalent might be useful for recovery of the wounds in the palate secondary to palatoplasty.

Keratinocytes modify fibroblast metabolism in hereditary gingival fibromatosis.

OBJECTIVES: Hereditary gingival fibromatosis (HGF) is a rare benign disorder characterized by progressive fibrous overgrowth of the gingiva. The proliferation and expression of growth factors of HGF keratinocytes are abnormal. However, the exact role of keratinocytes in HGF pathogenesis is still unknown. The present study aimed to clarify the interactions between HGF keratinocytes and underlying fibroblasts in the pathogenesis of HGF. DESIGN: Gingival tissues, fibroblasts and keratinocytes from three Chinese HGF patients and three healthy subjects were collected. Histological analyses were performed by histochemical and immunohistochemical staining (Ki-67). Gingival fibroblasts were cocultured with gingival keratinocytes in an in vitro coculture system. The mRNA levels of type I collagen, MMP-1, MMP-3, and TIMP-1 were analysed in the cocultured gingival fibroblasts by reverse-transcription polymerase chain reaction (RT-PCR). The production of type I collagen and TIMP-1 was examined by ELISA. RESULTS: The number of Ki-67-positive keratinocytes in tissue sections from patients was higher than in those from controls. HGF fibroblasts cocultured with HGF keratinocytes showed an increased expression of type I collagen and TIMP-1. Transmission electron microscopy showed increased rough endoplasmic reticulum and ribosomes in cocultured HGF fibroblasts. CONCLUSIONS: These results suggest that HGF keratinocytes have an important role in HGF pathogenesis by inducing extracellular matrix (ECM) accumulation by fibroblasts.