Double-faced CX3CL1 enhances lymphangiogenesis-dependent metastasis in an aggressive subclone of oral squamous cell carcinoma.

Because cancer cells have a genetically unstable nature, they give rise to genetically different variant subclones inside a single tumor. Understanding cancer heterogeneity and subclone characteristics is crucial for developing more efficacious therapies. Oral squamous cell carcinoma (OSCC) is characterized by high heterogeneity and plasticity. On the other hand, CX3C motif ligand 1 (CX3CL1) is a double-faced chemokine with anti- and pro-tumor functions. Our study reported that CX3CL1 functioned differently in tumors with different cancer phenotypes, both in vivo and in vitro. Mouse OSCC 1 (MOC1) and MOC2 cells responded similarly to CX3CL1 in vitro. However, in vivo, CX3CL1 increased keratinization in indolent MOC1 cancer, while CX3CL1 promoted cervical lymphatic metastasis in aggressive MOC2 cancer. These outcomes were due to double-faced CX3CL1 effects on different immune microenvironments indolent and aggressive cancer created. Furthermore, we established that CX3CL1 promoted cancer metastasis via the lymphatic pathway by stimulating lymphangiogenesis and transendothelial migration of lymph-circulating tumor cells. CX3CL1 enrichment in lymphatic metastasis tissues was observed in aggressive murine and human cell lines. OSCC patient samples with CX3CL1 enrichment exhibited a strong correlation with lower overall survival rates and higher recurrence and distant metastasis rates. In conclusion, CX3CL1 is a pivotal factor that stimulates the metastasis of aggressive cancer subclones within the heterogeneous tumors to metastasize, and our study demonstrates the prognostic value of CX3CL1 enrichment in long-term monitoring in OSCC.

Sturge-Weber syndrome with cemento-ossifying fibroma in the maxilla and giant odontoma in the mandible: A case report.

Sturge-Weber syndrome (SWS) is a neurocutaneous syndrome with vascular lesions of the cerebral meninges, port wine spots on the face, and glaucoma of the eyes; it is a congenital, non-genetic disease whose etiology and mechanisms are unknown. In this report, we describe a rare case of SWS with unilateral large odontogenic tumors in the maxilla and mandible. The histopathological diagnosis of the maxillary bone lesion on biopsy was juvenile psammomatoid ossifying fibroma, which is considered a type of ossifying fibroma of craniofacial bone origin. However, the final pathological diagnosis of the excision was cemento-ossifying fibroma derived from periodontal ligament cells, and we discuss the histopathology in detail. In addition, the mandibular lesion was one of the largest odontomas reported to date. Furthermore, in this case, we suggest the possibility that the maxillary and mandibular bone lesions are not separate lesions, but a series of lesions related to SWS.

Changes in Quality of Life After Secondary Closure of Palatal Defects: Prosthetic Obturation Versus Surgical Reconstruction.

BACKGROUND: The closure of palatal defects after tumor resection or irradiation is performed with either a prosthesis or autogenous tissue; however, there are no clear criteria regarding selection of the method. Thus, this study aimed to investigate the real-world situation and problems of palatal closure using prostheses, and examined patient opinion on how palatal closure using autogenous tissue improved their postoperative quality of life (QOL). METHODS: In 5 patients whose palatal defects resulted from treatment for head and neck cancer and were closed with a prosthesis, the palate was closed secondarily with autogenous tissue; a questionnaire on daily life was administered pre- and post-operatively. RESULTS: Functional improvements in terms of speech and eating were achieved in all and in 4 of 5 cases, respectively. In all cases, the QOL was better for palatal closure with autogenous tissue than with the prosthesis. CONCLUSIONS: As postoperative QOL was considered to be better when reconstructing the palate with autogenous tissue than with the prosthesis, we recommend to actively select autogenous tissue for palate reconstruction.

Morphological Changes and Durability of Skin and Mucosal Flaps in Intraoral and Pharyngeal Reconstructions: Long-term Follow-up and Literature Review for Potential Second Carcinomas.

The long-term changes in tissues implanted in the oral cavity and pharynx after head and neck reconstruction have not been fully evaluated. This study aimed to clarify the morphological changes, long-term durability, and potential for secondary carcinogenesis in such tissues. In our single-center study, the rough morphological changes in 54 cases of intraoral and pharyngeal skin and mucosal flaps were evaluated more than 10 years after flap transfer. In addition, the literature on the development of second carcinomas from skin flaps was reviewed. The mean follow-up period for transferred flaps was 148 months. The reconstruction areas and the probability of morphological changes were significantly correlated (p=0.006), especially in cases with tongue, lower gingiva, and buccal mucosal reconstruction. Free jejunal flap surfaces were well maintained, whereas tubed skin flaps showed severe morphological changes in cases with pharyngeal reconstruction. None of the flaps in our series developed second primary carcinomas. Skin flaps generally had good durability for > 10 years in intraoral environments, while mucosal flaps had better durability for pharyngeal reconstruction. Second squamous carcinomas arising from skin flaps are extremely rare; however, surgeons should take this possibility into consideration and conduct meticulous and long-term follow-up.

Epidemiological Correlations Between Head and Neck Cancer and Hepatitis B Core Antibody Positivity.

BACKGROUND/AIM: Hepatitis B core (HBc) antibody positivity indicates a history of hepatitis B virus (HBV) infection and latent infection. PATIENTS AND METHODS: We conducted a retrospective case-control study of 512 and 495 head and neck cancer (HNC) and non-HNC patients treated at the Okayama University Hospital, Head and Neck Cancer Center from 2008-2017. Demographic data and risk factors that might affect HNC diagnosis were analyzed to assess their effects. RESULTS: Cancer diagnosis was found to correlate with HBc antibody positivity [odds ratio (OR)=1.50, 95% confidence interval (CI)=1.09-2.08], smoking (OR=3.03, 95%CI=2.16-4.25), and a previous history of cancer (OR=4.12, 95%CI=2.79-6.09). The HBs antigen positivity rate in both groups was very close to that observed in the general Japanese population. The HBc antibody positivity rate was very high only in the HNC group. CONCLUSION: HBc antibody positivity and HNC are epidemiologically correlated.

Relationship Between Renal Dysfunction and Oral Mucositis in Patients Undergoing Concurrent Chemoradiotherapy for Pharyngeal Cancer: A Retrospective Cohort Study.

BACKGROUND/AIM: The aim of this retrospective cohort study was to investigate the association between renal dysfunction (RD) and the development of oral mucositis (OM) in patients undergoing concurrent chemoradiotherapy (CCRT) for pharyngeal cancer including radiation to the oral cavity. PATIENTS AND METHODS: Of 130 patients diagnosed as having pharyngeal cancer who received CCRT at the Okayama University Hospital Head and Neck Cancer Center, 44 were finally selected. RESULTS: During the observation period, 24 (54.5%) patients experienced severe OM (grade 3). The Cox proportional hazards regression model demonstrated that RD (hazard ratio(HR)=2.45, 95% confidence interval(CI)=1.067-6.116, p=0.035) and nasopharynx/oropharynx as center of the irradiated area (HR=2.56, 95% CI=1.072-5.604, p=0.034) were significantly associated with the incidence of severe OM (grade 3). CONCLUSION: In patients with pharyngeal cancer treated with CCRT including radiation to the oral cavity, RD at baseline can be a risk factor for developing severe OM.

The Incidence of Oral and Oropharyngeal Cancers in Betel Quid-Chewing Populations in South Myanmar Rural Areas.

Oral cancer is a very common disease in South and Southeast Asia. Betel quid (BQ)- chewing and tobaccosmoking habits are etiological factors for oral cancer patients in these regions. We conducted an oral cancer screening in BQ-chewing endemic rural areas in South Myanmar for the early detection of oral cancer in BQ-chewing and smoking individuals. We examined 105 subjects who were at high risk of oral cancer due to their oral habits (BQ users and/or smokers). Three carcinoma cases were detected, and there were 8 dysplasia cases. The carcinoma detection rate was 2.9%, and the carcinoma and precancerous lesion detection rate was 10.5%. In Myanmar, oral cancer screening has been conducted sporadically on a voluntary basis, and nationwide surveys have never been performed. There are also few reports of oral cancer screening for high-risk groups among the general population in Myanmar. Our present findings highlight the need for further screening and surveys. Education on betel quid chewing- and tobacco- related oral diseases and screening for the early detection of oral cancer are of the utmost importance in the control and prevention of oral cancer.

Structure of a New Palatal Plate and the Artificial Tongue for Articulation Disorder in a Patient with Subtotal Glossectomy.

A palatal augmentation prosthesis (PAP) is used to facilitate improvement in the speech and swallowing functions of patients with tongue resection or tongue movement disorders. However, a PAP's effect is limited in cases where articulation disorder is severe due to wide glossectomy and/or segmental mandibulectomy. In this paper, we describe speech outcomes of a patient with an articulation disorder following glossectomy and segmental mandibulectomy. We used a palatal plate (PP) based on a PAP, along with an artificial tongue (KAT). Speech improvement was evaluated by a standardized speech intelligibility test consisting of 100 syllables. The speech intelligibility score was significantly higher when the patient wore both the PP and KAT than when he wore neither (p＝0.013). The conversational intelligibility score was significantly improved with the PP and KAT than without PP and KAT (p＝0.024). These results suggest that speech function can be improved in patients with hard tissue defects with segmental mandibulectomy using both a PP and a KAT. The nature of the design of the PP and that of the KAT will allow these prostheses to address a wide range of tissue defects.

Surrounding cells affect the gene expression pattern of human beta-defensins in squamous cell carcinoma in vitro.

BACKGROUND/AIM: Defensins are basic peptides involved in non-immune bio-defense mechanisms in a normal epithelium. Human oral squamous cell carcinoma cells (OSCC) also produce human beta-defensins (HBDs), although their exact function is not clear. This study aimed to analyze the variation in gene expression levels of hBDs in co-cultures of OSCC with murine cells. MATERIALS AND METHODS: Two OSCC cell lines (HSC-3, HSC-4) were co-cultured with mouse embryonic fibroblasts, NIH/3T3 or a mouse chondrogenic cell line derived from teratocarcinoma, ATDC5, for 1.5 days. Expression patterns of the hBD genes were investigated by real-time polymerase chain reaction (RT-PCR). RESULTS: hBD1 expression increased when co-cultured with NIH/3T3 but decreased when co-cultured with ATDC5. Expression of hBD2 and hBD4 tended to decrease. OSCC cells formed colonies when co-cultured with NIH/3T3 but were scattered when co-cultured with ATDC5. CONCLUSION: hBDs expression in OSCC is dependent on the type of co-cultured cells and differences in gene expression may be responsible for the morphological differences observed. OSCC may produce HBDs for purposes other than bio-defense by surrounding cells.

Effects of radiation therapy on postoperative complications and adverse events in patients with head and neck reconstruction with flaps.

BACKGROUND: Radiation therapy is an essential treatment for head and neck cancer. However, the condition of the operative field is entirely altered after radiation therapy. This study aimed to examine the effects of preoperative radiation therapy on complications in patients who underwent head and neck reconstruction with flaps. METHODS: We retrospectively reviewed 252 instances of head and neck reconstruction with flaps in 240 patients between October 2000 and May 2011 at Okayama University Hospital. Of the participants, 51 had preoperative radiation exposure (21.3%) and 189 had no radiation exposure (78.7%). Postoperative complications were divided into three categories: minor complications that healed with conservative medical treatment within 4 weeks without a need for surgery; major complications requiring reoperation within 1 week after surgery (reoperation); and major complications needing additional operation later than 1 week after surgery (additional operation). RESULTS: Preoperative radiation therapy was only associated with major complications requiring reoperation later than 1 week after surgery (P < 0.001), open cervical wounds (P = 0.0030), and skin grafting for cervical skin necrosis (P = 0.0031) when compared to no radiation exposure. The results of flap failure were not significantly different between both groups (P = 0.3820). CONCLUSIONS: Minor complications and reoperation in the early postoperative period were not influenced by radiation exposure. The complications of radiation tend to be protracted and associated with additional operation later than 1 week after the initial surgery. It was thought that shortening of the duration of treatment was successful when we needed to perform early additional operations.

Secondary head and neck reconstruction using free flap to improve the postoperative function or appearance of cancer survivors.

BACKGROUND: Secondary reconstructive operations are needed when patients with head and neck cancers have complications such as tumor recurrence after initial treatment. These reconstructive procedures are also performed to improve the function and appearance of the head and neck region for many cancer survivors. We reviewed the patients who underwent secondary head and neck reconstruction to improve function and appearance and considered the significance of this procedure for cancer survivors, as well as its associated problems. METHODS: Among the secondary reconstruction patients, 20 patients underwent reconstruction to improve their function and/or appearance. The goal of reconstruction for the patients was functional improvement in eight cases, appearance improvement in ten cases, and both function and appearance in two cases. Chi-square analyses were performed between the secondary and primary reconstructive groups with regard to the incidence of postoperative complications. RESULTS: All transferred flaps survived completely. We performed a small postoperative modification procedure in four cases. Minor complications not requiring surgical correction occurred in 2 of 20 patients. Additional operations were required owing to major postoperative complications in 2 of 20 patients. No significant associations were identified between the secondary and primary reconstructive groups with regard to postoperative complications. CONCLUSION: The outcomes of the present report suggest that secondary reconstructive surgery is a relatively safe procedure. The decision to perform adaptation operations depends on various factors after sufficient discussion with patients.

Toluene Exposure Leads to a Change in Expression Patterns of ß Defensins in the Mouse Tracheal Epithelium.

Defensins are generally implicated in the quick resistance of epithelial surfaces to microbials; however, recent reports have indicated that defensins also have unknown purposes in relation to noninfectious diseases. In this study, the localization patterns of anti-microbial peptides, ß defensins (BDs), in the tracheal epithelium of male C3H mice under exposure to toluene were analyzed by immunohistochemistry. Mice were exposed one to ten times to toluene for 30 min by nose-only inhalation. Expression of BDs was revealed by immunohistochemistry in serial sections of trachea after the final exposure. Expression of BD-1 was usually observed at almost the same levels in all exposure groups, and expression of BD-2 was observed in the control group; however, the signals for BD-2 decreased gradually with frequency of exposure. In the group exposed ten times, expression of BD-2 decreased to far lower than that of the control group. No expression of BD-3 was detected in any groups. Interestingly, expression of BD-4 increased to the maximum in the group exposed four times and decreased to a level lower than that of the control in the group exposed ten times. The results of the present study indicated that toluene gas might change the expression pattern of BDs in the tracheal epithelial cells. The oscillation of expression of BD-4 was quite characteristic and might contribute to morphological damage in on the epithelial cells.

Prevention points for plate exposure in the mandibular reconstruction.

INTRODUCTION: The rate of complications for mandibular reconstruction after segmental mandibulectomy is higher with reconstruction plates than with vascularised bone grafts. We have experience of over 100 patients using reconstructive plates for reconstruction immediately after segmental mandibulectomy and have considered factors contributing to plate exposure. PATIENTS AND METHODS: Seventeen cases utilised our prevention methods in which reconstructive plates were used for mandibular reconstruction were reviewed. The flaps used with reconstruction plates were rectus abdominis myocutanenous flaps in 10 cases, anterolateral thigh flaps combined vastus lateralis muscle in four cases, and the omentum in one case; no flap was transferred in two cases. RESULTS: In only one of 17 cases was a plate exposed at 3 months postoperatively. No plate exposure occurred during the follow-up period in the other 16 cases. Because no flap had been transferred in the patient with plate exposure, a possible contributing factor was the persistence of dead space beneath the plate. CONCLUSION: This series suggests that factors other than flap selection contribute to the exposure of reconstructive plates. Use of a reconstruction plate is a useful reconstructive method, especially for patients who cannot tolerate transfer of a vascularised bone graft.

Bone quality and quantity of the anterior maxillary trabecular bone in dental implant sites.

OBJECTIVES: The aim of this study was to investigate the characteristics of implant sites on the edentulous alveolar ridge in the anterior maxilla. We studied the bone quantity and quality of implant sites at the anterior maxilla using CT images for the 33 implant sites on patients who underwent dental implant therapy in our Department since 2006. MATERIALS AND METHODS: Computed tomography (CT) images of 33 patients (20 women: 13 men) encompassing 33 implant sites were chosen and examined. The recipient sites for implant placement were determined based on CT data using an implant planning software (Simplant 11.0). The mean bone density values in Hounsfield unit (HU) were recorded using Simplant for both the simulated implant areas and the trabecular bone width. We classified the edentulous alveolar ridge and bone quality according to a classification based on Lekholm and Zarb (1985). RESULTS: Incisors had higher bone densities than canines. Women had lower bone densities than men. Canines displayed greater trabecular bone density and alveolar bone widths than incisors. No maxillary sites were judged to have a bone quality of 1 in this group. Quality 3 accounted for 69.7% of the total samples. CONCLUSIONS: An assessment of bone quality in the anterior alveolar ridge may well reflect age-related systemic pathological conditions and should be used in dental implant treatment planning to avoid associated risk factors.

Quantitative analysis of free flap volume changes in head and neck reconstruction.

BACKGROUND: The purpose of this study was to determine whether free flap volume decreases or increases in the long-term postoperative period. METHODS: We used a retrospective analysis of 17 patients to measure muscle and fat volume in free flap with 3-dimensional (3D) images using the AZE Virtual Place Lexus64. RESULTS: Seventeen patients underwent free flap reconstruction with rectus abdominis musculocutaneous flaps (n = 11) or anterolateral thigh flaps (n = 6). Median follow-up was 28.9 months (range, 2.1-48.4 months). Total flap volume was significantly decreased in flaps including ≥40% muscle (p = .011). Mean final muscle volume was 50% at an average of 12 months. Final fat volume was significantly higher for cases with no evidence of disease (mean, 116.7%) than for died-of-the-disease cases (mean, 70.3%; p = .007). CONCLUSION: Use of free flaps with a high ratio of fat to muscle is sustainable and can gain volume over time, as transplanted fat can increase depending on host condition.

Basic fibroblast growth factor supports expansion of mouse compact bone-derived mesenchymal stem cells (MSCs) and regeneration of bone from MSC in vivo.

Some progress has been made in development of methods to regenerate bone from cultured cells, however no method is put to practical use. Here, we developed methods to isolate, purify, and expand mesenchymal stem cells (MSCs) from mouse compact bone that may be used to regenerate bone in vivo. These cells were maintained in long-term culture and were capable of differentiating along multiple lineages, including chondrocyte, osteocyte, and adipocyte trajectories. We used standard cell isolation and culture methods to establish cell cultures from mouse compact bone and bone marrow. Cultures were grown in four distinct media to determine the optimal composition of culture medium for bone-derived MSCs. Putative MSCs were subjected to flow cytometry, alkaline phosphatase assays, immunohistochemical staining, and several differentiation assays to assess cell identity, protein expression, and developmental potential. Finally, we used an in vivo bone formation assay to determine whether putative MSCs were capable of regenerating bone. We found that compact bone of mice was a better source of MCSs than the bone marrow, that growth in plastic flasks served to purify MSCs from hematopoietic cells, and that MSCs grown in basic fibroblast growth factor (bFGF)-conditioned medium were, based on multiple criteria, superior to those grown in leukemia inhibitory factor-conditioned medium. Moreover, we found that the MSCs isolated from compact bone and grown in bFGF-conditioned medium were capable of supporting bone formation in vivo. The methods and results described here have implications for understanding MSC biology and for clinical purpose.

Relationship between serum albumin concentration and periodontal condition in patients with head and neck cancer.

BACKGROUND: Serum albumin concentration is known to be an independent predictor of survival in head and neck cancer. The previous studies suggested relationships between serum albumin concentration and oral health status in populations without serious systemic disorders. However, these relationships remain unclear in patients with head and neck cancer. The purpose of this study is to investigate the relationship between serum albumin concentration and oral health status in patients with head and neck cancer. METHODS: Fifty individuals diagnosed with primary head and neck cancer, 25 individuals with normal serum albumin concentration (≥3.85 g/dL), and 25 age- and sex-matched individuals with lower serum albumin concentration (<3.85 g/dL) were analyzed. General status, including cancer stage, body mass index, drinking and smoking habits, and biochemical serum markers, were evaluated. Oral health status, including periodontal condition and occlusion tooth pairs, were also evaluated. RESULTS: Mean clinical attachment level (CAL) and C-reactive protein in the lower serum albumin concentration group were greater than those in the normal serum albumin concentration group (P = 0.009 and P = 0.002, respectively). However, there were no significant differences in any other oral and/or serum parameters between the two serum albumin groups. A logistic regression model showed that mean CAL was significantly associated with high or low levels of serum albumin concentration (odds ratio = 9.752; 95% confidence interval = 1.702 to 55.861; P = 0.011). CONCLUSIONS: This study suggests an association between periodontal disease and serum albumin concentration in patients with head and neck cancer. Longitudinal studies are necessary to examine the causal relationship between serum albumin concentration and periodontal condition.

Localization of antimicrobial peptides human beta-defensins in minor salivary glands with Sjögren's syndrome.

Sjögren's syndrome is a common systemic autoimmune disease associated with inflammatory cells that infiltrate exocrine glands. The antimicrobial peptides human beta-defensin-1, human beta-defensin-2, and human beta-defensin-3 are expressed in various human epithelial cells and in normal salivary glands. Antimicrobial peptides provide local protection against infection and participate in inflammatory responses. Because of the presence of inflammation, we hypothesized that human beta-defensin expression in minor salivary glands may be increased in subjects with Sjögren's syndrome. However, the expression of human beta-defensins 1 and 2 was decreased in salivary glands affected by Sjögren's syndrome in comparison with the human beta-defensin expression patterns in salivary glands from normal subjects. In addition, the reduction in expression of human beta-defensin-2 was greater than the reduction in expression of human beta-defensin-1. The aforementioned result suggests that the reduction in expression of human beta-defensin-2 may occur earlier than the reduction in expression of human beta-defensin-1, which may lead to a greater decrease in human beta-defensin-2 than in human beta-defensin-1 during disease progression.

Immobilized recombinant human bone morphogenetic protein-2 enhances the phosphorylation of receptor-activated Smads.

Bone morphogenetic protein (BMP)-2 plays an important role in bone growth and regeneration; however, BMP-2 is easily lost by diffusion through body fluid and has some inhibitory pathways. To address this problem, we previously immobilized recombinant human BMP-2 (rhBMP-2) on succinylated type I atelocollagen. Here, we examined the effect of immobilized rhBMP-2 in vitro and vivo. In ST2, MC3T3-E1, and C2C12 cells, alkaline phosphatase activity, which is a marker of osteoblast differentiation, was enhanced more by immobilized than nonimmobilized rhBMP-2. In addition, the phosphorylation of receptor-activated Smads, part of the signaling pathway activated by BMP-2, was prolonged by immobilized rhBMP-2 in these cells. Furthermore, implantation of immobilized rhBMP-2 into the backs of rats promoted the formation of mature bone-like structure. These results demonstrate that immobilized rhBMP-2 has higher bioactivity than nonimmobilized rhBMP-2, and, therefore, immobilization of rhBMP-2 can prolong BMP signaling.