Porphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by stimulating the release of neutrophil extracellular traps in the tumor immune microenvironment.

BACKGROUND: The aim of this study was to investigate the impact of Porphyromonas gingivalis (P. gingivalis) on the progression of oral squamous cell carcinoma (OSCC) through neutrophil extracellular traps (NETs) in the tumor immune microenvironment. METHODS: The expression of NETs-related markers was identified through immunohistochemistry, immunofluorescence, and Western blotting in different clinical stages of OSCC samples. The relationship between NETs-related markers and clinicopathological characteristics in 180 samples was analyzed using immunohistochemistry data. Furthermore, the ability to predict the prognosis of OSCC patients was determined by ROC curve analysis and survival analysis. The effect of P. gingivalis on the release of NETs was identified through immunofluorescence and immunohistochemistry, both in vitro and in vivo. CAL27 and SCC25 cell lines were subjected to NETs stimulation to elucidate the influence of NETs on various cellular processes, including cell proliferation, migration, invasion, and metastasis in vitro. Furthermore, the impact of NETs on the growth and metastatic potential of OSCC was assessed using in vivo models involving tumor-bearing mice and tumor metastasis mouse models. RESULTS: Immunochemistry analysis revealed a significant correlation between the NETs-related markers and clinical stage, living status as well as TN stage. P. gingivalis has demonstrated its ability to effectively induce the release of NETs both in vivo and in vitro. NETs have the potential to facilitate cell migration, invasion, and colony formation. Moreover, in vivo experiments have demonstrated that NETs play a pivotal role in promoting tumor metastasis. CONCLUSION: High expression of NETs-related markers demonstrates a strong correlation with the progression of OSCC. Inhibition of the NETs release process stimulated by P. gingivalis and targeted NETs could potentially open up a novel avenue in the field of immunotherapy for patients afflicted with OSCC.

Effect of concentrated growth factor (CGF) on postoperative sequel of completely impacted lower third molar extraction: a randomized controlled clinical study.

BACKGROUND: The surgical extraction of impacted third molars is one of the most common procedures in oral and maxillofacial surgery, which associated with several postoperative complications. The aim of this clinical trial was to estimate the implication of concentrated growth factor (CGF) on postoperative sequelae after the completely impacted lower third molar extraction. MATERIALS AND METHODS: A total of 74 sides of 37 participants who had completely bilateral impacted lower third molars were enrolled in this split-mouth, randomized single­blind, clinical trial. Surgical extraction was undertaken on both sides of the mandible. Randomization was achieved by opaque, sealed envelopes. The postoperative outcomes including wound healing, swelling and pain were clinically assessed at different-time intervals(1st, 3rd and 7th days). A p-value < 0.05 was considered statistically significant. RESULTS: The wound healing index was significantly better in the test sides (P = 0.001). Regarding the facial swelling, the test sides had significantly less values than the control sides, particularly on the 1st (1.01 ± .57 vs. 1.55 ± .56) and 3rd days (1.42 ± 0.8 vs. 2.63 ± 1.2) postoperatively. Nonetheless, the swelling was disappeared within the 7th day in both sides. The pain scores of visual analog scale were no a statistically significant difference between both sides on the 1st day, meanwhile, the pain scores were significantly lower in the test sides compared with the control sides, especially on the 3rd (P = 0.001) and 7th days (P < 0.001) postoperatively. CONCLUSION: The application of CGF following the surgical extraction of lower third molar has accelerated the healing of soft tissues as well as reduced postoperative sequelae such as swelling and pain. Therefore, the CGF could be promoted among clinicians during the lower third molar surgical extraction. TRIAL REGISTRATION: This study was registered with the TCTR identification number TCTR20210325002 on 25/03/2021 at Thai Clinical Trials Register-Medical Research Foundation of Thailand (MRF). Also it was ethically approved from the institutional ethics committee at the Hospital of Stomatology, Xian Jiaotong University, Xian, China (No: 032), and has been conducted in accordance to the guidelines of the declaration of Helsinki. Written informed consent was obtained from all participants in the study.

Positional Changes of Mandibular Canal Before and After Decompression of Cystic Lesions in the Mandible.

PURPOSE: Significant displacement of the mandibular canal (MC), which occurs frequently in extensive mandibular cystic lesion cases, may raise the risk of inferior alveolar neurovascular bundle injury in surgery. The aim of the present study was to measure the association between positional changes of the MC and the direction (in the coronal plane) of bone expansion of cystic lesions in the mandible. PATIENTS AND METHODS: We performed a retrospective study of patients who had undergone decompression and enucleation surgery from January 2014 to December 2018. Based on coronal planes of cone-beam computerized tomography, the centroids of the expanded mandibles were calculated and considered markers for evaluation of the directions of bone expansion. In addition, the changes in the position of the MC before decompression and enucleation were measured and compared. A Cartesian coordinate system was introduced in this study to illustrate the relationship of positional changes between the displacement of the MC and expansion of the mandible in a straightforward manner. Statistical analysis was performed using the paired t test, unpaired t test, one-way analysis of variance or linear regression as appropriate. RESULTS: Thirty-six patients with an average age of 29.8 years (14 men, 22 women) who received treatment of decompression and enucleation for mandibular cystic lesions were included in this study. The MCs were displaced in the direction toward the lower edge of the mandible and opposite to the direction of mandibular expansion. In addition, the MCs were relocated close to their original location by 1.67 ± 1.45 mm (mean ± standard deviation) approximately 1 year after decompression, accounting for 22.66% of the total displacement. CONCLUSIONS: In mandibular cystic lesion cases, the MCs tend to displace opposite to the direction of mandibular expansion and relocate less after decompression.

An Automated Method of 3D Facial Soft Tissue Landmark Prediction Based on Object Detection and Deep Learning.

BACKGROUND: Three-dimensional facial soft tissue landmark prediction is an important tool in dentistry, for which several methods have been developed in recent years, including a deep learning algorithm which relies on converting 3D models into 2D maps, which results in the loss of information and precision. METHODS: This study proposes a neural network architecture capable of directly predicting landmarks from a 3D facial soft tissue model. Firstly, the range of each organ is obtained by an object detection network. Secondly, the prediction networks obtain landmarks from the 3D models of different organs. RESULTS: The mean error of this method in local experiments is 2.62±2.39, which is lower than that in other machine learning algorithms or geometric information algorithms. Additionally, over 72% of the mean error of test data falls within ±2.5 mm, and 100% falls within 3 mm. Moreover, this method can predict 32 landmarks, which is higher than any other machine learning-based algorithm. CONCLUSIONS: According to the results, the proposed method can precisely predict a large number of 3D facial soft tissue landmarks, which gives the feasibility of directly using 3D models for prediction.

Alveolar ridge preservation in post-extraction sockets using concentrated growth factors: a split-mouth, randomized, controlled clinical trial.

Aim: The aim of this clinical trial was to assess the impact of autologous concentrated growth factor (CGF) as a socket-filling material and its ridge preservation properties following the lower third molar extraction. Materials and methods: A total of 60 sides of 30 participants who had completely symmetrical bilateral impacted lower third molars were enrolled. The primary outcome variables of the study were bone height and width, bone density, and socket surface area in the coronal section. Cone beam computed tomography images were obtained immediately after surgery and three months after surgery as a temporal measure. Follow-up data were compared to the baseline using paired and unpaired t-tests. Results: CGF sites had higher values in height and width when compared to control sites (Buccal wall 32.9 ± 3.5 vs 29.4 ± 4.3 mm, Lingual wall 25.4 ± 3.5 vs 23.1 ± 4 mm, and Alveolar bone width 21.07 ± 1.55vs19.53 ± 1.90 mm, respectively). Bone density showed significantly higher values in CGF sites than in control sites (Coronal half 200 ± 127.3 vs -84.1 ± 121.3 and Apical half 406.5 ± 103 vs 64.2 ± 158.6, respectively). There was a significant difference between both sites in the reduction of the periodontal pockets. Conclusion: CGF application following surgical extraction provides an easy, low-cost, and efficient option for alveolar ridge preservation. Thus, the use of CGF by dentists during dental extractions may be encouraged, particularly when alveolar ridge preservation is required. Clinical trial registration: TCTR identification, TCTR20221028003.

Proteomic analysis of mitochondria from infantile hemangioma endothelial cells treated with sodium morrhuate and its liposomal formulation.

Hemangioma is the most common benign tumor of infancy. The aim of this study is to evaluate the biological effects of sodium morrhuate (SM) and its liposomal formulation on infantile hemangioma endothelial cells (IHECs). Morphological analysis revealed that exposure to liposomal sodium morrhuate (LSM) preferentially caused apoptotic death in IHECs, manifested as shrunken configuration and formation of apoptotic bodies. In contrast, necrotic death was prominent in IHECs treated with an equal concentration of SM. By means of proteomic analysis and confirmation experiments, we revealed that the apoptosis-inducing effects of LSM were associated with an upregulation of a set of genes involved in mitochondrial death pathway, including apoptosis-inducing factor, cytochrome c1, caspase-8, and lamin B1. In conclusion, our data highlight the proapoptotic activity of LSM in IHECs through the mitochondrial apoptotic pathway and may provide a promising avenue to treat hemangiomas of infancy.

Cartilage tissue engineering with demineralized bone matrix gelatin and fibrin glue hybrid scaffold: an in vitro study.

To develop a cartilage-like tissue with hybrid scaffolds of demineralized bone matrix gelatin (BMG) and fibrin, rabbit chondrocytes were cultured on hybrid fibrin/BMG scaffolds in vitro. BMG scaffolds were carefully soaked in a chondrocyte-fibrin suspension, which was polymerized by submerging the constructs into thrombin-calcium chloride solution. Engineered cartilage-like tissue grown on the scaffolds was characterized by histology, immunolocalization, scanning electron microscopy, biochemical assays, and analysis of gene expression at different time points of the in vitro culture. The presence of proteoglycan in the fibrin/BMG hybrid constructs was confirmed by positive toluidine blue and alcian blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrices. Chondrogenic properties were further demonstrated by the expression of gene-encoded cartilage-specific markers, collagen type II, and aggrecan core protein. The glycosaminoglycan production and hydroxyproline content of tissue grown on the fibrin/BMG hybrid scaffolds were higher than that of the BMG group. In conclusion, the fibrin/BMG hybrid scaffolds may serve as a potential cell delivery vehicle and a structural basis for cartilage tissue engineering.

[Study of immunolipo-sodium morrhuate applied on human hemangioma endothelial cells in vitro].

PURPOSE: To explore the method of preparing immunolipo-sodium morrhuate and evaluate its effect on human hemangioma endothelial cells in vitro. METHODS: Using SPDP((N-Succinimidyl-3-(2-pyridyldithio)) propionate) as cross-linker, anti-VEGFR2/KDR monoclone antibody was combined to the liposome surface to prepare immunolipo-sodium morrhuate by extruding method, and then its effect on human hemangioma endothelial cells in vitro was observed by laser scanning confocal microscope, inverted microscope, Gimsa staining, transmission electron microscope, MTT and flow cytometry. RESULTS: The average diameter of the immunoliposome was 122.9 nm, which had a very good stability when compared with normal liposome, it had stronger and faster combining ability, its potential to induce apoptosis was much more prominent, and its toxic effect on human hemangioma endothelial cells was gentle, which was similar to normal liposome. CONCLUSION: We have prepared immunolipo-sodium morrhuate successfully, which has very good specific initiative targetting ability in vitro and can induce pervasive apoptosis of human hemangioma endothelial cells.

[The experimental study of lipo-sodium morrhuate promoting apoptosis of ECV-304 cells].

OBJECTIVE: To evaluate the effect of lipo-sodium morrhuate on ECV-304 cell line. METHODS: The effect lipo-sodium morrhuate was evaluated by toxicology trial (MTT), electron microscope, DNA electrophoresis and flow cytometer. RESULTS: The toxicology results showed, that the number of vital cells in lipo-sodium morrhuate group decreased slowly. The electron microscope exhibited apoptosis in the lipo-sodium morrhuate group. And there were typical DNA ladder in DNA electrophoresis and typical apoptosis peak in flow cytometer. The apoptosis rate was 22.23%. CONCLUSIONS: Unlike the normal preparation of sodium morrhuate, lipo-sodium morrhuate could induce apoptosis of ECV-304 cell line.

[Apoptotic study on the effect of fluorine and selenium on the human hair follicle in vitro].

OBJECTIVE: The aim of this study was to observe the human hair follicle apoptosis status affected by fluorine and the antagonism effect by selenium in vitro. METHODS: The single hair follicles were separated and cultured, then they were added in different concentrations of sodium fluoride and sodium selenite. Chosen the appropriate concentrations, they were divided into 7 groups. The TUNEL was used to investigate the apoptotic cells of different parts. The morphous of hair follicles was observed consecutively and electron microscope was used. RESULTS: We found that in 1 mmol/L and 10 mmol/L sodium fluoride groups, when the human hair follicles in vitro were cultured on the 5th day, the apoptotic cells of outer root sheath (ORS), dermal sheath and hair papilla, hair bulb were obviously increased. But 0.01 mmol/L sodium selenite weakened the toxicity of 1 mmol/L sodium fluoride at the outer root sheath and hair bulb (P < 0.05). CONCLUSIONS: Different concentrations of sodium fluoride had different effect on the growth of human hair follicle in vitro which were cultured on 5th day. Sodium fluoride of certain concentration could accelerate the apoptosis of human hair follicle in vitro. Sodium selenite of certain concentration could act antagonism to the toxicity of sodium fluoride.

[Stress distribution analysis of the surface of condyle during simulated fracture healing of the neck of condyle by miniplate osteosynthesis].

OBJECTIVE: To find the reasons why patients always have temporomandibular disorders (TMD) after condylar fracture by analyzing the stress distribution change of the condylar surface whose subcondylar fracture ware fixed by miniplate during the whole healing process. METHODS: Analyzing the stress distribution change of the condylar surface whose subcondylar fracture were fixed by miniplate during the whole healing process by three-dimension finite element method (3D FEM). RESULTS: During the whole healing process of the fracture, the miniplate osteosynthesis was helpful to the biomechanics environment rehabilitation of the condylar local, but it still had difference with normal after fixing 12 weeks long. CONCLUSION: The difference of stress distribution of condylar surface that fractured and fixed by miniplate with normal may be part of the reason of TMD after the subcondylar fracture miniplate osteosynthesis.