Application of gel based on medical image inspection in total closure resection of oral cancer patients.

Oral cancer is a common malignant tumor, and total closed resection is a common treatment. However, it has always been a challenge to determine the exact extent of excision during surgery. The application of medical image examination in surgery can provide important reference information, but the current methods still have some limitations. This study explored the application of gels based on medical image examination in the total closed resection of oral cancer patients to improve the accuracy of resection range and surgical treatment effect. The study collected medical image data of patients with oral cancer for image enhancement and determination of resection boundaries. By comparing the results of the experimental group and the control group, the application effect of gel in operation was evaluated. Through the application of medical image inspection technology, the determination of surgical resection boundary is more accurate, and the positive incisal margin of patients is effectively avoided. Gel technology improves the success rate and efficacy of surgery, and this method helps to improve the accuracy of surgery and the certainty of the scope of resection, which is of great significance for improving the surgical treatment effect and the survival rate of patients.

The study on the effect of amino acid porphyrin conjugate-mediated antimicrobial photodynamic therapy on Streptococcus mutans biofilm.

Antimicrobial Photodynamic Therapy (aPDT) based on the action of visible light and photosensitizers has emerged as a promising microbial reduction and alternative to antibiotics resistance to cariogenic pathogens. The present research aims to evaluate the antimicrobial effect of aPDT mediated by a new photosensitizer (amino acid porphyrin conjugate 4i) on Streptococcus mutans (S. mutans) biofilm. Qualitative morphologic characteristics of S. mutans biofilms are shown by scanning electron microscopy (SEM). The colony plate counting method is used to measure the dark toxicity and the phototoxicity of different concentrations of 4i-aPDT to S. mutans biofilms. MTT assay is conducted to investigate the effect of 4i mediated aPDT on the metabolic activity of S. mutans biofilm. Changes in structure morphology, bacterial density and extracellular matrix of S. mutans biofilm are observed by SEM. The distribution of living and dead bacteria in biofilm is detected using Confocal laser microscopy (CLSM). The results indicate that single laser irradiation has no antibacterial effect on S. mutans biofilms. With the increase of 4i concentration or the prolongation of laser irradiation time, the antibacterial effect of 4i-mediated aPDT on S. mutans biofilm is more statistically significant compared to the control. When the concentration of 62.5 µmol/L 4i is continuously illuminated for 10 min, the logarithm of the colonies in the biofilm shows a reduction of 3.4 log10. MTT assay detected absorbance values of biofilm by 4i-mediated aPDT are the lowest, indicating a significant decrease in biofilm metabolic activity. SEM analysis shows that 4i mediated aPDT reduced the quantity and density of S. mutans. A dense red fluorescence image of the 4i-aPDT treated biofilm is observed under CLSM, indicating that the dead bacteria are widely distributed.

The impact of IGF-1 on alveolar bone remodeling and BMP-2 expression in orthodontic tooth movement in diabetic rats.

BACKGROUND: Orthodontic tooth movement is linked to alveolar bone reconstruction. OBJECTIVES: As a regulator of cell proliferation, insulin-like growth factor 1 (IGF-1) plays an important role in osteoporotic fracture healing. This study aims to investigate the effect of IGF-1 on alveolar bone remodeling in diabetic rats. MATERIAL AND METHODS: Sprague Dawley (SD) rats were randomly divided into 3 groups, including a control group, a model group established with streptozotocin (STZ) injection to prepare the diabetic rats (type 1 diabetes), and an IGF-1 group of diabetic rats receiving daily intraperitoneal injections of 1.0 mg/kg IGF-1. Nickel-titanium coil springs were used to pull the first molar forward to establish the model. The maxillary first to third molars and the surrounding alveolar bone were collected to measure tooth movement distance. Hematoxylin and eosin (H&E) staining was applied to detect the pathological changes in the periodontal tissue. Real-time polymerase chain reaction (PCR) and western blot were adopted to measure bone morphogenetic protein 2 (BMP-2) mRNA and protein expression. Enzyme-linked immunosorbent assays (ELISAs) were used to measure interleukin-1α (IL-1α) levels in the serum. RESULTS: The tooth movement distance was significantly decreased, BMP-2 expression was downregulated, and IL-lα levels were enhanced in the model group compared to the control group (p < 0.05). However, the tooth movement distance was increased, BMP-2 expression was increased, and IL-lα levels were reduced in the IGF-1 group compared to the model group (p < 0.05). Hematoxylin and eosin staining showed that alveolar bone destruction was attenuated in the IGF-1 group, while the new bone was not active in the model group. CONCLUSIONS: Diabetes can damage alveolar bone remodeling in orthodontic tooth movement. The IGF-1 promotes alveolar bone remodeling by inhibiting inflammation and upregulating BMP-2 expression.

Effect of super activated platelet lysate on cell proliferation, repair and osteogenesis.

BACKGROUND: Platelet lysate (PL) is considered as an alternative to fetal bovine serum (FBS) and facilitates the proliferation and differentiation of mesenchymal cells. OBJECTIVE: The aim of this study is to explore whether super activated platelet lysate (sPL), a novel autologous platelet lysate, has the ability to inhibit inflammation and promote cell proliferation, repair and osteogenesis as a culture medium. METHODS: Different concentrations of sPL on human fetal osteoblastic 1.19 cell line (hFOB1.19) proliferation and apoptotic repair were investigated; And detected proliferative capacity, inflammatory factor expressions and osteogenic differentiation of human dental pulp cells (hDPCs) stimulated by LPS under 10% FBS and 5% sPL mediums. RESULTS: sPL promoted hFOB1.19 proliferation and had repairing effects on apoptotic cells. No significant difference in proliferation and IL-1α, IL-6 and TNF-α expressions of hDPCs in FBS and sPL medium stimulated by LPS. hDPCs in sPL osteogenic medium had higher osteogenic-related factor expressions and ALP activity. LPS promoted osteogenic-related factor expressions and ALP activity of hDPCs in FBS osteogenic medium, but opposite effect showed in sPL medium. CONCLUSION: sPL promoted osteoblast proliferation and had restorative effects. Under LPS stimulation, sPL did not promote hDPCs proliferation or inhibit inflammation. sPL promotes osteogenic differentiation of hDPCs.

Effects of Super-Activated Platelet Lysate on Early Healing of Tooth Extraction Sockets in Rats.

Purpose: To evaluate the effect of super-activated platelet lysate (sPL) on wound healing of tooth extraction sockets in rats. Methods: Rat models of the tooth extraction socket were established. Thirty-six rats were divided into control and sPL groups and sacrificed on days 7, 14, and 28 after tooth extraction. Bone formation in tooth extraction sockets were observed by microscopic computed tomography (micro-CT) and hematoxylin and eosin (HE) staining; osteoprotegerin (OPG), receptor activator of nuclear factor kappa-Β ligand (RANKL), interleukin 6(IL-6), and tumor necrosis factor-alpha (TNF-α) proteins were detected by immunohistochemistry; and chemokine and osteogenic gene expressions were detected by polymerase chain reaction (PCR). Results: sPL accelerated soft tissue wound healing in the extraction socket of rats. Micro-CT showed that the amount of bone formation and bone volume fraction were higher in the sPL group than the control 14 days after extraction. HE staining showed promotion of the formation of bony trabeculae by sPL in the apical third of the extraction socket 7 days after extraction and more mature and organized bony trabeculae in the sPL group than the control 14 days after extraction; mature bony trabeculae filling most of the fossa with lesser bone porosity in the socket in the sPL group than the control 28 days after extraction. Immunohistochemistry showed that sPL induced OPG expressions 7 and 14 days after tooth extraction but did not affect the RANKL expression while transiently promoting the IL-6 expression 7 days after extraction. PCR showed that sPL promoted chemokine expressions 7 and 14 days after extraction. The expressions of osteogenesis-related factors were higher in the sPL group than the control 7 and 28 days after extraction, while the opposite trend was observed 14 days after extraction. Conclusion: sPL has a transient pro-inflammatory effect and promotes soft tissue healing and bone formation during early wound healing of extraction sockets in rats.

CD25+ B cells produced IL-35 and alleviated local inflammation during experimental periodontitis.

BACKGROUND AND OBJECTIVE: Host immunity is crucial during periodontal inflammations. B cells are considered to have a function of immunoregulation, and TLRs are considered to be crucial in this process. The present study illustrates the potential roles and rules of CD25+ B cells during periodontitis, especially its effect on regulating host IL-35 level and Th1, Th17, and Treg differentiation. MATERIAL AND METHODS: The proportion of local and systemic CD25+ B cell subpopulations from periodontitis models were identified by flow cytometry. To illustrate further mechanism, B cells were cultured with a different type of TLR activators. Expression of IL-10, IL-35, and TGF-ß was detected by ELISA and real-time PCR. We also set adoptive transfer models by using CD25+ B cells. Alveolar bone erosion, proportion of Th1, Th17, and Tregs, and levels of IFN-γ, TNF-α, IL-1ß, and IL-17 were identified. RESULT: Periodontitis induces more CD25+ B cell subpopulations and promotes their IL-10, IL-35, and TGF-ßproduction. TLR activators enhanced Breg proliferation and function. LPS+CpG obviously induced more CD25+ B cell differentiation and production of IL-10, IL-35, and TGF-ß. Adoptive transfer of CD25+ B cells reduces alveolar bone destruction and local Tregs, proportion, especially the local level of IFN-γ and IL-17. In addition, adoptive transfer of CD25+ B cells remedies the pathological change in the proportion of IL-1ß and Th1/Th17 in local lesions. We did not find any significant difference in peripheral blood, regardless of group and detected items. CONCLUSION: Results of the present study clarify that CD25+ B cells enlarged and produced more IL-10, IL-35 and TGF-ß during periodontitis, activation of TLR4 and TLR9 played crucial roles in this process. Also, CD25+ B cells alleviated periodontal inflammation and alveolar bone resorption. Our findings further expanded the potential of B cells during periodontitis.

Antimicrobial photodynamic therapeutic effects of cationic amino acid-porphyrin conjugate 4i on Porphyromonas gingivalis in vitro.

BACKGROUND: Porphyromonas gingivalis (P. gingivalis) is considered to be among the principal pathogens in periodontal disease. The present study aimed to investigate the effect of antimicrobial photodynamic therapy (aPDT) mediated by cationic amino acid-porphyrin conjugate 4i on P. gingivalis METHODS: The uptake of 4i by P. gingivalis over different times of incubation was evaluated by optical density using a microplate reader. Laser radiation at λ=650nm-660nm with I =50 mW/cm2 at doses of 0, 3.0, 6.0, 9.0, and 12 J/cm2 was used for aPDT. A colony-counting method and confocal laser scanning microscopy (CLSM) were used to observe the neutralization of P. gingivalis. The fluorescent molecular probe 3'(p-hydroxyphenyl)-fluorescein and the reagent Singlet Oxygen Sensor Green were used to measure the quantities of •OH and 1O2 produced by 4i after irradiation with different light energies. RESULTS: The 4i conjugate was absorbed gradually by P. gingivalis, reaching a maximum at 30 min. A clear cytotoxic effect on P. gingivalis was observed with aPDT using 62.5 µM 4i, with colony counts dropping by a factor of 3.35 log10, indicating a sterilization rate of 99.95%. Light irradiation resulted principally in the production of • OHby 4i. A live/dead viability assay demonstrated substantial red fluorescence in P. gingivalis treated with aPDT. CONCLUSIONS: The results suggest that 4i-aPDT caused substantial cytotoxicity in P. gingivalis.

Exploring the Imbalance of Periodontitis Immune System From the Cellular to Molecular Level.

Periodontitis is a common chronic inflammatory disease of periodontal tissue, mostly concentrated in people over 30 years old. Statistics show that compared with foreign countries, the prevalence of periodontitis in China is as high as 40%, and the prevalence of periodontal disease is more than 90%, which must arouse our great attention. Diagnosis and treatment of periodontitis currently rely mainly on clinical criteria, and the exploration of the etiologic criteria is relatively lacking. We, therefore, have explored the pathogenesis of periodontitis from the perspective of immune imbalance. By predicting the fraction of 22 immune cells in periodontitis tissues and comparing them with normal tissues, we found that multiple immune cell infiltration in periodontitis tissues was inhibited and this feature can clearly distinguish periodontitis from normal tissues. Further, protein interaction network (PPI) and transcription regulation network have been constructed based on differentially expressed genes (DEGs) to explore the interaction function modules and regulation pathways. Three functional modules have been revealed and top TFs such as EGR1 and ETS1 have been shown to regulate the expression of periodontitis-related immune genes that play an important role in the formation of the immunosuppressive microenvironment. The classifier was also used to verify the reliability of periodontitis features obtained at the cellular and molecular levels. In conclusion, we have revealed the immune microenvironment and molecular characteristics of periodontitis, which will help to better understand the mechanism of periodontitis and its application in clinical diagnosis and treatment.

Influence of photodynamic therapy on the periodontitis-induced bone resorption in rat.

This study aimed to evaluate the effects of toluidine blue-mediated photodynamic therapy (TB-PDT) on the periodontitis-induced bone resorption in periodontitis in rats. Periodontal disease was induced by cotton ligature around the right second maxillary molar in 64 rats. After 4 weeks, the rats were randomly divided into four groups: sterile saline solution (control group); laser therapy (laser group); TB (100 µg/mL); TB plus laser (0.15 W/cm2) irradiation every other day for 240 s (PDT group). All rats were euthanized at 15 days postoperatively. Eight gingival tissue samples were collected from each group. The expressions of receptor activator of nuclear factor kappa-Β ligand (RANKL) and osteoprotegerin (OPG) in gingival tissue samples were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The maxillae from the rest of the rats were taken for histological examination. In the PDT group, the analysis revealed less bone loss than in the control treatment (P < 0.05). No significant difference was found among the control group, TB group, and laser group (P > 0.05). Significantly higher and lower expressions of RANKL and OPG were revealed in the PDT group than that in control group, respectively (P < 0.01). When compared with the control group, the expression of RANKL was significantly reduced by 40.0% in periodontitis in rats treated with TB-PDT for 15 days (P < 0.01). The expression of OPG was increased in the PDT group with TB-PDT for 15 days, when compared with the control group (P < 0.05). TB-PDT treatment significantly reverses the abnormal expression of RANKL and OPG in periodontitis in rats.

Antimicrobial effect of photodynamic therapy using sinoporphyrin sodium and 390-400 nm light-emitting diode on Porphyromonas gingivalis in vitro.

This study aims to investigate the effect of antimicrobial photodynamic therapy (a-PDT) using a novel combination of sinoporphyrin sodium (DVDMS) and light-emitting diode (LED) with a wavelength of 390-400 nm on Porphyromonas gingivalis in vitro. Absorption spectrum of DVDMS was determined by spectrometer for selecting suitable wavelength light source. The uptake of DVDMS by P. gingivalis was evaluated according to fluorescence intensity detected by a spectrometer. Then effects of DVDMS alone, 390-400 nm LED alone, and photodynamic therapy produced by 10, 20, 40, and 80 µg/mL DVDMS and 390-400 nm LED on the suspension of P. gingivalis were evaluated by counting the number of colony forming units (CFU) after incubation. In the experiment, the LED illumination time was 30, 60, 90, 120, 180, 240, and 360 s, respectively, and the corresponding energy density was 1, 2, 3, 4, 6, 8, and 12 J/cm2, respectively. According to the absorption spectrum of DVDMS, the 390-400-nm light emitted by the LED was selected as the light source. The fluorescence intensity of DVDMS on P. gingivalis increased significantly at 5 min, and with the extension of time, it decreased at 30 min. DVDMS alone did not produce a significant toxicity on P. gingivalis compared with PBS (p = 0.979). While 390-400 nm LED alone had a certain bactericidal effect on P. gingivalis, the bactericidal effect was more obvious as the light dose increased (p < 0.001). The effect of a-PDT produced by 20, 40, and 80 µg/mL DVDMS and 390-400 nm LED were significantly better than that of 390-400 nm LED alone (p < 0.05). Both DVDMS concentration and light dose could enchance the bactericidal effect. The strongest photo-killing effect was generated by 80 µg/mL DVDMS with 360 s illumination (energy density is 12 J/cm2), and the log reduction of bacteria was 5.69 ± 1.70. a-PDT using the combination of DVDMS with 390-400 nm LED shows promise as a new treatment modality for pathogens elimination in periodontal therapy.

Super Activated Platelet Lysate, a Novel Autologous Platelet Lysate, Regulates the Expression of Inflammasome and Cytokine in the Experimental Periodontitis in Rats.

PURPOSE: The aim of the present study was to evaluate the expression of inflammasome and cytokine on experimental periodontitis with super activated platelet lysate (SPL) in rats. METHODS: Periodontitis was induced by submerging cotton ligatures on the right side of the maxillary second molar in 36 Wistar rats. The rats were divided into 3 groups randomly: the rats received no treatment (control group); local injection with sterile saline (ligature+saline group) and local injection with SPL (ligature+SPL group). After treatments, the alveolar bone level and inflammation of periodontal tissue were evaluated by micro-computed tomography (micro-CT) scanning and histological examination, respectively. The expression of inflammasome and cytokine was evaluated by real-time quantitative polymerase chain reaction (RT-qPCR) assay. RESULTS: Compared with the control group, the bone loss significantly increased by 0.9 mm in the ligature+saline group and 0.4 mm in the ligature+SPL group (P < 0.001). 0.5 mm reduction in the bone loss was founded in the ligature+SPL group compared with the ligature+saline group (P < 0.001). The gene expression of CCL2, CXCL2, IL-6, IL-18, IL-1α, IL-1ß, CXCL10, CXCL16, CCL5 was significantly reduced in the ligature+SPL group compared with the ligature+saline group (P < 0.05). Compared with the ligature+saline group, the expression for inflammasome NLRP3, AIM2, CASP1 was both downregulated in the ligature+SPL group (P < 0.05). CONCLUSION: Our present study demonstrated local injection of SPL regulated the expression of inflammasome and cytokine and had a visible effect of relieving inflammation in the experimental periodontitis in rats.

The penetration effect of HMME-mediated low-frequency and low-intensity ultrasound against the Staphylococcus aureus bacterial biofilm.

BACKGROUND: The purpose of this study was to observe the effect of hematoporphyrin monomethyl ether (HMME)-mediated low-frequency and low-intensity ultrasound on mature and stable Staphylococcus aureus (S. aureus) biofilms under different ultrasound parameters. METHODS: The biofilm was formed after 48-h culture with stable concentration of bacterial solution. Different types of ultrasound and time were applied to the biofilm, and the ultrasonic type and time of our experiments were determined when the biofilm was not damaged. The penetration effects of low-frequency and low-intensity ultrasound were decided by the amount of HMME that penetrated into the biofilm which was determined by fluorescence spectrometry. RESULTS: The destruction of biofilms by pulse waveform was the strongest. Sinusoidal low-frequency and low-intensity ultrasound can enhance the biofilm permeability. For a period of time after the ultrasound was applied, the biofilm permeability increased, however, changes faded away over time. CONCLUSIONS: Low-frequency and low-intensity sinusoidal ultrasound significantly increased the permeability of the biofilms, which was positively correlated with the time and the intensity of ultrasound. Simultaneous action of ultrasound and HMME was the most effective way to increase the permeability of the biofilms.

Hematoporphyrin monomethyl ether mediated sonodynamic antimicrobial chemotherapy on porphyromonas gingivalis in vitro.

This study aimed to evaluate the efficacy of hematoporphyrin monomethyl ether (HMME)-mediated sonodynamic antimicrobial chemotherapy (SACT) on Porphyromonas gingivalis (P. gingivalis). P. gingivalis (ATCC 33277) was used in the present study. The bacterial suspension was randomly divided into five groups: Group 1 was incubated for 2 h in the dark with HMME in various concentrations (10, 20, 30 and 40 µg/mL). Then exposed to 1 MHz ultrasound frequency with 3 W/cm2 ultrasound intensity for 10 min. Group 2 was incubated with 40 µg/mL HMME and then irradiated with 2, 4, 6, 8 and 10 min ultrasonic time. Group 3 received different HMME concentration (10, 20, 30 and 40 µg/mL) treatment alone with no ultrasound as the HMME control group. Group 4 received ultrasound treatment alone in different ultrasonic time (2, 4, 6, 8 and 10 min) with no HMME as the ultrasound control group. Group 5 received no treatment as the no treatment control group. After the SACT, the bactericidal effect was determined by the colony forming unit assay. The intracellular content of reactive oxygen species (ROS) was detected using the laser scanning confocal microscope based on DCFH-DA. 4.7 lg reduction in CFU, When P. gingivalis was treated with ultrasound (3 W/cm2 for 10 min) at 40 µg/mL HMME concentration (P < 0.01). The intracellular ROS in SDT group had a significant difference in comparison with the no treatment control group (P < 0.01). HMME mediated SACT can be a potential antibacterial therapy to significantly inhibit P. gingivalis growth.

Hematoporphyrin monomethyl ether-mediated sonodynamic therapy induces A-253 cell apoptosis.

It has been found that >90% of oral cancer patients suffer from squamous cell carcinoma (SCC). The 5-year survival rate of SCC is ~50%, despite the availability of different treatments. Sonodynamic therapy (SDT) has been developed as a novel therapy for cancer, resisting bacterial infection and inhibiting atherosclerotic plaque progression. The present study investigated the efficacy of hematoporphyrin monomethyl ether (HMME)-mediated SDT on the A-253 epidermoid cancer cell line. The cytotoxicity of HMME and the survival rate of cells following SDT were examined by the MTT assay. Apoptosis and necrosis of cells were detected using flow cytometry with Annexin V and propidium iodide (PI) staining, and fluorescence microscopy with Hoechst 33258 and PI staining. Intracellular reactive oxygen species (ROS) and Ca2+ levels were measured using a fluorescence microscope based on 2',7'-dichlorofluorescein diacetate and fluo-3/acetoxymethylester, respectively. Results of the MTT assay demonstrated that a lower concentration (<10 µg/ml) of HMME had no significant effect on the A-253 cells, but SDT combined with ultrasonic treatment for 1 min and 10 µg/ml HMME decreased the cell survival rate by 27%. Flow cytometry analysis revealed that A-253 cells in the SDT group had a higher rate of late apoptosis compared with the control group. Furthermore, fluorescence quantitation of apoptotic A-253 cells demonstrated that the percentages of apoptotic cells were increased in the ultrasound and SDT group compared with those in the control group. In the present study, the ROS level in the SDT group was elevated compared with that in the control group. The Ca2+ levels were increased to 181.2 and 268.7% in the ultrasound and SDT groups, respectively, relative to the control group. Taken together, the findings of the present study demonstrated that HMME-SDT significantly induces the apoptosis of A-253 cells together with intracellular ROS generation and Ca2+ overload. Thus, HMME-SDT may be a promising treatment option for patients with SCC.

Research progress of sonodynamic therapy in the field of stomatology / 口腔疾病防治

@#Sonodynamic therapy (SDT) is a cutting-edge method for the biological effects of ultrasound combined with sound-sensitive agents. In recent years, SDT has been a concern of experts and scholars in the oral field, and a series of experimental studies has been carried out. We will introduce the progress of SDT in the field of stomatology from three aspects: the therapeutic mechanism of SDT, the application of SDT in the oral field, and the current situation and future of SDT in the treatment of oral diseases. It is currently believed that singlet oxygen theory, cavitation effects, and induction of apoptosis are the main therapeutic mechanisms. The research of SDT on oral disease prevention mainly focuses on oral tumors (especially squamous cell carcinoma and osteosarcoma) and infectious diseases (such as periodontitis, maxillofacial infection, and oral mucosal disease). Although the current research is still in the experimental stage, with the continuous exploration of sonosensitizers and nanotechnology, SDT will provide great help for the clinical prevention and treatment of oral diseases in the future.

Epidermal growth factor receptor signaling suppresses αvß6 integrin and promotes periodontal inflammation and bone loss.

In periodontal disease (PD), bacterial biofilms cause gingival inflammation, leading to bone loss. In healthy individuals, αvß6 integrin in junctional epithelium maintains anti-inflammatory transforming growth factor-ß1 (TGF-ß1) signaling, whereas its expression is lost in individuals with PD. Bacterial biofilms suppress ß6 integrin expression in cultured gingival epithelial cells (GECs) by attenuating TGF-ß1 signaling, leading to an enhanced pro-inflammatory response. In the present study, we show that GEC exposure to biofilms induced activation of mitogen-activated protein kinases and epidermal growth factor receptor (EGFR). Inhibition of EGFR and ERK stunted both the biofilm-induced ITGB6 suppression and IL1B stimulation. Furthermore, biofilm induced the expression of endogenous EGFR ligands that suppressed ITGB6 and stimulated IL1B expression, indicating that the effects of the biofilm were mediated by autocrine EGFR signaling. Biofilm and EGFR ligands induced inhibitory phosphorylation of the TGF-ß1 signaling mediator Smad3 at S208. Overexpression of a phosphorylation-defective mutant of Smad3 (S208A) reduced the ß6 integrin suppression. Furthermore, inhibition of EGFR signaling significantly reduced bone loss and inflammation in an experimental PD model. Thus, EGFR inhibition may provide a target for clinical therapies to prevent inflammation and bone loss in PD.

Effects of palmatine hydrochloride mediated photodynamic therapy on oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a common malignant tumor, accounting for about 7% of all malignant tumors. Palmatine hydrochloride (PaH) is the alkaloid constituent of Fibraurea tinctoria Lour. The present study aims to investigate the antitumor effect of photodynamic therapy (PDT) with PaH (PaH-PDT) on human OSCC cell lines both in vitro and in vivo. The results indicate that PaH-PDT exhibited a potent phototoxic effect in cell proliferation and produced cell apoptosis. PaH-PDT increased the percentage of cells in the G0/G1 phase and decreased the CDK2 and Cyclin E1 protein level. In addition, PaH-PDT markedly increased the generation of intracellular ROS, which can be suppressed using the ROS scavenger N-acetylcysteine (NAC). Furthermore, PaH-PDT increased the expression of p53 protein in vitro and in vivo. In vivo experiments revealed that the PaH-PDT resulted in an effective inhibition of tumor growth and prolonged the survival time of tumor-bearing mice. Moreover, no obvious signs of side effects or a drop in body weight was observed. These results suggested that PaH was a promising sensitizer that can be combined with light to produce significant anti-tumor effects in oral squamous cell carcinoma via enhanced ROS production and up-regulated expression of p53.

MEHP promotes the proliferation of oral cancer cells via down regulation of miR-27b-5p and miR-372-5p.

Mono-2-ethyhexyl phthalate (MEHP), an environmental xenoestrogen, is widely used in the production of polyvinyl chloride materials and can be easily accumulated into human body. Emerging evidences showed that MEHP can regulate the progression of various cancers. Oral cancer cells could be directly exposed to MEHP during food and water digestion, while the roles of MEHP on the progression of oral cancer were rarely investigated. Our present study found that MEHP can trigger the proliferation of oral squamous cell carcinoma (OSCC) cells and increase the expression of proliferating cell nuclear antigen (PCNA). We checked the expression of various miRNAs which can target the 3'UTR of PCNA. Specifically, MEHP can decrease the expression of miR-27b-5p and miR-372-5p, which can directly bind with the 3'UTR of PCNA to inhibit its expression. Over expression of miR-27b-5p and miR-372-5p can abolish MEHP induced cell proliferation and expression of PCNA in OSCC cells. Further, MEHP can induce the expression of c-Myc, which can suppress the transcription of miR-27b-5p in OSCC cells. In vivo xenograft study on the basis of SCC-4 cells confirmed that MEHP can trigger the growth of OSCC and suppress the expression of miR-27b-5p and miR-372-5p. Collectively, our present study suggested that MEHP can promote the growth and progression of OSCC via down regulation of miR-27b-5p and miR-372-5p.

Inflammasome and cytokine expression profiling in experimental periodontitis in the integrin ß6 null mouse.

Epithelial αvß6 integrin participates in immune surveillance in many organs, including the gastrointestinal track. Expression of αvß6 integrin is reduced in the junctional epithelium of the gingiva in periodontal diseases, and mutations in the ITGB6 gene are associated with these diseases in humans and mice. The aim of this study was to unravel potential differences in the inflammatory responses in the periodontal tissues of FVB wild-type (WT) and ß6 integrin-null (Itgb6-/-) mice, using a ligature-induced periodontitis model and assessing inflammation, bone loss and expression profiles of 34 genes associated with periodontal disease. Using micro-CT and histology, we demonstrated more advanced inflammation and bone loss in the control and ligatured Itgb6-/- mice compared to the WT animals. Neutrophil and macrophage marker genes were significantly upregulated by ligation in both WT and Itgb6-/- mice while the expression of T-cell and B-cell markers was downregulated, suggesting acute-type of inflammation. Expression of inflammasome NLRP3-related genes Nlpr3 and Il1b was also significantly increased in both groups. However, the expression of Il18 was significantly lower in non-ligatured Itgb6-/- mice than in the WT mice and was further downregulated in both groups by the ligatures. IL-18 mediates many effects of the AIM2 inflammasome, including regulation of the microbiome. Interestingly, expression of Aim2 was significantly lower in both control and ligatured Itgb6-/- mice than in WT animals. Overall, ligature-induced periodontitis was associated with increased expression of pro-inflammatory cytokines, chemokines and osteoclastogenic regulatory molecules. Another significant difference between the Itgb6-/- and WT mice was that mRNA expression of the anti-inflammatory cytokine IL-10 was increased in ligatured WT mice but reduced in the Itgb6-/- mice. In conclusion, αvß6 integrin in junctional epithelium of the gingiva appears to positively regulate the expression of the AIM2 inflammasome and anti-inflammatory IL-10, thus providing protection against periodontal inflammation.