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The early inflammatory phase, which is impacted by both local and systemic reactions to noxious stimuli, is crucial for bone regeneration. One component of Garcinia mangostana, called αmangostin, offers several advantages, including acting as an anti-inflammatory. With the production of related transcription factor-2 (RUNX2), which in turn causes the appearance of various osteogenesis markers, the WNT signaling pathway is a mechanism for the formation of osteogenesis through inflammation. To analyze the effect of α-mangostin on the LPS-induced inflammatory process on osteocalcin and interleukin-17A gene expression in osteoblast cell line culture. This study examines the expression of the genes for IL-17A and OCN in osteoblast cell culture using a post-test-only control group design. A total of 4 groups of cell cultures were created: group K with osteogenic medium, group P1 with osteogenic medium and lipopolysaccharide, group P2 with osteogenic medium and α-mangostin, and group P3 with osteogenic medium and a combination of lipopolysaccharide and α-mangostin. Using the same concentration for all concentrates, the therapy was administered over three days in osteogenic media. The sample will subsequently be prepared and subjected to RT-PCR analysis. Result is significant upregulation of IL-17A and OCN gene expression was observed in the group combination of lipopolysaccharide and α-mangostin. In conclusion α-mangostin can boost osteocalcin gene expression in osteoblast cell line culture after LPS induction, however, it was unable to diminish IL-17A gene expression in osteoblast cell line culture. [ABSTRACT FROM AUTHOR]

Reconstruction of maxillofacial defect still remains a challenge for oral and maxillofacial surgeons. Autograft, which has been the gold standard, has limitations and donor site morbidity. Tissue engineering combining cells, scaffold, and growth factors is expected to produce ideal bone without morbidity, with Human umbilical cord mesenchymal stem cells (HUCMSC) and bovine scaffold are promising topics. HUCMSC therapy is influenced by biomolecules including cytokines, growth factors, and microRNAs, known as secretome. This study aims to analyze the osteoinduction potential of the HUCMSC secretome in the FDBBX scaffold through the expression of Runt Related Transcription Factor-2 (RUNX2) and Osteocalcin (OCN) markers. Objectives to analyze the increased expression of RUNX2 and OCN of MC3T3-E1 preosteoblast cells that were seeded into FDBBX scaffold after application of the HUCMSC secretome compared to scaffold without secretome. The study was an in vitro laboratory experiment. The first group was FDBBX scaffold after HUCMSC secretome application and the second group was FDBBX without secretome. The analysis of the RUNX2 and OCN expression of MC3T3-E1 pre-osteoblast cells that attached to the scaffold was conducted on days 7, 14 and 21 using RT-PCR method. Independent T test was conducted as a Statistical test to analyze the differences between groups, and One way ANOVA test was conducted to analyze the difference between time. The difference analysis was considered significant when p<0,05. The Independent T test result showed that the expression of RUNX2 and OCN in FDBBX scaffold after HUCMSC secretome application was significantly higher than scaffold without secretome (P=0,039; P=0,025). The expression of RUNX2 and OCN of MC3T3-E1 pre-osteoblast cells in the FDBBX scaffold after HUCMSC secretome application was higher than the scaffold without secretome application. [ABSTRACT FROM AUTHOR]

Bone regeneration in dentistry happens in the jawbone after simple tooth extraction or bone fracture. The bone regeneration process consists of a continuous and directed phase and takes place in three phases, which are inflammation, proliferation, and remodelling. a-Mangostin has been known to have potential anti-inflammatory effects and is most likely a potential therapeutic agent to inhibit bone resorption caused by the inflammatory process. Objectives to investigate the effect of a-mangostin exposure on osteoblast cell migration and viability during inflammatory induction. In vitro laboratory experimental study on 7F2 cell line culture. Cell cultures were divided into 5 groups, those were the cell culture group with osteogenic media, with lipopolysaccharide, with a-mangostin, a combination of lipopolysaccharide and a-mangostin, and a combination of lipopolysaccharide and a-mangostin after 24 hours. The sample will undergo 5 replicates of each of the MTT and scratch wound assays. The One-Way Anova test is used to assess the data results, and a significance level of 0.05 is required. There was a significant difference in viability and migration between the experimental groups (p=0.000). There was a correlation between viability and migration of osteoblast cell postinflammation (p=0.000). Exposure to a-mangostin had an impact on the migration and viability of osteoblast cells after inflammation. [ABSTRACT FROM AUTHOR]

Freeze-Dried Bovine Bone Scaffold (FDBB) and Decellularized Freeze-Dried Bovine Bone (DCFDBB) are promising new alternative of xenograft material in bone tissue engineering. However, their biocompatibility is still unknown. To investigate whether FDBB and DC-FDBB scaffolds are biocompatible, able to induce cell proliferation attachment in vitro. Human umbilical cord mesenchymal stem cells (hUC-MSCs) culture was exposed to FDBB, DC-FDBB, Deproteinized Bovine Bone Matrix (DBBM) scaffold (as positive control) conditioned medium for 24, 48 and 72 hours (n=8). MTT assay was then performed to measure the number of viable cells at each observation time. Normality (Shapiro-Wilk) and homogeneity (Levene) test were done on the results of the MTT assay, then ANOVA test was performed. To support the finding, further SEM observation was performed. hUC-MSCs were seeded on the surface of each scaffold type (n=3) and incubated for 24, 48 and 72 hours, then SEM observation was done on the scaffold surface to analyse their surface cell attachment. The mean percentage of living cells in FDBB group was the highest among all groups. There were significant differences (p<0.05) in each treatment group at each observation time, except between FDBB and DC-FDBB (48 hours), DC-FDBB and DBBM (48 hours), FDBB and DC-FDBB (72 hours) (p>0.05). SEM examination showed the same results, as the highest number of cell colonization was found on FDBB group at each observation time. FDBB and DC-FDBB scaffolds have good biocompatibility characteristics that can be used as a bone substitute in bone tissue engineering. [ABSTRACT FROM AUTHOR]

Defects in the mandible cause deformities, functional disturbances and facial esthetics that cause a decrease in a person's quality of life. Autograft is the gold standard because it has osteogenic, osteoinductive, osteoconductive properties and does not cause an immune response. Allograft as an alternative to autograft is the use of scaffold. The common bovine scaffolds that have been widely studied are DBBM (Deproteinized Bovine Bone Material), FDBB (Freeze-dried bovine bone xenograft) and dc-FDBB (Decellularized freeze-dried bovine bone xenograft). Permeability is a parameter that quantitatively measures the ability of a porous medium to flow liquids and depends on a combination of porosity, pore size, pore distribution and tortuosity. High viscosity can result in higher permeability. The porosity of decellularized cartilage bovine scaffold was significantly increased compared to fresh bovine cartilage. Objectives to find out whether the level of permeability of dc-FDBB qualifies as a bone tissue engineering scaffold. Scaffold FDBB, dc-FDBB, DBBM were divided into 2 groups, namely the group with distilled water and the group with 30% glycerol media. The permeability test was carried out in each group by measuring the liquid discharge every minute for 10 minutes and then calculating the permeability value. Then the permeability comparison between scaffolds is carried out. The maximum volume of fluid flow per minute was found on the DBBM scaffold, both in distilled water and 30% glycerol media. In the group with distilled water and 30% glycerol group, the highest permeability values were obtained in the DBBM scaffold test group, and the lowest in the dc-FDBB test group. The permeability of the dc-FDBB scaffold in this study shows a value that falls within the range of human cancellous bone permeability values so that it can be concluded that the dc-FDBB scaffold in this study meets the scaffold permeability standards for tissue engineering. [ABSTRACT FROM AUTHOR]

Deproteinized Bovine Bone Mineral (DBBM) has been widely used as tissue engineering scaffold, because of its not antigenic potential, although non-resorbable. Freeze-Dried Bovine Bone (FDBB) scaffold was developed to overcome the limitations of DBBM but it's still suspected to have antigenic potential. Therefore, to overcome the issues of antigenic potential, decellularization of FDBB was carried out (dc-FDBB). Objective: To analyze antigenic potential of FDBB scaffold with process decellularization. FDBB and dc-FDBB scaffolds compared to DBBM scaffold as gold standard. Ten samples for each group of scaffold FDBB, dc-FDBB, and DBBM. The groups were observed for residual osteoblasts and osteocytes with HE staining using a light microscope with 40 x and 400 x magnification. DNA concentration analysis was carried out on the three types of scaffolds using a spectrophotometer. Statistical analysis was performed using IBM SPSS version 25. Data were statistically analyzed using ANOVA and Post Hoc Games-Howell tests. There were degradation of osteocytes without nucleus and satisfactory condition of extracellular matrix geometrical structure in the FDBB scaffold. There were no osteoblasts and osteocytes with damaged extracellular matrix geometrical structures were seen in the dc-FDBB scaffold. There were no osteoblasts and osteocytes with satisfactory condition of extracellular matrix geometrical structure in the DBBM scaffold. Mean values of DNA concentration: FDBB, dc-FDBB, and DBBM were of (19.75 ng/µL ± 4.80), (16.84 ng/µL ± 6.55), and (8.72 ng/µL ± 0.65) respectively. Comparative p value FDBB and dc-FDBB scaffolds were 0,509 (p>0,05), there were no statistically significant difference between FDBB and dc-FDBB scaffolds. FDBB and dc-FDBB scaffolds can be used because it has low antigenic potential. [ABSTRACT FROM AUTHOR]

To improve the osteoconductivity of apatite cement (AC) for reconstruction of bone defects after oral maxillofacial surgery, we previously fabricated AC containing atelocollagen (AC(ate)). In the present study, we examined the initial attachment, proliferation and differentiation of mouse osteoblastic cells (MC3T3-E1 cells) on the surface of conventional AC (c-AC), AC(ate) and a plastic cell dish. The number of osteoblastic cells showing initial attachment to AC(ate) was greater than those attached to c-AC and similar to the number attached to the plastic cell wells. We also found that osteoblastic cells were well spread and increased their number on AC(ate) in comparison with c-AC and the wells without specimens, while the amount of procollagen type I carboxy-terminal peptide (PIPC) produced in osteoblastic cells after three days on AC(ate) was greater as compared to the others. There was no significant difference in regard to alkaline phosphatase (ALP) activity and osteocalcin production by osteoblastic cells among the three surface types after three and six days. However, after 12 days, ALP activity and the produced osteocalcin were greater with AC(ate). In conclusion, AC(ate) may be a useful material with high osteoconductivity for reconstruction of bone defects after oral maxillofacial surgery. [ABSTRACT FROM AUTHOR]

Background: CD44 and aldehyde dehydrogenase 1 (ALDH1) have been shown to be useful markers for identification of cancer stem cells (CSCs). We previously reported that glycogen synthase kinase 3β (GSK3β) is involved in regulation of the self-renewal ability of head and neck squamous cell carcinoma (HNSCC) CSCs. The purpose of the present study was to clarify the role of GSK3β in CD44(high) /ALDH1(high) HNSCC cells. Methods: Cells with greater expression of CD44 and higher ALDH1 enzymatic activity were FACS sorted from the OM-1 HNSCC cell line. The self-renewal ability of CD44(high) /ALDH1(high) cells was then examined using a tumor sphere formation assay. mRNA expressions of the stem cell markers Sox2, Oct4, and Nanog, as well as GSK3β were evaluated by real-time RT-PCR. Results: CD44(high) /ALDH1(high) cells exhibited higher tumor sphere forming ability and increased expression of stem cell markers as compared with CD44(high) /ALDH1(low) cells. Interestingly, spindle-shaped cells positive for vimentin were found in the CD44(high) /ALDH1(high) but not the CD44(high) /ALDH1(low) cell population. In addition, the ALDH1 activity and sphere forming ability of CD44(high) /ALDH1(high) cells was significantly inhibited by GSK3β knockdown. On the other hand, CD44(high) /ALDH1(low) cells exhibited high epidermal growth factor receptor (EGFR) expression and increased cell growth. Conclusions: Our results show that GSK3β plays a major role in maintenance of stemness of CD44(high) /ALDH1(high) HNSCC cells. Additionally, they indicate a close relationship between CSC and mesenchymal characteristics in HNSCC. [ABSTRACT FROM AUTHOR]

The present study reviewed the records of 58 patients who underwent orthognathic surgery [sagittal split ramus osteotomy (SSRO), Le Fort I osteotomy, genioplasty, anterior maxillary alveolar osteotomy] between 2010 and 2015. To investigate the influence of preoperative oral health care on postoperative inflammation, infection and length of hospital stay in those patients, white blood cell (WBC) count and C-reactive protein (CRP) levels were compared between patients who received and did not receive preoperative oral care. The mean CRP level throughout the postoperative term was lower in the oral care group as compared to the non-oral care group. By contrast, the oral care group had a lower occurrence of postoperative infectious complications (surgical site infection, anastomotic leak) (13.6 vs. 20.8%) and a shorter average length of hospital stay (16.2±3.8 vs. 21.2±7.4 days). These results suggest that preoperative professional oral health care decreases the duration of hospital stay following orthognathic surgery by inhibiting inflammation and infectious complications during the postoperative stage. [ABSTRACT FROM AUTHOR]

Background: Loss of permanent teeth after tooth extraction without replacement of missing teeth can result in impaired masticatory, esthetic, phonetic functions, and impaired balance of the masticatory organ in the mouth. Therefore, a method is needed to inhibit the alveolar bone resorption process so that the dimensions of the tooth socket can be maintained vertically or horizontally until the time of implant placement, which is called the socket preservation procedure. α-mangostin is known to have a potential anti-inflammatory effect and most likely can be used as a potential therapeutic agent to inhibit bone resorption caused by posttooth extraction inflammatory processes., Aims: The aim of the study was to determine the effect on the inflammatory process and osteogenesis on osteoblast cell line culture by induction with lipopolysaccharide (LPS) and α-mangostin., Materials and Methods: This was an in vitro laboratory experimental study on mouse osteoblast cell line culture. The treatment was given with LPS, α-mangostin, and combination on osteogenic medium, using the same concentration for all concentrates. The sample will then be processed and analyzed using the real-time polymerase chain reaction., Results: The highest interleukin-11 (IL-11) gene expression was found in α-mangostin treatment, but there was no significant difference in IL-11 expression between the study groups. The highest runt-related transcription factor-2 (RUNX-2) gene expression was found in a group that received induction with LPS and α-mangostin, and from these results, it was found that there was a significant difference in RUNX-2 expression between the study groups., Conclusions: LPS and α-mangostin can increase osteogenesis in osteoblast cell culture in the osteogenic medium., Competing Interests: There are no conflicts of interest., (Copyright: © 2023 Contemporary Clinical Dentistry.)

Our previous study revealed that treatment with a combination of fibroblast growth factor‑2 and melatonin (MEL) synergistically augmented osteogenic activity and mineralization of MC3T3‑E1 mouse preosteoblast cells. Thus, the objective of the present study was to assess the effect of MEL on osteogenetic characteristics in human osteoblastic cells. Human jawbone‑derived osteoblastic (hOB) cells were isolated from mandibular bone fragments. RUNX family transcription factor 2 (Runx2) expression, alkaline phosphatase (ALP) enzyme activity and the mineralization ability of hOB cells in the presence of MEL were evaluated. Microarray analysis was also performed to assess the expression of MEL‑induced microRNAs (miRNAs/miRs) in hOB cells. Treatment with MEL significantly enhanced Runx2 expression, ALP activity and mineralization staining. However, this effect was significantly reduced following transforming growth factor‑β1 treatment. In total, 124 miRNAs were differentially expressed in MEL‑treated hOB cells, compared with untreated cells. Of the upregulated miRNAs, miR‑181c‑5p exhibited the largest fold change. Runx2 mRNA expression and mineralization staining in the presence of MEL were significantly reduced following transfection with a miR‑181c‑5p inhibitor. In addition, transfection with miR-181c-5p mimics significantly increased Runx2 expression and mineralization staining. These results suggested that MEL‑induced miR‑181c‑5p was involved in osteogenic differentiation and mineralization of hOB cells. Using TargetScan, a putative miR‑181c‑5p binding site was identified in the Notch2 gene. Moreover, Notch2 mRNA and protein expression levels in hOB cells were significantly reduced following transfection with miR‑181c‑5p mimics, confirming Notch2 as a target gene for miR‑181c‑5p. Notch2 siRNA knockdown significantly increased Runx2 expression and mineralization staining, which suggested that Notch2 may negatively regulate osteogenic differentiation of hOB cells by downregulating Runx2. In conclusion, MEL‑induced expression of miR‑181c‑5p enhanced osteogenic differentiation and calcification of hOB cells.

We previously found that CD44 high /ESA low head and neck squamous cell carcinoma (HNSCC) cells harboring high dihydropyrimidine dehydrogenase (DPD) expression exhibited potent resistance to 5-fluorouracil (5-FU)-induced apoptosis. In addition, susceptibility of HNSCC cells to 5-FU was compromised in the presence of cyclooxygenase 2 (COX2)-derived prostaglandin E 2 (PGE 2 ). In this study, we examined 5-FU-induced apoptosis in sorted cell populations (i.e., CD44 high /ESA low , CD44 high /ESA high , and CD44 low cells from the HNSCC cell line A-253) to clarify the anti-apoptotic effect of PGE 2 on CD44 high cells. Notably, CD44 high /ESA low cells upregulated PGE 2 , compared with other populations. To investigate the effect of CD44 high /ESA low cell-derived PGE 2 on CD44 high /ESA high cells, direct and indirect co-culture assays were performed. The percentage of apoptotic cells in a culture of CD44 high /ESA high cells was significantly reduced when they were directly and indirectly co-cultured with CD44 high /ESA low cells. Furthermore, 5-FU-induced apoptosis of CD44 high /ESA high cells was significantly increased in the presence of an inhibitor of the PGE 2 receptors (EP1/EP2) when CD44 high /ESA high cells were co-cultured with CD44 high /ESA low cells. These results suggest that CD44 high /ESA low cell-derived PGE 2 may contribute to the inhibition of 5-FU-induced apoptosis in CD44 high /ESA high cells. Additionally, NR4A2 knockdown enhances 5-FU-induced apoptosis in CD44 high /ESA high cells, suggesting that PGE 2 attenuates 5-FU-induced apoptosis in an NR4A2-dependent manner in CD44 high /ESA high cells. In conclusion, CD44 high /ESA low cells contribute to induction of resistance to 5-FU in CD44 high /ESA high cells through provision of PGE 2 . CD44 high /ESA low cell-targeted therapy may be effective in treatment of HNSCC., Competing Interests: None., (IJCEP Copyright © 2018.)

To improve the osteoconductivity of apatite cement (AC) for reconstruction of bone defects after oral maxillofacial surgery, we previously fabricated AC containing atelocollagen (AC(ate)). In the present study, we examined the initial attachment, proliferation and differentiation of mouse osteoblastic cells (MC3T3-E1 cells) on the surface of conventional AC (c-AC), AC(ate) and a plastic cell dish. The number of osteoblastic cells showing initial attachment to AC(ate) was greater than those attached to c-AC and similar to the number attached to the plastic cell wells. We also found that osteoblastic cells were well spread and increased their number on AC(ate) in comparison with c-AC and the wells without specimens, while the amount of procollagen type I carboxy-terminal peptide (PIPC) produced in osteoblastic cells after three days on AC(ate) was greater as compared to the others. There was no significant difference in regard to alkaline phosphatase (ALP) activity and osteocalcin production by osteoblastic cells among the three surface types after three and six days. However, after 12 days, ALP activity and the produced osteocalcin were greater with AC(ate). In conclusion, AC(ate) may be a useful material with high osteoconductivity for reconstruction of bone defects after oral maxillofacial surgery.

The present study reviewed the records of 58 patients who underwent orthognathic surgery [sagittal split ramus osteotomy (SSRO), Le Fort I osteotomy, genioplasty, anterior maxillary alveolar osteotomy] between 2010 and 2015. To investigate the influence of preoperative oral health care on postoperative inflammation, infection and length of hospital stay in those patients, white blood cell (WBC) count and C-reactive protein (CRP) levels were compared between patients who received and did not receive preoperative oral care. The mean CRP level throughout the postoperative term was lower in the oral care group as compared to the non-oral care group. By contrast, the oral care group had a lower occurrence of postoperative infectious complications (surgical site infection, anastomotic leak) (13.6 vs. 20.8%) and a shorter average length of hospital stay (16.2±3.8 vs. 21.2±7.4 days). These results suggest that preoperative professional oral health care decreases the duration of hospital stay following orthognathic surgery by inhibiting inflammation and infectious complications during the postoperative stage.

Malignant salivary gland tumors are rare and exhibit a broad spectrum of phenotypic heterogeneity. The objective of this study was to investigate prognostic factors in patients with salivary gland carcinomas and review the results in light of other reports. We retrospectively reviewed 40 patients with primary salivary gland carcinomas who were diagnosed and treated at our institution between 1991 and 2014. Of the 40 tumors, 19 (47.5%) were mucoepidermoid carcinomas, 11 (27.5%) were adenoid cystic carcinomas, 7 (17.5%) were acinic cell carcinomas, 2 (5.0%) were myoepithelial carcinomas and 1 (2.5%) was a squamous cell carcinoma. Clinically positive lymph nodes were present in 4 patients (10.0%). As regards clinical stage, 15 cases (37.5%) were stage I, 13 (32.5%) were stage II, 1 (2.5%) was stage III and 11 (27.5%) were stage IVA. The majority of the patients (97.5%) were treated with surgery, of whom 25 (62.5%) received surgery alone and 14 (35.0%) underwent surgery in combination with chemotherapy or chemotherapy and radiotherapy. The median follow-up time for all the patients was 48 months. The disease-specific survival rate at 5 years was 87.1%. We identified a significant correlation between poor survival rate and histological grade (intermediate/high), tumor size (T3/T4), lymph node metastasis (node-positive) and clinical stage (III/IV) using the Kaplan-Meier method (P<0.05 for each). In addition, the Cox proportional hazards regression analysis confirmed that lymph node metastasis and tumor size were independent prognostic factors for disease-specific survival (hazard ratio = 18.7 and 15.1, respectively; P=0.023 and 0.037, respectively). Furthermore, tumor size was found to be a predictive factor regarding recurrence in the multivariate logistic regression analysis (odds ratio = 8.35; P=0.025). Our results suggest that lymph node metastasis and tumor size are significant prognostic factors for patients with salivary gland carcinomas.