Effect of the secretome of mesenchymal stem cells overexpressing BMP-9 on osteoblast differentiation and bone repair.

The secretome present in the conditioned medium (CM) of mesenchymal stem cells (MSCs) is a promising tool to be used in therapies to promote bone regeneration. Considering the high osteogenic potential of the bone morphogenetic protein 9 (BMP-9), we hypothesized that the secretome of MSCs overexpressing BMP-9 (MSCsBMP-9 ) enhances the osteoblast differentiation of MSCs and the bone formation in calvarial defects. CM of either MSCsBMP-9 (CM-MSCsBMP-9 ) or MSCs without BMP-9 overexpression (CM-MSCsVPR ) were obtained at different periods. As the CM-MSCsBMP-9 generated after 1 h presented the highest BMP-9 concentration, CM-MSCsBMP-9 and CM-MSCsVPR were collected at this time point and used to culture MSCs and to be injected into mouse calvarial defects. The CM-MSCsBMP-9 enhanced the osteoblast differentiation of MSC by upregulating RUNX2, alkaline phosphatase (ALP) and osteopontin protein expression, and ALP activity, compared with CM-MSCsVPR . The CM-MSCsBMP-9 also enhanced the bone repair of mouse calvarial defects, increasing bone volume, bone volume/total volume, bone surface, and trabecular number compared with untreated defects and defects treated with CM-MSCsVPR or even with MSCsBMP-9 themselves. In conclusion, the potential of the MSCBMP-9 -secretome to induce osteoblast differentiation and bone formation shed lights on novel cell-free-based therapies to promote bone regeneration of challenging defects.

Effect of Mesenchymal Stem Cells Overexpressing BMP-9 Primed with Hypoxia on BMP Targets, Osteoblast Differentiation and Bone Repair.

Bone formation is driven by many signaling molecules including bone morphogenetic protein 9 (BMP-9) and hypoxia-inducible factor 1-alpha (HIF-1α). We demonstrated that cell therapy using mesenchymal stem cells (MSCs) overexpressing BMP-9 (MSCs+BMP-9) enhances bone formation in calvarial defects. Here, the effect of hypoxia on BMP components and targets of MSCs+BMP-9 and of these hypoxia-primed cells on osteoblast differentiation and bone repair was evaluated. Hypoxia was induced with cobalt chloride (CoCl2) in MSCs+BMP-9, and the expression of BMP components and targets was evaluated. The paracrine effects of hypoxia-primed MSCs+BMP-9 on cell viability and migration and osteoblast differentiation were evaluated using conditioned medium. The bone formation induced by hypoxia-primed MSCs+BMP-9 directly injected into rat calvarial defects was also evaluated. The results demonstrated that hypoxia regulated BMP components and targets without affecting BMP-9 amount and that the conditioned medium generated under hypoxia favored cell migration and osteoblast differentiation. Hypoxia-primed MSCs+BMP-9 did not increase bone repair compared with control MSCs+BMP-9. Thus, despite the lack of effect of hypoxia on bone formation, the enhancement of cell migration and osteoblast differentiation opens windows for further investigations on approaches to modulate the BMP-9-HIF-1α circuit in the context of cell-based therapies to induce bone regeneration.

Genetic polymorphisms are associated with alterations in anxiety levels and vital signs in patients undergoing third molar extractions.

OBJECTIVE: The objective of this study was to evaluate the association between SLC6A4 (rs1042173 and rs3813034), DRD2 (rs6275 and rs6276), ANKK1 (rs1800497), and COMT (rs174675) genetic polymorphisms and alterations in anxiety levels and vital signs in individuals undergoing third molar extractions. STUDY DESIGN: One hundred sixty-eight individuals were evaluated at the pre-, trans-, and postoperative periods by checking systolic blood pressure, diastolic blood pressure, heart rate, respiratory rate, and body temperature. Anxiety levels were assessed using the State-Trait Anxiety Inventory (STAI). Buccal mucosa cells were collected for genetic evaluation using real-time polymerase chain reaction. Statistical analysis was performed at a significance level of 5%. RESULTS: The level of anxiety was associated with rs1800497 for STAI-Trait (P = .031) and rs174675 for STAI-State (P = .007). Considering the vital signs, there was a significant difference between the values of respiratory rate and rs1042173 (P = .029), rs3813034 (P = .024), and rs6275 (P = .025). The diastolic blood pressure values differed significantly for rs1042173 (P = .042), and the body temperature values differed significantly for rs174675 (P = .016). CONCLUSIONS: Polymorphisms in SLC6A4, DRD2, ANKK1, and COMT genes could be associated with alterations in anxiety levels and vital signs in individuals undergoing third molar extractions.

Single Nucleotide Polymorphisms in Runt-related Transcription Factor 2 and Bone Morphogenetic Protein 2 Impact on Their Maxillary and Mandibular Gene Expression in Different Craniofacial Patterns - A Comparative Study.

Introduction: This study aimed to evaluate if single nucleotide polymorphisms (SNPs) in runt-related transcription factor 2 (RUNX2) and bone morphogenetic protein 2 (BMP2) are associated with different craniofacial patterns. Furthermore, we also investigated if RUNX2 and BMP2 expression in the maxilla and mandible are differently expressed according to facial phenotypes and influenced by the SNPs in their encoding genes. Orthognathic patients were included. Materials and Methods: Lateral cephalometric radiographs were used to classify facial phenotypes based on Steiner's ANB and Ricketts' NBa-PtGn angles. Bone samples from 21 patients collected during orthognathic surgery were used for the gene expression assays. DNA from 129 patients was used for genotyping the SNPs rs59983488 and rs1200425 in RUNX2 and rs235768 and rs1005464 in BMP2. The established alpha was 5%. Results: A statistically significant difference was observed in the relative BMP2 expression in the mandible between Class I and III participants (P = 0.042). Homozygous GG (rs59983488) had higher RUNX2 expression (P = 0.036) in the mandible. In maxilla, GG (rs1200425) had a higher BMP2 expression (P = 0.038). Discussion: In conclusion, BMP2 is expressed differently in the mandible of Class I and Class III participants. Genetic polymorphisms in RUNX2 and BMP2 are associated with their relative gene expression.

Effect of different factors on patient perception of surgical discomfort in third molar surgery.

The aim of this study was to evaluate patient perception of surgical discomfort in third molar surgery and the association with clinical variables and polymorphisms associated with the FKBP5, SLC6A4, and COMT genes. This cross-sectional observational study was carried out on 196 participants aged between 18 and 64 years at the Federal University of Paraná in 11 months. The intensity of surgical discomfort was assessed using the QCirDental questionnaire. Data on surgical and individual procedures were also cataloged. The oral health related quality of life was assessed by the Oral Health Impact Profile questionnaire (OHIP-14). The DNA sample was obtained from cells of the oral mucosa. Five markers of the FKBP5, SLC6A4, and COMT genes were genotyped. The data were submitted to statistical analysis with a significance level of 5%. Women reported greater intensity of discomfort associated with third molar surgery compared to men (p = 0.001). In the recessive model, the AA genotype of the rs3800373 marker was associated with greater surgical discomfort (p = 0.026). Therefore, women and individuals of the AA genotype for the rs3800373 marker in the FKBP5 gene reported greater surgical discomfort associated with third molar surgery.

Association between gender, estrogen receptors genes and anxiety levels in patients undergoing orthognathic surgery.

Orthognathic surgery is a procedure that is performed for the correction of dentofacial deformities and can lead to a change in an individual's anxiety levels. Anxiety is a multifactorial condition in which hormones and genes play an important role. This study aimed to evaluate if gender and genetic polymorphisms in estrogen receptor alpha (ESR1) and beta (ESR2) are associated with anxiety levels in patients undergoing orthognathic surgery. In this longitudinal observational study, 44 patients were included. Anxiety level assessments were performed at three time periods: 2 days before the surgical procedure and 1 and 6 months postoperatively, using the State-Trait Anxiety Inventory Scale. Gender, age, and facial profile were also evaluated. Additionally, a saliva sample from each individual was collected for the genotypic evaluation of ESR1 (rs2234693 and rs9340799) and ESR2 (rs1256049 and rs4986938) using real time polymerase chain reaction. Data were analyzed with a significance level of 0.05. There was a decrease in trait-anxiety and state-anxiety when comparing the preoperative measurements with those obtained 1 and 6 months postoperatively (p < 0.05). Females were more anxious than males at each time point during the study (p < 0.05). The genetic polymorphism rs9340799 in ESR1 was associated with state-anxiety during the preoperative period (p = 0.046). In conclusion, an individual's gender and genetic polymorphism in ESR1 are associated with anxiety in orthognathic surgery patients.

Effect of different factors on patient perception of surgical discomfort in third molar surgery

Abstract: The aim of this study was to evaluate patient perception of surgical discomfort in third molar surgery and the association with clinical variables and polymorphisms associated with the FKBP5, SLC6A4, and COMT genes. This cross-sectional observational study was carried out on 196 participants aged between 18 and 64 years at the Federal University of Paraná in 11 months. The intensity of surgical discomfort was assessed using the QCirDental questionnaire. Data on surgical and individual procedures were also cataloged. The oral health related quality of life was assessed by the Oral Health Impact Profile questionnaire (OHIP-14). The DNA sample was obtained from cells of the oral mucosa. Five markers of the FKBP5, SLC6A4, and COMT genes were genotyped. The data were submitted to statistical analysis with a significance level of 5%. Women reported greater intensity of discomfort associated with third molar surgery compared to men (p = 0.001). In the recessive model, the AA genotype of the rs3800373 marker was associated with greater surgical discomfort (p = 0.026). Therefore, women and individuals of the AA genotype for the rs3800373 marker in the FKBP5 gene reported greater surgical discomfort associated with third molar surgery.