Cannabidiol and Vitamin D3 Impact on Osteogenic Differentiation of Human Dental Mesenchymal Stem Cells.

Background and objective: The aim of the present study was to establish a new differentiation protocol using cannabidiol (CBD) and vitamin D3 (Vit. D3) for a better and faster osteogenic differentiation of dental tissue derived mesenchymal stem cells (MSCs). Materials and methods: MSCs were harvested from dental follicle (DFSCs), dental pulp (DPSCs), and apical papilla (APSCs) of an impacted third molar of a 17-year old patient. The stem cells were isolated and characterized using flow cytometry; reverse transcription polymerase chain reaction (RT-PCR); and osteogenic, chondrogenic, and adipogenic differentiation. The effects of CBD and Vit. D3 on osteogenic differentiation of dental-derived stem cell were evaluated in terms of viability/metabolic activity by alamar test, expression of collagen1A, osteopontin (OP), osteocalcin (OC), and osteonectin genes and by quantification of calcium deposits by alizarin red assay. Results: Stem cell characterization revealed more typical stemness characteristics for DFSCs and DPSCs and atypical morphology and markers expression for APSCs, a phenotype that was confirmed by differences in multipotential ability. The RT-PCR quantification of bone matrix proteins expression revealed a different behavior for each cell type, APSCs having the best response for CBD. DPSCs showed the best osteogenic potential when treated with Vit. D3. Cultivation of DFSC in standard stem cell conditions induced the highest expression of osteogenic genes, suggesting the spontaneous differentiation capacity of these cells. Regarding mineralization, alizarin red assay indicated that DFSCs and APSCs were the most responsive to low doses of CBD and Vit. D3. DPSCs had the lowest mineralization levels, with a slightly better response to Vit. D3. Conclusions: This study provides evidence that DFSCs, DPSCs, and APSCs respond differently to osteoinduction stimuli and that CBD and Vit. D3 can enhance osteogenic differentiation of these types of cells under certain conditions and doses.

Advanced glycation end products as biomarkers in systemic diseases: premises and perspectives of salivary advanced glycation end products.

Advanced glycation end products (AGEs) are glycated proteins associated with high dry temperature food processing, coloring and flavor modification of food products. Previous studies on diet-related disease support the role of the glycation products as biomarkers in local and general proinflammatory response. Exogenous and endogenous AGEs are involved in chronic low-level inflammation, which underlies the onset of metabolic syndrome influenced by food intake, there by demonstrating their implication in diet-related pathologies. Although studies have revealed a strong association between the accumulation of AGEs and the occurrence/worsening of metabolic diseases, their routine use for the diagnosis or monitoring of local and general disease has not yet been reported.

Histopathological and clinical expression of periodontal disease related to the systemic inflammatory response.

Periodontal disease with its systemic implications is highly prevalent among the population, and this correlation could have an impact on the quality lives of many humans. The purpose of this study was to assess the clinical and histopathological changes of the periodontium correlated with the systemic inflammatory response in periodontal disease. An experimental study was performed on male Wistar rats which were subjected to a procedure of periodontitis induction through placing silk thread ligatures around the lower incisors, under general anesthesia. Clinically, the changes of the periodontal tissue induced by the periodontitis progression were daily assessed. Two blood samples were obtained from each animal, at baseline and on completion of the experiment. The plasma level of the cytokine IL-6 and haematological parameters such as leukocytes, neutrophils, lymphocytes, monocytes, and platelets were determined. After seven days the animals were sacrificed, and samples were prepared for histological evaluation. Clinical manifestations such as changes in the color, contour and consistency of the gingival tissue and the bleeding on probing were registered. Histopathological analysis showed an intense inflammatory cell infiltration, the presence of osteoclasts and an obvious bone resorption activity. A significant increase in IL-6 values during the progression of periodontitis in rats (p<0.001) was also observed. The results of this research demonstrated that the clinical and histological changes in the rat's periodontium are correlated with a notable systemic inflammatory response. Therefore, periodontitis control can be inserted as part of the programs of systemic disorders prevention, in clinical practice.

Evaluation of the Biocompatibility of New Fiber-Reinforced Composite Materials for Craniofacial Bone Reconstruction.

This study aims to assess the biocompatibility of new advanced fiber-reinforced composites (FRC) to be used for custom-made cranial implants. Four new formulations of FRC were obtained using polymeric matrices (combinations of monomers bisphenol A glycidylmethacrylate [bis-GMA], urethane dimethacrylate [UDMA], triethylene glycol dimethacrylate [TEGDMA], hydroxyethyl methacrylate [HEMA]) and E-glass fibers (300 g/mp). Every FRC contains 65% E-glass and 35% polymeric matrix. Composition of polymeric matrices are: bis-GMA (21%), TEGDMA (14%) for FRC1; bis-GMA (21%), HEMA (14%) for FRC2; bis-GMA (3.5%), UDMA (21%), TEGDMA (10.5%) for FRC3, and bis-GMA (3.5%), UDMA (21%), HEMA (10.5%) for FRC4. Cytotoxicity test was performed on both human dental pulp stem cells and dermal fibroblasts. Viability was assessed by tetrazolium dye colorimetric assay. Subcutaneous implantation test was carried out on 40 male Wistar rats, randomly divided into 4 groups, according to the FRC tested. Each group received subcutaneous dorsal implants. After 30 days, intensity of the inflammatory reaction, tissue repair status, and presence of the capsule were the main criteria assessed. Both cell populations showed no signs of cytotoxicity following the FRC exposures. In terms of cytotoxicity, the best results were obtained by FRC3 followed by FRC2, FRC4, and FRC1. FRC3 showed also the mildest inflammatory reaction and this correlated both with the noncytotoxic behavior and the presence of a well-organized capsule. The composite biomaterials developed may constitute an optimized alternative of the similar materials used for the reconstruction of craniofacial bone defects. According to authors' studies, the authors conclude that FRC3 is the best formulation regarding the biological behavior.

Tracing CD34+ Stromal Fibroblasts in Palatal Mucosa and Periodontal Granulation Tissue as a Possible Cell Reservoir for Periodontal Regeneration.

The aim of the present research was to trace CD34+ stromal fibroblastic cells (CD34+ SFCs) in the palatal connective tissue harvested for muco-gingival surgical procedures and in granulation tissues from periodontal pockets using immunohistochemical and transmission electron microscopy. Immunohistochemical analysis targeted the presence of three antigens: CD31, α-smooth muscle actin (α-SMA), and CD34. In the palate, CD31 staining revealed a colored inner ring of the vessels representing the endothelium, α-SMA+ was located in the medial layer of the vasculature, and CD34 was intensely expressed by endothelial cells and artery adventitial cells (considered to be CD34+ SFCs). Granulation tissue showed the same pattern for CD31+ and α-SMA, but a different staining pattern for CD34. Ultrastructural examination of the palatal tissue highlighted perivascular cells with fibroblast-like characteristics and pericytes in close spatial relationship to endothelial cells. The ultrastructural evaluation of granulation tissue sections confirmed the presence of neovasculature and the inflammatory nature of this tissue. The present study traced the presence of CD34+ SFCs and of pericytes in the palatal connective tissue thus highlighting once more its intrinsic regenerative capabilities. The clinical and systemic factors triggering mobilization and influencing the fate of local CD34+SCFs and other progenitors are issues to be further investigated.

Lip Forces and Chewing Efficiency in Children with Peripheral Facial Paralysis.

Peripheral facial paralysis is accompanied by facial motor disorders and also, by oral dysfunctions. The aim of this study was to evaluate the lip forces and chewing efficiency in a group of children with peripheral facial paralysis. The degree of peripheral facial paralysis in the study group (n 11) was assessed using the House-Brackmann scale. The control group consisted of 21 children without facial nerve impairment. To assess lip forces, acrylic vestibular plates of three sizes were used: large (LVP), medium (MVP) and small (SVP). The lip force was recorded with a force transducer coupled with the data acquisition system. Masticatory efficiency was evaluated by the ability to mix two differently colored chewing gums. The images were processed with Adobe Photoshop CS3 (Delaware Corporation, San Jose, California, United States) and the number of pixels was quantified with the Image J software (DHHS/NIH/NIMH/RSB, Maryland, United States). For statistical analysis, the following statistical analysis were used: Pearson or Spearman correlation coefficient, multiple linear regression analysis, multiple logistic regression analysis, and optimal cutoff values for muscular dysfunction. There were statistically significant differences between lip forces in the following three groups: p=0.01 (LVP), p=0.01 (MVP), and p=0.008 (SVP). The cutoff values of lip forces in the study group were as follows: 7.08 N (LVP), 4.89 N (MVP), and 4.24 N (SVP). There were no statistically significant differences between the masticatory efficiency in the two groups (p=0.25). Lip forces were dependent on the degree of peripheral facial paralysis and age, but not on gender. In peripheral facial paralysis in children, a significant decrease of lip forces, but not masticatory efficiency, occurs.

Management of patients with facial paralysis in the dental office: A brief review of the literature and case report.

In the dental office, the dentist may have to examine patients with facial asymmetry and functional disorders caused by facial paralysis (FP). Following clinical examination, it is important for the dental practitioner to establish whether FP was caused by injury to the facial nerve, and to focus on the site of the lesion and potential risk factors. The risks of dental treatment in a patient with FP should also be assessed. Through dental or surgical procedures, the dentist may cause transient or permanent FP. Interdisciplinary collaboration is required for the confirmation of diagnosis and etiology, and for the complex treatment of FP. This article aims to examine the role of the dentist within the multidisciplinary medical team and to present two cases with transient FP following intraoral anesthesia in the dental office.