Search

In the realm of regenerative medicine and therapeutic applications, stem cell research is rapidly gaining traction. Dental pulp stem cells (DPSCs), which are present in both deciduous and permanent teeth, have emerged as a vital stem cell source due to their accessibility, adaptability, and innate differentiation capabilities. DPSCs offer a readily available and abundant reservoir of mesenchymal stem cells, showcasing impressive versatility and potential, particularly for regenerative purposes. Despite their promise, the main hurdle lies in effectively isolating and characterizing DPSCs, given their representation as a minute fraction within dental pulp cells. Equally crucial is the proper preservation of this invaluable cellular resource. The two predominant methods for DPSC isolation are enzymatic digestion (ED) and outgrowth from tissue explants (OG), often referred to as spontaneous growth. This protocol concentrates primarily on the enzymatic digestion approach for DPSC isolation, intricately detailing the steps encompassing extraction, in-lab processing, and cell preservation. Beyond extraction and preservation, the protocol delves into the differentiation prowess of DPSCs. Specifically, it outlines the procedures employed to induce these stem cells to differentiate into adipocytes, osteoblasts, and chondrocytes, showcasing their multipotent attributes. Subsequent utilization of colorimetric staining techniques facilitates accurate visualization and confirmation of successful differentiation, thereby validating the caliber and functionality of the isolated DPSCs. This comprehensive protocol functions as a blueprint encompassing the entire spectrum of dental pulp stem cell extraction, cultivation, preservation, and characterization. It underscores the substantial potential harbored by DPSCs, propelling forward stem cell exploration and holding promise for future regenerative and therapeutic breakthroughs.

Recently, astaxanthin, a red lipophilic pigment belonging to the xanthophyllic family of carotenoids, has shown the feasibility of its uses in tissue engineering and regenerative medicine, due to its excellent antioxidant activities and its abilities to enhance the self-renewal potency of stem cells. In this study, we demonstrate the influence of astaxanthin on the proliferation of adipose-derived mesenchymal stem cells in tissue-engineered constructs. The tissue engineered scaffolds were fabricated using photopolymerizable gelatin methacryloyl (GelMA) with different concentrations of astaxanthin. The effects of astaxanthin on cellular proliferation in two-dimensional environments were assessed using alamar blue assay and reverse transcription polymerase chain reaction (RT-PCR). Then, rheological properties, chemical structures and the water absorption of the fabricated astaxanthin-incorporated GelMA hydrogels were characterized using NMR analysis, rheological analysis and a swelling ratio test. Finally, the influence in three-dimensional environments of astaxanthin-incorporated GelMA hydrogels on the proliferative potentials of adipose-derived stem cells was assessed using alamar blue assay and the confocal imaging with Live/dead staining. The experimental results of the study indicate that an addition of astaxanthin promises to induce stem cell potency via proliferation, and that it can be a useful tool for a three-dimensional culture system and various tissue engineering applications.

Background: Enhancement of the bone regenerative capacity of the bone substitutes could be achieved by incorporating bioactive agents such as proteins, and different drugs. Simvastatin, an inhibitor of cholesterol synthesis, stimulates bone formation by enhancing the expression of Bone Morphogenetic Protein-2 (BMP-2) in osteoblasts., Objective: The objective of the study is to evaluate bone regeneration following simvastatin loaded nano-hydroxyapatite scaffold in the bone defect created on the femoral condyle of rabbits., Methods: Twelve adult, New Zealand white rabbits were used in the study. Twenty-four defects of size 5x8 mm were created on the lateral aspect of the femoral condyle. The defects were filled with either Nano-Hydroxyapatite (nHA) particles alone or nHA with Simvastatin (SIM). The condyles were retrieved after 8 weeks and analyzed using micro CT and histology., Results: The Bone Mineral Density (BMD) was significantly higher for the defects filled with SIM loaded nHA compared to the nHA site. Micro CT showed a significantly higher bone volume in the defects filled with Simvastatin loaded site compared to the control site. Quantitative analysis of the histologic sections also showed significantly higher bone volume in the defects filled with SIM loaded nHA (57.2±4.8) compared to nHA alone (50.1±5.5)., Conclusion: Based on the results, it can be concluded that local delivery of simvastatin enhanced the bone regeneration in rabbit femoral condyle. Simvastatin could be used as an activator to enhance bone regeneration in bone defects along with hydroxyapatite ceramics., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Bone marrow mesenchymal stem cells (BMSCs) are adult multipotent stem cells that can differentiate into mesodermal lineage cells, including adipocytes and osteoblasts. However, the epigenetic mechanisms governing the lineage-specific commitment of BMSCs into adipocytes or osteoblasts are under investigation. Herein, we investigated the epigenetic effect of romidepsin, a small molecule dual inhibitor targeting HDAC1 and HDAC2 identified through an epigenetic library functional screen. BMSCs exposed to romidepsin (5 nM) exhibited enhanced adipocytic and osteoblastic differentiation. Global gene expression and signaling pathway analyses of differentially expressed genes revealed a strong enrichment of genes involved in adipogenesis and osteogenesis in romidepsin-treated BMSCs during induction into adipocytes or osteoblasts, respectively. Pharmacological inhibition of FAK signaling during adipogenesis or inhibition of FAK or TGF β signaling during osteogenesis diminished the biological effects of romidepsin on BMSCs. The results of chromatin immunoprecipitation combined with quantitative polymerase chain reaction indicated a significant increase in H3K9Ac epigenetic markers in the promoter regions of peroxisome proliferator-activated receptor gamma (PPAR γ ) and KLF15 (related to adipogenesis) or SP7 (Osterix) and alkaline phosphatase (ALP) (related to osteogenesis) in romidepsin-treated BMSCs. Our data indicated that romidepsin is a novel in vitro modulator of adipocytic and osteoblastic differentiation of BMSCs.

Objectives: To assess the prevalence of periodontal disease among patients undergoing renal dialysis., Methods: Sixty hemodialysis patients (30 males, 30 females) with a mean age of 44.4±9.5 years comprised the study group. Periodontal parameters such as plaque index (PI), bleeding on probing (BOP), probing pocket depth (PPD) and clinical attachment level (CAL) were measured in these patients and 60 age and sex-matched control subjects. The data was tabulated and analyzed., Results: The mean duration of dialysis was 5.50±3.02 years. The plaque index (PI) bleeding on probing (BOP), Periodontal probing depth (PPD) and clinical attachment level (CAL) were significantly higher in patients undergoing hemodialysis than in control subjects. The mean clinical attachment level was significantly higher among the dialysis patients (2.78±0.83 mm) than the control subjects (1.97±0.53 mm). The plaque index and bleeding on probing also showed a similar pattern in patients undergoing hemodialysis compared to control patients. The prevalence and severity of periodontal disease seems to be higher in patients undergoing dialysis., Conclusion: From the observations of this study, it can be concluded that patients undergoing hemodialysis are more prone to periodontal diseases. Further studies with a larger population and a comparison with the duration of dialysis may further substantiate the current findings.

Stem cell biology has become an important field in regenerative medicine and tissue engineering therapy since the discovery and characterization of mesenchymal stem cells. Stem cell populations have also been isolated from human dental tissues, including dental pulp stem cells, stem cells from human exfoliated deciduous teeth, stem cells from apical papilla, dental follicle progenitor cells, and periodontal ligament stem cells. Dental stem cells are relatively easily obtainable and exhibit high plasticity and multipotential capabilities. The dental stem cells represent a gold standard for neural-crest-derived bone reconstruction in humans and can be used for the repair of body defects in low-risk autologous therapeutic strategies. The bioengineering technologies developed for tooth regeneration will make substantial contributions to understand the developmental process and will encourage future organ replacement by regenerative therapies in a wide variety of organs such as the liver, kidney, and heart. The concept of developing tooth banking and preservation of dental stem cells is promising. Further research in the area has the potential to herald a new dawn in effective treatment of notoriously difficult diseases which could prove highly beneficial to mankind in the long run., Competing Interests: Declaration of conflicting interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

The role of bone marrow adipocytes (BMAs) in overall energy metabolism and their effects on bone mass are currently areas of intensive investigation. BMAs differentiate from bone marrow stromal cells (BMSCs); however, the molecular mechanisms regulating BMA differentiation are not fully understood. In this study, we investigated the effect of CUDC-907, identified by screening an epigenetic small-molecule library, on adipocytic differentiation of human BMSCs (hBMSCs) and determined its molecular mechanism of action. Human bone marrow stromal cells exposed to CUDC-907 (500 nM) exhibited enhanced adipocytic differentiation (∼2.9-fold increase, P < 0.005) compared with that of control cells. Global gene expression and signaling pathway analyses of differentially expressed genes revealed a strong enrichment of genes involved in adipogenesis, cell cycle, and DNA replication. Chromatin immune precipitation combined with quantitative polymerase chain reaction showed significant increase in H3K9ac epigenetic marker in the promoter regions of AdipoQ, FABP4, PPARγ, KLF15, and CEBPA in CUDC-907-treated hBMSCs. Follow-up experiments corroborated that the inhibition of histone deacetylase (HDAC) activity enhanced adipocytic differentiation, while the inhibition of PI3K decreased adipocytic differentiation. In addition, CUDC-907 arrested hBMSCs in the G0-G1 phase of the cell cycle and reduced the number of S-phase cells. Our data reveal that HDAC, PI3K, and cell cycle genes are important regulators of BMA formation and demonstrate that adipocyte differentiation of hBMSCs is associated with complex changes in a number of epigenetic and genetic pathways, which can be targeted to regulate BMA formation.

Background/aims: 5-Fluorouracil (5-FU) is widely used in the treatment of patients with colorectal cancer (CRC). However, the efficacy of 5-FU as a single agent is limited, with multiple undesired side effects. Therefore, the aim of the current study was to assess the efficacy of CUDC-907 (a dual inhibitor of histone deacetylase and phosphatidylinositide 3-kinase) in combination with 5-FU against CRC cells., Materials and Methods: Cell viability was determined using AlamarBlue and colony formation assays. Acridine orange/ethidium bromide staining and flow cytometry were used to measure apoptotic and necrotic events, as well as cell cycle progression. Immunoblotting was used to assess acetylation of histone H3 and phosphorylation of AKT., Results: Our data revealed enhanced toxicity of CUDC-907 against HCT116, RKO, COLO-205, and HT-29 CRC cells when combined with 5-FU. Similarly, the colony formation capability of HCT116 cells was suppressed by the combination treatment. Cells treated with CUDC-907 and 5-FU underwent apoptosis and necrosis, and exhibited increased polyploidy. Furthermore, CRC cells treated with CUDC-907 exhibited a higher degree of histone H3 lysine 9 acetylation (H3K9ac) and reduced AKT phosphorylation (Ser473)., Conclusion: Our data revealed, for the first time, the enhanced inhibitory effect of CUDC-907 against CRC cells when combined with 5-FU, supporting the application of this combination as a potential therapeutic strategy in CRC treatment.

Background: Papillon-Lefèvre syndrome is a rare autosomal recessive disorder characterized by palmoplantar hyperkeratosis and aggressively progressing periodontitis leading to premature loss of deciduous and permanent dentition. The etiopathogenesis of the syndrome is relatively obscure, and immunologic, genetic, or possible bacterial etiologies have been proposed., Case Presentation: A series of five cases of Papillon-Lefèvre syndrome among the siblings in a family is presented here: a 3-year-old Arab girl, a 4-year-old Arab boy, a 11-year-old Arab boy, a 12-year-old Arab boy, and a 14-year-old Arab boy. The patients presented with severe gingival inflammation and mobility of teeth. The clinical manifestations were typical of Papillon-Lefèvre syndrome and the degree of involvement of the oral and skin conditions varied among them., Conclusions: This case series stresses the consanguinity in the family as an etiologic factor. All siblings in the family were affected with Papillon-Lefèvre syndrome which makes this a rare case. A multidisciplinary approach with the active participation of a dental surgeon, dermatologist, and pediatrician is essential for the management of cases of Papillon-Lefèvre syndrome.

Objectives: The purpose of this in vitro study is to compare the compressive strength (CS) and diametral tensile strength (DTS) of the zirconia-reinforced restorative material (Zirconomer®) with conventional glass ionomers (Fuji 1X) and amalgam., Materials and Methods: Specimens (n = 120) were fabricated from silver amalgam, reinforced glass ionomer cement (GIC) (glass ionomer, Fuji 1X GC Corp.), and zirconia-reinforced glass ionomer (Zirconomer, Shofu Inc.) for testing the CS and DTS. The results were analyzed using analysis of variance, followed by a Tukey post hoc test., Results: Both CS and DTS were found to be significantly higher for the zirconia-reinforced GIC and silver amalgam compared with GIC (p < 0.001)., Conclusion: A newer class of restorative material like Zirconomer helps to overcome the potential hazard of mercury, but retains the strength and durability of amalgam as well as the sustained high-fluoride release of GICs. Furthermore, long-term studies are required to confirm its use as an alternative to the currently available posterior restorative material.

Objective: To study the prevalence of anterior dental trauma and its associated factors among 800 preschool children aged 3 to 5 years in Jaipur City, Rajasthan, India., Materials and Methods: A cross-sectional survey was conducted among children aged 3-5 years, who were enrolled in various private and public schools in Jaipur. Parents were asked to fill a form addressing socio-demographic data and clinical examinations were performed by a single dentist. Traumatic dental injuries (TDI) were assessed and recorded based on Andreasen's classification. Associated factors such as sex, socioeconomic status (SES), and the type of injury were also analyzed. The data were analyzed statistically using Statistical Package for the Social Sciences software (version 20)., Results: An overall 10.2% prevalence of TDI was observed among the study population. TDI were reported to be more among male children (11.87%) compared to female children (8.14%). Enamel fractures (69%) were the most prevalent type of anterior dental trauma. Upper central incisors were the most frequently affected. The SES of the parents had little influence on the prevalence of TDI., Conclusions: The prevalence rate of dental trauma among children aged 3-5 years was 10.2%. Associated factors, such as SES, were observed to be not significantly correlated to dental trauma among the studied preschoolers.

Dental plaque is a biofilm that forms naturally on the surfaces of exposed teeth and other areas of the oral cavity. It is the primary etiological factor for the most frequently occurring oral diseases, such as dental caries and periodontal diseases. Specific, nonspecific, and ecologic plaque hypothesis explains the causation of dental and associated diseases. Adequate control of biofilm accumulation on teeth has been the cornerstone of prevention of periodontitis and dental caries. Mechanical plaque control is the mainstay for prevention of oral diseases, but it requires patient cooperation and motivation; therefore, chemical plaque control agents act as useful adjuvants for achieving the desired results. Hence, it is imperative for the clinicians to update their knowledge in chemical antiplaque agents and other developments for the effective management of plaque biofilm-associated diseases. This article explores the critical analysis of various chemical plaque control strategies and the current trends in the control and prevention of dental plaque biofilm.

Background: Oral cancers account for approximately 2% of all cancers diagnosed each year; however, the vast majority (80%) of the affected individuals are smokers whose risk of developing a lesion is five to nine times greater than that of non-smokers. Tobacco smoke contains numerous carcinogens that cause DNA damage, including oxidative lesions that are removed effectively by the base-excision repair (BER) pathway, in which poly (ADP-ribose) polymerase 1 (PARP-1), plays key roles. Genetic variations in the genes encoding DNA repair enzymes may alter their functions. Several studies reported mixed effects on the association between PARP-1 variants and the risk of cancer development. Till now no reported studies have investigated the association between PARP-1 variants and oral squamous cell carcinoma (OSCC) risk in an Indian population., Materials and Methods: In the present case control study 100 OSCC patients and 100 matched controls were genotyped using PARP1 single nucleotide peptides (SNP's) rs1136410 and rs3219090 using TaqMan assays., Results: The results indicated significantly higher risk with PARP1 rs1136410 minor allele "C" (OR=1.909; p=0.02942; CI, 1.060- 3.439). SNP rs1136410 also showed significantly increased risk in patients with smoking habit at C/C genotype and at minor allele C., Conclusions: The PAPR-1 Ala762Val polymorphism may play a role in progression of OSCC. Larger studies with a greater number of samples are needed to verify these findings.

Nano-Hydroxyapatite (nHA) was isolated from salmon bone by alkaline hydrolysis. The resulting nHA was characterized using several analytical tools, including thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM), to determine the purity of the nHA sample. The removal of organic matter from the raw fish was confirmed by TGA. FT-IR confirmed the presence of a carbonated group and the similarities to synthetic Sigma HA. XRD revealed that the isolated nHA was amorphous. Microscopy demonstrated that the isolated nHA possessed a nanostructure with a size range of 6-37 nm. The obtained nHA interacted with mesenchymal stem cells (MSCs) and was non-toxic. Increased mineralization was observed for nHA treated MSCs compared to the control group. These results suggest that nHA derived from salmon is a promising biomaterial in the field of bone tissue engineering.

Objective: The purpose of the current in-vivo study was to assess the effect of using 0.12% chlorhexidine (CHX) mouth rinse, before bonding, on shear bond strength of polycarbonate brackets bonded with composite adhesive., Subjects and Methods: Eighteen orthodontic patients with a mean age 21.41 ± 1.2 years, who were scheduled to have 2 or more first premolars extracted, were included in this study. Patients were referred for an oral prophylaxis program which included, in part, the use of a mouth rinse. Patients were divided into 2 groups, a test group of 9 patients who used 0.12% CHX gluconate mouth rinse twice daily and a control group of 9 patients who used a mouth rinse without CHX, but with same color. After 1 week, polycarbonate brackets were bonded to first premolars with Transbond XT composite adhesive. Premolars were extracted after 28 days and tested for shear bond strength on a universal testing machine. Student's t-test was used to compare shear bond strengths of both groups., Results: No statistically significant difference was found in bond strengths' values between both groups. The test group (with CHX) has mean shear bond strength of 14.21 ± 2.42 MPa whereas the control group (without CHX) revealed a mean strength of 14.52 ± 2.31 MPa., Conclusion: The use of 0.12% CHX mouth rinse, for one week before bonding, did not affect the shear bond strength of polycarbonate brackets bonded with Transbond composite. Furthermore, these brackets showed clinically acceptable bond strength.

Hepatocyte growth factor (HGF) production by oral fibroblasts is enhanced by various molecules that are induced during inflammatory conditions including periodontitis. HGF plays an important role in the progression of periodontitis, by stimulating intense growth of epithelial cells and preventing regeneration of connective tissue attachments. Smokers have a greater risk factor in the pathogenesis and progression of periodontal disease. The objective of the study was to estimate the level of HGF in saliva and gingival crevicular fluid (GCF) in smokers with periodontitis and to compare these levels with that of nonsmokers with periodontitis and healthy controls. The HGF levels were found to be significantly high in the saliva and GCF of smokers with periodontitis compared to both never-smokers with periodontitis and the healthy control group. The elevated levels of HGF in the saliva and GCF in the study population could explain the intrinsic mechanism triggering the severity of the periodontitis in smokers. Further studies are necessary to validate the current observations and to establish a sensitive marker to predict periodontal disease activity.