Fluid flow-induced modulation of viability and osteodifferentiation of periodontal ligament stem cell spheroids-on-chip.

Developing physiologically relevant in vitro models for studying periodontitis is crucial for understanding its pathogenesis and developing effective therapeutic strategies. In this study, we aimed to integrate the spheroid culture of periodontal ligament stem cells (PDLSCs) within a spheroid-on-chip microfluidic perfusion platform and to investigate the influence of interstitial fluid flow on morphogenesis, cellular viability, and osteogenic differentiation of PDLSC spheroids. PDLSC spheroids were seeded onto the spheroid-on-chip microfluidic device and cultured under static and flow conditions. Computational analysis demonstrated the translation of fluid flow rates of 1.2 µl min-1 (low-flow) and 7.2 µl min-1 (high-flow) to maximum fluid shear stress of 59 µPa and 360 µPa for low and high-flow conditions, respectively. The spheroid-on-chip microfluidic perfusion platform allowed for modulation of flow conditions leading to larger PDLSC spheroids with improved cellular viability under flow compared to static conditions. Modulation of fluid flow enhanced the osteodifferentiation potential of PDLSC spheroids, demonstrated by significantly enhanced alizarin red staining and alkaline phosphatase expression. Additionally, flow conditions, especially high-flow conditions, exhibited extensive calcium staining across both peripheral and central regions of the spheroids, in contrast to the predominantly peripheral staining observed under static conditions. These findings highlight the importance of fluid flow in shaping the morphological and functional properties of PDLSC spheroids. This work paves the way for future investigations exploring the interactions between PDLSC spheroids, microbial pathogens, and biomaterials within a controlled fluidic environment, offering insights for the development of innovative periodontal therapies, tissue engineering strategies, and regenerative approaches.

MicroRNA Identification, Target Prediction, and Validation for Crop Improvement.

Micro-RNAs (mi-RNAs) are regulatory elements that play a vital role in the growth, development, and metabolic regulation of plants. In current research, the isolation of miRNAs is a tedious and difficult task using in vitro methods. However, recent exploration into the remarkably highly conserved nature of nucleotide sequences of miRNAs assists in the identification of miRNAs in plant species through homologous approaches. Here, we describe the in silico-based method for identification of miRNAs from the EST database which is emerging as a faster and more reliable approach along with the development of miRNA-SSR markers. This approach has the potential to accelerate research into the regulation of gene expression in various plant species such as tea, potato, tomato, tobacco, and orphan crops like cluster bean.

Unboxing the black box-one step forward to understand the soil microbiome: A systematic review.

Soil is one of the most important assets of the planet Earth, responsible for maintaining the biodiversity and managing the ecosystem services for both managed and natural ecosystems. It encompasses large proportion of microscopic biodiversity, including prokaryotes and the microscopic eukaryotes. Soil microbiome is critical in managing the soil functions, but their activities have diminutive recognition in few systems like desert land and forest ecosystems. Soil microbiome is highly dependent on abiotic and biotic factors like pH, carbon content, soil structure, texture, and vegetation, but it can notably vary with ecosystems and the respective inhabitants. Thus, unboxing this black box is essential to comprehend the basic components adding to the soil systems and supported ecosystem services. Recent advancements in the field of molecular microbial ecology have delivered commanding tools to examine this genetic trove of soil biodiversity. Objective of this review is to provide a critical evaluation of the work on the soil microbiome, especially since the advent of the NGS techniques. The review also focuses on advances in our understanding of soil communities, their interactions, and functional capabilities along with understanding their role in maneuvering the biogeochemical cycle while underlining and tapping the unprecedented metagenomics data to infer the ecological attributes of yet undiscovered soil microbiome. This review focuses key research directions that could shape the future of basic and applied research into the soil microbiome. This review has led us to understand that it is difficult to generalize that soil microbiome plays a substantiated role in shaping the soil networks and it is indeed a vital resource for sustaining the ecosystem functioning. Exploring soil microbiome will help in unlocking their roles in various soil network. It could be resourceful in exploring and forecasting its impacts on soil systems and for dealing with alleviating problems like rapid climate change.

Characterization of silver diamine fluoride cytotoxicity using microfluidic tooth-on-a-chip and gingival equivalents.

OBJECTIVE: This study aims to characterize the cytotoxicity potential of silver diamine fluoride (SDF) on dental pulp stem cells (DPSC) and gingival equivalents. METHODS: DPSC cultured on 96-well plates was exposed directly to SDF (0.0001-0.01%) and cell viability (IC50) quantified. Effect of SDF on DPSC viability under flow (with dentin barrier) conditions was evaluated using a custom-designed microfluidic "tooth-on-a-chip". Permeability of dentin discs (0.5-1.5 mm thickness) was evaluated using lucifer yellow permeation assay. Dentin discs were treated with 38% SDF (up to 3 h), and cell viability (live/dead assay) of the DPSC cultured in the inlet (unexposed) and outlet (exposed) regions of the pulp channel was evaluated. To assess the mucosal corrosion potential, gingival equivalents were treated with 38% SDF for 3 or 60 min (OECD test guideline 431) and characterized by MTT assay and histomorphometric analysis. RESULTS: DPSC exposed directly to SDF showed a dose-dependent reduction in cell viability (IC50: 0.001%). Inlet channels (internal control) of the tooth-on-a-chip exposed to PBS and SDF-exposed dentin discs showed> 85% DPSC viability. In contrast, the outlet channels of SDF-exposed dentin discs showed a decreased viability of< 31% and 0% (1.5 and ≤1.0 mm thick dentin disc, respectively) (p < 0.01). The gingiva equivalents treated with SDF for 3 and 60 min demonstrated decreased epithelial integrity, loss of intercellular cohesion and corneal layer detachment with significant reduction in intact epithelial thickness (p < 0.05). SIGNIFICANCE: SDF penetrated the dentin (≤1 mm thick) inducing significant death of the pulp cells. SDF also disrupted gingival epithelial integrity resulting in mucosal corrosion.

Spatiotemporal Regulation of CRISPR/Cas9 Enables Efficient, Precise, and Heritable Edits in Plant Genomes.

CRISPR/Cas-mediated editing has revolutionized crop engineering. Due to the broad scope and potential of this technology, many studies have been carried out in the past decade towards optimizing genome editing constructs. Clearly, the choice of the promoter used to drive gRNA and Cas9 expression is critical to achieving high editing efficiency, precision, and heritability. While some important considerations for choosing a promoter include the number and nature of targets, host organism, mode of transformation and goal of the experiment, spatiotemporal regulation of Cas9 expression using tissue-specific or inducible promoters enables higher heritability and efficiency of targeted mutagenesis with reduced off-target effects. In this review, we discuss specific studies that highlight the prospects and trade-offs associated with the choice of promoters on genome editing and emphasize the need for inductive exploration and discovery to further advance this area of research in crop plants.

Multidimensional approaches of biogas production and up-gradation: Opportunities and challenges.

The expanding interest towards biogas generation from biowaste via complex anaerobic digestion (AD) opened new avenues in the improvement of biogas production processes and their up-gradation. The adsorption/removal of impurities particularly hydrogen sulfide (H2S) and carbon dioxide (CO2) from the biogas stream will significantly improve the efficiency of biogas for its further use as a renewable energy fuel. The production and up-gradation of biogas rely upon the types of feedstocks, AD condition, microbial diversity, purification methods along with the application of various additives. In that context, this review aims to emphasize the current state of the art in the field of biogas production via AD using diverse bio-waste. Further, this review will critically explore the biogas up-gradation technologies adopted so far and their pros and cons. Finally, techno-economic and environmental impact assessment of the biogas production process will be underlined to make the process cost-effective and environmentally sustainable.

Constructed wetland for wastewater reuse: Role and efficiency in removing enteric pathogens.

Water stress has become a perennial concern in most of the developing countries due to rapid urbanization and population growth. As the growing population requires more fresh water and better ways for wastewater disposal, the demand for wastewater reclamation has increased drastically in recent years. Wastewater, either raw or treated, is being widely used for agricultural irrigation in developing countries, which cause a serious threat to human health mainly because of its pathogenic content. One of the alternative methods to treat wastewater and make it reusable for agricultural irrigation is to implement constructed wetland (CW); a sustainable and cost-effective technology that is applicable for the elimination of both pollutants and pathogens from wastewater. Despite its wide application, the role of macrophytes that form an integral part of CW and specific mechanisms involved in pathogen removal by them is still barely understood due to complexities involved and influencing factors. This has, therefore, attracted various scientific studies to reveal further functional mechanisms involved in vegetated CW to increase its proficiencies. This review paper illustrates the comparative studies of different CW and their pathogen removal efficiencies with major emphasis on macrophytes involved and factors influencing related mechanism. Further, the paper also covers detailed information on the enteric pathogens present in wastewater and the associated health risks involved in its reuse. The ultimate objective is to further clarify the role of CW in enteric pathogen removal and its efficiency for wastewater purification in perspective with safe reuse in agriculture.

Use of paracrine factors from stem cells to treat local radiation burns in rats.

BACKGROUND: Mesenchymal stem cells based paracrine bioactive factors that deploy their task as an essential mechanism, but their efficiency for skin regeneration still requires clarification. METHODS: The mesenchymal stem cell-based paracrine factors were administered by subcutaneous injection of 0.5 mL peptides (general protein 8 mg/mL). These were performed after radiation on different days like the first, third, sixth, eighth, and 10th. To determine the consequences, we performed photography, planimetry, and preclinical test each week after 15 days of radiation. MSC-based peptides were injected into a rat that had radiation burns, and its observation encouraged cell-free therapeutic remedies to regenerate skin. Both control and experimental groups were exposed to 110 Gy of X-rays, which resulted in the formation of localized radiation burns on the skin (S=6 cm2) 15 days later. Thirty days after radiation, the wound stabilized (surface of the wound was S=2.2±0.2 cm2) and fluctuated throughout the course of the pathological process. RESULTS: The wounded area on the skin from the 15th to the 29th day after radiation was practically the same in both groups. The wounded area gradually reduced by 6.1±0.4 cm2 (experimental group) and 5.9±0.6 cm2 (control group) 15 days after radiation up to 2.2±0.3 cm2 (in both control and experimental groups) on the 29th day after radiation. However, starting from the 36th day, there was a constant reduction in the burn area in the experimental group up to 0.2±0.1 cm2 till the 71st day after radiation. CONCLUSION: In the control group, the area of the lesion ranged from 1.4±0.6 cm2 on the 50th day to 1.9±0.8 cm2 on the 71st day. During the 57th to the 71st day, the difference between the affected area in the experimental and control groups was 1:8. The experimental group has a significantly higher level of skin regeneration and significant decrease in the level of leukocyte infiltration, thereby reducing necrosis.

Oral health awareness in school-going children and its significance to parent's education level.

BACKGROUND: The oral health imparts a major impact on one's quality of life. Since last decades, there is an increase in the prevalence of dental afflictions which can be avoided by the knowledge of simple preventive measures. AIM: The aim of this study was to assess the oral health status of children and to analyze its association with the education level of their parents. MATERIALS AND METHODS: Two randomly schools were selected and all the children from 5-15 years willing to participate in the present cross-sectional study were assessed for oral health status using the World Health Organization for the oral health awareness in children, 2004. Both child and parents were directly interviewed to avoid any bias in understanding the questionnaire. Statistical Package for the Social Sciences (16.0) was used for data analysis. Chi-square/Fisher's exact test was used to compare the dichotomous variables. RESULTS: A total of 210 (133 males and 77 females) children were assessed along with their parents for the awareness of child's oral health condition. Totally 134 (63.3%) children were of age group 5-10 years, while 76 (36.7%) children were of age group 11-15 years. Children with uneducated parents have often experienced toothache, brushes once a daily and frequently visits the dentist for toothache compared to children of educated parents, and this association was observed to be statistically significant (<0.001). CONCLUSION: The oral health status of children was average. However, education level and awareness of parents regarding oral health preventive measures play a key role in determining the oral health of children.

The incidence of facial injuries in children in Indian population: A retrospective study.

OBJECTIVE: To determine the incidence and pattern of facial fracture in children of age 0-16 years of age. MATERIAL & METHODS: Patients with trauma reporting to department of paediatric and preventive dentistry and department of oral & maxillofacial surgery from January 2011 to December 2011 were selected for the present study. The records of patients treated for maxillofacial injuries were retrospectively retrieved and analyzed for prevalence, pattern, etiology, and management of maxillofacial trauma. The data collected were subjected to statistically analysis. SPSS software version 16.0 was used for the data analysis. RESULTS: Total 5049 patients were screened and fifty one was reported to have facial injury. The incidence of facial trauma was 1.01% for the present study. The age wise distribution of the fracture amongst groups (I, II and III) were found 17.65%, 54.9% and 27.45% respectively. The sex-wise distribution of facial fracture was reported twice in male than females. The mandibular fracture was found the most common fracture (0.59%) followed by dento alveolar (0.30%) and midface (0.12%) fractures. Most of the mandibular fractures were found in the para symphysis region. Compound fractures seem to be commonest in the mandible. CONCLUSION: The fall was the predominant cause for most of the facial fractures in children followed by road traffic accident. The pattern of facial injury is influenced by the age and the growth of facial skeleton.

Obstructive sleep apnea risk for driving license applicants in India - A community based study.

OBJECTIVES: To determine the risk of obstructive sleep apnea (OSA) for male permanent driving license (DL) applicants of Lucknow, India. MATERIAL AND METHODS: In this cross-sectional community based, study body mass index, waist-hip ratio, blood pressure of each subject were determined as an anthropometric parameter along with the history of habit of smoking, tobacco chewing, alcohol consumption. STOP-Bang (Snoring, Tired or sleepy, Observed apnea, high blood Pressure, Body mass index, Age, Neck, Gender) Questionnaire - a scoring risk assessment tool - was applied for assessment of OSA risk (high OSA risk defined by score ≥ 3) for 542 male DL recipients at 2 Regional Transport Office (RTO) centers in Lucknow, India. The statistical software SPSS 17.0 was applied to the testing. RESULTS: In total 23% (N = 125) of participants were found with the risk of OSA. High blood pressure (≥ 140/90 mm Hg) was found for the maximum number of participants (40.5%) followed by neck circumference > 40 cm (17.1%), age (> 50 years old) (15.3%), snoring (12.3%) and tired/sleepy (10.5%). Mean values of age, anthropometric measurements and blood pressure were observed significantly higher (p < 0.001) for participants with the OSA risk. In this population the risk of OSA risk (STOP-Bang score ≥ 3) was observed for 6.7% of young (< 35 years old), 34% of middle (35-45 years old) and 73% of elder age adults (> 45 years old). CONCLUSIONS: In view of findings of this study a high number of male driving license applicants were observed with the risk of OSA. Therefore efforts should be made to develop a national screening guideline/protocol for the OSA risk assessment for driving license applicants in India. This may reduce the possibility of road traffic accidents due to the OSA-associated fatigue and drowsiness behind the wheels. Int J Occup Med Environ Health 2018;31(1):25-36.

Sexual dimorphism, pattern of third molar and mandibular second premolar agenesis in Indian paediatric orthodontic patients.

OBJECTIVE: To determine and compare the prevalence and pattern of agenesis of third molar and mandibular second premolar in paediatric orthodontic patients of age group 9-15 for sexual dimorphism. METHODS: The digital orthopantograph was obtained from the archive record of patients of age group 9-15 year. Radiographs of 301 patients were evaluated after taking exclusion criteria into account and were assessed for the presence/absence of third molars and mandibular second premolar. Tooth development evaluation followed the method of Demirjian et al., based on eight stages of tooth formation. The agenesis of third molar in maxilla and mandible between age groups and gender was compared using Chi-squared test. RESULTS: The rate of agenesis of third molars was observed 36.8% in the present study. Twenty-four (24.3%) percentage of the study population showed agenesis of all the four third molars. The agenesis of third molars was found to be higher among males than females (p > 0.05). Prevalence of agenesis of mandibular second premolar was 4.7-5%. CONCLUSIONS: Agenesis of third molars was more commonly seen in the maxilla, having male predilection. Maxillary right third molar was the most commonly missing tooth irrespective of gender.

Antibacterial effect and physical properties of chitosan and chlorhexidine-cetrimide-modified glass ionomer cements.

AIMS: To compare antibacterial effect and physical properties of chitosan (CH) modified glass ionomer cement (GIC) (10% v/v), chlorhexidine-cetrimide (CHX-CT) modified GIC (2.5/2.5% w/w) and conventional GIC. MATERIALS AND METHODS: A total of fifty healthy children of age 7-12 years were selected and randomly assigned to class A and B for in vivo analysis. Slabs of CH modified GIC (Group II) along with slabs of conventional GIC (Group I, control) were cemented on buccal surfaces of maxillary molars (split-mouth technique) for class A children. Similarly, slabs of CHX-CT modified GIC (Group III) were cemented against control (Group I, control) in class B children. Slabs were assessed after 48 h for microbial load of Streptococcus mutans and Lactobacillus (LB) on mitis salivarius-bacitracin and Man Rogosa Sharpe agar media, respectively. Agar diffusion test was done to access the antibacterial effect of each group against Streptococcus muatns and LB. Slabs and cylinders of GICs were made for in vitro evaluation of compressive and flexure strength in each group. RESULTS: Comparison was done by nonparametric Kruskal-Wallis analysis followed by Dunn's multiple comparison test. Categorical groups were compared by Chi-square test. The increase in antibacterial activity (Group II > III > I) (P < 0.001) and marked increase in compressive and flexure strength (Group II > I > III) were observed. CONCLUSIONS: In the view of findings, it is concluded that CH modified GIC would be effective in inhibiting the bacteria associated with dental caries along with improved physical properties when compared with CHX-CT modified GIC and conventional GIC.

Sleeping Habits among School Children and their Effects on Sleep Pattern.

Introduction: Sleep problems can occur at any age. Inadequate sleep affects the physiological as well as psychological well-being of an individual. Thus, the objective of the present study is, to determine the pre sleep habits, duration and pattern of sleep among school children and to determine association between their sleep schedules and sleep habits. Methods: This cross-sectional study comprised of 1050 children attending the government school. Based on inclusion and exclusion criteria children were from three age groups: 4-5 years, 6-10 years and 11-15 years of age. A questionnaire about demographical data, sleep problems and habits, was duly filled by the parents. The parents of children were questioned for bed time, wakeup time, sleep time and sleep duration during both weekdays and weekends. Results: Total sleep time during weekdays was 8.9 (1.2) hours and 10.7 (1.1) hours during weekends. The wakeup time was significantly delayed during weekends in all age groups. Moreover, total sleep time increased significantly during weekends in all age groups. Children using media after 8 pm and sleeping alone are also in significant sleep debt. Screen activities such as TV, internet and cellular phones in a child's bedroom had a negative effect on children's sleep/wake patterns and duration of sleep. Children in higher grades are sleep debt compared to younger ones. Practices such as co sleeping and sharing bed with parents significantly improve the duration and quality of sleep. Conclusion: The sleep durations reported in the present study were lower than recommended sleep duration for children.

Molar Incisor Hypomineralization: An Epidemiological Study with Prevalence and Etiological Factors in Indian Pediatric Population.

AIMS: To determine the prevalence of molar incisor hypomineralization (MIH) in Indian children and to analyze the possible etiological factors. MATERIALS AND METHODS: First permanent molars and all permanent incisors were examined in 1,369 children aged 8 to 12 years. Examinations were performed by two calibrated observers. The subjects were evaluated using judgment criteria proposed by Weerheijm et al in 2003. The parents accompanying children were given a questionnaire regarding pre- and postnatal history of the children. RESULTS: A total of 191 children were diagnosed with MIH with a prevalence of 13.9%. Chi-square/Fisher exact test was used to compare the dichotomous variables. The relative risk with its 95% confidence interval was calculated to find the risk of clinical infections, such as chicken pox, jaundice, renal disorders, cardiac disorders, and affected molars with sex and type of delivery. Pre- and postnatal history of infection in a child was significantly correlated with the prevalence of MIH. CONCLUSION: The prevalence of MIH was 13.9% in the age group of 8 to 12 years. Prenatal and postnatal infections play an important role in hypomineralization of molars and incisors. How to cite this article: Mishra A, Pandey RK. Molar Incisor Hypomineralization: An Epidemiological Study with Prevalence and Etiological Factors in Indian Pediatric Population. Int J Clin Pediatr Dent 2016;9(2):167-171.

Fibro-epithelial polyps in children: A report of two cases with a literature review.

A fibro-epithelial polyp is the most common epithelial benign tumor of the oral cavity. Such a polyp is of mesodermal origin and it is a pink, red, or white knob-like painless growth that is sessile or pedunculated. A fibro-epithelial polyp commonly occurs on buccal mucosa, the tongue, or the gingiva. A fibro-epithelial polyp is an inflammatory hyperplastic lesion in response to chronic irritation due to calculus, sharp tooth edges, irregular denture borders, or overhanging restorations. Such a polyp rarely occurs before the fourth decade of life and its prevalence is not sex-specific. The current paper presents two cases where an intraoral fibro-epithelial polyp was successfully managed in children. Conservative surgical excision was performed in both cases. A follow-up at 3 months revealed uneventful healing of the site without reoccurrence of the lesion.

Witkop's syndrome: A case report.

Witkop's syndrome is a rare autosomal dominant disorder characterized by tooth and nail abnormalities. It is a variant of ectodermal dysplasia, which is thought to occur in approximately 1 in 100,000 live births. Thus, such a low occurrence and paucity of clearly delineating features make its identification in general population a real challenge. This case report aims to add to the existing literature a commonsense approach of identifying the uncommon thing.

The significance of gtf genes in caries expression: a rapid identification of Streptococcus mutans from dental plaque of child patients.

AIM: Rapid phylogenetic and functional gene (gtfB) identification of S. mutans from the dental plaque derived from children. MATERIAL AND METHODS: Dental plaque collected from fifteen patients of age group 7-12 underwent centrifugation followed by genomic DNA extraction for S. mutans. Genomic DNA was processed with S. mutans specific primers in suitable PCR condtions for phylogenetic and functional gene (gtfB) identification. The yield and results were confirmed by agarose gel electrophoresis. RESULTS: 1% agarose gel electrophoresis depicts the positive PCR amplification at 1,485 bp when compared with standard 1 kbp indicating the presence of S. mutans in the test sample. Another PCR reaction was set using gtfB primers specific for S. mutans for functional gene identification. 1.2% agarose gel electrophoresis was done and a positive amplication was observed at 192 bp when compared to 100 bp standards. CONCLUSION: With the advancement in molecular biology techniques, PCR based identification and quantification of the bacterial load can be done within hours using species-specific primers and DNA probes. Thus, this technique may reduce the laboratory time spend in conventional culture methods, reduces the possibility of colony identification errors and is more sensitive to culture techniques.

A pedoprosthetic rehabilitation in patients with severe early childhood caries (S-ECC).

Early childhood caries (ECC) is a rapid form of dental caries. ECC is the most common chronic childhood disease that can compromise a child's self-esteem, nutritional intake, oral development and quality of life, subsequently leading to malocclusion and psychological problems. The restoration of severely decayed primary teeth is often a difficult procedure that offers a great challenge to paediatric dentists. The present case series document the clinical management of patients suffering from severe early childhood caries using removable partial prosthesis restoring their masticatory function and aesthestics. The ultimate aim of the treatment was to improve psychological and physiological development leading to better functioning of the stomatognathic system.