Effect of Incorporating Titanium Dioxide Nanoparticles into White Portland Cement, White Mineral Trioxide Aggregate, and Calcium Enriched Mixture Cement on the Push-out Bond Strength to Furcal Area Dentin.

Statement of the Problem: Bond strength of furcation repair materials is an essential factor in clinical success. Studies on the effect of adding titanium dioxide (TiO2) nanoparticles on the push-out bond strength of commonly used endodontic cements for furcation perforation repair is limited. Purpose: This study aimed to evaluate the effect of adding TiO2 nanoparticles to white Portland cement (PC), white mineral trioxide aggregate (MTA), and calcium enriched mixture cement (CEM) on their push-out bond strengths. Materials and Method: In this in vitro study, 120 endodontically treated molars were assigned to six groups (n=20) based on the material used to repair the perforation. In three groups, the cements (white PC, white MTA, and CEM) were placed in pure form, and in the three remaining groups, 1 weight % of TiO2 was added. The push-out bond strength was measured using a universal testing machine at a strain rate of 0.5 mm/min. Data were analyzed using one-way ANOVA and post hoc Games-Howell test (p< 0.05). Results: One-way ANOVA showed significant differences in the mean bond strength values between the six groups (p= 0.002). The post hoc Games-Howell test showed that the bond strengths in MTA+TiO2 and PC+TiO2 groups were significantly higher than those in MTA and PC groups, respectively. However, there was no significant difference in the bond strength between CEM and CEM+ TiO2 groups. Conclusion: The incorporation of TiO2 into MTA and PC increased their push-out bond strength. However, it did not affect the push-out bond strength of CEM cement.

The Effect of Low-Frequency Pulsed Electromagnetic Fields on the Differentiation of Permanent Dental Pulp Stem Cells into Odontoblasts.

Introduction: Exposure to pulsed electromagnetic field (PEMF) has been revealed to affect the differentiation and proliferation of human mesenchymal stem cells derived from dental pulp multipotent stromal stem cells (DP-MSCs). This study aimed to investigate the differentiation effect of electromagnetic fields (EMFs) on the DP-MSC. Materials and Methods: PEMF was produced by a system comprising a multi-meter autotransformer, solenoid coils, and teslameter. This study included 10 groups of DP-MSCs which underwent different electromagnetic radiation time and beam intensity. Three samples tested for each group. The effect of PEMF with the intensity of 0.5 and 1 mT (mili Tesla) and 50 Hz on the proliferation rate of DP-MSC was evaluated at 20 and 40 minutes per day for seven days. MTT assay was applied to determine the growth and proliferation of DP-MSC. Gene expression of DMP1 for differentiation of DPSCs to odontoblasts was confirmed by Real Time PCR., ANOVA statistical analysis and Kruskal-Wallis test were used to analyze the data. Results: The survival in all exposure groups was significantly higher than that in control except in the group of 40 minutes, 1 mT (P<0.05). In 20 minutes, 0.5 mT exposure, the survival intensity is significantly more than others (P<0.05). In general, the intensity of survival was recorded, 20, 0.5 mT≥20, 1 mT≥40, 0.5 mT≥40, 1 mT respectively. Therefore, according to the obtained results, ELF-EMF increases the survival of cells except for one case (40 minutes, 1 mT), even though the effective underlying mechanisms in this process are still unclear. Conclusions: The results obtained promise that in the future, by placing an important part of the pulp next to the electromagnetic field, the lost part of the pulp can be reconstructed and the dentin barrier can be created.

Effect of Silver Nanoparticles of Herbal Origin on the Compressive and Push-out Bond Strengths of Mineral Trioxide Aggregate.

Introduction: The purpose of this in vitro study was to investigate the effect of incorporating silver nanoparticles (AgNPs) of herbal origin into mineral trioxide aggregate (MTA) on the push-out bond strength (PBS) and compressive strength (CS) in simulated furcal area perforations. Materials and Methods: In this in vitro study, simulated furcal area perforations (1.3 mm in diameter and 2 mm in depth) were created in 40 extracted human lower molar teeth, which were divided into two groups (n=20): MTA alone and MTA combined with AgNPs (2% wt). Using a universal testing machine, PBS was evaluated by performing push-out tests, while CS was assessed using cylindrical specimens. The normal distribution of data was checked using the Kolmogorov-Smirnov test, and statistical analysis was performed using two-way ANOVA. Results: The CS results showed no significant difference between the MTA group at 4 and 21 days (P=0.297), but a significant difference was observed in the nanosilver/MTA group (P=0.013). However, there was no significant difference in the push-out bond strength among the study groups (P>0.05). Conclusion: The incorporation of herbal origin silver nanoparticles did not significantly affect the PBS or CS of MTA.

Synthesis, characterization, and evaluation of Hesperetin nanocrystals for regenerative dentistry.

Hesperetin (HS), a metabolite of hesperidin, is a polyphenolic component of citrus fruits. This ingredient has a potential role in bone strength and the osteogenic differentiation. The bone loss in the orofacial region may occur due to the inflammation response of host tissues. Nanotechnology applications have been harshly entered the field of regenerative medicine to improve the efficacy of the materials and substances. In the current study, the hesperetin nanocrystals were synthesized and characterized. Then, the anti-inflammatory and antioxidative effects of these nanocrystals were evaluated on inflamed human Dental Pulp Stem Cells (hDPSCs) and monocytes (U937). Moreover, the osteoinduction capacity of these nanocrystals was assessed by gene and protein expression levels of osteogenic specific markers including RUNX2, ALP, OCN, Col1a1, and BSP in hDPSCs. The deposition of calcium nodules in the presence of hesperetin and hesperetin nanocrystals was also assessed. The results revealed the successful fabrication of hesperetin nanocrystals with an average size of 100 nm. The levels of TNF, IL6, and reactive oxygen species (ROS) in inflamed hDPSCs and U937 significantly decreased in the presence of hesperetin nanocrystals. Furthermore, these nanocrystals induced osteogenic differentiation in hDPSCs. These results demonstrated the positive and effective role of fabricated nanocrystal forms of this natural ingredient for regenerative medicine purposes.

Injectable thermosensitive chitosan/gelatin hydrogel for dental pulp stem cells proliferation and differentiation.

Introduction: Biocompatible and biodegradable scaffolds based on natural polymers such as gelatin and chitosan (CS) provide suitable microenvironments in dental tissue engineering. In the present study, we report on the synthesis of injectable thermosensitive hydrogel (PNIPAAm-g-CS copolymer/gelatin hybrid hydrogel) for osteogenic differentiation of human dental pulp stem cells (hDPSCs). Methods: The CS-g-PNIPAAm was synthesized using the reaction of carboxyl terminated PNIPAAm with CS, which was then mixed with various amounts of gelatin solution in the presence of genipin as a chemical crosslinker to gain a homogenous solution. The chemical composition and microstructures of the fabricated hydrogels were confirmed by FT-IR and SEM analysis, respectively. To evaluate the mechanical properties (e.g., storage and loss modulus of the gels), the rheological analysis was considered. Calcium deposition and ALP activity of DPSCs were carried out using alizarin red staining and ALP test. While the live/dead assay was performed to study its toxicity, the real-time PCR was conducted to investigate the osteogenic differentiation of hDPSCs cultured on prepared hydrogels. Results: The hydrogels with higher gelatin incorporation showed a slightly looser network compared to the other ones. The hydrogel with less gelatin indicates a rather higher value of G', indicating a higher elasticity due to more crosslinking reaction of amine groups of CS via a covalent bond with genipin. All the hydrogels contained viable cells with negligible dead cells, indicating the high biocompatibility of the prepared hydrogels for hDPSCs. The quantitative results of alizarin red staining displayed a significant rise in calcium deposition in hDPSCs cultured on prepared hydrogels after 21 days. Further, hDPSCs cultured on hydrogel with more gelatin displayed the most ALP activity. The expression of late osteogenic genes such as OCN and BMP-2 were respectively 6 and 4 times higher on the hydrogel with more gelatin than the control group after 21 days. Conclusion: The prepared PNIPAAm-g-CS copolymer/gelatin hybrid hydrogel presented great features (e.g., porous structure, suitable rheological behavior, and improved cell viability), and resulted in osteogenic differentiation necessary for dental tissue engineering.

Salivary biomarkers in cancer.

In recent years, the comprehensive analysis of saliva, i.e., salivaomics, has played an increasing role in biomarker discovery for disease detection in general and cancer specifically. Saliva is a readily accessible, non-invasive and low-cost specimen that can be used to detect biomarkers of clinical relevance. Saliva-based "omics" technologies, which include proteomics, transcriptomics, metabolomics and microbiomics, have rapidly evolved and may be applicable in point-of-care detection, liquid biopsy and nanoscience. Advances in analytical methods has increased the scope and application of salivaomics from solely the oral cavity to the entire physiologic system, and accordingly to personalized medicine. In this chapter, we highlight recent advances in analytical approaches to identify and detect biomarkers in saliva and their potential use as diagnostic, prognostic and therapeutic markers with a focus on cancer.

Effect of three intracanal medicaments used in pulp regeneration on the push-out bond strength of mineral trioxide aggregate and calcium-enriched mixture: An in vitro study.

Background. The bond strength of the materials used as a cervical barrier in the pulp regeneration is essential for the success of treatment. This study aimed to evaluate the effects of triple antibiotic paste (TAP), double antibiotic paste (DAP), and simvastatin as intracanal medicaments on the dislodgement resistance of mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM). Methods. A total of 160 extracted human single-rooted teeth were selected, and root canal preparation was carried out. The teeth in each group were randomly divided into four subgroups: TAP, DAP, simvastatin, and the control group (without intracanal medicament). Four weeks after placing the medicaments, it was removed by sodium hypochlorite, and MTA and CEM were placed in the coronal third of the root canals. After a week, 2-mm-thick dentin disks were prepared from the coronal third of the roots, and the push-out test was performed using a universal testing machine. The data were analyzed using two-way ANOVA and independent t-test at a significance level of 0.05. Results. Regardless of the intracanal medicament, there was no significant difference between the overall bond strength of MTA (59.3±10 MPa) and CEM (55.8±11 MPa) (P=0.6). Furthermore, there were no significant differences in bond strength between the two intracanal medicament groups and the control group (P>0.05). Conclusion. Under the limitations of the current study, DAP, simvastatin, and TAP, as intracanal medicaments, did not adversely affect the push-out bond strength of CEM and MTA.

Enhancing the function of PLGA-collagen scaffold by incorporating TGF-ß1-loaded PLGA-PEG-PLGA nanoparticles for cartilage tissue engineering using human dental pulp stem cells.

Since cartilage has a limited capacity for self-regeneration, treating cartilage degenerative disorders is a long-standing difficulty in orthopedic medicine. Researchers have scrutinized cartilage tissue regeneration to handle the deficiency of cartilage restoration capacity. This investigation proposed to compose an innovative nanocomposite biomaterial that enhances growth factor delivery to the injured cartilage site. Here, we describe the design and development of the biocompatible poly(lactide-co-glycolide) acid-collagen/poly(lactide-co-glycolide)-poly(ethylene glycol)-poly(lactide-co-glycolide) (PLGA-collagen/PLGA-PEG-PLGA) nanocomposite hydrogel containing transforming growth factor-ß1 (TGF-ß1). PLGA-PEG-PLGA nanoparticles were employed as a delivery system embedding TGF-ß1 as an articular cartilage repair therapeutic agent. This study evaluates various physicochemical aspects of fabricated scaffolds by 1HNMR, FT-IR, SEM, BET, and DLS methods. The physicochemical features of the developed scaffolds, including porosity, density, degradation, swelling ratio, mechanical properties, morphologies, BET, ELISA, and cytotoxicity were assessed. The cell viability was investigated with the MTT test. Chondrogenic differentiation was assessed via Alcian blue staining and RT-PCR. In real-time PCR testing, the expression of Sox-9, collagen type II, and aggrecan genes was monitored. According to the results, human dental pulp stem cells (hDPSCs) exhibited high adhesion, proliferation, and differentiation on PLGA-collagen/PLGA-PEG-PLGA-TGFß1 nanocomposite scaffolds compared to the control groups. SEM images displayed suitable cell adhesion and distribution of hDPSCs throughout the scaffolds. RT-PCR assay data displayed that TGF-ß1 loaded PLGA-PEG-PLGA nanoparticles puts forward chondroblast differentiation in hDPSCs through the expression of chondrogenic genes. The findings revealed that PLGA-collagen/PLGA-PEG-PLGA-TGF-ß1 nanocomposite hydrogel can be utilized as a supportive platform to support hDPSCs differentiation by implementing specific physio-chemical features.

Novel nanocomposite scaffold based on gelatin/PLGA-PEG-PLGA hydrogels embedded with TGF-ß1 for chondrogenic differentiation of human dental pulp stem cells in vitro.

In the current study, a novel nanocomposite hydrogel scaffold comprising of natural-based gelatin and synthetic-based (poly D, L (lactide-co-glycolide) -b- poly (ethylene glycol)-b- poly D, L (lactide-co-glycolide) (PLGA-PEG-PLGA) triblock copolymer was developed and loaded with transforming growth factor- ß1 (TGF-ß1). Synthesized scaffolds' chemical structure was examined by 1H NMR and ATR-FTIR. Scanning electron microscopy (SEM) confirmed particle size and morphology of the prepared nanoparticles as well as the scaffolds. The morphology analysis revealed a porous interconnected structure throughout the scaffold with a pore size dimension of about 202.05 µm. The swelling behavior, in vitro degradation, mechanical properties, density, and porosity were also evaluated. Phalloidin/DAPI staining was utilized for confirming the extended cytoskeleton of the chondrocytes. Alcian blue staining was conducted to determine cartilaginous matrix sulfated glycosaminoglycan (sGAG) synthesis. Eventually, over a period of 21 days, a real-time RT-PCR analysis was applied to measure the mRNA expression of chondrogenic marker genes, type-II collagen, SOX 9, and aggrecan, in hDPSCs cultured for up to 21 days to study the influence of gelatin/PLGA-PEG-PLGA-TGF-ß1 hydrogels on hDPSCs. The findings of the cell-encapsulating hydrogels analysis suggested that the adhesion, viability, and chondrogenic differentiation of hDPSCs improved by gelatin/PLGA-PEG-PLGA-TGF-ß1 nanocomposite hydrogels. These data supported the conclusion that gelatin/PLGA-PEG-PLGA-TGF-ß1 nanocomposite hydrogels render the features that allow thein vitrofunctionality of encapsulated hDPSCs and hence can contribute the basis for new effective strategies for the treatment of cartilage injuries.

Application of Collagen and Mesenchymal Stem Cells in Regenerative Dentistry.

Collagen is an important macromolecule of Extracellular Matrix (ECM) in bones, teeth, and temporomandibular joints. Mesenchymal Stem Cells (MSCs) interact with the components of the ECM such as collagen, proteoglycans, Glycosaminoglycans (GAGs), and several proteins on behalf of variable matrix elasticity and bioactive cues. Synthetic collagen-based biomaterials could be effective scaffolds for regenerative dentistry applications due to mimicking of host tissues' ECM. These biomaterials are biocompatible, biodegradable, readily available, and non-toxic to cells whose capability promotes cellular response and wound healing in the craniofacial region. Collagen could incorporate other biomolecules to induce mineralization in calcified tissues like bone and tooth. Moreover, the addition of these molecules or other polymers to collagen-based biomaterials could enhance mechanical properties, which is important in load-bearing areas such as the mandible. A literature review was performed via a reliable internet database (mainly PubMed) based on MeSH keywords. This review first describes the properties of collagen as a key protein in the structure of hard tissues. Then, it introduces different types of collagens, the correlation between collagen and MSCs, and the methods used to modify collagen in regenerative dentistry, including recent progression on the regeneration of periodontium, dentin-pulp complex, and temporomandibular joint by applying collagen. The prospects and challenges of collagen-based biomaterials in the craniofacial region are pointd out.

Management of an infected immature tooth with a talon cusp using regenerative endodontic treatment: A case report.

OBJECTIVE: Regenerative endodontic treatment (RET) has been considered a successful approach to manage infected immature teeth; however, cases associated with dental anomalies, i.e., talon cusp, need to be further investigated. CASE REPORT: A 7-year-old girl with local swelling associated with the permanent maxillary right central incisor was referred; cone-beam computed tomography revealed a talon cusp, an immature root and two well-defined endodontic lesions. The treatment consisted of regenerative endodontic treatment (RET); the canal was chemically cleaned and a modified triple antibiotic paste was used as intracanal medication. In the next appointment, RET was performed through the creation of blood clot scaffold in the canal covered/sealed with calcium-enriched mixture (CEM) cement. RESULTS: In the 7-day recall session, clinical examination showed that the swelling had completely resolved. At 24-month recall, the treated tooth was asymptomatic and functional. CBCT images demonstrated evidence of maturation in the apical third of the root, healing of two large endodontic lesions and complete dentinal bridge formation beneath CEM cement. CONCLUSION: RET for an infected immature tooth with a dental anomaly, i.e., talon cusp, may be a desirable treatment option and result in the resolution of endodontic lesions as well as regeneration of new vital tissues; allowing continuous root maturation.

Effects of Intracanal Curcumin and Aloe vera on pH Changes in Simulated Root Surface Resorption Defects: An In vitro Study.

Introduction: Commonly used medicaments in the treatment of external inflammatory root resorption (EIRR) have shown adverse effects; resulting in an increasing tendency to employ natural and/or herbal medication. The present in vitro study aimed to evaluate the effects of curcumin and aloe vera, as two natural medicaments, on the changes of pH in external root surface defects; and compare their outcomes with the results obtained from the application of calcium hydroxide, as a conventional medicament used in endodontic treatments. Materials and Methods: In the current investigation, 92 permanent teeth, with a single root canal, were randomly divided into four groups. Similar cavities were created on the buccal surfaces of roots, 5 mm from their apices. The root canals in each of the study groups were filled with curcumin, aloe vera, calcium hydroxide or normal saline. The pH was measured after 20 min (i.e. the baseline), 1, 7, 14, 21, and 28 days using a digital pH meter. The data were analysed using repeated-measures ANOVA and the statistical significance was set at P<0.05. Results: At the baseline, day 1 and day 7, the mean pH of both curcumin and aloe vera groups was higher than the mean pH of calcium hydroxide and normal saline groups (P<0.05). On day 14, the mean pH of aloe vera group was higher than that of calcium hydroxide and normal saline groups (P<0.05). On days 21 and 28, the mean pH of aloe vera group was higher than the mean pH of all the other groups (P<0.05). All other intergroup differences were not statistically significant at each time point (P>0.05). Conclusion: The current in vitro study demonstrated that aloe vera was more alkaline than curcumin; nevertheless, both groups exhibited more alkalinity than calcium hydroxide.

Microhardness of Calcium-enriched Mixture Cement and Covering Glass Ionomers after Different Time Periods of Application.

Introduction: Various studies have recommended using calcium-enriched mixture (CEM) cement in different endodontic treatments, including vital pulp therapy. However, possible reciprocal effects of the covering glass ionomer cement (GIC) on their mechanical properties have not been yet investigated in detail. The current research aimed to experimentally evaluate the surface microhardness of CEM cement and the covering GICs after different application/testing times. Materials and Methods: Using stainless steel moulds (8×4×4 mm), CEM cement samples were prepared (n=120) and randomly divided into 12 experimental groups (n=10). CEM cement with thickness of 4 mm was inserted into the moulds, and the remaining spaces were filled with self-cured or light-cured resin-modified GICs at three-time intervals; immediate, in 15 min and after 24 h. Then, the samples were incubated for one and seven days. Using a Vickers microhardness tester, the microhardness of CEM and GICs was measured. The data were analyzed using two-way ANOVA and Tukey's test, and the significance level was set at 5% (P<0.05). Results: The reciprocal effects of the type/time of application of GICs on the surface microhardness of CEM cement or GICs were statistically significant (P<0.001). The surface microhardness of CEM cement and both covering GICs significantly increased over time and in seven-day samples was significantly higher than in one-day samples (P<0.05). Conclusions: Low surface microhardness of CEM/GICs in short-term (24 h) seems transient; and appears to be compensated over a longer period (i.e. 7-day). Therefore, using GICs adjacent to CEM cement in single-visit restorative treatments may be advocated.

Bioactive chitosan biguanidine-based injectable hydrogels as a novel BMP-2 and VEGF carrier for osteogenesis of dental pulp stem cells.

Nowadays, vascularization and mineralization of bone defects is the main bottleneck in the bone regeneration field that is needed to be overcome and developed. Here, we prepared novel in-situ formed injectable hydrogels based on chitosan biguanidine and carboxymethylcellulose loaded with vascular endothelial growth factor (VEGF) and recombinant Bone morphogenetic protein 2 (BMP-2) and studied its influence on osteoblastic differentiation of dental pulp stem cells (DPSCs). The sequential release behavior of the VEGF and BMP-2 from hydrogels adjusted with the pattern of normal human bone growth. MTT assay exhibited that these hydrogels were non-toxic and significantly increased DPSCs proliferation. The Real-time PCR and Western blot analysis on CG11/BMP2-VEGF showed significantly higher gene and protein expression of ALP, COL1α1, and OCN. These results were confirmed by mineralization assay by Alizarin Red staining and Alkaline phosphatase enzyme activity. Based on these evaluations, these hydrogel holds potential as an injectable bone tissue engineering platform.

Electrochemical immunoplatform to assist in the diagnosis of oral cancer through the determination of CYFRA 21.1 biomarker in human saliva samples: Preparation of a novel portable biosensor toward non-invasive diagnosis of oral cancer.

In this study, a novel, low-cost, and flexible paper-based electrochemical immunosensor was developed for the bioanalysis of Cyfra 21.1 biomarker in human saliva samples by using stabilization of synthesis Ag nano-ink on the surface of paper using pen-on-paper technology. The employed electrochemical techniques for the evaluation of immunoplatform performance were differential pulse voltammetry and chronoamperometry. Also, the prepared immunosensor showed great ability in the determination of Cyfra21.1 in human saliva specimens. Under the optimized conditions, the obtained linear range was from 0.0025 to 10 ng/mL, and the obtained LLOQ was 0.0025 ng/mL. The developed immunosensor is easy to prepare, sensitive, cost-effective, portable, and simple. So proposed immunoplatform can be an accomplished biodevice in clinical laboratories. The proposed paper-based immunosensor could be a hopefully new and cheap tool for the diagnosis of other biomarkers. Also, the prepared immunosensor showed great ability in the determination of Cyfra21.1 biomarker in human saliva specimens.

Early Osteogenic Differentiation Stimulation of Dental Pulp Stem Cells by Calcitriol and Curcumin.

Curcumin, as a natural phenolic substance, is extracted from the rhizome of Curcuma longa (turmeric), which is effective in bone healthfulness. Calcitriol is an effective hormone in regulating bone remodeling and mineral homeostasis and immune response. Mesenchymal stem cells (MSCs) are found in most dental tissues and resemble bone marrow-derived MSCs. In this work, we investigated the effect of combination and individual treatment of curcumin and calcitriol on early osteogenic differentiation of dental pulp stem cells (DPSCs). Early osteogenic differentiation was evaluated and confirmed by the gene expression level of ALP and its activity. Curcumin individually and in combination with calcitriol increased ALP activity and osteoblast-specific mRNA expression of ALP when DPSCs were cultured in an osteogenic medium. Calcitriol alone increased the enzyme more than in combination with curcumin. These findings demonstrate that curcumin can induce early osteogenic differentiation of DPSCs like calcitriol as a potent stimulant of osteogenesis.

Chemical binding of pyrrolidinyl peptide nucleic acid (acpcPNA-T9) probe with AuNPs toward label-free monitoring of miRNA-21: A novel biosensing platform for biomedical analysis and POC diagnostics.

miRNAs are attractive factors in cancer research studies due to their important roles for regulating of gene expression. Because of miRNA-21 expression surplus in many types of cancers, so accurate identification is important. Increasing efforts have caused different methods to improve the sensitivity and specificity of detection. Present study is an attempt to report a new electrochemical label-free PNA-based bioassay for detection of miRNA-21. In this study, gold electrode was modified by gold nanoparticles to improve a functional PNA-based biosensor. The EDS and field emission scanning electron microscope (FE-SEM) were used to detect fabrication of biosensor. The electrochemical behavior of sensor was evaluated after inserting of acpcPNA probes and miRNA-21 on the stucture of electrode and analyzed essential parameters such as various concentration of target miRNA, hybridization time, reproducibility, stability, and applicability. The results of study demonstrated that engineered biosensor was successfully fabricated. The findings showed the highest amount of current in 5 minutes hybridization time, with suitable reproducibility and stability. This innovative miRNA-based biosensor presents a sensitive and specific method in fast and may be lab-on chip assay in future.

Flexible paper-based label-free electrochemical biosensor for the monitoring of miRNA-21 using core-shell Ag@Au/GQD nano-ink: a new platform for the accurate and rapid analysis by low cost lab-on-paper technology.

miRNA-21 is one of the most famous and prominent microRNAs that is important in the development and emergence of cancers. So, the sensitive and selective monitoring of miRNA-21 as a very common biomarker in cancer treatment is necessary. In this work, a novel paper-based electrochemical peptide nucleic acid (PNA) sensor was developed for the detection of miRNA-21 in human plasma samples by using Ag@Au core-shell nanoparticles electrodeposited on graphene quantum dots (GQD) conductive nano-ink (Ag@Au core-shell/GQD nano-ink), which was designed directly by writing pen-on paper technology on the surface of photographic paper. This nano-ink has a great surface area for biomarker immobilization. The prepared paper-based biosensor is very small and cheap, and also has high stability and sensitivity. Hybridization of PNA was measured using various electrochemical techniques, such as cyclic voltammetry (CV), square wave voltammetry (SWV) and chronoamperometry (ChA). FE-SEM (Field Scanning Electron Microscope), TEM (Transmission Electron Microscope), EDS and DLS (Dynamic Light Scattering) tests were performed to identify the engineering safety sensor. Under optimal conditions, the linear range for the calibration curve was from 5 pM to 5 µM, and the achieved LLOQ was 5 pM. The obtained results recommended that the proposed bioassay might be suitable for an early diagnosis of cancer based on the inhibition of the expression of miRNA-21, which activates the enzyme caspase and accelerates apoptotic proteins and death in tumor cells.

A Comprehensive Review of Detection Methods for SARS-CoV-2.

Recently, the outbreak of the coronavirus disease 2019 (COVID-19), caused by the SARS-CoV-2 virus, in China and its subsequent spread across the world has caused numerous infections and deaths and disrupted normal social activity. Presently, various techniques are used for the diagnosis of SARS-CoV-2 infection, with various advantages and weaknesses to each. In this paper, we summarize promising methods, such as reverse transcription-polymerase chain reaction (RT-PCR), serological testing, point-of-care testing, smartphone surveillance of infectious diseases, nanotechnology-based approaches, biosensors, amplicon-based metagenomic sequencing, smartphone, and wastewater-based epidemiology (WBE) that can also be utilized for the detection of SARS-CoV-2. In addition, we discuss principles, advantages, and disadvantages of these detection methods, and highlight the potential methods for the development of additional techniques and products for early and fast detection of SARS-CoV-2.

The osteogenic differentiation of human dental pulp stem cells in alginate-gelatin/Nano-hydroxyapatite microcapsules.

BACKGROUND: Microcapsule is considered as a promising 3D microenvironment for Bone Tissue Engineering (BTE) applications. Microencapsulation of cells in an appropriate scaffold not only protected the cells against excess stress but also promoted cell proliferation and differentiation. Through the current study, we aimed to microcapsulate the human Dental Pulp Stem Cells (hDPSCs) and evaluated the proliferation and osteogenic differentiation of those cells by using MTT assay, qRT-PCR, Alkaline phosphatase, and Alizarine Red S. RESULTS: The SEM results revealed that Alg/Gel microcapsules containing nHA showed a rough and more compact surface morphology in comparison with the Alg/Gel microcapsules. Moreover, the microencapsulation by Alg/Gel/nHA could improve cell proliferation and induce osteogenic differentiation. The cells cultured in the Alg/Gel and Alg/Gel/nHA microcapsules showed 1.4-fold and 1.7-fold activity of BMP-2 gene expression more in comparison with the control group after 21 days. The mentioned amounts for the BMP-2 gene were 2.5-fold and 4-fold more expression for the Alg/Gel and Alg/Gel/nHA microcapsules after 28 days. The nHA, addition to hDPSCs-laden Alg/Gel microcapsule, could up-regulate the bone-related gene expressions of osteocalcin, osteonectin, and RUNX-2 during the 21 and 28 days through the culturing period, too. Calcium deposition and ALP activities of the cells were observed in accordance with the proliferation results as well as the gene expression analysis. CONCLUSION: The present study demonstrated that microencapsulation of the hDPSCs inside the Alg/Gel/nHA hydrogel could be a potential approach for regenerative dentistry in the near future.