Confocal Laser Microscopy Studies Research Articles

Evaluation of 3D-Printed Solid Microneedles Coated with Electrosprayed Polymeric Nanoparticles for Simultaneous Delivery of Rivastigmine and N-Acetyl Cysteine.

In the current study, coated microneedle arrays were fabricated by means of digital light processing (DLP) printing. Three different shapes were designed, printed, and coated with PLGA particles containing two different actives. Rivastigmine (RIV) and N-acetyl-cysteine (NAC) were coformulated via electrohydrodynamic atomization (EHDA), and they were incorporated into the PLGA particles. The two actives are administered as a combined therapy for Alzheimer's disease. The printed arrays were evaluated regarding their ability to penetrate skin and their mechanical properties. Optical microscopy and scanning electron microscopy (SEM) were employed to further characterize the microneedle structure. Confocal laser microscopy studies were conducted to construct 3D imaging of the coating and to simulate the diffusion of the particles through artificial skin samples. Permeation studies were performed to investigate the transport of the drugs across human skin ex vivo. Subsequently, a series of tape strippings were performed in an attempt to examine the deposition of the APIs on and within the skin. Light microscopy and histological studies revealed no drastic effects on the membrane integrity of the stratum corneum. Finally, the cytocompatibility of the microneedles and their precursors was evaluated by measuring cell viability (MTT assay and live/dead staining) and membrane damages followed by LDH release.

Targeted Delivery of 5-Fluorouracil and Sonidegib via Surface-Modified ZIF-8 MOFs for Effective Basal Cell Carcinoma Therapy.

The therapeutic effectiveness of the most widely used anticancer drug 5-fluorouracil (5-FU) is constrained by its high metabolism, short half-life, and rapid drug resistance after chemotherapy. Although various nanodrug delivery systems have been reported for skin cancer therapy, their retention, penetration and targeting are still a matter of concern. Hence, in the current study, a topical gel formulation that contains a metal-organic framework (zeolitic imidazole framework; ZIF-8) loaded with 5-FU and a surface modified with sonidegib (SDG; acting as a therapeutic agent as well as a targeting ligand) (5-FU@ZIF-8 MOFs) is developed against DMBA-UV-induced BCC skin cancer in rats. The MOFs were prepared using one-pot synthesis followed by post drug loading and SDG conjugation. The optimized MOFs were incorporated into hyaluronic acid-hydroxypropyl methyl cellulose gel and further subjected to characterization. Enhanced skin deposition of the 5-FU@ZIF-8-SDG MOFs was observed using ex vivo skin permeation studies. Confocal laser microscopy studies showed that 5-FU@ZIF-8-SDG MOFs permeated the skin via the transfollicular pathway. The 5-FU@ZIF-8-SDG MOFs showed stronger cell growth inhibition in A431 cells and good biocompatibility with HaCaT cells. Histopathological studies showed that the efficacy of the optimized MOF gels improved as the epithelial cells manifested modest hyperplasia, nuclear pleomorphism, and dyskeratosis. Additionally, immunohistochemistry and protein expression studies demonstrated the improved effectiveness of the 5-FU@ZIF-8-SDG MOFs, which displayed a considerable reduction in the expression of Bcl-2 protein. Overall, the developed MOF gels showed good potential for the targeted delivery of multifunctional MOFs in topical formulations for treating BCC cancer.

Protocol on comparative evaluation of the effect of application of 38% silver diamine fluoride on sealing ability of Biodentine and MTA Angelus as a perforation repair material: a confocal laser microscopic study

Background: Perforation can occur while performing root canal treatment. Biodentine has been made utilizing MTA (mineral trioxide aggregate) as a base, and claims to improve its properties in aspects such as physical property, durability, and handling, as well as its many other applications in restorative dentistry. The chemical components of SDF (silver diamine fluoride) have been suggested to have the following benefits: fluoride aids in remineralization and prevention, silver salts induce dentin sclerosis/calcification, and exhibit germicidal actions. It works by occluding dentinal tubules thus helping in preventing microleakage. This study aims to focus on the use of SDF on Biodentine and MTA as perforation repair material using a confocal microscope. Methods: Mandibular molars will be selected for the study. The study will be divided into 4 groups: Group 1: Biodentine without application of 38% SDF; Group 2: Biodentine with the application of 38% SDF; Group 3: MTA Angelus without application of 38% SDF; and Group 4: MTA Angelus along with the application of 38% SDF. An artificial perforation 2 mm in diameter will be made directly in the middle of the pulpal floor. 38% silver diamine fluoride (FAMAGAIN) will be applied on the walls of the perforation according to the manufacturer’s instructions for the experimental groups (Groups 2 and 4). In the four groups, Biodentine will be applied in groups 1 and 2 and MTA will be applied in 3 and 4 (perforation repair material). To evaluate the sealing capacity of perforation material in this study, we will use the average values of coronal dye leakage at the perforation site and Escobar’s criteria, which are used to assess the infiltration proportions. Conclusions: It is expected that application of 38% of silver diamine fluoride on MTA Angelus and Biodentine may show better sealing ability than MTA Angelus and Biodentine alone.

Effect of continuous chelation on the dentinal tubule penetration of a calcium silicate–based root canal sealer: a confocal laser microscopy study

BackgroundThis study aimed to evaluate the effect of various irrigation protocols on the penetration depth of a calcium silicate–based sealer into dentinal tubules using confocal laser scanning microscopy (CLSM).MethodsTwenty single-rooted mandibular premolars were endodontically prepared and divided into the following two groups according to the irrigation protocol used (n = 10): Group I: NaOCl + EDTA and Group II: continuous chelation (NaOCl/Dual Rinse). Obturation was performed with the warm vertical compaction technique using TotalFill HiFlow bioceramic sealer mixed with a fluorophore dye. Samples were observed using CLSM at 10× to measure the percentage of sealer penetration and its maximum depth into the dentinal tubules. Data were analysed using one-way ANOVA followed by Tukey’s post-hoc test. The significance level was set at p < 0.05 within all tests.ResultsComparing the overall results of all sections tested, no statistically significant differences existed between the groups regarding the percentage of sealer penetration (p = 0.612) and maximum depth of penetration (p > 0.05).ConclusionsWith both types of irrigation used, dentinal tubule penetration was higher in the coronal section than in the apical section. Continuous chelation using NaOCl/Dual Rinse HEDP performed better in the coronal segments, while irrigation using NaOCl + EDTA promoted a higher percentage of sealer penetration in the apical segment.

Fabrication of Polymeric Microneedles using Novel Vacuum Compression Molding Technique for Transdermal Drug Delivery.

To demonstrate the feasibility of vacuum compression molding as a novel technique for fabricating polymeric poly (D, L-lactic-co-glycolic acid) microneedles. First, polydimethylsiloxane molds were prepared using metal microneedle templates and fixed in the MeltPrep® Vacuum Compression Molding tool. Poly (D, L-lactic-co-glycolic acid) (EXPANSORB® DLG 50-5A) was added, enclosed, and heated at 130°C for 15 min under a vacuum of -15 psi, cooled with compressed air for 15 min, followed by freezing at -20°C for 30 min, and stored in a desiccator. The microneedles and microchannels were characterized by a variety of imaging techniques. In vitro permeation of model drug lidocaine as base and hydrochloride salt was demonstrated across intact and microporated dermatomed human skin. Fabricated PLGA microneedles were pyramid-shaped, sharp, uniform, and mechanically robust. Scanning electron microscopy, skin integrity, dye-binding, histology, and confocal laser microscopy studies confirmed the microchannel formation. The receptor delivery of lidocaine salt increased significantly in microporated (270.57 ± 3.73 μg/cm2) skin as compared to intact skin (142.19 ± 13.70 μg/cm2) at 24 h. The receptor delivery of lidocaine base from microporated skin was significantly higher (312.37 ± 10.57 μg/cm2) than intact skin (169.68 ± 24.09 μg/cm2) up to 8 h. Lag time decreased significantly for the base (2.24 ± 0.17 h to 0.64 ± 0.05 h) and salt (4.76 ± 0.31 h to 1.47 ± 0.21 h) after microporation. Vacuum compression molding was demonstrated as a novel technique to fabricate uniform, solvent-free, strong polymer microneedles in a short time.

Comparative Evaluation of Microleakage of Flowable Composite Resin Using Etch and Rinse, Self-Etch Adhesive Systems, and Self-Adhesive Flowable Composite Resin in Class V Cavities: Confocal Laser Microscopic Study.

The essential factor in determining the preservation of restoration is the marginal seal. Restoring cervical lesions with a resin composite has always been a challenge. Composite resins with various viscosities and different bonding systems are being researched to reduce the microleakage. Confocal laser scanning microscopy (CLSM) is the latest non-destructive technique for visualizing the microleakage. Objectives: To evaluate and compare the microleakage of Universal Flo composite resin (G-aenial) using etch and rinse adhesive system ER-2 steps (Adper Single Bond 2), self-etch adhesive system SE-1 step (G-Bond), and self-adhesive flowable composite resin (Constic) in Class V cavities using a confocal laser scanning microscope. Materials and Method: Class V cavities were prepared on 27 caries-free human extracted premolar teeth on the buccal and lingual surfaces with standardized dimensions of 2 mm height, width 4 mm, and a depth of 2 mm. After the cavity preparation, all teeth were randomly divided into three groups, namely Group-I: G-aenial Universal Flo with Single Bond 2 (n = 9 teeth); Group-II: G- aenial Universal Flo with G-Bond (n = 9 teeth), and Group-III: Constic (n = 9 teeth). The prepared and restored specimens were then subjected to thermocycling for 500 cycles in a water bath at 5 °C and 55 °C with a dwelling time of 30 s. The specimens were placed in 0.6% aqueous rhodamine dye for 48 h. Sectioning was carried out bucco-lingually and specimens were evaluated for microleakage under a confocal laser scanning microscope. Results: There was a significant difference (p = 0.009) in microleakage when comparing total etch and rinse, specifically between Adper Single Bond 2 ER-2 steps (fifth generation) and self-adhesive flowable composite resin, which is Constic. There was more microleakage in the self-etch bonding agent, particularly G-Bond, SE-1 step (seventh generation), when compared to ER-2 steps (fifth generation bonding agent); however, the results were not statistically significant (p = 0.468). The self-adhesive flowable composite resin showed more microleakage than SE-1 step and ER-2 steps. Conclusions: None of the adhesive systems tested were free from microleakage. However, less microleakage was observed in the total etch and rinse, especially Adper Single Bond 2 (ER-2 steps), than the self-etch adhesive system SE-1 step and self-adhesive flowable composite resin. Clinical significance: Constant research and technological advancements are taking place in dentin adhesives to improve the marginal seal. This has led to the evolution of total acid-etching dentin bonding agents termed as etch and rinse (ER)-2 steps (fifth generation dentin bonding agents) and self-etching (SE) 2 steps, and SE-1 step dentin bonding agents termed as the sixth and seventh generation bonding agents, respectively.

Inter-Genera Colonization of Ocimum tenuiflorum Endophytes in Tomato and Their Complementary Effects on Na+/K+ Balance, Oxidative Stress Regulation, and Root Architecture Under Elevated Soil Salinity.

Endophytic bacilli of ethano-botanical plant Ocimum tenuiflorum were screened for salt stress-alleviating traits in tomato. Four promising O. tenuiflorum endophytes (Bacillus safensis BTL5, Bacillus haynesii GTR8, Bacillus paralicheniformis GTR11, and Bacillus altitudinis GTS16) were used in this study. Confocal scanning laser microscopic studies revealed the inter-genera colonization of O. tenuiflorum endophytes in tomato plants, giving insights for widening the applicability of potential endophytes to other crops. Furthermore, in a pot trial under 150 mM NaCl concentration, the inoculated endophytes contributed in reducing salt toxicity and improving recovery from salt-induced oxidative stress by different mechanisms. Reduction in reactive oxygen species (ROS) (sub-cellular H2O2 and superoxide) accumulation was observed besides lowering programmed cell death and increasing chlorophyll content. Endophyte inoculation supplemented the plant antioxidant enzyme system via the modulation of enzymatic antioxidants, viz., peroxidase, ascorbate peroxidase, superoxide dismutase, and catalase, apart from increasing proline and total phenolics. Antioxidants like proline have dual roles of antioxidants and osmoregulation, which might also have contributed to improved water relation under elevated salinity. Root architecture, viz., root length, projection area, surface area, average diameter, tips, forks, crossings, and the number of links, was improved upon inoculation, indicating healthy root growth and enhanced nutrient flow and water homeostasis. Regulation of Na+/K+ balance and water homeostasis in the plants were also evident from the modulation in the expression of abiotic stress-responsive genes, viz., LKT1, NHX1, SOS1, LePIP2, SlERF16, and SlWRKY39. Shoot tissues staining with light-excitable Na+ indicator Sodium GreenTM Tetra (tetramethylammonium) salt showed low sodium transport and accumulation in endophyte-inoculated plants. All four endophytes exhibited different mechanisms for stress alleviation and indicated complementary effects on plant growth. Furthermore, this could be harnessed in the form of a consortium for salt stress alleviation. The present study established inter-genera colonization of O. tenuiflorum endophytes in tomato and revealed its potential in maintaining Na+/K+ balance, reducing ROS, and improving root architecture under elevated salinity.

Development of carboplatin loaded bovine serum albumin nanoparticles and evaluation of its effect on an ovarian cancer cell line

Ovarian cancer has the highest mortality rate among gynecological cancers. It requires urgent therapy, but standard treatments are not sufficient to cure ovarian cancer completely. Chemotherapy has critical side effects due to non-specific toxicity. In this regard, nanoparticulate drug delivery systems, one of the most promising therapeutic approaches, have been developed for cancer therapy. The main objective of the present research was to develop novel bovine serum albumin (BSA) nanoparticle formulation encapsulating carboplatin (CRB) and to evaluate its anti-cancer activity against A2780 human ovarian cancer cell line. CRB-loaded BSA nanoparticles were prepared using a desolvation method and characterized by several physicochemical properties, including encapsulation efficiency, particle size and size distribution, surface charge, particle morphology, thermal behavior. Different CRB-loaded BSA nanoparticles were prepared using 33 factorial design in which varying the BSA concentration, ethanol/BSA solution ratio and glutaraldehyde/BSA ratio as three factors. The effects of these factors on the particle size, polydispersity index and encapsulation efficiency were evaluated to produce an optimized nanoformulation. Based on dependent variables, the formulation including 3% BSA concentration, 4/1 (v/v) ethanol/BSA solution ratio (v/v), 7.80 μg/mg glutaraldehyde/BSA ratio was selected as optimum. It was found to have spherical shapes with negative surface charges and about 253 nm particle size with a low polydispersity index indicating a narrow particle size distribution. According to the confocal laser microscopy and flow cytometer studies, it exhibited a high uptake into A2780 cells. In addition, cytotoxicity experiments revealed that CRB-loaded albumin nanoparticles showed more potent anti-cancer effect on A2780 ovarian cancer cell line with approximately 2-fold lower IC50 value, compared with free CRB.

Brucine-loaded transliposomes nanogel for topical delivery in skin cancer: statistical optimization, in vitro and dermatokinetic evaluation.

The aim of the present study was to develop, optimize brucine-loaded transliposomes (BRC-TL) formulation for dermal delivery of brucine for skin cancer. The BRC-TL formulations were evaluated for vesicle size, entrapment efficiency, and in vitro drug release. The optimized formulation was further evaluated for skin penetration by confocal laser microscopy and dermatokinetic study. The optimized BRC-TL formulation presented sealed lamellar shaped vesicles, with vesicles size, polydispersity index, entrapment efficiency, and in vitro drug release of 136.20 ± 2.87nm, 0.354 ± 0.02, 86.01 ± 1.27%, and 83.09 ± 2.07%, respectively. Ex vivo permeation study showed that, developed BRC-TL formulation had a 2.4-fold increment in permeation as compared to BRC suspension. Texture analysis showed that the BRC-TL gel presented firmness of 158.91g, consistency of 615.03g/s, cohesiveness of - 115.26g and a viscosity index of - 472.05g/s. The confocal images of rat skin clearly showed the deeper penetration of rhodamine B-loaded TL formulation as compared to the Rhodamine B-hydro alcoholic solution. The optimized BRC-TL formulation demonstrated significantly higher cytotoxicity than placebo liposome and BRC suspension (P < 0.05). Further, the BRC-TL nanogel treated rat skin showed a substantial increase in CSkin max and AUC0-8 in comparison to rat skin treated with BRC conventional gel (P < 0.05). The data revealed that the developed TLs formulation could be a promising drug nanocarrier for brucine dermal delivery in the treatment of skin cancer.

Synthesis and characterization of chitosan silver nanoparticle decorated with benzodioxane coupled piperazine as an effective anti-biofilm agent against MRSA: A validation of molecular docking and dynamics

Biomaterial research has improved the delivery and efficacy of drugs over a wide range of pharmaceutical applications. The objective of this study was to synthesize benzodioxane coupled piperazine decorated chitosan silver nanoparticle (Bcp*C@AgNPs) against methicillin-resistant Staphylococcus aureus (MRSA) and to assess the nanoparticle as an effective candidate for antibacterial and anti-biofilm care. Antibacterial activity of the compound was examined and minimum inhibitory concentration (MIC) was observed at (10.21 ± 0.03 ZOI) a concentration of 200 μg/mL. The Bcp*C@AgNPs interferes with surface adherence of MRSA, suggesting an anti-biofilm distinctive property that is verified for the first time by confocal laser microscopic studies. By ADMET studies the absorption, distribution, metabolism, excretion and toxicity of the compound was examined. The interaction solidity and the stability of the compound when surrounded by water molecules were analyzed by docking and dynamic simulation analysis. The myoblast cell line (L6) was considered for toxicity study and was observed that the compound exhibited less toxic effect. This current research highlights the biocidal efficiency of Bcp*C@AgNPs with their bactericidal and anti-biofilm properties over potential interesting clinical trial targets in future.

Zwitterionic poly(carboxybetaine) microgels for enzyme (chymotrypsin) covalent immobilization with extended stability and activity

AbstractThere is emerging evidence that biocompatible zwitterionic materials can prevent nonspecific interactions within protein systems and increase protein stability. Here, a zwitterionic microgel was synthesized from poly (carboxybetaine methyl methacrylate) (pCB) using an inverse emulsion, free radical polymerization reaction technique. The microgel was loaded with a model enzyme, α‐chymotrypsin (ChT), using a post‐fabrication loading technique. A reaction scheme was developed and studied for covalent immobilization of ChT within the microgel. Confocal laser microscopy studies showed that immobilized ChT (i‐ChT) was distributed within the hydrogel. The enzyme‐immobilized microgels showed excellent reusability (72% of its initial activity after 10 uses) and could undergo several freezing/drying/rehydration cycles while retaining enzymatic activity. The i‐ChT activity, half‐life, and conformational stability were studied at varying pH and temperatures with results compared to free ChT in buffer. ChT immobilized within pCB hydrogel showed increased enzymatic stability as observed by a 13°C increase in the temperature at which i‐ChT loses activity compared to free ChT. Furthermore, enzyme half‐life increased up to seven‐fold for the pCB immobilized ChT, and the increased stability resulted in higher activity at elevated pH. The i‐ChT was most active at pH of 8.5 and was partially active up to the pH of 10.2.

Comparison between Allogenic and Xenogenic Bone Blocks on the Osteogenic Potential of Cultured Human Periodontal Ligament Stem Cells: Confocal Laser and Scanning Electron Microscopy Study.

Aim:The aim of this study was to compare between equine and human bone blocks in the osteogenic differentiation of cultured human periodontal ligament stem cells (hPDLSCs) at 14 and 21 days of culture, using confocal laser microscopy and scanning electron microscopy.Materials and Methods:In vitro cultures of commercially obtained hPDLSCs were seeded onto equine and human bone blocks. At 14 days and 21 days of culture, confocal laser microscope images were obtained to assess cellular differentiation and adhesion, and scanning electron microscope images were obtained to validate the osteogenic differentiation by showing the morphological characteristics of the new bone cells.Results:Both equine and human bone blocks showed positive staining for newly formed bone cells through the confocal laser microscope analysis, however, a higher signal intensity was expressed at 21 days of culture. These findings indicate the biocompatibility of hPDLSC with both types of bone blocks, cellular differentiation, and adhesion. Scanning electron microscopy images validated the osteogenic differentiation by showing the common characteristics of bone cells as flattened, polygonal morphology with multiple extending cytoplasmic processes.Conclusion:Both equine and human bone blocks were able to confirm the osteogenic capability of seeded human PDLSC. There was no significant difference between equine and human bone blocks on the human PDLSC differentiation. Superior osteogenic differentiation of cultured hPDLSCs was evident at 21 days in comparison to 14 days.

Formulation and evaluation of thymoquinone niosomes: application of developed and validated RP-HPLC method in delivery system

A rapid, accurate, and sensitive reverse phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated for the estimation of Thymoquinone (TMQ) in API as well as in noisome. The chromatograms were developed with the mobile phase – water: 2-propanol: methanol (50:45:5 v/v/v) as a solvent system at 254 nm. The method was validated as per ICH guidelines for different parameters and the recovery of TMQ was calculated in developed niosomes. Further, TMQ loaded niosomes (TMQNIOS) were prepared and evaluated for different parameters. The optimized TMQNIOS (F3) was further evaluated for surface morphology, in vitro drug release, permeation study, and confocal laser scanning microscopic (CLSM) study. The method showed linearity range between 6.25 and 100 µg/ml with low detection limit and quantitation limit with a value of 2.08 and 6.25 µg/ml. The developed formulations showed the vesicle size and encapsulation efficiency in the range of 157.32 ± 3.15 to 211.44 ± 5.23 nm and 59.32 ± 4.87 to 83.21 ± 3.55%, respectively. The drug release result showed the significant higher release from TMQNIOS in compared to TMQDIS, and the release kinetics data showed Higuchi's equation with highest regression coefficient values. The permeation study and the confocal laser microscopy study further confirmed the enhancement in permeation of TMQ in the intestinal mucosa.

Investigations on Polymeric Nanoparticles for Ocular Delivery

In the present investigation, an attempt was made to formulate timolol maleate (TML) loaded polymeric nanoparticles of flax seed gum (FX) and chitosan (CH) for ocular delivery using ionic gelation method. The process of nanoparticle preparation was optimized using 2-factor, 3-level central composite experimental design. The optimal concentration of FX and CH that yielded nanoparticles with minimum particle size (267.06 ± 8.65 nm) and maximum encapsulation efficiency (74.96 ± 4.78%) was found to be 0.10% w/v and 0.08% w/v, respectively. The formulated nanoparticles revealed considerable bioadhesive strength and exhibited sustained release of drug in in vitro diffusion studies. The ex vivo transcorneal penetration study revealed higher corneal penetration of TML compared to marketed eye drops. The confocal scanning laser microscopy (CSLM) studies also confirmed the ability of nanoparticles to penetrate into deeper layers of cornea. The histopathological studies revealed corneal biocompatibility of nanoparticles. The nanoparticles were found to reduce the intra ocular pressure (IOP) in rabbits for prolonged period when compared to conventional eye drops. The results of the present study suggested a promising role of polymeric nanoparticles for ocular drug delivery in treatment of glaucoma.

Improved bioavailability of raloxifene hydrochloride using limonene containing transdermal nano-sized vesicles

The present investigation was carried out with the aim of optimization of raloxifene hydrochloride-entrapped, limonene containing transfersomes for transdermal delivery. Several formulations were prepared by varying independent variables such as Phospholipid, sodium cholate and limonene while vesicles size, entrapment efficiency and polydispersity index were the dependent variables for Box-Behnken Design. Further, bioavailability assessment of raloxifene in Wistar rats was evaluated after transdermal application and compared with its oral formulation. It was observed that the optimized raloxifene loaded transfersomes formulation showed vesicle size of 107.8 nm, polydispersity index of 0.252 and entrapment efficiency of 90.2% which is close to the predicted values of 114.45 nm, 0.251 and 82.82% respectively generated by the Design Expert software. The TEM image shows the outline and the core of the well identified sealed spherical structure. Confocal laser microscopy study demonstrated that the prepared transfersomes formulation was capable to increase the permeation of probe dye into deeper layer of rat's skin in comparison to the control dye solution. The transdermal flux presented by the transfersomes gel formulation was found to be 13.20 μg/cm2/h whereas the control gel showed the transdermal flux of 2.35 μg/cm2/h across rat skin. Further the in vivo pharmacokinetic study revealed that the relative bioavailability following application of transfersomes gel in Wistar rats was found be increased by 2.71 times as compared to the oral suspension of raloxifene. It was concluded that the prepared transfersomes formulation was found to be a potentially useful drug carrier for the enhancement of raloxifene bioavailability in rats.

Lamotrigine encapsulated intra-nasal nanoliposome formulation for epilepsy treatment: Formulation design, characterization and nasal toxicity study.

The purpose of this study was to develop lamotrigine nanoliposomes (LTG-NLs) for the treatment in seizures. The formulation was prepared using thin film hydration and rehydration method using the phospholipon 90 G, cholesterol and tween 80 as main ingredients. The nanoliposomes were optimized by plucket burman design (PBD) and response surface methodology (RSM) optimization techniques. The optimized LTGNLopt was further characterized for surface morphology, in-vitro release, stability study, confocal laser scanning microscopic (CLSM) study and naso toxicity study. The results showed that LTGNLopt shown nano size with high entrapment and drug release. The ex-vivo permeation study and confocal laser microscopy study confirmed the enhancement in permeation across the goat nasal mucosa. From the study, it was concluded that the independent variables used to optimize the NLs shown significant effect on the dependent variables and consider effective lipid carrier system for intranasal delivery.

Antimicrobial effect of bioceramic cements on multispecies microcosm biofilm: a confocal laser microscopy study

To assess the viability of multispecies microcosm biofilm after contact with NeoMTA Plus, Biodentine, and MTA Angelus. Fifty-four human dentin blocks (4 × 5 × 4mm) were allocated to Hawley retainers, worn by six volunteers for 72h. The blocks were then individually incubated in BHI broth for 21days at 37°C. At the end of experimental time for biofilm growth, the samples were randomly divided into four groups (n = 12): NeoMTA Plus, Biodentine, MTA Angelus, and negative control. The materials were placed in contact with the blocks. All samples were placed in cell-culture plate wells and incubated in BHI broth for 7days at 37°C. One sample from each volunteer (n = 6) was analyzed by SEM to describe the biofilm morphology. CLSM was performed to determine the percentage of viable biofilm biovolume. The data were statistically analyzed by one-way ANOVA and Tukey's multiple comparison test (α = 5%). SEM showed biofilm formed by spherical and rod-shaped bacteria surrounded by an extracellular matrix. No material was able to kill all biofilm cells, and all groups had more than 50% of viable bacteria. NeoMTA Plus was significantly different from the negative control group (P < .05). All tested materials were not effective against multispecies microcosm biofilm. NeoMTA Plus, Biodentine, and MTA Angelus were not effective against multispecies microcosm biofilm. It is essential to understand that these bioceramic cements are indicated for infected clinical situations. Thus, complementary disinfection procedures should be conducted prior to filling with these materials.

Corncob as an effective, eco-friendly, and economic biosorbent for removing the azo dye Direct Yellow 27 from aqueous solutions.

The corncob is an agricultural waste generated in huge quantities during corn processing. In this paper, we tested the capacity of corncob particles for water purification by removing the azo dye Direct Yellow 27 (DY27) via biosorption. The biosorption process was investigated in terms of the kinetics, equilibria, and thermodynamics. Batch biosorption studies showed that the biosorption performance has strong inverse correlations to the solution pH and the corncob particle size, and it increases quickly with increasing contact time and initial dye concentration. The pseudo-second-order kinetic model provides the best fit to the experimental data, whereas the Redlich-Peterson isotherm model is most suitable for describing the observed equilibrium biosorption. The biosorption process is exothermic, spontaneous, and physisorption in character. Fourier transform infrared (FTIR) spectroscopy and confocal scanning laser microscopy (CSLM) studies suggest that lignocellulose and proteins play key roles in the biosorption of DY27 from aqueous solutions by corncob. Furthermore, after biosorption onto the corncob, the dye can be effectively desorbed using 0.1 M NaOH solution. Therefore, the corncob can be used as a promising biosorbent to remediate DY27-contaminated water and wastewater.

Qualitative and quantitative analysis of lateral diffusion of drugs in human skin

This study aimed to qualitatively and quantitatively analyze lateral diffusion of drugs in dermatomed human skin. Lateral diffusion of calcein and methylene blue dyes in skin was investigated using confocal laser microscopy, calcein imaging, and histology studies. In in vitro permeation studies, two linear microdialysis probes were inserted into the dermis of untreated, poly lacto-glycolic acid microneedle-treated, and ablative laser-treated skin such that one was in the center of the diffusion area and the other was parallel, at 8 mm from the central probe. Skin was mounted on Franz cells, sandwiched between donor containing diclofenac sodium solution and receptor containing phosphate buffered saline, pH 7.4. Qualitative techniques revealed faster lateral diffusion of the dyes in microneedle-treated skin than laser-treated skin. Rate of drug diffusion in the central probe in the microneedle-treated skin (11.8 ± 2.5 μg/h) was significantly higher than untreated and laser-treated skin (p < 0.05). Rate of lateral diffusion in untreated group (0.7 ± 0.1 μg/h) was significantly lower than microneedle and laser-treated skin (p < 0.05). Overall, in vitro microdialysis was demonstrated as a novel and valuable tool that can be employed for quantitative investigation of rate of vertical and lateral diffusion of drugs in intact and microporated skin.

Flow Behaviour and Microstructure of a \u03b2-Glucan Concentrate

The extensional viscosity is an important rheological characteristic of polymer melts. It is however not as frequently reported on as the shear viscosity. The extensional viscosity is of special interest when considering polymeric materials for foaming and film blowing processes. Here, the extensional (and shear) viscosity along with the melt strength and the tensile properties of the corresponding solid film of a β-glucan concentrate are reported on. A capillary viscometer equipped with a hyperbolic die, yielding a contraction flow, was used to assess the extensional viscosity of the aqueous β-glucan compound at room temperature and at elevated temperatures (110 and 130 °C). In general, the extensional viscosity as well as the shear viscosity decreased with increasing deformation rate. The influence of two different amounts of added water (40 and 50%) was also examined. As expected, both types of viscosities decreased with increasing temperature. It is suggested that gelatinization of the starch fraction in the concentrate at 110 and 130 °C contributes to temperature dependence of the viscosity. To some extent, this is supported by light microscopy and confocal scanning laser microscopy studies of the microstructure of the materials. The results reported here indicate that the β-glucan concentrate might, after some modifications, be used as a complement to fossil-based polymers and processed by conventional manufacturing techniques.