L929 Mouse Fibroblasts Research Articles

Methanolic leaf extract of Catharanthus roseus reveals wound healing activity on mouse fibroblast L929 cell lines

Methanolic leaf extract of Catharanthus roseus reveals wound healing activity on mouse fibroblast L929 cell lines

Improvement in photodynamic activity by a porphyrin-fullerene composite system in lipid membranes.

Porphyrin-fullerene composite systems are attracting great attention as photodynamic agents; however, water-soluble derivatives are still scarce. Herein, we prepared noncovalently a lipid membrane-incorporated porphyrin-fullerene composite system with relative stability in aqueous solution. As in the case of porphyrin-fullerene composite systems in nonpolar solvents, efficient formation of singlet oxygen occurred via photoinduced energy transfer between porphyrin and fullerene as the predominant pathway in the photodynamic activity under the hydrophobic conditions of the lipid membranes, resulting in enhanced photodynamic activity toward Colon26 and HeLa cells compared with the individual porphyrin and fullerene components. Furthermore, the porphyrin-fullerene composite system exhibited high selectivity toward HeLa cells over normal mouse fibroblast L929 cells.

Comparison of Antibacterial Activity, Cytotoxicity, and Fluoride Release of Glass Ionomer Restorative Dental Cements in Dentistry.

BACKGROUND This study aimed to compare the antibacterial activity, cytotoxicity, and fluoride release of 4 different glass ionomer cements (GIC). MATERIAL AND METHODS A total of 200 samples were prepared: Riva Silver, a silver-reinforced GIC; Equia Forte HT, a glass hybrid GIC; ChemFil Rock, a zinc-added GIC; and Ketac™ Molar Easymix. Using the agar diffusion test for antibacterial activity, 30 samples from each group were analyzed for Streptococcus mutans, Lactobacillus acidophilus, and Actinomyces naeslundii. The areas around the samples where no bacterial growth occurred were digitally measured and recorded. For cytotoxicity analysis, the WST-1 test was performed on 10 samples from each group using the L929 mouse fibroblast cell line. The fluoride release property was evaluated using an ion-selective electrode method on 10 samples from each group. RESULTS The group that used Ketac™ Molar Easymix showed the lowest antibacterial activity against S. mutans, L. acidophilus, and A. naeslundii. In all 3 days of cytotoxicity testing, the group that used Riva Silver was found to be the least toxic material, while the group that used ChemFil Rock did not have viable cells after the 1st day. In all 4 materials, fluoride release values gradually increased since the first day, with Ketac™ Molar Easymix having the highest fluoride release. CONCLUSIONS Of all the GICs tested, Ketac™ Molar Easymix demonstrated the least antibacterial activity despite having the highest fluoride release, while Riva Silver was found to be the least cytotoxic material.

Insights into the remarkable attenuation of hen egg white lysozyme amyloid fibril formation mediated by biogenic gold nanoparticles stabilized by quercetin-functionalized tara gum

Aberrant protein misfolding and/or aggregation and fibrillation has been linked to the pathogenesis of several debilitating chronic diseases including Alzheimer's and Parkinson's disease. Inhibiting protein amyloidogenesis has been proposed as a viable strategy to prevent or ameliorate associated disorders. Herein, we investigated the anti-amyloidogenic properties of biogenic gold nanoparticles (QTG-GNP) prepared via a simple green chemistry route and stabilized by quercetin-functionalized tara gum (QTG). The synthesized QTG-GNP was extensively characterized for its physicochemical attributes via UV–visible spectroscopy, TEM, FESEM, EDX, DLS/Zeta potential, FTIR, RAMAN, XRD, XPS, and TGA analyses, as well as for its biological properties. The results revealed that small-sized (5.01 ± 1.17 nm), well-dispersed, highly stable and round-shaped biogenic gold nanoparticles were successfully synthesized at room temperature with QTG as the sole reductant /stabilizer. Importantly, QTG-GNP demonstrated potent anti-aggregation and fibrillation inhibitory effects against amyloidogenic hen egg white lysozyme (HEWL). Also, QTG-GNP was able to dissociate pre-formed HEWL amyloid fibrils. Furthermore, the constructed nanoparticles exhibited potent anti-radical activities against DPPH and ABTS+ and were cytocompatible with mouse L929 fibroblast cells. On the basis of these findings, it was established that QTG-GNP holds strong prospects for further development as an agent for countering protein aggregation and associated disease conditions.

Temperature responsive poly(N-isopropylacrylamide-co-styrene) nanofilms for non-enzymatic cell sheet harvesting

In this study, temperature-responsive cell culture substrates with non-enzymatic cell sheet detachment attributes in response to temperature reduction were prepared. Poly(N-isopropylacrylamide) (PNIPAAm) with a physiologically proximal lower critical solution temperature (LCST) induces a temperature responsivity to the substrate, and polystyrene was used to regulate the hydrophobicity of the substrate. Different compositions of poly(NIPAAm-co-styrene) were synthesized by free radical polymerization. The resulting copolymers were characterized using Fourier transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (1HNMR) spectroscopy. The copolymer composition was calculated from 1HNMR results. The molecular weight and molecular weight distribution were measured by gel permeation chromatography (GPC). The nanofilms of the prepared copolymers were physically coated by spin-coating technique on tetramethyl orthosilicate (TMOS) pre-coated glass slides. Attenuated total reflection-FTIR (ATR-FTIR) and field emission scanning electron microscopy (FESEM) analyses showed that the spin coating procedure was performed successfully, and the change in the water contact angle (WCA) in response to temperature alteration indicated the temperature-responsive behavior of the polymer layers. L929 mouse fibroblast cell line was cultured on nanofilm-coated glass slides until a confluent cell sheet covered the entire surface. An almost integrated cell sheet was harvested by temperature reduction from 37 °C to 20 °C. The results revealed that copolymer composition is vital in harvesting contiguous cell sheets. This fabrication method, compared to others such as plasma polymerization and surface-initiated atom transfer radical polymerization (SI-ATRP), is very cost-effective and applicable for large-scale cell culture investigations.

TiO2/HA and Titanate/HA Double-Layer Coatings on Ti6Al4V Surface and Their Influence on In Vitro Cell Growth and Osteogenic Potential.

Hydroxyapatite (HA) layers are appropriate biomaterials for use in the modification of the surface of implants produced inter alia from a Ti6Al4V alloy. The issue that must be solved is to provide implants with appropriate biointegration properties, enabling the permanent link between them and bone tissues, which is not so easy with the HA layer. Our proposition is the use of the intermediate layer ((IL) = TiO2, and titanate layers) to successfully link the HA coating to a metal substrate (Ti6Al4V). The morphology, structure, and chemical composition of Ti6Al4V/IL/HA systems were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectrometry (EDS). We evaluated the apatite-forming ability on the surface of the layer in simulated body fluid. We investigated the effects of the obtained systems on the viability and growth of human MG-63 osteoblast-like cells, mouse L929 fibroblasts, and adipose-derived human mesenchymal stem cells (ADSCs) in vitro, as well as on their osteogenic properties. Based on the obtained results, we can conclude that both investigated systems reflect the physiological environment of bone tissue and create a biocompatible surface supporting cell growth. However, the nanoporous TiO2 intermediate layer with osteogenesis-supportive activity seems most promising for the practical application of Ti6Al4V/TiO2/HA as a system of bone tissue regeneration.

Cytotoxicity and Antibacterial Efficacy of AgCu and AgFe NanoAlloys: A Comparative Study.

Although Ag nanoparticles (NPs) have been widely applied in daily life and in biomedical and industrial fields, there is a demand for Ag-based bimetallic nanoalloys (NAs), such as AgCu and AgFe, due to their enhanced antibacterial efficacy and reduced Ag consumption. In this work, we present a comparison study on the antibacterial efficacy and cytotoxicity rates of Ag NPs and AgCu and AgFe NAs to L929 mouse fibroblast cells using the CCK-8 technique based on the relative cell viability. The concept of the minimum death concentration (MDC) is introduced to estimate the cytotoxicity to the cells. It is found that the minimum inhibitory concentrations (MICs) of the NPs against E. coli and S. aureus decrease with the addition of both Cu and Fe. There is a strong correlation between the MDC and MIC, implying that the mechanisms of both antibacterial efficacy and cytotoxicity are similar. The enhanced antibacterial efficacy to bacteria and cytotoxicity toward the cell are attributed to Ag+ release. The following order is found for both the MIC and MDC: AgFe < AgCu < Ag NPs. However, there is no cytotoxicity to the L929 cells for AgFe and AgCu NAs at their MIC Ag concentrations against S. aureus.

Cytotoxicity and anti-biofilm properties of novel hybrid-glass-based caries infiltrant.

To assess the cytotoxicity of an experimental hybrid-glass-based infiltrant and its effect on biofilm attachment, growth and metabolic activity, and to compare it to the resin-based infiltrant Icon. Cytotoxicity of hybrid-glass-based material (EXP) and resin-based infiltrant Icon (Icon) was tested in direct contact tests on freshly cured (direct_mat) and on materials kept for 24h in cell culture medium (direct_exmat), and extract test with materials 24-h extracts (extract). Cell viability of L929 mouse fibroblast cell line was measured with MTT assay, according to ISO10993-5:2009. Biofilm attachment (5h), growth (24h and 48h) and lactic-acid production (24h and 48h) on glass-disk specimens coated with EXP or Icon, or uncoated (control), were assessed using a microcosm biofilm model and Amsterdam Active Attachment system. At indicated time points, biofilms were harvested, plated, and CFU counts were determined, while lactic-acid production was measured colorimetrically. Cell viability reduction by EXP was below 30%-threshold in direct contact tests, while in extract test an increased cell viability was observed. Icon reduced cell viability substantially in all three tests. Significantly less bacteria attached to the surface of EXP after 5h compared to Icon and control. Biofilm growth was significantly lower on EXP than on Icon and control after 24h, but this difference was smaller and statistically insignificant after 48h. There was no difference in lactic-acid production among groups. Novel hybrid-glass-based infiltrant seems to have a better biocompatibility and accumulates on its surface less bacteria than resin-based infiltrant, which makes it an attractive resin-free alternative.

In vitro evaluation of 2-pyrazoline derivatives as DPP-4 inhibitors

Abstract Objectives In this study, the synthesis of three pyrazoline derivatives and the evaluation of their inhibitory effects on dipeptidyl peptidase (DPP-4) were aimed. Methods Pyrazoline-based compounds (1–3) were obtained via the reaction of 1-(2-furyl)-3-(1,3-benzodioxol-5-yl)-2-propen-1-one with 4-substituted phenylhydrazine hydrochloride. The DPP-4 inhibitory effects of compounds 1–3 were determined with a fluorometric assay using Gly-Pro-aminomethylcoumarin as the fluorogenic substrate. The cytotoxicity of compounds 1–3 on L929 mouse fibroblast (healthy) cell line was evaluated using MTT assay. Results 1-(4-Methylsulfonylphenyl)-3-(2-furyl)-5-(1,3-benzodioxol-5-yl)-2-pyrazoline (2) exhibited the highest DPP-4 inhibitory activity (IC50=5.75 ± 0.35 µM). Moreover, compound 2 exerted no significant cytotoxicity against L929 cells (IC50=34.33 ± 7.09 µM). Conclusions Target compounds exhibited moderate DPP-4 inhibitory activity and compound 2 was identified as the most active compound.

Bacteria-Adhesive Nitric Oxide-Releasing Graphene Oxide Nanoparticles for MRPA-Infected Wound Healing Therapy.

A bacteria-infected wound can lead to being life-threatening and raises a great economic burden on the patient. Here, we developed polyethylenimine 1.8k (PEI1.8k) surface modified NO-releasing polyethylenimine 25k (PEI25k)-functionalized graphene oxide (GO) nanoparticles (GO-PEI25k/NO-PEI1.8k NPs) for enhanced antibacterial activity and infected wound healing via binding to the bacterial surface. In vitro antibacterial activity and in vivo wound healing efficacy in an infected wound model were evaluated compared with NO-releasing NPs (GO-PEI25k/NO NPs). Surface modification with PEI1.8k can enhance the ability of nanoparticles to adhere to bacteria. GO-PEI25k/NO-PEI1.8k NPs released NO in a sustained manner for 48 h and exhibited the highest bactericidal activity (99.99% killing) against methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Pseudomonas aeruginosa (MRPA) without cytotoxicity to L929 mouse fibroblast cells at 0.1 mg/mL. In the MRPA-infected wound model, GO-PEI25k/NO-PEI1.8k NPs showed 87% wound size reduction while GO-PEI25k/NO NPs showed 23% wound size reduction at 9 days postinjury. Masson trichrome and hematoxylin and eosin staining revealed that GO-PEI25k/NO-PEI1.8k NPs enhanced re-epithelialization and collagen deposition, which are comparable to healthy mouse skin tissue. GO-PEI25k/NO-PEI1.8k NPs hold promise as effective antibacterial and wound healing agents.

The Design of 3D-Printed Polylactic Acid-Bioglass Composite Scaffold: A Potential Implant Material for Bone Tissue Engineering.

Bio-based and patient-specific three-dimensional (3D) scaffolds can present next generation strategies for bone tissue engineering (BTE) to treat critical bone size defects. In the present study, a composite filament of poly lactic acid (PLA) and 45S5 bioglass (BG) were used to 3D print scaffolds intended for bone tissue regeneration. The thermally induced phase separation (TIPS) technique was used to produce composite spheres that were extruded into a continuous filament to 3D print a variety of composite scaffolds. These scaffolds were analyzed for their macro- and microstructures, mechanical properties, in vitro cytotoxicity and in vivo biocompatibility. The results show that the BG particles were homogeneously distributed within the PLA matrix and contributed to an 80% increase in the mechanical strength of the scaffolds. The in vitro cytotoxicity analysis of PLA-BG scaffolds using L929 mouse fibroblast cells confirmed their biocompatibility. During the in vivo studies, the population of the cells showed an elevated level of macrophages and active fibroblasts that are involved in collagen extracellular matrix synthesis. This study demonstrates successful processing of PLA-BG 3D-printed composite scaffolds and their potential as an implant material with a tunable pore structure and mechanical properties for regenerative bone tissue engineering.

Effect of Currently Available Nanoparticle Synthesis Routes on Their Biocompatibility with Fibroblast Cell Lines.

Nanotechnology has acquired significance in dental applications, but its safety regarding human health is still questionable due to the chemicals utilized during various synthesis procedures. Titanium nanoparticles were produced by three novel routes, including Bacillus subtilis, Cassia fistula and hydrothermal heating, and then characterized for shape, phase state, size, surface roughness, elemental composition, texture and morphology by SEM, TEM, XRD, AFM, DRS, DLS and FTIR. These novel titanium nanoparticles were tested for cytotoxicity through the MTT assay. L929 mouse fibroblast cells were used to test the cytotoxicity of the prepared titanium nanoparticles. Cell suspension of 10% DMEM with 1 × 104 cells was seeded in a 96-well plate and incubated. Titanium nanoparticles were used in a 1 mg/mL concentration. Control (water) and titanium nanoparticles stock solutions were prepared with 28 microliters of MTT dye and poured into each well, incubated at 37 °C for 2 h. Readings were recorded on day 1, day 15, day 31, day 41 and day 51. The results concluded that titanium nanoparticles produced by Bacillus subtilis remained non-cytotoxic because cell viability was >90%. Titanium nanoparticles produced by Cassia fistula revealed mild cytotoxicity on day 1, day 15 and day 31 because cell viability was 60–90%, while moderate cytotoxicity was found at day 41 and day 51, as cell viability was 30–60%. Titanium nanoparticles produced by hydrothermal heating depicted mild cytotoxicity on day 1 and day 15; moderate cytotoxicity on day 31; and severe cytotoxicity on day 41 and day 51 because cell viability was less than 30% (p < 0.001). The current study concluded that novel titanium nanoparticles prepared by Bacillus subtilis were the safest, more sustainable and most biocompatible for future restorative nano-dentistry purposes.

Fabrication of a Patterned Scaffold Using Soft Lithography Technique to be Used in Cell Growth Applications.

In recent years, within tissue engineering, cell growth on patterned surfaces have gained significant attention. Growing cells in patterns is important to manufacture polymeric tissues that can be used within the medical field. For this reason, the main focus of this study was to prepare patterned scaffolds using Titanium (Ti) and polyvinyl chloride (PVC) covered on microscope lamellas and examine their liability for cell growth. A polydimethylsiloxane stamp was initially prepared which was then used to transfer a predefined pattern onto PVC- and Ti-covered surfaces. Cell growth experiments were performed on the prepared materials by seeding L929 mouse fibroblasts. The growth of cells seeded on the surface of the scaffolds were spectroscopically followed using Neutral Red uptake assay. The results showed cell proliferation on both patterned surfaces, however, it was higher on Ti-covered samples. In addition, three different alkanethiols were tested for cell adhesion on patterned surfaces. A higher number of cell proliferation was observed with undecanethiol, which has a shorter alkane group among them. The morphological properties of the samples before and after cell-seeding were analyzed via scanning electron microscope and optical microscopy. Significant amount of cell proliferation was observed on all of the prepared samples.

In vitro cytotoxic effects, in silico studies, some metabolic enzymes inhibition, and vibrational spectral analysis of novel β-amino alcohol compounds

In this study, an efficient single-step method for the preparation of β-amino alcohols (1–3) in aqueous media was applied. The aim was to investigate the cytotoxic activity of Compounds 1, 2 and 3 in neuroblastoma SH-SY5Y cell line and mouse fibroblast l-929 cell lines. Cytotoxic activities of compounds 1, 2 and 3 in this cell lines were also determined by MTT method. Cells were incubated with different concentrations of Compound 3 showed the highest cytotoxic activity in SHY5Y cells at an IC50 dose of 13.01±0.87 µM at 72 h compared to other compounds. Compound 3 was determined to have lower cytotoxic activity in l-929 cells. The chemical activities of the molecules against the B3LYP, HF, M062X level 3–21 g, 6–31 g, and SDD basis set with the Gaussian package program and biologically against the adenosine A(2A) receptor (PDB ID: 3PWH and 5NM4) proteins for neuroblastoma tumors cell with the Maestro Molecular modeling platform by Schrödinger were compared. Both experimental and theoretical NMR, UV–vis, and IR spectra of the studied molecules were compared. ADME/T analysis was performed to examine the drug properties of the molecules. Finally, these assayed for their activities against metabolic enzymes acetylcholinesterase and α-glucosidase. The most potent compounds against AChE were order compounds 3, 2 and 1 with Ki values of 35.88±6.61, 43.75±8.28, and 45.34±3.50 µM against AChE, respectively. The results indicated that all the synthesized compounds exhibited excellent inhibitory activities against mentioned enzymes as compared with standard inhibitors. These inhibitors may be candidates for drug design.

Immobilization of Gelatin on Fibers for Tissue Engineering Applications: A Comparative Study of Three Aliphatic Polyesters.

Immobilization of cell adhesive proteins on the scaffold surface has become a widely reported method that can improve the interaction between scaffold and cells. In this study, three nanofibrous scaffolds obtained by electrospinning of poly(caprolactone) (PCL), poly(L-lactide-co-caprolactone) (PLCL) 70:30, or poly(L-lactide) (PLLA) were subjected to chemical immobilization of gelatin based on aminolysis and glutaraldehyde cross-linking, as well as physisorption of gelatin. Two sets of aminolysis conditions were applied to evaluate the impact of amine group content. Based on the results of the colorimetric bicinchoninic acid (BCA) assay, it was shown that the concentration of gelatin on the surface is higher for the chemical modification and increases with the concentration of free NH2 groups. XPS (X-ray photoelectron spectroscopy) analysis confirmed this outcome. On the basis of XPS results, the thickness of the gelatin layer was estimated to be less than 10 nm. Initially, hydrophobic scaffolds are completely wettable after coating with gelatin, and the time of waterdrop absorption was correlated with the surface concentration of gelatin. In the case of all physically and mildly chemically modified samples, the decrease in stress and strain at break was relatively low, contrary to strongly aminolyzed PLCL and PLLA samples. Incubation testing performed on the PCL samples showed that a chemically immobilized gelatin layer is more stable than a physisorbed one; however, even after 90 days, more than 60% of the initial gelatin concentration was still present on the surface of physically modified samples. Mouse fibroblast L929 cell culture on modified samples indicates a positive effect of both physical and chemical modification on cell morphology. In the case of PCL and PLCL, the best morphology, characterized by stretched filopodia, was observed after stronger chemical modification, while for PLLA, there was no significant difference between modified samples. Results of metabolic activity indicate the better effect of chemical immobilization than of physisorption of gelatin.

Comparison of physicochemical and functional properties of elemental sulfur, sulfur nanoparticles, and sulfur quantum dots for sustainable biological applications

Elemental sulfur (ES) and sulfur nanoparticles (SNPs) have been widely used as antibacterial ingredients, especially in dermatological formulations. However, their use in biological systems is greatly limited because of their inability to form hydrophilic and stable aqueous suspensions. On the other hand, sulfur quantum dots (SQDs) are surface-passivated nano-sulfur particles with less than 10 nm, are hydrophilic, and exhibit superior biological activity to ES or SNPs. It is also a sustainable material produced from sublimated sulfur, obtained as a by-product in huge quantities from petroleum refineries. However, systematic comparisons of these sulfur particles' physicochemical and functional properties have not yet been performed, resulting in knowledge gaps. The present work has characterized and compared the physicochemical and functional properties of ES, SNPs, and SQDs. SQDs had the highest zeta potential (−11.9 mV) of these three sulfur forms and were most stable in aqueous suspension. Surface-passivated SQDs with polyethylene glycol were more biologically compatible without cytotoxicity to L929 mouse fibroblasts. Unlike ES or SNPs, SQDs showed high antioxidant activity, showing 100% oxidative free radical scavenging activity in an aqueous medium. In addition, SQDs showed potent antibacterial activity against pathogenic bacteria such as Listeria monocytogenes and Escherichia coli and the fungal strains such as Penicillium chrysogenum and Aspergillus niger. With its aqueous stability and diverse biological activities, SQDs are expected to have tremendous potential as multifunctional components in various biological and related applications such as biomedical engineering and food packaging.

Development of antimicrobial nanocomposite scaffolds via loading CZTSe quantum dots for wound dressing applications

The antimicrobial properties of scaffolds designed for use in wound healing are accepted as an important factor in the healing process to accelerate the wound healing process without causing inflammation. For this purpose, chitosan–polyvinyl alcohol composite membranes loaded with Cu2ZnSnSe4 quantum dots (CZTSe QDs) as an antibacterial and cytocompatible biomaterial to regulate the wound healing process were produced. CZTSe QDs particles were synthesized under hydrothermal conditions. Polymer-based nanocomposites with different concentrations of the synthesized nanoparticles were produced by the solvent casting method. After detailed physicochemical and morphological characterizations of CZTSe QDs and composite membranes, antibacterial activities and cell viability were extensively investigated against gram-positive and gram-negative bacterial and yeast strains, and L929 mouse fibroblast cells lines, respectively. The results show that the preparation of composite scaffolds at a QDs concentration of 3.3% by weight has the best antimicrobial activity. Composite scaffold membranes, which can be obtained as a result of an easy production process, are thought to have great potential applications in tissue engineering as wound dressing material due to their high mechanical properties, wettability, strong antibacterial properties and non-toxicity.

Some Heterocyclic Hydrazone Compounds: Synthesis, Spectral Characterization and Anticancer Activity Study

Cancer is currently ongoing to be a significant health problem threatening human health. Hydrazone compounds constitute a popular class of organic compounds used in novel drug discovery studies in therapy of cancer. In the current study, the preparation and structural characterization of some heterocyclic hydrazone compounds (7-12) and their anticancer capacities against HeLa cervical cancer and MCF-7 breast cancer cell lines were reported. The target compounds were characterized by some spectroscopic techniques (1H NMR, 13C NMR and FT-IR). The in vitro cytotoxic potentials of the target molecules were assessed by using MTT assay against two cancer cell lines. L929 mouse fibroblast cell lines were employed as normal cell. The results displayed that some of the tested molecules had varying anticancer activities. Among the tested compounds, compound 8 indicated anticancer activity against HeLa cells with IC50 value of 34.38 µM. On the other hand, of these tested compounds, compound 11 (IC50 = 26.84 µM) displayed anticancer activity against MCF-7 cells.

EVALUATION OF TOXICITY AND ANTIBACTERIAL ACTIVITIES OF Andrographis paniculata HERBAL MOUTHWASH AGAINST ORAL PATHOGENS

Andrographis paniculata (AP) is an ancient herb known for its medicinal and therapeutic values. In this study, we aimed to evaluate the antibacterial activity and cytotoxicity of AP herbal mouthwash. Aqueous extract of AP was used to prepare the herbal mouthwash. The product was tested against selected oral pathogens namely Actinomyeces viscosus, Staphylococcus aureus, Streptococcus mutans, Streptococcus sobrinus, and Porphyromonas gingivalis for its antibacterial activity using the agar well diffusion method. Toxicity analysis was carried out and subjected to cytotoxicity screening using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, in vivo study using brine shrimp lethality bioassay, and detection of heavy metals using atomic absorption spectroscopy (AAS). Five AP herbal mouthwash concentrations (62.5, 125, 250, 500, 1000 mg/mL) were developed. AP herbal mouthwash exhibited the strongest antibacterial activity against A. viscosus. AP herbal mouthwash (125 to 1000 mg/mL) showed inhibition against P. gingivalis, S. mutans and S. sobrinus, except for S. aureus which did not reflect any sign of antibacterial activity. In brine shrimp lethality bioassay, AP herbal mouthwash has LC50 3255.064 µg/mL. In vitro cytotoxic evaluation was done on L929 mouse fibroblast cell lines using MTT assay with IC50 43.55 mg/mL denotes the mouthwash is non-toxic. Heavy metals content in AP herbal mouthwash falls within the permissible range of plumbum (2.50 ppm) followed by arsenic (1.875 ppm), mercury (0.15 ppm), and cadmium (0.06 ppm). Thus, verifying AP herbal mouthwash is effective in common oral pathogens and has a non-toxic effect.

Development of Lipidic Nanoplatform for Intra-Oral Delivery of Chlorhexidine: Characterization, Biocompatibility, and Assessment of Depth of Penetration in Extracted Human Teeth.

Microorganisms are the major cause for the failure of root canal treatment, due to the penetration ability within the root anatomy. However, irrigation regimens have at times failed due to the biofilm mode of bacterial growth. Liposomes are vesicular structures of the phospholipids which might help in better penetration efficiency into dentinal tubules and in increasing the antibacterial efficacy. Methods: In the present work, chlorhexidine liposomes were formulated. Liposomal chlorhexidine was characterized by size, zeta potential, and cryo-electron microscope (Cryo-EM). Twenty-one single-rooted premolars were extracted and irrigated with liposomal chlorhexidine and 2% chlorhexidine solution to evaluate the depth of penetration. In vitro cytotoxicity study was performed for liposomal chlorhexidine on the L929 mouse fibroblast cell line. Results: The average particle size of liposomes ranged from 48 ± 4.52 nm to 223 ± 3.63 nm with a polydispersity index value of <0.4. Cryo-EM microscopic images showed spherical vesicular structures. Depth of penetration of liposomal chlorhexidine was higher in the coronal, middle, and apical thirds of roots compared with plain chlorhexidine in human extracted teeth when observed under the confocal laser scanning microscope. The pure drug exhibited a cytotoxic concentration at which 50% of the cells are dead after a drug exposure (IC50) value of 12.32 ± 3.65 µg/mL and 29.04 ± 2.14 µg/mL (on L929 and 3T3 cells, respectively) and liposomal chlorhexidine exhibited an IC50 value of 37.9 ± 1.05 µg/mL and 85.24 ± 3.22 µg/mL (on L929 and 3T3 cells, respectively). Discussion: Antimicrobial analysis showed a decrease in colony counts of bacteria when treated with liposomal chlorhexidine compared with 2% chlorhexidine solution. Nano-liposomal novel chlorhexidine was less cytotoxic when treated on mouse fibroblast L929 cells and more effective as an antimicrobial agent along with higher penetration ability.