Cell Viability Test Research Articles

Viability Test of Osteoblast Cells After Application of Hydroxyapatite Nanoparticle from Unam Snail’s Shells In-vitro Examination

The bone defect reconstruction process can use hydroxyapatite is osteoconductive and can retain the original biocompatible shape to enhance hydroxyapatite with osteogenic proteins. To analyze the most appropriate concentration of hydroxyapatite nanoparticles using the MTT Assay method and to test the viability of osteoblast cells after being given hydroxyapatite nanoparticle (nHA) derived from unam snail shells. The fabrication of hydroxyapatite nanoparticles from unam snail shells using a mechanical-chemical combination method. Osteoblast cells are obtained from Calvaria rats after being cultured in DMEM. Viability tests of osteoblast cells were done using the MTT Assay method and repeated three times, and then results were measured using an Elisa reader. Viability of osteoblast cells in nHA 1,25 mg/ml (164,60 % ± 0,096), nHA 1,5 mg/ml (151,72 % ± 0,176), nHA 1,75 mg/ml (90,55 % ± 0,243), nHA 2 mg/ml (74,23 % ± 0,301) respectively. ANOVA test shows p < 0,05. IC50 value of hydroxyapatite nanoparticle from the unam snail’s shells to viability osteoblast cells is 2,23 mg/ml. Less concentration of hydroxyapatite nanoparticles tends to increase the viability of osteoblast cells. 1,75 mg/ml and below hydroxyapatite nanoparticles derived from unam snail shells are not toxic to osteoblast cells.

Future-Oriented Nanosystems Composed of Polyamidoamine Dendrimer and Biodegradable Polymers as an Anticancer Drug Carrier for Potential Targeted Treatment.

Background/Objectives: Camptothecin (CPT) is a well-known chemical compound recognized for its significant anticancer properties. However, its clinical application remains limited due to challenges related to CPT's high hydrophobicity and the instability of its active form. To address these difficulties, our research focused on the development of four novel nanoparticulate systems intended for either oral or intravenous administration. Methods: These nanosystems were based on a poly(amidoamine) (PAMAM) dendrimer/CPT complex, which had been coated with biodegradable homo- and copolymers, designed with appropriate physicochemical properties and chain microstructures. Results: The resulting nanomaterials, with diameters ranging from 110 to 406 nm and dispersity values between 0.10 and 0.67, exhibited a positive surface charge and were synthesized using biodegradable poly(L-lactide) (PLLA), poly(L-lactide-co-ε-caprolactone) (PLACL), and poly(glycolide-co-ε-caprolactone) (PGACL). Biological assessments, including cell viability and hemolysis tests, indicated that all polymers demonstrated less than 5% hemolysis, confirming their hemocompatibility for potential intravenous use. Furthermore, fibroblasts exposed to these matrices showed concentration-dependent viability. The entrapment efficiency (EE) of CPT reached up to 27%, with drug loading (DL) values as high as 17%. The in vitro drug release studies lasted over 400 h with the use of phosphate buffer solutions at two different pH levels, demonstrating that time-dependent processes allowed for a gradual and controlled release of CPT from the developed nanosystems. The release kinetics of the active compound at pH 7.4 ± 0.05 and 6.5 ± 0.05 followed near-first-order or first-order models, with diffusion and Fickian/non-Fickian transport mechanisms. Importantly, the nanoparticulate systems enabled the stabilization of the pharmacologically active form of CPT, while providing protection against hydrolysis, even in physiological environments. Conclusions: In our opinion, these results underscore the promising future of biodegradable nanosystems as effective drug delivery systems (DDSs) for targeted cancer treatment, offering stability and efficacy over short, medium, and long-term applications.

Impact and mechanism study of dioscin on biological characteristics of colorectal cancer cells.

Colorectal cancer (CRC) is a considerable global health issue. Dioscin, a compound found in several medicinal plants, has shown potential anticancer effects. To find the relationship between CRC cells (HCT116) and diosgenin and clarified their mechanisms of action. CRC cell line HCT116 was cultured by dividing cells into control and dioscin groups (dioscin + Jagged 1 group; Jagged 1 group, 5 μg/mL; and dioscin group, 2.5 μg/mL). The dioscin groups were given different concentrations of dioscin. Cell Counting Kit-8 was chosen for testing cell viability in different groups. Flow cytometry was established to undiscover the apoptosis rate of human liver cancer cell line 11. Real-time PCR as well as Western blot analyses were applied to reveal the expression levels of caspase-3, Notch, and other proteins. Transwell and scratch experiments were conducted to assess cell migration and invasion abilities. This study indicated that dioscin restricted the growth of HCT116 cells, boosted cell apoptosis, and rose the Bax/Bcl-2 ratio as well as the expression of Caspase-3. Dioscin also inhibited physiological activities, for instance cell migration, and significantly reduced the expression levels of proteins for instance Notch1 (P < 0.05). Dioscin partially reversed the effects of Jagged 1. Dioscin exerts a certain inhibitory effect on HCT116, and its mechanism of action may be linked, with the inhibition of the Notch1 signaling pathway.

FRESH 3D printing of zoledronic acid-loaded chitosan/alginate/hydroxyapatite composite thermosensitive hydrogel for promoting bone regeneration

The aim of this study was to develop a composite thermosensitive hydrogel for bone regeneration applications. This hydrogel consisted of chitosan, alginate and hydroxyapatite, and was loaded with zoledronic acid as a model drug. The feasibility of three-dimensional (3D) printing of the thermosensitive hydrogel using the extrusion based technique was investigated. The 3D printing technique called Freeform Reversible Embedded Suspended Hydrogel (FRESH) printing was employed for this purpose. To characterize the composite hydrogels, several tests were conducted. The gelation time, rheological properties, and in vitro drug release were analyzed. Additionally, the cell viability test on human osteosarcoma MG-63 cells for the composite hydrogel was assessed using an MTT assay. The results of the study showed that the zoledronic acid-loaded composite thermosensitive hydrogel was successfully printed using the FRESH 3D printing technique which was not possible otherwise i.e., by using traditional 3D printing techniques. Further examination of the printed constructs using a Scanning Electron Microscope revealed the presence of porous and layered structures. The gelation times of the composite thermosensitive hydrogel was determined to be 10 and 20 min, respectively for scaffolds with and without HA, indicating the successful formation of the gel within a reasonable time to the FRESH technique. The flow behavior of the hydrogel was found to be pseudoplastic, following a non-Newtonian flow pattern with Farrow’s constant (N) values of 1.708 and 1.853 for scaffolds with and without hydroxyapatite, respectively. In terms of drug release, scaffolds prepared with and without hydroxyapatite reached nearly 100% of zoledronic acid release in 360 h and 48 h, respectively. The cell viability test on human osteosarcoma MG-63 cells using MTT assay has shown increased cell viability % in the case of composite hydrogel, indicating biocompatibility of the scaffold. Overall, this study successfully developed a composite thermosensitive hydrogel loaded with zoledronic acid for bone regeneration applications and was 3D printed using the FRESH 3D printing technique. The results of this study provide valuable insights into the potential use of this composite hydrogel for future biomedical applications.

Highly porous sildenafil loaded polylactic acid/polyvinylpyrrolidone based 3D printed scaffold containing forsterite nanoparticles for craniofacial reconstruction

Tissue engineering has emerged as a promising substitute for traditional tissue repair methods. Nowadays, advancements in 3D printing technology have enabled the fabrication of customized scaffolds to support tissue regeneration. In the present study, a polylactic acid-polyvinylpyrrolidone 3D-printed scaffold containing 10 % forsterite was fabricated. Subsequently, lyophilized fucoidan microstructures loaded with sildenafil were filled the channels of this 3D-printed scaffold. The fabricated scaffold loaded with sildenafil was thoroughly characterized, revealing that 97.46 % of the loaded sildenafil was released in a sustained manner over 28 days. Furthermore, the biocompatibility of MG63 was evaluated through cell viability and adhesion tests. The findings indicated a direct and favorable influence on cell behavior. Based on the chicken chorioallantoic membrane assay, the fabricated scaffold significantly increases angiogenesis due to the sustained release of sildenafil. Moreover, in-vivo studies conducted on a rat model demonstrated that the 3D-printed scaffold was able to stimulate and accelerate the repair of calvarial defects within 8 weeks, and the amount of new bone tissue formation was significantly higher than that of other experimental groups. Based on the comprehensive in-vitro and in-vivo assessments, the scaffold with a macro- and microporous structure combined with the ability to release sildenafil is suggested as a potential candidate for repairing bone tissue, especially in the context of skull defects.

In-vitro studies of B2O3 – Li2O – P2O5 – CaO – ZrO2 nano glasses for biomedical applications

The current work focuses on developing 40B2O3-(20-X)Li2O-15P2O5-20CaO-XZrO2 (X= 0, 1 and 2 mol %) bioactive nano glasses by a novel microemulsion method and conducting in-vitro studies. CaO derived from fresh water snail shells has been used to produce cost effective glasses. DTA, FTIR, XRD, FESEM, degradation, pH, BET and cell viability tests have been carried out for these glasses. The glasses show good bioactivity and cell viability. ZrO2 reduces the formation of hydroxyapatite and decreases cell viability. The role of ZrO2 on the glass structure and in-vitro properties has been clearly discussed.

Sustained release of heparin from PLLA micropartricles for tissue engineering applications

Heparin holds promise for cardiac tissue engineering, but challenges such as hematoma or bleeding and accumulation in tissue caused by excessive release, and short half-life persist. The present study aimed to introduce a reliable mechanism for the prolonged heparin release from a biocompatible polymer carrier. The designed system must ensure that heparin retains its bioactivity over time while preventing premature release. Heparin was encapsulated within poly (L-lactic acid) microparticles using the double emulsion method, with polyvinyl alcohol employed as the stabilizer. The encapsulation efficiency of heparin in the microparticles was calculated as 25.56 %. The functionality of the design was evaluated using Attenuated Total Reflection Fourier Transform Infrared Spectroscopy, Scanning Electron Microscopy, and Energy Dispersive X-ray Spectroscopy. Drug release and microparticle degradation studies were conducted alongside cell viability tests. The particle sizes ranged from 5 to 10 ± 2.53 μm, with evidence suggesting that heparin promotes the smaller particle formation. The system demonstrated a consistent drug release profile over six weeks with a release rate of 54 % by week two and 97.65 % by week six. The degradation of heparin-loaded microparticles reached less than 50 % by week six, and the loading of heparin did not significantly affect the degradation behavior of the PLLA microparticles in PBS. Furthermore, heparin concentrations between 200 and 400 μg/ml enhanced the viability of Placenta-derived Mesenchymal Stem Cells and H9c2. These findings suggest that the system could be considered as an effective vehicle for sustained heparin delivery across a spectrum of biological applications, particularly in cardiac tissue engineering.

A novel approach to regulate glucose uptake in an anaplastic thyroid cancer cell line.

Curcumin's function in affecting cancer metabolic reprogramming remains poorly understood. Herein, we aimed to elucidate a novel link between Curcumin and the glucose uptake metabolism and glucose transporters (GLUTs) status in SW1736 cell line derived from anaplastic thyroid cancer. TheMTT test and flow cytometry was employed to test cell viability and cell death. For glucose uptake detection, ''GOD-PAP'' enzymatic colorimetric assay was applied to measure the direct glucose levels inside of the cells. Determination of GLUT1 and GLUT3 mRNA and protein expression in SW1736 cells was performed by qRT-PCR and western blotting. Also, the scratch wound healing assay was conducted for cell migration. The data indicated that Curcumin-induced cell death is independent of apoptosis in this type of thyroid cancer cell line. Furthermore, significantly reduced GLUT1 and GLUT3 expression was observed after treatment with Curcumin, resulting in the inhibition of glucose uptake (p < 0.05). Scratch assay indicated the inhibition of cell migration in SW1736 cells treated by Curcumin (p < 0.05). It can be concluded that GLUTs as metabolic targets can be blocked specifically by Curcumin for thyroid cancer prevention. Curcumin, as a promising anti-cancer agent, inhibits the growth of SW1736 anaplastic thyroid cancer cell line by regulating glucose uptake pathway and cell death. Altogether, these results suggest that the glucose pathway may be an important target for therapeutic intervention to sensitize tumor cells to cell death process by inhibition of glucose transporters.

Bioactive dextran-based scaffolds from emulsion templates co-stabilized by poly(lactic-co-glycolic acid) nanocarriers

Porous polymer scaffolds are widely investigated as temporary implants in regenerative medicine to repair damaged tissues. While biocompatibility, degradability, mechanical properties comparable to the native tissues and controlled porosity are prerequisite for these scaffolds, their loading with pharmaceutical or biological active ingredients such as growth factors, in particular proteins, opens up new perspective for tissue engineering applications. This implies the development of scaffold loading strategies that minimize the risk of protein denaturation and allow to control their release profile. This work reports on a straightforward method for preparing bioactive dextran-based scaffolds from high internal phase emulsion (HIPE) templates containing poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) serving both as co-stabilizers for the emulsion and nanocarriers for drug or therapeutic protein models. Scaffold synthesis are achieved by photocuring of methacrylated dextran located in the external phase of a HIPE stabilized by the NPs in combination or not with a non-ionic surfactant. Fluorescent labelling of the NPs highlights their integration in the scaffold. The introduction of NPs, and even more so when combined with a surfactant, increases the stability and mechanical properties of the scaffolds. Cell viability tests demonstrate the non-toxic nature of these NPs-loaded scaffolds. The study of the release of a model protein from the scaffold, namely lysozyme, shows that its encapsulation in nanoparticles decreases the release rate and provides additional control over the release profile.

Enhancing Surgical Care: Development of Biocompatible, Superabsorbent Alternatives to Cotton Gauze Using Chia Mucilage and Poly(vinylpyrrolidone).

Cotton gauze bandages have traditionally played a pivotal role in wound care and surgical procedures, absorbing fluids, including blood, and protecting against infection. However, their limited liquid absorption capacity raises concern about potential post-surgery complications if inadvertently retained. In response, resorbable and biocompatible polymers have emerged as a promising alternative to enhance surgical outcomes and mitigate inflammation. This study aims to develop a biocompatible, highly absorbent, and preferably resorbable substitute for cotton gauze, utilizing natural polysaccharides from chia seeds' mucilage alongside the synthetic polymer poly(vinylpyrrolidone) (PVP). Incorporating tranexamic acid, an antifibrinolytic agent, into the PVP solution enhances its efficacy in controlling blood flow. The polymer solution is then processed into nonwoven materials via solution blow spinning. UV-C radiation cross-linking is employed to bolster the nonwovens' performance and durability during liquid absorption and swelling. Results demonstrate that nonwoven samples comprising PVP and chia mucilage, cross-linked for 60 min with UV-C radiation, exhibit exceptional swelling capacity, absorbing approximately 3291% of their dry weight in saline solution. Microfiber analysis indicates alterations in fiber characteristics due to cross-linking duration. Cell viability tests affirm the biocompatibility of the produced materials. With their remarkable fluid absorption properties and potential for resorption, PVP/chia mucilage compositions supplemented with tranexamic acid offer a promising avenue for effectively managing surgical bleeding without adverse effects. Furthermore, these materials can safely remain within the surgical site, eventually undergoing natural resorption by the body owing to their resorbable nature.

Development and optimization of hydrogel-forming microneedles fabricated with 3d-printed molds for enhanced dermal diclofenac sodium delivery: a comprehensive in vitro, ex vivo, and in vivo study.

With the developing manufacturing technologies, the use of 3D printers in microneedle production is becoming widespread. Hydrogel-forming microneedles (HFMs), a variant of microneedles, demonstrate distinctive features such as a high loading capacity and controlled drug release. In this study, the conical microneedle master molds with approximately 500μm needle height and 250μm base diameter were created using a Stereolithography (SLA) 3D printer and were utilized to fabricate composite HFMs containing diclofenac sodium. Using Box-Behnken Design, the effects of different polymers on swelling index and mechanical strength of the developed HFMs were evaluated. The optimum HFMs were selected according to experimental design results with the aim of the highest mechanical strength with varying swelling indexes, which was needed to use 20% Gantrez S97 and 0.1% (F22), 0.42% (F23), and 1% (F24) hyaluronic acid. The skin penetration and drug release properties of the optimum formulations were assessed. Ex vivo studies were conducted on formulations to determine drug penetration and accumulation. F24, which has the highest mechanical strength and optimized swelling index, achieved the highest drug accumulation in the skin tissue (17.70 ± 3.66%). All optimum HFMs were found to be non-cytotoxic by the MTT cell viability test (> 70% cell viability). In in vivo studies, the efficacy of the F24 was assessed for the treatment of xylene-induced ear edema by contrasting it to the conventional dosage form. It was revealed that HFMs might be an improved replacement for conventional dosage forms in terms of dermal diseases such as actinic keratosis.

Preservative-free electrospun nanofibrous inserts for sustained delivery of ceftazidime; design, characterization and pharmacokinetic investigation in rabbit's eye

Ocular drug delivery presents significant challenges, attributed to the various anatomical and physiological barriers, as well as the limitations associated with conventional ocular formulations including low bioavailability, necessitating frequent dosing. The objective of the essay was to design sustained release nanofibrous inserts loaded with ceftazidime (CAZ), an antibiotic effective against gram-negative and gram-positive microorganisms, for the treatment of ocular infections. These nanofibers were fabricated using the electrospinning technique, employing biodegradable polymers such as polyvinyl alcohol (PVA), polycaprolactone (PCL) and Eudragit® (EUD). The nanofibrous inserts exhibited adequate mechanical strength for ocular use with an average diameter < 250 nm. In the initial 12-h period, a burst drug release was observed, followed by a controlled release for 120 h. Cell viability test confirmed the non-toxicity and safety of the nanofibers. The in vivo study demonstrated that the inserts sustain a drug concentration exceeding the minimum inhibitory concentration (MIC) of Pseudomonas aeruginosa and Staphylococcus aureus for 4 and 5 days, respectively. The AUC0–120 for CAZ-PVA-PCL was reported 11,882.81 ± 80.5 μg·h/ml and for CAZ-PVA-EUD was 9649.39 ± 86.84 μg·h/ml. The nanofibrous inserts' extended drug release maintains effective antimicrobial concentrations, avoids the fluctuations of eye drops, and, by being preservative-free, eliminates cytotoxicity.

Dual-Action Gemcitabine Delivery: Chitosan–Magnetite–Zeolite Capsules for Targeted Cancer Therapy and Antibacterial Defense

Cancer and infectious diseases are two of the world’s major public health problems. Gemcitabine (GEM) is an effective chemotherapeutic agent against several types of cancer. In this study, we developed macrocapsules incorporating GEM into a chitosan matrix blended with magnetite and zeolite by ionic gelation. Physicochemical characterization was performed using HRTEM-ED, XRD, FESEM–EDS, FT-IR, TGA, encapsulation efficiency (%E.E.), and release profiles at pHs 7.4 and 5.0. Cell viability tests against A549 and H1299 cell lines, and microbiological properties against staphylococcal strains were performed. Our results revealed the successful production of hemispherical capsules with an average diameter of 1.22 mm, a rough surface, and characteristic FT-IR material interaction bands. The macrocapsules showed a high GEM encapsulation efficiency of over 86% and controlled release over 24 h. Cell viability assays revealed that similar cytotoxic effects to free GEM were achieved with a 45-fold lower GEM concentration, suggesting reduced dosing requirements and potentially fewer side effects. Additionally, the macrocapsules demonstrated potent antimicrobial activity, reducing Staphylococcus epidermidis growth by over 90%. These results highlight the macrocapsules dual role as a chemotherapeutic and antimicrobial agent, offering a promising strategy for treating lung cancer in patients at risk of infectious diseases or who are immunosuppressed.

3D Coaxial Printing of Small‐Diameter Artificial Arteries

As a treatment for the widely spread cardiovascular diseases (CVD), bypass vascular grafts have room for improvement in terms of mechanical property match with native arteries. A 3D‐printed nozzle is presented, featuring unique internal structures, to extrude artificial vascular grafts with a flower‐mimicking geometry. The multilayer‐structured graft wall allows the inner and outer layers to interfere sequentially during lateral expansion, replicating the nonlinear elasticity of native vessels. Both experiment and simulation results verify the necessity and benefit of the flower‐mimicking structure in obtaining the self‐toughening behavior. The gelation study of natural polymers and the utilization of sacrificial phase enables the smooth extrusion of the multiphase conduit, where computer‐assisted image analysis is employed to quantify manufacturing fidelity. The cell viability tests demonstrate the cytocompatibility of the gelatin methacryloyl (GelMA)/sodium alginate grafts, suggesting potential for further clinical research with further developments. This study presents a feasible approach for fabricating bypass vascular grafts and inspires future treatments for CVD.

Phytochemical Extract from Syzygium cumini Leaf: Maximization of Compound Extraction, Chemical Characterization, Antidiabetic and Antibacterial Activity, and Cell Viability

This work aimed to obtain a phytochemical extract from jambolan leaf using a hydroethanolic solvent and ultrasound-assisted extraction. For this purpose, an experimental design was applied to analyze the effect of process variables related to temperature (30–60 °C), time (10–30 min), and solvent to leaf ratio (5–15 mL g−1), on the extraction mass yield (EMY) and on the yield of phenolic compounds (PCY). The effect of extractor solvent, AE (absolute ethanol), 75E (75% v·v−1 ethanol) and 50E (50% v·v−1), on the chemical characterization of the extracts, antidiabetic and antimicrobial activity, and cell viability, were also evaluated. The application of the highest values of process variables resulted in obtaining the maximum of the response variables (EMY = 9.94 wt% and PCY = 13.01 mg GAE g−1 leaf). A higher content of phenolic compounds and flavonoids was obtained with 50E, which is mainly composed of sinapic, vanillic, trans-caffeic, and quinic acids, which were responsible for the greatest antioxidant potential, antibacterial activity (against Staphylococcus aureus and Pseudomonas aeruginosa), and inhibition of α-amylase. On the other hand, the use of AE allowed us to obtain extracts with higher concentrations of squalene, α-tocopherol, β-sitosterol, and friedelin. From cell viability tests, the extracts are not considered toxic at the concentration tested (100 µg mg−1).

Exploring Thermal Stability, Vibrational Properties, and Biological Assessments of Dichloro(l-histidine)copper(II): A Combined Theoretical and Experimental Study.

Dichloro(l-histidine)copper(II) crystal ([Cu(l-His)Cl2] complex) was obtained by the slow evaporation method and characterized concerning its thermal stability, phase transformations, and electronic and vibrational properties. X-ray diffraction (XRPD) confirmed that this complex crystallizes with an orthorhombic structure (P212121 space group). Thermal analyses (TG and DTA) demonstrate stability from ambient temperature up to 460 K, followed by a phase transition from the orthorhombic structure to the amorphous form around 465 K, as confirmed by temperature-dependent XRPD studies. The active modes in Fourier transform infrared (FT-IR) and Raman spectroscopy spectra were suitably assigned via density functional theory (DFT) calculations. Additionally, Hirshfeld surface analysis uncovered the prominence of Cl···H, O···H, and H···H interactions as the primary intermolecular forces within the crystal structure. The antimicrobial activity of the [Cu(l-His)Cl2] complex was investigated, demonstrating significant efficacy against Gram-positive bacteria (Staphylococcus aureus), Gram-negative bacteria (Pseudomonas aeruginosa), and fungi (Candida albicans). The minimum inhibitory concentration and cell viability tests showed that the complex inhibits the growth of S. aureus bacteria at a concentration of 1.5 μM without causing damage to the human cell line. The pharmacokinetic parameters corroborate the other tested parameters and highlight the [Cu(l-His)Cl2] complex as a promising alternative for future clinical trials and medicinal applications. The alignment of the pharmacokinetic parameters with other tested criteria highlights the potential of the [Cu(l-His)Cl2] complex as a promising candidate for future clinical studies.

Polyvinyl alcohol assisted citrate based reduction of gold(III) ions: Theoretical design and experimental study on green synthesis of spherical and biocompatible gold nanoparticles

In this study, we demonstrate the synthesis of small gold nanoparticles (typically 8–10 nm) through a green synthesis approach. This method involves utilizing chloroauric acid as the gold precursor, trisodium citrate as a mild reducing and co-capping agent, and polyvinyl alcohol (PVA), as a co-capping and shape-directing agent. This novel synthesis method stands alone as a protocol that involves just mixing the reagents at room temperature and allowing the reaction to proceed at ambient temperature without any disturbance. In the absence of PVA, spherical particles are not formed and the reaction mixture slowly turns black followed by precipitation.The synthesis process was meticulously tracked using time-resolved monitoring of the growth of the localized surface plasmon resonance (LSPR) by UV-vis spectroscopy and transmission electron microscopy. The as-prepared colloidal gold solution was bright red, attributed to the small size (≤10 nm) and spherical geometry of nanoparticles. This colloidal solution could be stored indefinitely at room temperature without precipitation or a change in its absorption profile. The nanoparticles remained stable in adverse conditions such as 100 mM NaCl solution, 1 × PBS and cell culture medium. Additionally, they could be easily loaded with drugs like doxorubicin by adsorption. The particles were thoroughly characterized using a range of techniques, including UV-vis spectroscopy, attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR), high-resolution transmission electron microscopy (HRTEM), hyperspectral-enhanced dark field microscopy (HEDFM), dynamic light scattering (DLS), and X-ray photoelectron spectroscopy (XPS). Cell viability tests conducted on 2D cell lines and 3D spheroids revealed negligible toxicity even at high concentrations. Furthermore, we demonstrated that an advanced version of conventional diffusion-limited aggregation (DLA) schemes, which allows individual clusters to move freely within a domain to form larger aggregates, can effectively replicate the intricate interactions between chloroauric acid, trisodium citrate, and PVA, the agents involved in the synthesis of gold nanoparticles.

Cold plasma irradiation of chitosan: A straight pathway to selective antitumor therapy

Plasma-activated chitosan (PAC) colloids for cancer treatment were obtained by using the cold atmospheric plasma technique. Chitosan solutions were irradiated by plasma ignited in argon gas and in a mixture of argon with nitrogen and oxygen gases in certain ratios. The structural modifications of chitosan and the chemical species generated in plasma were investigated by EPR, LC-MS/MS, XRD, DLS, and TGA methods. The cell viability test showed a selective cytotoxic effect on human breast carcinoma cells (MCF-7), while the human mammary epithelial cells (MCF-10A) were left unharmed. The cytotoxic effect was attributed mainly to chitooligosaccharides, but also to a synergistic effect with other compounds generated in very low concentrations in plasma, such as glyceric acid, ethyl acetate, or tricarballylic acid. The plasma irradiation improved the antioxidant activity and mucoadhesivity, while not affecting the hemocompatibility investigated by a standard hemolysis ex vivo test on mice blood. Moreover, the in vivo biocompatibility investigation at intraperitoneal administration of PAC in mice showed no statistically significant changes in the hematologic, biochemical, and immune system parameters, and no morphologic alterations of the liver and kidney. All these data indicate the cold plasma activation of chitosan as a straight method to produce biocompatible, antitumor systems.

Formulation-Property Effects in Novel Injectable and Resilient Natural Polymer-Based Hydrogels for Soft Tissue Regeneration

The development of injectable hydrogels for soft tissue regeneration has gained significant attention due to their minimally invasive application and ability to conform precisely to the shape of irregular tissue cavities. This study presents a novel injectable porous scaffold based on natural polymers that undergoes in situ crosslinking, forming a highly resilient hydrogel with tailorable mechanical and physical properties to meet the specific demands of soft tissue repair. By adjusting the formulation, we achieved a range of stiffness values that closely mimic the mechanical characteristics of native tissues while maintaining very high resilience (&gt;90%). The effects of gelatin, alginate, and crosslinker concentrations, as well as porosity, on the hydrogel’s properties were elucidated. The main results indicated a compression modulus range of 2.7–89 kPa, which fits all soft tissues, and gelation times ranging from 5 to 30 s, which enable the scaffold to be successfully used in various operations. An increase in gelatin and crosslinker concentrations results in a higher modulus and lower gelation time, i.e., a stiffer hydrogel that is created in a shorter time. In vitro cell viability tests on human fibroblasts were performed and indicated high biocompatibility. Our findings demonstrate that these injectable hydrogel scaffolds offer a promising solution for enhancing soft tissue repair and regeneration, providing a customizable and resilient framework that is expected to support tissue integration and healing with minimal surgical intervention.

PLGA-PEG Nanoparticles Loaded with Cdc42 Inhibitor for Colorectal Cancer Targeted Therapy.

Background/Objectives: An inhibitor of small Rho GTPase Cdc42, CASIN, has been shown to reduce cancer cell proliferation, migration, and invasion, yet it has several limitations, including rapid drug elimination and low bioavailability, which prevents its systemic administration. In this study, we designed and characterized a nanoparticle-based delivery system for CASIN encapsulated within poly(lactide-co-glycolide)-block-poly(ethylene glycol)-carboxylic acid endcap nanoparticles (PLGA-PEG-COOH NPs) for targeted inhibition of Cdc42 activity in colon cancer. Methods: We applied DLS, TEM, and UV-vis spectroscopy methods to characterize the size, polydispersity index, zeta potential, encapsulation efficiency, loading capacity, and in vitro drug release of the synthesized nanoparticles. The CCK-8 cell viability test was used to study colorectal cancer cell growth in vitro. Results: We showed that CASIN-PLGA-PEG-COOH NPs were smooth, spherical, and had a particle size of 86 ± 1 nm, with an encapsulation efficiency of 66 ± 5% and a drug-loading capacity of 5 ± 1%. CASIN was gradually released from NPs, reaching its peak after 24 h, and could effectively inhibit the proliferation of HT-29 (IC50 = 19.55 µM), SW620 (IC50 = 9.33 µM), and HCT116 (IC50 = 10.45 µM) cells in concentrations ranging between 0.025-0.375 mg/mL. CASIN-PLGA-PEG-COOH NPs demonstrated low hemolytic activity with a hemolytic ratio of less than 1% for all tested concentrations. Conclusion: CASIN-PLGA-PEG-COOH NPs have high encapsulation efficiency, sustained drug release, good hemocompatibility, and antitumor activity in vitro. Our results suggest that PLGA-PEG-COOH nanoparticles loaded with CASIN show potential as a targeted treatment for colorectal cancer and could be recommended for further in vivo evaluation.