Human Fibroblast Cells Research Articles

Antibacterial gelatin/tragacanth gum films containing galbanum essential oil for in vitro scratch-healing

Natural substances and bioactive agents possess great potential in wound care based on their ability to promote healing and prevent infection. This study focused on the fabrication of antibacterial wound dressings by combining gelatin (G), tragacanth gum (TG), and galbanum essential oil (GEO) as a loaded drug. TG addition resulted in more elastic and flexible films besides enabling encapsulation of the hydrophobic GEO into the biopolymeric matrix. GEO was utilized as an antibacterial and a wound-healing enhancer for open wounds such as incisions. Field emission scanning electron microscopy (FE-SEM) analysis revealed a porous film structure after GEO incorporation. Higher GEO concentration caused reduced swelling and slower degradation. Water vapor transfer rate varied from 596 to 894 g/m2.day, making the films suitable for wound dressings. GEO release exhibited a two-phase profile with prolonged diffusion-controlled release for a higher content of GEO. The films demonstrated dose-dependent antimicrobial activity against S. aureus and E. coli strains. Effectiveness and noteworthy application of this research were approved by scratch assay on human dermal fibroblast cells, and films with 3 % GEO showed 79.42 % wound closure, which is significantly higher than the control sample (55.15 %), indicating promoted cell migration and promising wound healing properties.

A Two-Photon-Active Zr-Based Metal-Organic Framework-Based Orthogonal Nanoprobe for Recognition of Cellular Senescence.

A luminescent nanoprobe capable of orthogonal sensing of two independent events is highly significant for unbiased disease-related detection such as the detection of senescent cells. Moreover, it is invaluable that the nanoprobe possesses a two-photon excitable characteristic that is highly suitable for imaging living cells and tissues. Herein, we present a two-photon-excitable multiluminescent orthogonal-sensing nanoprobe (OS nanoprobe) capable of detecting both pH elevation and β-galactosidase (β-gal) overexpression in senescent cells. In the design, Zr-based dual-emissive metal-organic frameworks prepared from two mixed amino linkers, referred to as NH2-MU, were used as the component for the ratiometric sensing of pH; additionally, fluorogenic resorufin-β-d-galactopyranoside, linked to the NH2-MU framework, enables β-gal detection. In the OS nanoprobe, the signals for pH and β-gal sensing remain independent while maintaining high colocalization. The two-photon excitable organic linkers of NH2-MU impart the OS nanoprobe with a bioimaging capability, allowing for the differentiation of senescent human foreskin fibroblast (HFF) cells from younger HFF cells or LacZ positive cells with the 800 nm laser excitation. This study marks the first instance of achieving the multiplexed orthogonal fluorescent sensing of cellular senescence using a two-photon excitation strategy, suggesting the potential of using versatile metal-organic framework (MOFs)-based fluorophores to realize the orthogonal multiplexing of disease-related biomarkers through multiphoton excitation.

β-Cyclodextrin as an elicitor of polyphenolic contents of barley (Hurdeum vulgare) callus with antioxidant and anti-aging properties on human skin fibroblast cells (HFF2)

The skin, as the largest organ of body, plays a crucial role in human beauty, yet is vulnerable to aging due to ultraviolet radiation (UV) exposure. Various methods, including intradermal injection, surgery, laser therapy, stem cell applications, and cosmetic treatments, are employed to combat skin aging. Phytochemicals, known for their antioxidant properties, are extensively studied for skincare. Barley (Hurdeum vulgare) contains diverse phenolic compounds, exhibiting strong antioxidant activity, potentially reducing cellular aging. Here, we examined the anti-aging properties of barley callus extract enriched with polyphenols in human skin fibroblast cells (HFF2) exposed to UVB radiation. β-cyclodextrin was used as an elicitor at concentrations of 2.5 and 5 ppm to enhance the production of polyphenolic compounds in the callus of black and yellow barley. A concentration of 5 ppm significantly increased the phenolic and flavonoid content in both black and yellow barley callus. Analysis of the extracts obtained from the callus showed that aqueous and ethanolic extracts of black barley callus, at a concentration of 0.1 μg/mL, enhanced cell viability and decreased matrix metalloproteinase-1 (MMP-1) levels while increasing sirtuin (SIRT1) production in UVB-exposed HFF2 cells. These extracts also exhibited high antioxidant effects and promoted cell migration, DNA integrity, and cell cycle advancement. The findings suggest the potential use of black barley callus extract in cosmetics to mitigate UV radiation effects and premature skin aging. Additionally, β-cyclodextrin is highlighted as a valuable elicitor for enhancing polyphenol content, contributing to the antioxidant and anti-aging properties of black barley callus for skincare applications.

Synthesis of new phenothiazine derivatives: Molecular docking, assessment of cytotoxic activity and oxidant-antioxidant properties on PCS-201-012, HT-29, and SH-SY5Y cell lines.

Phenothiazine (PTZ) derivatives have been acknowledged as versatile compounds with significant implications across various areas of medicine, particularly, in cancer research. The cytotoxic effects of synthesized compounds on both normal and cancerous cells, along with their oxidant-antioxidant properties, are pivotal factors in cancer treatment strategies. In the current study, eight new PTZ derivatives were synthesized and the compounds' cytotoxic activities were assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay while the oxidant-antioxidant properties were evaluated by oxidative stress index (OSI) calculation in SH-SY5Y (a human neuroblastoma cell line), HT-29 (a human colorectal adenocarcinoma cell line), and PCS-201-012 (a human primary dermal fibroblast cell line) cells. Consequently, the half-maximal inhibitory concentration (IC50) values of compound 3a were determined to be 218.72, 202.85, and 227.86 μM while the IC50 values of compound 3b were defined to be 227.42, 199.27, and 250.11 μM in PCS-201-012, HT-29, and SH-SY5Y cells, respectively. Additionally, it was determined that the synthesized compounds demonstrated the lowest OSI in PCS-201-012 cells as compared to the other cell lines.

Synthesis and characterization of 7-diethylamino-4-Chloromethyl coumarin: Spectroscopic analysis, molecular docking, and anticancer activity on large intestine carcinoma cells

Cancer, characterized by uncontrolled cell growth and metastasis, poses a significant global health burden, ranking as a leading cause of mortality worldwide. Colorectal cancer (CRC) specifically accounts for a substantial portion of cancer cases, with increasing incidence projected over the coming decades. While conventional treatments exist, they often entail adverse effects and limited efficacy, driving interest in natural remedies like coumarin derivatives due to their diverse biological activities and perceived safety profile. This study focuses on the synthesis and characterization of a novel compound, 7-diethylamino-4-chloromethyl coumarin (referred to as 7D4C), derived from coumarin. Structural elucidation employed Fourier transform infrared spectroscopy (FT-IR), proton and carbon-13 nuclear magnetic resonance spectroscopy (1H and 13C NMR), and mass spectrometry (MALDI-TOF-MS). Molecular docking studies were conducted to explore potential biological interactions. Furthermore, the anti-cancer potential of 7D4C was assessed using human epithelial adenocarcinom (LoVo) and healthy fibroblast (CCD-18Co) cell lines. Viability analysis, comet assay for DNA damage, and evaluation of cancer biomarkers including apoptosis, intracellular reactive oxygen species (iROS) levels, mitochondrial membrane potential (MMP), intracellular glutathione (GSH) concentration, and intracellular calcium (iCa2+) levels were performed. The synthesis of 7D4C was successfully completed, and its structure was confirmed. Molecular docking results indicate that 7D4C exhibits strong binding affinity to the p53 protein, highlighting its potential as a novel modulator of p53 activity. Subsequent investigations revealed that the synthesized compound induced apoptosis in cancer cells by reducing MMP and triggering DNA damage through the production of iROS. The promising anti-cancer activity of 7D4C in the LoVo cell line highlights its importance in coumarin-based therapies. Introducing 7D4C could significantly enhance future research in this area, leveraging insights from in vitro coumarin studies.

Electrospun zeolitic imidazole framework-8 loaded silk fibroin/polycaprolactone nanofibrous scaffolds for biomedical application

The development of electrospun nanofibrous scaffolds (NFSs) have aroused much attraction in the field of biomedical engineering, due to their small fiber diameter, high specific surface area, and excellent extracellular matrix comparability. The main focus of this study is to design and fabricate novel zeolitic imidazole framework-8 (ZIF-8)-loaded silk fibrin/polycaprolactone (SF/PCL) nanofiber composite scaffolds by using the electrospinning strategy. Firstly, ZIF-8 was synthesized and characterized, which showed remarkable features in terms of shape, size, chemical and physical properties. Then, three different amounts of ZIF-8 were encapsulated into SF/PCL nanofibers during electrospinning, to investigate how the addition of ZIF-8 affected the morphology, and structure, as well as physical, mechanical, and biological properties of the nanofiber composite scaffolds. It was found that the addition of ZIF-8 didn't change the nanofibrous morphology of the composite scaffold, and no bead-like structure were found for the SF/PCL composite scaffolds loading with or without ZIF-8. The appropriate addition of ZIF-8 could significantly increase the mechanical properties of SF/PCL NFSs. The SF/PCL NFS containing 5% ZIF-8 showed high ultimate stress and initial modulus, which were 40.31 ± 2.31 MPa, and 569.19 ± 21.38 MPa, respectively. Furthermore, the MTT assay indicated that the pure SF/PCL scaffold and one with 1% ZIF-8 exhibited nearly identical cell compatibility toward human dermal fibroblast (HDF) cells, but some obvious cytotoxicity was observed with the increase of ZIF-8 content. However, the incorporation of ZIF-8 into SF/PCL NFSs was found to have excellent antibacterial rate against both E. coli and S. aureus. In all, the incorporation of 1% ZIF-8 could impart the SF/PCL NFS with balanced bio-function, making it a promising candidate for diverse biomedical applications such as tissue engineering and wound healing.

Design and Synthesis of Six‐Armed Antimicrobial Polymer to Battle Against Nosocomial Pathogens

ABSTRACTThe world is under threat from microorganisms that have evolved resistance to currently available medicinal drugs and multidrug resistance among bacterial pathogens is becoming to be a serious public health issue worldwide. It is imperative to find or develop new antibiotics against Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species (ESKAPE) pathogens because they exhibit multidrug resistance and virulence through a variety of mechanisms. ESKAPE infections are often associated with increased global healthcare costs, morbidity, and illnesses. As a result, the utilization of naturally occurring bioactive agents offers a solution since these molecules are widely available and safe to employ during their use. Ricinoleic acid, an extract of seed oil that contains an unsaturated omega‐9 fatty acid and hydroxy acid, has garnered increasing interest due to its wide‐range antibacterial properties. Herein, we report the synthesis of polyethyleneimine‐ricinoleic acid‐based polymer (PEIRA) to battle against Methicillin‐resistant Staphylococcus aureus (MRSA), Acinetobacter baumannii (one among ESKAPE) and Staphylococcus aureus. The synthesized polymer showed efficient activity against MRSA, Acinetobacter baumannii, and Staphylococcus aureus. A noticeable percentage inhibition against MRSA (74%), A. baumannii (73%), and S. aureus (81%) was achieved at 2.5 mg/mL. The time‐kill assay revealed that the PEIRA polymer exhibited a bacteriostatic effect against all the tested pathogens. Cytotoxicity assay on human dermal fibroblast (HDF) cell lines revealed the PEIRA has an IC 50 of 31.7 μg/mL and exhibited above 75% cell viability at the concentration 5 and 10 μg/mL were deemed nontoxic on HDF. Based on the investigation, the synthesized PEIRA polymer may have promising applications in the biomedical sector and hospital‐acquired infections (HAIs).

Folate-Functionalized Chitosan-PLGA Nanoparticles: A Novel approach for targeted osthole delivery in pancreatic cancer

Efficient drug delivery systems targeting cancer cells are crucial for enhancing cancer therapy. In this study, we developed PLGA nanoparticles coated with folate-conjugated chitosan (osthole-PLGA-NPs/CS-FA) to deliver osthole to cancer cells and investigated its inhibitory and molecular signaling mechanisms in the PANC-1 pancreatic cancer cell line. Field emission scanning electron microscopy (FESEM) revealed that osthole-PLGA-NPs/CS-FA had a spherical structure with a uniform size distribution. Dynamic light scattering (DLS) analysis showed an average size of 171.76 nm, a dispersion index 0.26, and a surface charge of + 33.08 mV, indicating stability and uniform dispersion. Fourier-transform infrared (FTIR) spectrum analysis confirmed the successful incorporation of osthole into the PLGA nanoparticles, with an encapsulation efficiency of 93.12 %. These physicochemical properties suggest efficient cellular uptake and targeted delivery. The antioxidant potential of osthole-PLGA-NPs/CS-FA was evaluated using the ABTS assay, showing concentration-dependent inhibition of free radicals with an IC50 value of 172.95 μg/mL. The anticancer properties were assessed using the MTT assay, demonstrating a significant and concentration-dependent cytotoxic effect on PANC-1 cells (IC50 = 31.2 μg/mL) with minimal impact on normal human foreskin fibroblast (HFF) cells. DAPI staining and flow cytometry analyses confirmed a concentration-dependent increase in apoptosis in PANC-1 cells. The nanoparticles induced upregulation of Bax and downregulation of Bcl2, indicating activation of the intrinsic mitochondrial apoptotic pathway. The anti-angiogenic activity of osthole-PLGA-NPs/CS-FA was evaluated using the chick chorioallantoic membrane (CAM) assay. The results showed significant inhibition of angiogenesis in a concentration-dependent manner, starting at 40 μg/mL and increasing up to 120 μg/mL. In conclusion, osthole-PLGA-NPs/CS-FA nanoparticles exhibit promising potential for targeted pancreatic cancer therapy by enhancing cellular uptake, inducing apoptosis, and inhibiting angiogenesis.

The In vitro evaluation of in silico-designed synthetic peptides AKVUAM-1 and AKVUAM-2 on human lung fibroblast cell line MRC5 and Mycobacterium tuberculosis isolates

Tuberculosis is a major global health problem caused by Mycobacterium tuberculosis and the increase in drug resistance is driving the need for new treatments. Today, various approaches are being applied in the development of drugs for the treatment of tuberculosis. Computer-aided drug design (CADD) enables the prediction of pharmacological efficacy for potential drug molecules during the design process. Thus, new therapeutic compounds can be developed that are more potent, less toxic and have fewer side effects than existing drugs. In this study, we investigated the in vitro activities of AKVUAM-1 and AKVUAM-2 synthetic peptides designed in silico by computer-aided drug design method to inhibit the interaction between M. tuberculosis outer membrane protein Cpn T and macrophage surface receptor CR-1 and Surfactant D protein. Notably, these synthetic peptides do not show cytotoxic effect on normal lung tissue and do not kill M. tuberculosis directly. The MIC values for AKVUAM-1 were higher than 512 μg/ml for all bacterial strains except IST-16 strain (128 μg/ml). According to our results, AKVUAM-1 and AKVUAM-2 synthetic peptides have the potential to be successful candidates for investigating their potential to block macrophage entry of bacilli as targeted.

Chromatin condensed domains revealed by AFM, and their transformation in mechanically deformed normal and malignant cell nuclei

It has been generally accepted that heterochromatin is represented by a regular, dense and closed structure, while euchromatin is open and sparse. Recent evidence indicates that chromatin is comprised of irregular nucleosome clutches compacted within the nucleus. Transcriptional events transform the chromatin architecture, resulting in appearance of 100–300 nm nucleosomal aggregates. Meanwhile, the current paradigm of chromatin architecture is largely fragmented. In this communication, we unraveled chromatin ultrastructure of normal and malignant cell nuclei through mechanical deformation of the nuclei and Atomic Force Microscopy (AFM) analysis of the resulting landscape. In human skin fibroblasts cell nuclei, nanodomains of about 16.5–33.5 nm were revealed. Hierarchical folding of the chromatin of normal nuclei was observed: the nanodomains formed irregular fiber-like structures that coalesced into the macroscale chromatin compartments. In fibrosarcoma cell nuclei DNA supercoiling domains (SDs) of about 66.3–113.0 nm, uniformly distributed within the nuclei, were revealed. Transformation of the morphology of the condensed chromatin domains through up- and downregulation of supercoiling was demonstrated.

Coenzyme Q10 and Vitamin E Regulate the Bioactivity of Human Corneal Fibroblast Cells.

Purpose: Corneal fibroblasts are involved in the wound healing of the cornea with proliferation, migration, and differentiation processes. Coenzyme Q10 (CoQ10) and vitamin E can enhance corneal wound healing when applied after a corneal lesion as an eye drop. Thus, this study was performed to determine the potential efficiency of a CoQ10 ophthalmical solution containing a CoQ10 and vitamin E D-α-tocopherol polyethylene glycol 1000 succinate (TPGS)-derived formulation in human corneal fibroblasts (HCFs) in vitro. Methods: Primary HCFs were obtained from cadaveric corneal tissue, and cell viability was determined using MTT assay at 24 and 72 h. Cell migration was evaluated using an in vitro wound healing assay, and mRNA expressions of collagen type I (COL-I), collagen type III (COL-III), lumican, hyaluronan, matrix metalloproteinase (MMP)-1, MMP-2, MMP-9, tissue inhibitors of MMP (TIMP)-1, TIMP-2, interleukin (IL)-1β, IL-6, IL-8, and IL-10 were assessed using reverse transcription polymerase chain reaction at 24 and 72 h. Results: At various concentrations of CoQ10 ophthalmical solution (CoQ10-os), cell viability and wound healing rates of HCFs increased compared with the control group. The expressions of COL-I, COL-III, lumican, and hyaluronan were increased by CoQ10-os, whereas those of MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 were not affected by CoQ10-os at 24 and 72 h. In treating HCFs with a CoQ10-os medium, IL-1β, IL-6, and IL-8 decreased, whereas IL-10 was significantly increased in a time- and dose-dependent manner. Conclusions: The findings indicate that CoQ10 and vitamin E-TPGS are potent regulators of the bioactivity of HCFs, thus supporting their potential application as ophthalmical solutions in therapies aimed at the fast regeneration of damaged cornea tissues.

Carob seeds as a source of bioactive flavonoid derivatives: isolation, network pharmacology-guided anti-cancer activity, and HPLC standardization.

Carob, Ceratonia siliqua L. (CS), is a legume well-known for its edible pod pulp. Its seeds are used almost exclusively as a source of the food additive E410. Although a variety of metabolites have been identified by HPLC and LC-MS analysis in CS,reports concerned with their isolation are scarce. Methodology:In this study, two flavonoid derivatives were isolated from the methanolic extract of CS seeds, namely, quercetin-3-O-rhamnosideand 4'-p-hydroxybenzoylisorhamnetin-3,7-di-O-rhamnoside.Network pharmacology was unusually used as a guide for estimation of the biological potential of the isolated compounds. Finally, the methanolic extract of CS seeds and its ethyl acetate fraction were standardized for their 4'-p-hydroxybenzoylisorhamnetin-3,7-di-O-rhamnoside content by HPLC. The identified isolates displayed the ability to interfere with the activity of several target proteins associated with renal and colon cancers.Their cytotoxic effect on renal and colorectal cancer cell lines was investigated in comparison to Doxorubicin. The selectivity of the isolated compounds was evaluated on normal human fetal fibroblast cell lines. The isolated 4'-p-hydroxybenzoylisorhamnetin-3,7-di-O-rhamnoside showed very potent cytotoxic activity against the tested cell lines with the highest selectivity. CS seeds can be used as a source of bioactive flavonoid derivatives that can be incorporated in pharmaceutical industries.

Molecular determinants of phospholipid treatment to reduce intracellular cholesterol accumulation in NPC1 deficiency

Niemann-Pick type C (NPC) disease, caused by mutations in the NPC1 or NPC2 genes, leads to abnormal intracellular cholesterol accumulation in late endosomes/lysosomes (LE/LY). Exogenous enrichment with lysobisphosphatidic acid (LBPA), also known as bis-monoacylglycerol phosphate or BMP, either directly or via the LBPA precursor phosphatidylglycerol (PG), has been investigated as a therapeutic intervention to reduce cholesterol accumulation in NPC disease. Here we report the effects of stereoisomer configuration and acyl chain composition of LBPA on cholesterol clearance in NPC1-deficient cells. We find that S,R, S,S, and S,R LBPA stereoisomers behaved similarly, with all 3 compounds leading to comparable reductions in filipin staining in two NPC1-deficient human fibroblast cell lines. Examination of several LBPA molecular species containing one or two mono- or polyunsaturated acyl chains showed that all LBPA species containing one 18:1 chain significantly reduced cholesterol accumulation, whereas the shorter chain species di-14:0 LBPA had little effect on cholesterol clearance in NPC1 deficient cells. Since cholesterol accumulation in NPC1 deficient cells can also be cleared by PG incubation, we used non-hydrolyzable PG analogues to determine whether conversion to LBPA is required for sterol clearance, or whether PG itself is effective. The results showed that non-hydrolyzable PG species were not appreciably converted to LBPA and showed virtually no cholesterol clearance efficacy in NPC1 deficient cells, supporting the notion that LBPA is the active agent promoting LE/LY cholesterol clearance. Overall these studies are helping to define the molecular requirements for potential therapeutic use of LBPA as an option for addressing NPC disease.

Effect of Metformin on Interleukin-6 Expression in Human Fibroblast Cell Aging Model

During skin aging, there is excessive secretion of inflammatory cytokines such as interleukin-6 (IL-6), which can make the ageing process more severe. Metformin has been known to have good anti-inflammatory activity through various pathways, one of which reduces the expression of inflammatory cytokines. This study aimed to analyze the effect of metformin treatment on IL-6 gene expression in an in vitro human fibroblast cell. Human fibroblast cells were obtained through foreskin isolation, and 72 cultured human fibroblast cells were divided into six groups based on cell passages, ranging from the third to the eighth. The expression of IL-6 was assessed in three treatments: negative control (normal cells), metformin 100 µM, and positive control (Vitamin E 50 µM). Treatment and measurement of IL-6 gene expression were carried out using qRT-PCR and calculated using the Livak-Schmittgen method. The results were then compared and analyzed using One-Way Analysis of Variance (ANOVA) and followed by post-hoc analysis. The negative control group had the lowest IL-6 gene expression compared to the metformin group and positive control group. The negative control IL-6 expression showed the highest value at passage 3 (0.166 ± 0.04) and the lowest at passage 6 (0.048 ± 0.04). Meanwhile, the expression of IL-6 positive control passage 4 (5.590 ± 3.34) showed the highest value and passage 7 (0.000 ± 0.00) showed the lowest value. In the metformin group, IL-6 gene expression was highest in passage 5 (0.836 ± 0.15) and lowest in passage 3 (0.078 ± 0.02). Based on treatment, there is a difference in IL-6 gene expression at passage 4 (p-value <0.05) and passage 5 (p-value <0.001). Metformin treatment passage 5 showed a significant difference with negative control (p-value <0.05) and with positive control (p-value <0.05). Our study concluded that the administration of metformin had no effect on IL-6 gene expression in human fibroblasts. However, metformin demonstrated anti-aging potential, as evidenced by a statistically significant difference in IL-6 expression between the negative control and positive control treatment groups.

Determination of Cytotoxic Activity of Aronia melanocarpa (Michx.) Elliot Fruit Extracts on Breast Cancer (MCF-7) and Cervical Cancer (HeLa) Cell Lines

Aronia (chokeberry) fruits are consumed as fresh fruit due to their high antioxidant activity, and are also preferred among the public in the production of natural medicines. Aronia melanocarpa (Michx.) Elliot, a species of the Rosaceae family, contains many phytochemical compounds such as flavonoids, phenolic compounds, lignans, terpenes, tocopherols, phospholipids, organic acids and high amounts of anthocyanins. In this study, it was aimed to determine the in vitro anticarcinogenic activities of A. melanocarpa (Michx.) Elliot fruit extracts prepared with 6 different solvents. In the study, the cytotoxic effects of the fruits were investigated using breast cancer cell line (MCF-7) and cervical cancer cell line (HeLa), and their effects on healthy cells were investigated using human endothelial cells (HUVEC) and mouse fibroblast cells (L929) by the MTT method. As a result of the study; It was determined that the highest cytotoxicity on the breast cancer (MCF-7) cell line was observed in the ethanol extract (IC50=111.44 µg/mL) and the lowest cytotoxicity was observed in the hexane extract (IC50=661.80 µg/mL). It was determined that the highest cytotoxicity on the cervical cancer (HeLa) cell line was observed in the ethanol extract (IC50=95.14 µg/mL) and the lowest cytotoxicity was observed in the ethyl acetate extract (IC50=319.51 µg/mL). According to these values; It was determined that all extracts of Aronia fruit had cytotoxic effects on MCF-7 and HeLa cell lines, selectivity index values were higher in HeLa cells, and they did not have cytotoxic effects on HUVEC and L929 healthy cell lines (IC50=411.25-663.27 µg/mL). Thus, it has been determined that the fruits of the Aronia melanocarpa species are promising in the development of new natural resources, new drugs and therapeutic agents in cancer treatment, thanks to their anticancer activities and low cytotoxicity. It is recommended that further research be conducted on the mechanisms of anticancer activity in the future

Biocompatibility and Enamel Remineralization Potential of Titanium-Doped Phosphate Glasses With/Without Diode Laser Irradiation

This study investigates the biocompatibility and enamel remineralization potential of titanium-doped phosphate glasses (TDPG), third-generation materials with promising applications in dentistry. The biocompatibility of TDPG paste and its components (TDPG powder and polyacrylic acid liquid) was assessed using human gingival fibroblast cells (hGF) and human dental pulp stem cells (hDPSC). Cell attachment and viability were examined at 1, 3, and 7 days, comparing results with Nupro® Prophy paste and Teethmate, commonly used desensitizers, and positive control cells in normal tissue culture medium. Furthermore, the remineralization potential of these glasses with or without the use of diode laser irradiation on white spot lesions in enamel was explored at 12- and 21-days using Raman Spectroscopy and Field Emission Scanning Electron Microscopy (FESEM). The remineralization potential of TDPG was compared with Nupro® Prophy paste and Teethmate. Furthermore, both sound and demineralized enamel were also used as other controls.Results indicated that cells exposed to TDPG extracts displayed similar morphology and viability to positive control cells. While all tested pastes did not show remineralizing action after 12 days, Nupro® Prophy and Teethmate exhibited such action after 21 days. However, laser application enhanced remineralization potential for all pastes at both time points, with TDPG showing comparable results to Nupro® Prophy paste and significantly higher potential than Teethmate when used with laser at 21 days.In summary, this study reveals the biocompatibility and enhanced remineralization potential of TDPG when combined with diode laser irradiation. These findings signify innovative strides in dental materials and treatment modalities, offering new possibilities for advanced dental applications.

Spontaneous Efficient Differentiation of Human Pluripotent Stem Cells (hPSC) Upon Co-culture of hPSCs with Human Neonatal Foreskin Fibroblasts in 3D.

Pluripotent stem cells (PSCs) form well-formed embryoid bodies (EBs) in 3D culture. These EBs are formed in culture media lacking leukemia inhibitory factor (LIF) or basic fibroblast growth factor (bFGF) in mouse and human PSCs, respectively. EBs are excellent technical tools for understanding developmental biology and inducing controlled differentiation in succeeding experimental steps. Technically speaking, EBs are spontaneously differentiated PSCs in 3D and exhibit all three lineages in a time-point/sequential manner. For example, ectoderm will form first, followed by mesoderm and endoderm. We have attempted to co-culture human neonatal foreskin-derived fibroblast cells in our laboratory with the PSCs first in 2D conditions followed by the induction of EBs (PSC+fibroblasts co-cultured) in low attachment dishes. We also performed spontaneous differentiation of such EBs (co-cultured with fibroblasts). We checked the presence of markers of various lineages, namely, ectoderm, mesoderm, and endoderm in days 6, 10, and 12day EBs. We have also compared the fibroblast co-cultured EBs, along with control EBs (derived from only PSCs). This co-culture system mimics the natural conditions of uterine implantation and the role of the endometrial fibroblasts in the induction of further embryonic development. The fibroblast co-cultured iPSC EBs had better roundness scores than the normal iPSC EBs and had a higher expression of lineage-specific markers.

Selective laser melting of Ti6Al4V alloy: Effects of graphene-TiO2 nanotubes composites corrosion and biocompatibility

This study investigates the effects of graphene amount on TiO2 nanotubes (TNT) synthesized on Ti6Al4V alloy via selective laser melting (SLM) to optimize corrosion resistance and biocompatibility. XRD analysis indicated the presence of anatase and rutile phases in TNTs, and the peak shifts indicated that graphene was successfully incorporated into the TNT structure. SEM images revealed that increasing the amount of graphene resulted in smaller nanotube diameters, increased contact angles, and imparted hydrophobic properties. Corrosion tests including Tafel polarization and electrochemical impedance spectroscopy (EIS) showed that graphene, especially C4-TNTs, exhibited superior corrosion resistance with high Ecorr and Rt values. Biocompatibility tests with human dermal fibroblast cells (HDFa) demonstrated cell viability with the incorporation of graphene into TNTs. The findings suggest that the optimum amount of graphene can significantly improve the corrosion resistance and biocompatibility of TiO2 nanotubes on Ti6Al4V alloy, making them more suitable for biomedical implants.

Reusable glassware for routine cell culture—a sterile, sustainable and affordable alternative to single-use plastics

Cell culture is a cornerstone of in vitro biological research. Whilst glassware was once commonplace in tissue culture facilities, in recent decades laboratories have moved towards a heavy reliance on single use plastics for routine procedures. Single use plastics allow for accessible, sterile, and often affordable equipment that comes at a high environmental cost. We developed a glassware preparation and cleaning process that allowed the comparison of “traditional” plastic-heavy, and adapted “sustainable,” cell culture practices, to empirically compare the sterility, viability, and proliferative capacity of cells cultured with differing techniques, by observing IL-6 production, morphology, and proliferation rate of cultured human pulmonary fibroblast cells. During which, we calculated the carbon footprint of traditional versus sustainable methods. We additionally endeavored to provide a realistic overview of the steps required to transition to more sustainable cell culture practices and make suggestions to ease the cost, labor, and time required to uptake similar practices in other laboratories. Cells cultured using reusable glassware did not show signs of contamination or stress compared to cells grown solely with plasticware, and glassware baked at 180°C for 120 min was sufficiently decontaminated and depyrogenated for culturing these cells. An individual researcher adopting the same methodology could reduce their carbon footprint by 105.92 kg of Carbon dioxide equivalent (CO2e) whilst also saving money (£408.78) over a 10-year period. We predict that these benefits would be greater if more researchers were to uptake these adapted practices. We intend for this paper to reassure researchers that viable, sterile, and sustainable routine cell culture can be achieved with little upfront cost to the researcher, with the prospective benefit of greatly reducing the cost to the environment. We additionally hope that increased uptake, and thus demand of more sustainable practices, encourages suppliers, policy makers, and funding bodies to make sustainable practices more accessible to individual researchers and institutions worldwide.

Enhanced Therapeutic Efficacy of Gold Nanoparticle-Enhanced Laser Therapy for Oral Cancer: A Promising Photothermal Approach.

Introduction: Laser therapy employs a concentrated and slender light beam to eliminate or eradicate cancerous cells and pre-cancerous abnormalities. The specific wavelength of 808 nm light is preferentially absorbed by tumor cells compared to healthy cells. This study aimed to assess the combined therapeutic impact of laser and gold, given that gold exhibits photothermal properties when exposed to laser radiation. Methods: In this in vitro study, two cell lines, namely healthy HuGu cells (human gingival fibroblast cells) and head and neck cancer cells (HN-5), were obtained from the Pasteur Institute. The effect of the laser diode with a density of 3 J/cm2 and wavelength of 808 nm on the proliferation and the survival rate of oral squamous cell carcinoma (HN-5) and human gingival fibroblast (HUGU) was assessed in 60seconds. MTT assay, DAPI test, and trypan blue staining were used to determine the growth and proliferation of HN-5 and HUGU cell lines. Results: Findings showed that the laser diode along with gold decreased the rate of proliferation and survival cells in HN-5 compared to healthy cells. The changes in the cell population treated with gold and laser diode 808 were insignificant. Conclusion: Findings reveal that using a low-power laser can effectively inhibit the growth of oral cancer cells. It seems that photothermal therapy is a novel approach to oral cancer therapy.