Improved Cell Proliferation Research Articles

Senescence and Stress Signaling Pathways in Corneal Cells After Nitrogen Mustard Injury

Mustard gas keratopathy (MGK), a complication of exposure to sulfur mustard, is a blinding ocular surface disease involving key cellular pathways, including apoptosis, oxidative stress, and inflammation. Recent studies indicate that cellular senescence contributes to the pathophysiology of mustard gas toxicity. This study aimed to assess senescence and stress-related pathways—particularly mitogen-activated protein kinase (MAPK) signaling—in nitrogen mustard (NM)-induced corneal injury. In vitro, primary human corneal epithelial (P-HCECs), primary human corneal mesenchymal stromal cells (hcMSCs), and human corneal–limbal epithelial cell (HCLE) lines were exposed to varying concentrations of NM. The results demonstrated a dose-dependent increase in cellular senescence, characterized by reduced Ki67 expression, elevated p16, and p21 mRNA levels, as well as activation of the MAPK pathway activation. Treatment with a selective p38-MAPK inhibitor significantly reduced senescence markers and improved cell proliferation following exposure to NM. Overall, these studies indicate that NM exposure triggers cellular senescence and stress-related MAPK signaling, while p38-MAPK inhibition mitigates these effects, suggesting a potential therapeutic strategy.

TCF4 promotes apoptosis and Wnt/β-catenin signaling pathway in acute kidney injury via transcriptional regulation of COX7A2L.

Acute kidney injury (AKI) is still a serious kidney illness with high morbidity and death rates, and it's crucial to comprehend the underlying molecular causes. Bioinformatics analysis was performed on GSE139061 and GSE30718 data sets, and COX7A2L was screened out. The role of COX7A2L in H/R-treated cells and its transcriptional regulation with TCF4 were assessed. In vitro experiments analyzed the regulation of COX7A2L and TCF4 on the proliferation, apoptosis, and Wnt/β-catenin signaling pathway of H/R-treated cells. COX7A2L as a hub gene was downregulated in AKI samples. In H/R-treated cells, COX7A2L overexpression inhibited apoptosis and promoted cell proliferation, while COX7A2L knockdown promoted apoptosis and inhibited cell proliferation. Notably, TCF4 exhibited a significant positive correlation with COX7A2L. TCF4 overexpression-induced apoptosis was lessened and improved cell proliferation was countered by COX7A2L knockdown, according to rescue study findings. Besides, we discovered that TCF4 overexpression increased the expression of proteins linked to the Wnt/β-catenin signaling pathway (c-myc, β-catenin, and cyclin D1), while underexpression of COX7A2L counteracted this effect. The study revealed the pivotal role of COX7A2L in AKI, which is regulated by TCF4 and modulates the Wnt/β-catenin signaling pathway, highlighting its potential as a therapeutic target.

ALKBH5 Protects Against Hepatic Ischemia-Reperfusion Injury by Regulating YTHDF1-Mediated YAP Expression.

Ischemia/reperfusion (I/R) injury with severe cell death is a major complication involved in liver transplantation and resection. The identification of key regulators improving hepatocyte activity may provide potential strategies to clinically resolve I/R-induced injury. N6-methyladenosine (m6A) RNA modification is essential for tissue homeostasis and pathogenesis. However, the potential involvement of m6A in the regulation of hepatocyte activity and liver injury has not been fully explored. In the present study, we found that hepatocyte AlkB homolog H5 (ALKBH5) levels were decreased both in vivo and in vitro I/R models. Hepatocyte-specific ALKBH5 overexpression effectively attenuated I/R-induced liver necrosis and improved cell proliferation in mice. Mechanistically, ALKBH5-mediated m6A demethylation improved the mRNA stability of YTH N6-methyladenosine RNA-binding protein 1 (YTHDF1), thereby increasing its expression, which consequently promoted the translation of Yes-associated protein (YAP). In conclusion, ALKBH5 is a regulator of hepatic I/R injury that improves hepatocyte repair and proliferation by maintaining YTHDF1 stability and YAP content. The ALKBH5-m6A-YTHDF1-YAP axis represents promising therapeutic targets for hepatic I/R injury to improve the prognosis of liver surgery.

Non-electrophilic NRF2 activators promote wound healing in human keratinocytes and diabetic mice and demonstrate selective downstream gene targeting

The transcription factor NRF2 plays an important role in many biological processes and is a promising therapeutic target for many disease states. NRF2 is highly expressed in the skin and is known to play a critical role in diabetic wound healing, a serious disease process for which treatment options are limited. However, many existing NRF2 activators display off-target effects due to their electrophilic mechanism, underscoring the need for alternative approaches. In this work, we investigated two recently described non-electrophilic NRF2 activators, ADJ-310 and PRL-295, and demonstrated their efficacy in vitro and in vivo in human keratinocytes and Leprdb/db diabetic mice. We also compared the downstream targets of PRL-295 to those of the widely used electrophilic NRF2 activator CDDO-Me by RNA sequencing. Both ADJ-310 and PRL-295 maintained human keratinocyte cell viability at increasing concentrations and maintained or improved cell proliferation over time. Both compounds also increased cell migration, improving in vitro wound closure. ADJ-310 and PRL-295 enhanced the oxidative stress response in vitro, and RNA-sequencing data showed that PRL-295 activated NRF2 with a narrower transcriptomic effect than CDDO-Me. In vivo, both ADJ-310 and PRL-295 improved wound healing in Leprdb/db diabetic mice and upregulated known downstream NRF2 target genes in treated tissue. These results highlight the non-electrophilic compounds ADJ-310 and PRL-295 as effective, innovative tools for investigating the function of NRF2. These compounds directly address the need for alternative NRF2 activators and offer a new approach to studying the role of NRF2 in human disease and its potential as a therapeutic across multiple disease states.

Enhancing Cellular Homeostasis: Targeted Botanical Compounds Boost Cellular Health Functions in Normal and Premature Aging Fibroblasts.

The human skin, the body's largest organ, undergoes continuous renewal but is significantly impacted by aging, which impairs its function and leads to visible changes. This study aimed to identify botanical compounds that mimic the anti-aging effects of baricitinib, a known JAK1/2 inhibitor. Through in silico screening of a botanical compound library, 14 potential candidates were identified, and 7 were further analyzed for their effects on cellular aging. The compounds were tested on both normal aged fibroblasts and premature aging fibroblasts derived from patients with Hutchinson-Gilford Progeria Syndrome (HGPS). Results showed that these botanical compounds effectively inhibited the JAK/STAT pathway, reduced the levels of phosphorylated STAT1 and STAT3, and ameliorated phenotypic changes associated with cellular aging. Treatments improved cell proliferation, reduced senescence markers, and enhanced autophagy without inducing cytotoxicity. Compounds, such as Resveratrol, Bisdemethoxycurcumin, Pinosylvin, Methyl P-Hydroxycinnamate, cis-Pterostilbene, and (+)-Gallocatechin, demonstrated significant improvements in both control and HGPS fibroblasts. These findings suggest that these botanical compounds have the potential to mitigate age-related cellular alterations, offering promising strategies for anti-aging therapies, particularly for skin health. Further in vivo studies are warranted to validate these results and explore their therapeutic applications.

The Effect of Platelet Fibrin Plasma (PFP) on Postoperative Refractory Wounds: Physiologically Concentrated Platelet Plasma in Wound Repair.

Surgical wounds that can't complete primary healing three weeks after surgery are called postoperative refractory wounds. Postoperative refractory wounds would bring great physical and life burdens to the patients and seriously affect their quality of life. To investigate the effect of platelet fibrin plasma (PFP) on postoperative refractory wound healing. The composition of PFP was analyzed using blood routine and blood biochemicals. Clinical data were collected that met the inclusion criteria after treatment with PFP, and the efficacy of PFP was evaluated by wound healing rate and days to healing. Next, growth factor content in PFP, PRP, and PPP was analyzed using ELISA, and PFP-treated cells were applied to investigate the effect of PFP on fibroblast and endothelial cell function. PFP component analysis revealed no statistical difference between platelet concentration in PFP and physiological concentration. Clinical statistics showed that PFP treatment was effective in the postoperative refractory wound (four-week wound healing rate > 90%), significantly better than continuous wound dressing. Meanwhile, our result also proved that PFP treatment significantly enhanced vascularization by upregulated the expression level of CD31 and improved granulation tissue thickness. Activated PFP, PRP, and PPP could continuously release growth factors in vitro and the amount of growth factors released by PRP and PFP was significantly higher than PPP. In vitro studies demonstrated that active PFP could improve cell proliferation, migration, adhesion, and angiogenesis in fibroblasts and endothelial cells. Physiologically concentrated platelet plasma promoted wound healing and improved related cellular functions. The modified PFP (responsible for accelerating wound healing and enhancing the migration and proliferation of fibroblasts and endothelial cells) was prepared and analyzed for its clinical effectiveness in postoperative refractory wounds. Physiologically concentrated platelet plasma promoted wound healing and improved related cellular functions. The preparation of PFP could significantly reduce the amount of prepared blood, with a good application value for postoperative wounds. PFP can be considered a treatment option, especially for postoperative refractory wounds.

Rejuvenation of human mesenchymal stem cells using a nonintegrative and conditionally removable Sendai virus vector

Human mesenchymal stem cells (hMSCs) with extended lifespan and differentiation potential that can recapitulate in vivo characteristics could significantly contribute to basic research, drug development, and cell therapy. Specifically, they could ensure a stable supply of specific cellular resources, and possibly extracellular vesicles. Here, we established a technology for extending the lifespan while maintaining differentiation potential, termed “rejuvenation,” of hMSCs (rej-hMSCs) using nonintegrative and conditionally removable temperature-sensitive Sendai virus (SeV) vectors. Various immortalizing factors (i.e., Bmi-1, hTERT, SV40T, and/or HPV E6/E7) were first introduced by the SeV vector into the cells. A combination of three SeVs with Bmi-1, hTERT, or SV40T conferred markedly improved cell proliferation and cloning ability while maintaining differentiation potential and a normal karyotype. An extended lifespan was also demonstrated in other cell types. The rejuvenation of long-passaged or aged hMSCs was also confirmed. SeV vectors were rapidly removed as a function of cell doubling by increasing the temperature from 35 °C to 37 °C or higher, while proliferative ability was maintained. Following FACS sorting, the complete removal of SeV vectors was confirmed by qPCR analyses. Therefore, our cell rejuvenation technology could contribute to research and clinical applications by enabling the supply of modified cells without damaging host chromosomes.

Bioactive surface modification of Ti–Nb alloy by alkaline treatment in potassium hydroxide solution

Abstract The aim of this work is to compare the responsive behaviour of titanium, niobium and titanium–niobium alloy during alkaline treatment in forming alkaline titanate layer and their resultant bioactive properties. Titanium and niobium powder mixture with composition in the beta region was pressed at 550 MPa and sintered at 1,200 °C for 2 h. The alloy was soaked in potassium hydroxide aqueous solution at 60 °C for 24 h with different concentrations of 0.5 M and 5 M. The effect of post sintering-heat treatment was investigated by annealing the treated alloy at 600 °C for 2 h. X-ray diffraction analysis and Fourier transform infrared spectroscopy analysis were used to evaluate the chemical composition and the functional group of material on the treated alloy surface respectively. Immersion in Hanks solution for 1 day resulted in traces of calcium and phosphate on alloy surfaces treated in different concentrations of alkali as well as post-heat treatment. The cell viability evaluation using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay on the new beta-Ti alloy with potassium-based titanate layer demonstrated potassium hydroxide treatment with a 5 M concentration after post-heat treatment significantly improved cell proliferation, which is a prerequisite for bone mineral apatite deposition.

Preparation and Biological Evaluation of Porous Tantalum Scaffolds Coated with Hydroxyapatite.

Orthopedic implants, such as porous scaffolds, are an effective way to repair bone defects. However, the lack of osseointegration and osteoinduction limits the achievement of an ideal therapeutic effect. This study aimed to prepare hydroxyapatite (HA) coatings for the surface of porous tantalum (Ta) scaffolds and to assess the effectively improved biological activities of the coated scaffolds. The porous Ta scaffolds were prepared by chemical vapor deposition, and then the porous Ta scaffolds were coated with HA via electrochemical deposition. The elements and phase compositions of the coatings were analyzed by energy-dispersive X-ray spectroscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The results showed that the coating covered the whole surfaces of porous Ta scaffolds with a uniform and compact distribution and did not exert any obvious effect on the porous structure. The biological activity of porous Ta scaffolds after surface modification increased and the water contact angle decreased, indicating that hydrophilicity was significantly improved. Cell live/dead staining, cytoskeletal fluorescence staining, and alkaline phosphatase immunofluorescence staining showed that the coating exhibited no cytotoxicity and notably improved cell proliferation, spreading, and osteogenic differentiation. In addition, in vivo experiments in animals have demonstrated that HA-coated porous Ta scaffolds contribute to bone formation. In conclusion, the HA coating notably improves the biological activities of the porous Ta scaffolds, achieving the goal of the present study. The HA coating presents great potential for the modification of porous Ta implants to improve their osteogenesis and osseointegration.

Pharmacological inhibition of ENT1 enhances the impact of specific dietary fats on energy metabolism gene expression

Medium chain fatty acids are commonly consumed as part of diets for endurance sports and as medical treatment in ketogenic diets where these diets regulate energy metabolism and increase adenosine levels. However, the role of the equilibrative nucleoside transporter 1 (ENT1), which is responsible for adenosine transport across membranes in this process, is not well understood. Here, we investigate ENT1 activity in controlling the effects of two dietary medium chain fatty acids (decanoic and octanoic acid), employing the tractable model system Dictyostelium. We show that genetic ablation of three ENT1 orthologues unexpectedly improves cell proliferation specifically following decanoic acid treatment. This effect is not caused by increased adenosine levels triggered by both fatty acids in the presence of ENT1 activity. Instead, we show that decanoic acid increases expression of energy-related genes relevant for fatty acid β-oxidation, and that pharmacological inhibition of ENT1 activity leads to an enhanced effect of decanoic acid to increase expression of tricarboxylicacid cycle and oxidative phosphorylation components. Importantly, similar transcriptional changes have been shown in the rat hippocampus during ketogenic diet treatment. We validated these changes by showing enhanced mitochondria load and reduced lipid droplets. Thus, our data show that ENT1 regulates the medium chain fatty acid-induced increase in cellular adenosine levels and the decanoic acid-induced expression of important metabolic enzymes in energy provision, identifying a key role for ENT1 proteins in metabolic effects of medium chain fatty acids.

Fiber/hydrogel hybrid wound dressing based on eggshell membrane containing postbiotic ingredients

Skin is the largest organ in body which has important functions. Therefore, to have a healthy skin is very essential, and wound dressings are specifically designed to promote the wound healing process. The aim of this study is to prepare and characterize a fiber-hydrogel wound dressing based on eggshell membrane (ESM) enriched with postbiotic compounds extracted from Lactobacillus plantarum NIMBB003 bacteria. For this purpose, ESM was effectively separated from eggshells through acidic treatment. Then, ultrasound was used for an optimal duration of 1.89 min at 95 % of device's power to expand the pore size of ESM from 6.89 to 10.84 μm to enhance hydrogel infiltration into ESM. The hydrogel (alginate and oxidized alginate) was then infiltrated into the ESM. ATR, SEM, and weight measurement of samples showed the proper infiltration of the hydrogel within the ESM structure. However, biostability analysis revealed that alginate hydrogel was more stable in the hybrid structure compared to oxidase alginate hydrogel. Alginate infiltration into ESM, improved the ultimate strength of the ESM to 1.89 ± 0.17 MPa and water uptake degree to 368.05 % ± 24.34 %. The water vapor transmission rate of the designed dressing was 34.14 ± 1.05 mg/cm2 after 72 h, which means the proper moist management in wound bed. Finally, addition of postbiotics at a concentration of 10 mg/ml into the hydrogel improved cell proliferation in five days. Furthermore, human dermal fibroblast cells adhered to the wound dressings properly and spread along the fibers of the ESM. In general, the developed wound dressing composed of natural biomaterials with extracellular matrix-like structure, can be used effectively to assist the wound healing process.

A Photothermal Polymeric Platform for Efficient and Safe Gene Transfection: When Polyethylenimine Collaborates with Indocyanine Green.

Gene transfection, defined by the delivery of nucleic acids into cellular compartments, stands as a crucial procedure in gene therapy. While branched polyethylenimine (PEI) is widely regarded as the "gold standard" for nonviral vectors, its cationic nature presents several issues, including nonspecific protein adsorption and notable cytotoxicity. Additionally, it often fails to achieve high transfection efficiency, particularly with hard-to-transfect cell types. To overcome these challenges associated with PEI as a vector for plasmid DNA (pDNA), the photothermal agent indocyanine green (ICG) is integrated with PEI and pDNA to form the PEI/ICG/pDNA (PI/pDNA) complex for more efficient and safer gene transfection. The negatively charged ICG serves a dual purpose: neutralizing PEI's excessive positive charges to reduce cytotoxicity and, under near-infrared irradiation, inducing local heating that enhances cell membrane permeability, thus facilitating the uptake of PI/pDNA complexes to boost transfection efficiency. Using pDNA encoding vascular endothelial growth factor as a model, our system shows enhanced transfection efficiency in vitro for hard-to-transfect endothelial cells, leading to improved cell proliferation and migration. Furthermore, in vivo studies reveal the therapeutic potential of this system in accelerating the healing of infected wounds by promoting angiogenesis and reducing inflammation. This approach offers a straightforward and effective method for gene transfection, showing potentials for tissue engineering and cell-based therapies.

Fibronectin Functionalization: A Way to Enhance Dynamic Cell Culture on Alginate/Hydroxyapatite Scaffolds.

Bone defects are a global health concern; bone tissue engineering (BTE) is the most promising alternative to reduce patient morbidity and overcome the inherent drawbacks of autograft and allograft bone. Three-dimensional scaffolds are pivotal in this field due to their potential to provide structural support and mimic the natural bone microenvironment. Following an already published protocol, a 3D porous structure consisting of alginate and hydroxyapatite was prepared after a gelation step and a freezing-drying step. Despite the frequent use of alginate in tissue regeneration, the biological inertness of this polysaccharide hampers proper cell colonization and proliferation. Therefore, the purpose of this work was to enhance the biological properties by promoting the interaction and adhesion between cells and biomaterial with the use of Fibronectin. This extracellular matrix protein was physically adsorbed on the scaffold, and its presence was evaluated with environmental scanning electron microscopy (eSEM) and the Micro-Bicinchoninic Acid (μBCA) protein assay. The MG-63 cell line was used for both static and dynamic (i.e., in bioreactor) 3D cell culturing on the scaffolds. The use of the bioreactor allowed for a better exchange of nutrients and oxygen and a better removal of cell catabolites from the inner portion of the construct, mimicking the physiological environment. The functionalized scaffolds showed an improvement in cell proliferation and colonization compared to non-functionalized ones; the effect of the addition of Fibronectin was more evident in the dynamic culturing conditions, where the cells clearly adhered on the surface of functionalized scaffolds.

Effects of ∆-9 tetrahydrocannabinol on the small intestine altered by high fructose diet: A Histopathological study

The consumption of fructose is increasing day by day. Understanding the impact of increasing fructose consumption on the small intestine is crucial since the small intestine processes fructose into glucose. ∆9-Tetrahydrocannabinol (THC), a key cannabinoid, interacts with CB1 and CB2 receptors in the gastrointestinal tract, potentially mitigating inflammation. Therefore, this study aimed to investigate the effects of the high-fructose diet (HFD) on the jejunum of rats and the role of THC consumption in reversing these effects. Experiments were conducted on Sprague–Dawley rats, with the experimental groups as follows: control (C), HFD, THC, and HFD + THC. The HFD group received a 10% fructose solution in drinking water for 12 weeks. THC groups were administered 1.5 mg/kg/day of THC intraperitoneally for the last four weeks. Following sacrification, the jejunum was evaluated for mucus secretion capacity. IL-6, JNK, CB2 and PCNA expressions were assessed through immunohistochemical analysis and the ultrastructural alterations via transmission electron microscopy. The results showed that fructose consumption did not cause weight gain but triggered inflammation in the jejunum, disrupted the cell proliferation balance, and increased mucus secretion in rats. Conversely, THC treatment displayed suppressed inflammation and improved cell proliferation balance caused by HFD. Ultrastructural examinations showed that the zonula occludens structures deteriorated in the HFD group, along with desmosome shrinkage. Mitochondria were found to be increased due to THC application following HFD. In conclusion, the findings of this research reveal the therapeutic potential of THC in reversing HFD-related alterations and provide valuable insights for clinical application.Graphical abstract

NAT10 Promotes Prostate Cancer Growth and Metastasis by Acetylating mRNAs of HMGA1 and KRT8.

N4-acetylcytidine (ac4C) is essential for the development and migration of tumor cells. According to earlier research, N-acetyltransferase 10 (NAT10) can increase messenger RNAs (mRNAs) stability by catalyzing the synthesis of ac4C. However, little is known about NAT10 expression and its role in the acetylation modifications in prostate cancer (PCa). Thus, the biological function of NAT10 in PCa is investigated in this study. Compared to paraneoplastic tissues, the expression of NAT10 is significantly higher in PCa. The NAT10 expression is strongly correlated with the pathological grade, clinical stage, Gleason score, T-stage, and N-stage of PCa. NAT10 has the ability to advance the cell cycle and the epithelial-mesenchymal transition (EMT), both of which raise the malignancy of tumor cells. Mechanistically, NAT10 enhance the stability of high mobility group AT-hook 1 (HMGA1) by acetylating its mRNA, thereby promoting cell cycle progression to improve cell proliferation. In addition, NAT10 improve the stability of Keratin 8 (KRT8) by acetylating its mRNA, which promotes the progression of EMT to improve cell migration. This findings provide a potential prognostic or therapeutic target for PCa.

A PLATZ transcription factor PhePLATZ8 from Moso bamboo (Phyllostachys edulis) plays a positive role in regulating growth and abiotic stress tolerance

Moso bamboo is recognized as a rapid-growing plant with abundant lignocellulosic content, making it a crucial non-timber forest resource on a global scale. The growth of Moso bamboo is influenced by the rapid division and elongation of its internode meristem cells, but the molecular mechanism behind this process is still unclear. Here, by combining RNA-seq data from Moso bamboo internode samples with gene co-expression network and gene functional annotation enrichment analysis, a key candidate gene named PhePLATZ8, which regulates cell division in the internodes of Moso bamboo, has been screened out. PhePLATZ8 is localized to the nucleus and cytoplasm. The phenotype of transgenic Arabidopsis thaliana plants showed that PhePLATZ8 positively regulated the organ development of transgenic plants, including hypocotyl length, plant height, petiole length, blade shape, and seed size. Overexpression of PhePLATZ8 also significantly improved cell proliferation activities in the hypocotyl calluses of transgenic plants. qRT-PCR and luciferase assays showed that PhePLATZ8 was involved in the regulation of cell proliferation by activating the expression of several GRFs in A. thaliana and Moso bamboo. Futhermore, PhePLATZ8 was significantly induced by abiotic stress, and overexpression of PhePLATZ8 increased the tolerance of transgenic A. thaliana plants to drought and salt stress. Overall, our current study initially explored the role of PhePLATZ8 in the process of rapid growth and stress response of Moso bamboo, providing genetic resources for future molecular breeding of Moso bamboo.

Development of Chitosan/Mg−Zn−HAP/ZrO2 Bilayer Coating on Ti Alloy for Biomedical Applications

AbstractTitanium alloys (Ti−6Al−4V) are being used in many biomedical applications due to their unique properties of bioactivity, excellent mechanical properties, low toxicity, biocompatibility, and long‐term implant application for their satisfactory corrosion resistance. We have developed a new two‐step fabrication process of zirconium oxide (ZrO2) and Chitosan/Mg−Zn−HAP (Chitosan/M−HAP) bilayer layer coating on Ti alloy for biomedical applications. Flower‐structured bilayers were coated by the electrochemical deposition method. Electrochemically deposited bilayer‐coated Ti alloy specimens were characterized by various analytical techniques like field‐emission scanning electron microscope (FE‐SEM), X‐ray diffraction (XRD), and EDS mapping analysis. Furthermore, the developed bilayer (Chitosan/Mg−Zn−HAP/ZrO2) coated Ti alloy samples improved mechanical properties such as adhesion strength (20.1±0.4 MPa) and hardness (446±4 Hv), compared to other coatings. An investigation of polarization curves showed a positive shift in the polarization values (Ecorr, and Icorr) of the Chitosan/M−HAP/ZrO2 bilayer coatings on Ti alloy. Bilayer‐modified Ti alloy samples show good antimicrobial response toward Gram‐negative and Gram‐positive bacteria. Meanwhile, the cell viability and proliferation of the bilayer‐coated samples were evaluated and the exhibited result improved cell proliferation, and bioactivity compared to other coated materials. From the results, it can be evident that the developed Chitosan/Mg−Zn−HAP/ZrO2 bilayer‐coated Ti alloy could be a good potential candidate for biomedical applications.

Electrical Stimulating Redox Membrane Incorporated with PVA/Gelatin Nanofiber for Diabetic Wound Healing.

Chronic wounds adversely affect the quality of life. Although electrical stimulation has been utilized to treat chronic wounds, there are still limitations to practicing it due to the complicated power system. Herein, an electrostimulating membrane incorporated with electrospun nanofiber (M-sheet) to treat diabetic wounds is developed. Through the screen printing method, the various alternate patterns of both Zn and AgCl on a polyurethane substrate, generating redox-mediated electrical fields are introduced. The antibacterial ability of the patterned membrane against both E. coli and S. aureus is confirmed. Furthermore, the poly(vinyl alcohol) (PVA)/gelatin electrospun fiber is incorporated into the patterned membrane to enhance biocompatibility and maintain the wet condition in the wound environment. The M-sheet can improve cell proliferation and migration in vitro and has an immune regulatory effect by inducing the polarization of macrophage to the M2 phenotype. Finally, when applied to a diabetic skin wound model, the M-sheet displays an accelerated wound healing rate and enhances re-epithelialization, collagen synthesis, and angiogenesis. It suggests that the M-sheet is a simple and portable system for the spontaneous generation of electrical stimulation and has great potential to be used in the practical wound and other tissue engineering applications.

Nanoclay Reinforced Integrated Scaffold for Dual-Lineage Regeneration of Cartilage and Subchondral Bone.

Tissue engineering is theoretically considered a promising approach for repairing osteochondral defects. Nevertheless, the insufficient osseous support and integration of the cartilage layer and the subchondral bone frequently lead to the failure of osteochondral repair. Drawing from this, it was proposed that incorporating glycine-modified attapulgite (GATP) into poly(1,8-octanediol-co-citrate) (POC) scaffolds via the one-step chemical cross-linking is proposed to enhance cartilage and subchondral bone defect repair simultaneously. The effects of the GATP incorporation ratio on the physicochemical properties, chondrocyte and MC3T3-E1 behavior, and osteochondral defect repair of the POC scaffold were also evaluated. In vitro studies indicated that the POC/10% GATP scaffold improved cell proliferation and adhesion, maintained cell phenotype, and upregulated chondrogenesis and osteogenesis gene expression. Animal studies suggested that the POC/10% GATP scaffold has significant repair effects on both cartilage and subchondral bone defects. Therefore, the GATP-incorporated scaffold system with dual-lineage bioactivity showed potential application in osteochondral regeneration.

Cytoprotective and Anti-inflammatory Effects of Derma Genie™-H002 against Paraphenylenediamine-induced Cytotoxicity and Inflammation in RAW 264.7 Cells

Purpose: This study aimed to establish an <i>in vitro</i> model to examine paraphenylenediamine (PPD)-induced cytotoxicity and inflammation within RAW 264.7 cells. Additionally, we investigated the cytoprotective and anti-inflammatory effects of Derma Genie™-H002 derived from extracts of five medicinal herbs. Methods: Various biochemical analyses, including water-soluble tetrazolium salt, crystal violet staining, and lactate dehydrogenase leakage assays, were employed to evaluate the impact of Derma Genie™-H002 on PPD-induced cytotoxicity and inflammation. Furthermore, apoptosis- and inflammation-related gene expression was analyzed using reverse transcription-polymerase chain reaction and Western blotting. Results: Derma Genie™-H002 exhibited no significant cytotoxicity in RAW 264.7 cells, whereas PPD induced notable cytotoxicity at concentrations >25 μM. Cotreatment with PPD and Derma Genie™-H002 not only improved cell proliferation rates but also alleviated cell viability in PPD-induced RAW 264.7 cells. Furthermore, findings demonstrated that Derma Genie™-H002 mitigates the cytotoxicity of PPD-induced RAW 264.7 cells by reducing the expression of the proapoptotic protein Bax and increasing the expression of the antiapoptotic protein Bcl-2. Moreover, cotreatment with PPD and Derma Genie™-H002 downregulated the mRNA expression of inflammasome-related genes, inflammatory cytokines, and allergic contact dermatitis-related genes, all elevated by PPD. The increased expression of nuclear factor-κB protein induced by PPD was also reduced upon cotreatment with Derma Genie™-H002, providing compelling evidence of its anti-inflammatory efficacy in the PPD-induced inflammation model. Conclusion: This study demonstrated that Derma Genie™-H002 effectively alleviates PPD-induced cytotoxicity and inflammation in RAW 264.7 cells. Therefore, the potential application of Derma Genie™-H002 as a naturally derived functional anti-inflammatory material has been validated.