Nonviral gene delivery systems, such as calcium phosphate (CaP) nanoparticles, offer a potentially safer alternative to viral vectors for multigene delivery. This study evaluated CaP nanoparticles as carriers for transferring a plasmid encoding Yamanaka factors and GFP to PBMC-derived monocytes, with a view to future nonviral iPSC production strategies. CaP nanoparticles were synthesized by microemulsion and characterized by dynamic light scattering before and after pDNA conjugation. pDNA loading efficiency was determined by measuring the unbound plasmid in the conjugation supernatant. Cytotoxicity was assessed by MTT assay for CaP/pDNA conjugate, naked pDNA, and blank nanoparticles for 24–72 h and over a concentration range. Transfection efficiency was monitored by GFP expression, and pluripotency-related marker expression (OCT-4, SSEA-4) and alkaline phosphatase (ALP) activity were evaluated in cells cultured on Matrigel. CaP1 nanoparticles (234.2 ± 10.2 nm, + 18.6 ± 4.8 mV) formed a stable conjugate with pDNA (loading efficiency: 81.3 ± 1.0
Osteoporosis (OP) is a major disease threatening worldwide public health. Mendelian randomization (MR) and bioinformatics analysis can provide novel insights for treatments. The object of our study is to identify oxidative stress-related genes associated with osteoporosis and explore their potential prognostic value by integrating bioinformatics, machine learning, and Mendelian randomization approaches. We first screened oxidative stress-related differentially expressed genes (DEGs) using GEO database data. To evaluate biological variables linked to these DEGs, we used GO biological function analysis and KEGG pathway enrichment analysis. Additionally, an algorithmic model was applied to identify highly specific genes for OP diagnosis. OP analysis was then conducted to confirm the causal relationship between PNOC (Prepronociceptin) and OP. Finally, a model was built to analyze immune cell infiltration and the PNOC-associated ceRNA network. From GEO (GSE13850, GSE35958), we retrieved DEGs between non-OP (High BMD) and OP (Low BMD) groups. Oxidative stress-related genes (OSRGs) were obtained from GENECARDS, and intersecting DEGs with OSRGs gave 37 differentially expressed OSRGs (DEOSRGs); 265 DEGs and 2069 OSRGs were identified in total. GO/KEGG showed DEOSRGs mainly participate in osteoclast differentiation. LASSO regression and SVM identified OP prognostic biomarkers: up-regulated PNPLA2, ATXN2L, ARRB2, and down-regulated PNOC, CAPN3 (AUC: 0.995–0.943). OP analysis supported PNOC-OP causality. GSEA/GSVA linked DEOSRGs to bone and immune cell differentiation (non-OP vs. OP). Further GO/KEGG connected DEOSRGs to immune cells, and PNOC-associated genes showed higher OP prognostic value. PNOC may be a highly specific prognostic biomarker and a potential therapeutic target for OP. Osteoporosis is a common condition in which bones become weak and break easily. It affects millions, so better detection and treatment are needed. Oxidative stress — an imbalance that damages cells, including bone cells — is thought to play a role. We compared gene activity in people with low and high bone density using public databases, then focused on genes linked to oxidative stress. Two computer-based methods identified five genes that distinguished people with osteoporosis from those without, with very high accuracy. One gene, PNOC, stood out: it was less active in osteoporosis, and a separate genetic analysis suggested low PNOC activity may directly contribute to the disease. These genes were also closely linked to the immune system and to bone-removing cells. PNOC therefore shows promise as a marker for identifying osteoporosis and as a possible target for new treatments, though further laboratory and clinical studies are needed to confirm these findings.
Oral thin films (OTFs) are an emerging drug delivery platform with the characteristics of high surface area, porous nanofibrous architecture, fast disintegrating and multi-drug loading ability. This review critically discusses the recent developments in OTFs, which are used for dual-drug delivery, with the major focus on cascade drug release systems, where spatially and temporally controlled drug delivery can enhance the therapeutic outcomes. A comprehensive analysis of the literature published over the past decade was conducted covering electrospinning technologies, polymer selection, drug–polymer compatibility, dual-drug loading strategies, nanofiber architecture, release mechanisms, and characterization techniques. Advanced electrospinning techniques are also emphasized, such as coaxial, triaxial, and side-by-side electrospinning, which can produce multifunctional nanofibers that deliver sequential or simultaneous delivery of two or more drugs, achieve high encapsulation efficiency, improve dissolution, and enable controlled release. The impact of formulation variables such as the nature of the polymer, the drug-polymer compatibility, the morphology of the fibers and the processing conditions on the stability and release of the drug was also addressed. There is a critical review of the key concerns that hamper the clinical translation, such as scale-up, process reproducibility, moisture sensitivity, formulation stability, quality control, regulatory compliance, and the development of strong in vitro–in vivo correlations. Lastly, new possibilities in the field of smart and stimuli-responsive polymers, AI-aided formulation design, continuous electrospinning methods, and personalized medicine are discussed. Thus, the combination of novel developments with the current challenges and future potential offer a broad perspective for both rational design and development of electrospun OTFs for dual-drug delivery and cascade drug release. Electrospun oral thin films (OTFs) are very thin, fast-dissolving films that can carry two or more medicines. They may offer a simple and convenient way to deliver medicines, especially when different drugs need to be released at different times.This review summarizes recent advances in using electrospun OTFs for dual-drug delivery and cascade drug release, where medicines are released together or in a planned sequence. This approach may improve treatment effectiveness, drug delivery, and patient convenience.The review also discusses how the materials and manufacturing methods used to prepare these films affect their quality, stability, and drug-release behaviour. However, challenges such as large-scale production, moisture sensitivity, storage stability, quality control, regulatory requirements, and confirmation of effectiveness in patients still need to be addressed.Future research involving artificial intelligence, advanced materials, improved manufacturing methods, and personalized medicine may help overcome these challenges.
In recent years, plant-derived exosomes have been proposed as novel bioactive nanocarriers to improve wound healing. This study aimed to investigate the therapeutic effect of orange exosomes loaded in carboxymethyl cellulose/gelatin/gentamicin (CMC/Gel/G) hydrogel on skin wound healing in an animal model. Exosomes were extracted from fresh orange juice by ultracentrifugation and identified using DLS, FE-SEM, and zeta potential analyses. After hydrogel preparation, biocompatibility (MTT) and microstructure (SEM) tests were performed. Then, full-thickness skin wounds were created in male rat and treated in three groups (control, hydrogel, Exosome-loaded hydrogel). On days 7 and 14, the extent of wound healing was evaluated macroscopically, histologically, and by measuring IL-1β and VEGF factors. The results showed that the exosome-loaded hydrogel significantly accelerated wound closure, improved epithelial regeneration, increased collagen deposition, and reduced inflammation compared to the control group and hydrogel alone. Also, the level of VEGF increased on day 7 and decreased on day 14, while IL-1β decreased significantly at both times. The use of orange exosomes in the hydrogel substrate, by modulating inflammation and enhancing angiogenesis, improves the wound healing process and can be considered as a safe, natural, and effective solution in regenerative medicine.
Triple-negative breast cancer (TNBC) is a serious clinical problem as it is a very aggressive cancer, that lacks targeted treatment options, and can develop a complex tumor microenvironment (TME) that promotes immune evasion, stromal desmoplasia, and hypoxia. In this review, the transformative capabilities of hyaluronic acid (HA)-based nanocapsules as a versatile nanoplatform for reprogramming the TNBC TME and addressing therapeutic resistance are discussed. These nanocapsules allow active tumor targeting and efficient delivery of intracellular immunomodulators, siRNA, and metabolic disruptors can be delivered into the tumor using their natural affinity of HA to CD44 receptors that are overexpressed in TNBC. We logically elaborate on how engineered HA nanocapsules can alter the immunosuppressive TME through tumor-associated macrophage repolarization, myeloid-derived suppressor cell elimination, and increased T-cell infiltration. In addition, we look at their effects of degrading fibrotic extracellular matrix, normalizing tumor vasculature and relieving hypoxia and acidosis, critical barriers to drug penetration and efficacy. The current preclinical innovations demonstrate synergetic interactions, including the simultaneous delivery of paclitaxel with Transforming Growth Factor Beta (for TGF-β) and Interleukin-12 (for IL-12), which leads to substantial tumor remission and metastatic inhibition. Although the outcomes have been promising, clinical translation has challenges such as off-target CD44 binding, scalability in production and long-term stability. This review summarizes the existing knowledge, discusses novel combinatoric approaches, and offers future perspectives on customized and biomarker-based HA nanocapsules as a next-generation, multi-mechanistic TNBC therapeutic. TNBC is a very aggressive form of breast tumor that tests negative for all three markers. TNBC is inherently heterogenous. TME in TNBC is immunosuppressed. HA-nanocapsules addresses therapeutic resistance and tumor regression. Lay Summary HA nanocapsules made from hyaluronic acid can target triple-negative breast cancer by binding CD44 receptors on tumor cells. They deliver drugs and immune modulators that reprogram the tumor microenvironment—repolarizing macrophages, degrading scar tissue, relieving hypoxia, and boosting T-cell activity. Preclinical studies show reduced tumors and metastasis. Though challenges like scalability and stability remain, these nanocapsules offer a promising, personalized treatment strategy for this aggressive cancer. Triple-Negative Breast Cancer (TNBC) pathophysiology. The left panel depicts the complex Tumor Microenvironment (TME) and associated drug resistance. The right panel demonstrates TME reprogramming and enhanced therapeutic response following targeted delivery via HA-nanocapsules. This graphical abstract was created by the authors using BioRender.com under a standard academic license and is original to this manuscript.
