Methods最新文献

Skin aging is driven by the progressive exhaustion of stem cell niches, epigenetic drift, and accumulation of senescent cells, which together promote both aesthetic decline and a pro-tumorigenic microenvironment. This review focuses on the emerging methodological theme of small-molecule-mediated reprogramming as a strategy to restore skin homeostasis. We evaluated the shift from traditional regenerative medicine toward targeted chemical modulation, focusing on the use of small-molecule cocktails to induce partial reprogramming and rejuvenate aged stem cell populations without erasing cellular identity. Central to this theme is the integration of high-throughput virtual screening and AI-driven predictive modeling to identify potent modulators of Wnt, Notch, and TGF-β pathways. We further bridge the gap between preclinical innovation and clinical application by analyzing "serious clinical studies" with proven efficacy, including randomized controlled trials of stem cell-derived secretomes and clinically validated small molecules, such as tretinoin and firming peptides. By contextualizing advanced delivery systems, including microneedles and stimuli-responsive nanoparticles, within this reprogramming framework, we demonstrate how spatially controlled interventions can optimize clinical outcomes. This review provides a unified perspective on how the intersection of computational drug discovery and niche-targeted pharmacology is moving small-molecule skin rejuvenation from theoretical potential to widespread clinical translation.

Establishing stable and dependable animal models of experimental periodontitis is critical in advancing our understanding of the etiology, clinical diagnosis, and treatment of periodontitis. However, conventional silk ligation methods for inducing periodontitis in rats have limitations, with silk thread detachment being a major concern. In this study, we established a reliable rat periodontitis model using a novel "double-ligature" technique combining silk sutures with orthodontic wires, augmented by a high-sugar diet. Thirty rats were randomized into five groups: control, wire-only, silk-only, wire+silk, and wire+silk+sugar. After 2 weeks, the wire+silk+sugar group demonstrated significantly elevated clinical indices (sulcus bleeding index, probing depth, plaque index) versus controls (*p* < 0.05), alongside severe alveolar bone resorption quantified by micro-CT. Molecular analyses revealed upregulated inflammatory gene expression (e.g., TNF-α, IL-1β) in double-ligature groups. Histology confirmed extensive immune infiltration and periodontal ligament disruption. The combined approach induced robust periodontitis with 103% greater CEJ-ABC distance versus controls, while TRAP staining revealed 5-fold increased osteoclast activity. By this "double-ligature" technique, the pathogenesis of periodontitis can be studied on the one hand, and the therapeutic effect of biomaterials on the alveolar bone defects caused by periodontitis can be verified on the other hand. This reproducible method overcomes traditional silk ligature limitations (e.g., thread detachment) and provides a foundation for translational periodontitis research.

Fluorescence-based analysis for protein detection has been widely adopted; however, they fall short in achieving ultra-sensitive detection because of their inherently low signal-to-noise ratio at sub-picomolar concentrations. To overcome this limitation, signal amplifying materials such as gold nanoparticles (AuNPs) have been integrated into various detection platforms to enhance fluorescence signals. However, fabrication of such AuNP integrated platforms remains complex, often requiring sophisticated fabrication steps, yet none approach the ultra-sensitive detection range of 10⁰ fg mL^-1. In this study, we enabled analysis of captured protein/antibody at 10⁰ fg mL^-1 concentrations through fluorescence signals utilizing freestanding gold nanoparticle integrated freestanding hydrogel platforms. The platform is fabricated by integrating AuNPs into the hydrogel matrix using a single-step photolithographic technique. The integrated AuNPs significantly enhanced the fluorescence signals compared to controls. Notably, at fg mL^-1 levels of fluorophore, the AuNP integrated hydrogels produced a robust optical signal in contrast to negligible responses observed in controls. The platform's versatility was validated using tumor necrosis factor-alpha antibody (TNF-α Ab), achieving detection at 10⁰ fg mL^-1. By synergizing the hydrogels' porous structure with AuNPs' signal amplification, this platform successfully achieves fluorescence-based ultra-sensitive protein/antibody detection.