RESUMO
Current cell grafting techniques are majorly dependent on seeding cells on a pre-formed scaffold. However, cells grow in a 2-dimensional (2D) space in such constructs, not mimicking the tissue's 3-dimensional (3D) architecture. The present study evaluated a unique poly-electrolyte complexation (PEC) based strategy for the 3D engraftment of cells in a porous polymeric scaffold. The scaffold was synthesized using a positively charged polysaccharide chitosan (CH) and negatively charged glycosaminoglycans chondroitin sulfate (CS) and hyaluronic acid (HA). Two different scaffolds were synthesized, one using CH and CS [CH-CS] and another using CH and CS + HA [CH-(CS-HA)]. The physicochemical characterization of both the PECs confirmed electrostatic interactions, leading to a porous and viscoelastic PEC formation. Fibroblast cells were grafted and seeded in both scaffolds to evaluate the effect of different scaffold compositions and the difference between seeded and grafted cells. Imaging studies confirmed that grafting of the fibroblast cells supports cellular proliferation. The qPCR studies demonstrated increased expression of functional markers TGF-ß, α-SMA, collagen-I, and fibronectin in the CH-(CS-HA) grafted cells. In summary, it was demonstrated that an in-situ forming PEC of CH, CS, and HA had good physicochemical properties for cell grafting and supported grafted cells with improved function.
Assuntos
Quitosana , Ácido Hialurônico , Ácido Hialurônico/química , Sulfatos de Condroitina/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Quitosana/química , Proliferação de CélulasRESUMO
None of the currently available wound dressings exhibit combined antibacterial and anti-inflammatory activity. Using polyelectrolyte complexation (PEC) between a cationic polysaccharide chitosan (CH) and an anionic glycosaminoglycan chondroitin sulfate (CS), we have developed a unique in-situ forming scaffold (CH-CS PEC), which develops at the wound site itself to influence the function of the wound bed cells. The current study demonstrated that CH-CS PEC could induce bacterial cell death through membrane pore formation and increased ROS production. Moreover, possibly due to its unique material properties including medium-soft viscoelasticity, porosity, and surface composition, CH-CS PEC could modulate macrophage function, increasing their phagocytic ability with low TNF-α and high IL-10 production. Faster wound closure and decreased CFU count was observed in an in-vivo infected wound model, with reduced NF-κB and increased VE-cadherin expression, indicating reduced inflammation and enhanced angiogenesis. In summary, this study exhibited that CH-CS PEC has substantial antibacterial and immunomodulatory properties.
Assuntos
Quitosana , Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Bandagens , Quitosana/farmacologia , Sulfatos de Condroitina/farmacologia , Sulfatos de Condroitina/uso terapêutico , Glicosaminoglicanos , Interleucina-10 , NF-kappa B , Polieletrólitos , Espécies Reativas de Oxigênio , Fator de Necrose Tumoral alfaRESUMO
Aim: Selenium nanoparticles (SeNPs) may have a potential role in treating dermal disorders due to its wide therapeutic properties, but there is a need to evaluate its toxicity in keratinocytes. The present study evaluated the molecular mechanism and mode of cell death induced by SeNPs on dermal keratinocytes. Materials & methods: SeNPs were synthesized, characterized and studied in human keratinocytes cells. Oxidative stress and mitochondrial membrane depolarization were evaluated by various techniques. Additionally, autophagy mediated apoptotic cell death was evaluated. Results: SeNPs induced oxidative stress and apoptotic cell death in keratinocytes by increasing autophagy through the formation of acidic lysosomes and autophagosomes. Conclusion: Overall, SeNPs induce the oxidative stress and autophagy mediated apoptotic cell death in human keratinocytes cells.