Browsing by Autor "Javier Cifuentes"

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Porcine small intestinal submucosa (SIS) and graphene oxide/reduced graphene oxide scaffolds for potential application in electrostimulation therapy: preliminary formulation and characterization
(2020) Juan C. Cruz; Javier Cifuentes; Julian A. Serna; Carolina Muñoz-Camargo
Chronic wounds are a health problem of enormous magnitude that affects millions of patients around the world. The most promising treatments for chronic wounds healing are the therapies related to the development of biomimetic technologies that successfully improve cell migration, growth and proliferation. The implementation of scaffolds or hydrogels, based on natural and biosynthetic extracellular matrix (ECM) or individual components of ECM, have shown to provide an adequate environment to enhance cellular migration, angiogenesis and regulation of wound healing processes. Additionally, electrostimulation therapies have gained attention in recent years due to their capability for simulating electric currents to direct cell migration, promote cell proliferation and increase oxygenated blood perfusion towards damaged tissues. In the present work, we propose innovative regenerative 3D scaffolds based on small intestinal submucosa (SIS) combined with graphene oxide (GO)/reduced graphene oxide (rGO) to improve their electrical conductivity such that they can be potentially applied in the healing of chronic wounds. To achieve this, decellularized SIS was obtained and mixed with GO flakes to make 3D scaffolds that were chemically crosslinked and reduced in-situ. GO and rGO were characterized by thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, atomic force microscopy (AFM) and the four-point probe conductivity method. These techniques confirmed the effective synthesis of GO, the reduction to rGO and the improvement of electrical conductivity. Crosslinked SIS, SIS-GO and SIS-rGO scaffolds were characterized by FTIR, TGA, SEM, Raman spectroscopy and liquid displacement method. In addition, the biocompatibility of scaffolds was carried out via hemolysis activity, platelet aggregation, and cytotoxicity in Vero cells. Experiments revealed high hemocompatibility, low cytotoxicity and no significant impact on platelet aggregation. Finally, microscopic structure characteristics and cell attachment abilities demonstrated the potential of the developed technology for multiple applications in tissue engineering and regenerative medicine.
Unlocking cellular barriers: silica nanoparticles and fullerenol conjugated cell-penetrating agents for enhanced intracellular drug delivery
(Frontiers Media, 2023) Eduardo Ravelo-Nieto; Javier Cifuentes; Paola Ruiz Puentes; Laura Rueda-Gensini; Valentina Quezada; C. Ostos; Carolina Muñoz-Camargo; Luis H. Reyes; Alvaro Duarte‐Ruiz; Juan C. Cruz
The limited delivery of cargoes at the cellular level is a significant challenge for therapeutic strategies due to the presence of numerous biological barriers. By immobilizing the Buforin II (BUF-II) peptide and the OmpA protein on magnetite nanoparticles, a new family of cell-penetrating nanobioconjugates was developed in a previous study. We propose in this study to extend this strategy to silica nanoparticles (SNPs) and silanized fullerenol (F) as nanostructured supports for conjugating these potent cell-penetrating agents. The same molecule conjugated to distinct nanomaterials may interact with subcellular compartments differently. On the obtained nanobioconjugates (OmpA-SNPs, BUF-II-PEG12-SNPs, OmpA-F, and BUF-II-PEG12-F), physicochemical characterization was performed to evaluate their properties and confirm the conjugation of these translocating agents on the nanomaterials. The biocompatibility, toxicity, and internalization capacity of nanobioconjugates in Vero cells and THP-1 cells were evaluated in vitro. Nanobioconjugates had a high internalization capacity in these cells without affecting their viability, according to the findings. In addition, the nanobioconjugates exhibited negligible hemolytic activity and a low tendency to induce platelet aggregation. In addition, the nanobioconjugates exhibited distinct intracellular trafficking and endosomal escape behavior in these cell lines, indicating their potential for addressing the challenges of cytoplasmic drug delivery and the development of therapeutics for the treatment of lysosomal storage diseases. This study presents an innovative strategy for conjugating cell-penetrating agents using silica nanoparticles and silanized fullerenol as nanostructured supports, which has the potential to enhance the efficacy of cellular drug delivery.