Radiosterilized Pig Skin, Silver Nanoparticles and Skin Cells as an Integral Dressing Treatment for Burns: Development, Pre-Clinical and Clinical Pilot Study
Por:
Ortega-Sanchez, Carmina, Perez-Diaz, Mario, Melgarejo-Ramirez, Yaaziel, Chopin-Doroteo, Mario, Silva-Bermudez, Phaedra, Martinez-Lopez, Valentin, Zacaula-Juarez, Noe, Zamudio-Cuevas, Yessica, Hernandez-Valencia, Carmen, Lopez-Jacome, Luis Esau, Carlos-Martinez, Alberto, Reyes-Medina, Naxieli, Tamez-Pedroza, Luis, Martinez-Pardo, Maria Esther, Reyes-Frías M.D.L., Lecona, Hugo, Baeza, Isabel, Martinez-Gutierrez, Fidel, Marquez-Gutierrez, Erik, Martinez-Castanon, Gabriel, Sanchez-Sanchez, Roberto
Publicada:
1 ene 2023
Categoría:
Pharmaceutical science
Resumen:
Radiosterilized pig skin (RPS) has been used as a dressing for burns since the 1980s. Its similarity to human skin in terms of the extracellular matrix (ECM) allows the attachment of mesenchymal stem cells, making it ideal as a scaffold to create cellularized constructs. The use of silver nanoparticles (AgNPs) has been proven to be an appropriate alternative to the use of antibiotics and a potential solution against multidrug-resistant bacteria. RPS can be impregnated with AgNPs to develop nanomaterials capable of preventing wound infections. The main goal of this study was to assess the use of RPS as a scaffold for autologous fibroblasts (Fb), keratinocytes (Kc), and mesenchymal stem cells (MSC) in the treatment of second-degree burns (SDB). Additionally, independent RPS samples were impregnated with AgNPs to enhance their properties and further develop an antibacterial dressing that was initially tested using a burn mouse model. This protocol was approved by the Research and Ethics Committee of the INRLGII (INR 20/19 AC). Transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis of the synthesized AgNPs showed an average size of 10 nm and rounded morphology. Minimum inhibitory concentrations (MIC) and Kirby–Bauer assays indicated that AgNPs (in solution at a concentration of 125 ppm) exhibit antimicrobial activity against the planktonic form of S. aureus isolated from burned patients; moreover, a log reduction of 1.74 ± 0.24 was achieved against biofilm formation. The nanomaterial developed with RPS impregnated with AgNPs solution at 125 ppm (RPS-AgNPs125) facilitated wound healing in a burn mouse model and enhanced extracellular matrix (ECM) deposition, as analyzed by Masson’s staining in histological samples. No silver was detected by energy-dispersive X-ray spectroscopy (EDS) in the skin, and neither by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) in different organs of the mouse burn model. Calcein/ethidium homodimer (EthD-1), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and scanning electron microscopy (SEM) analysis demonstrated that Fb, Kc, and MSC could attach to RPS with over 95% cell viability. Kc were capable of releasing FGF at 0.5 pg above control levels, as analyzed by ELISA assays. An autologous RPS-Fb-Kc construct was implanted in a patient with SDB and compared to an autologous skin graft. The patient recovery was assessed seven days post-implantation, and the patient was followed up at one, two, and three months after the implantation, exhibiting favorable recovery compared to the gold standard, as measured by the cutometer. In conclusion, RPS effectively can be used as a scaffold for the culture of Fb, Kc, and MSC, facilitating the development of a cellularized construct that enhances wound healing in burn patients. © 2023 by the authors.
Filiaciones:
Ortega-Sanchez, Carmina:
Laboratorio de Biotecnología, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Perez-Diaz, Mario:
Laboratorio de Biotecnología, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Laboratorio de Biomembranas, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, 07738, Mexico
Melgarejo-Ramirez, Yaaziel:
Laboratorio de Biotecnología, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Chopin-Doroteo, Mario:
Laboratorio de Tejido Conjuntivo, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Silva-Bermudez, Phaedra:
Unidad de Ingeniería de Tejidos Terapia Celular y Medicina Regenerativa, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Martinez-Lopez, Valentin:
Unidad de Ingeniería de Tejidos Terapia Celular y Medicina Regenerativa, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Zacaula-Juarez, Noe:
Laboratorio de Biotecnología, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Zamudio-Cuevas, Yessica:
Laboratorio de Líquido Sinovial, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Hernandez-Valencia, Carmen:
Departamento de Alimentos y Biotecnología, Facultad de Química, Universidad Nacional Autónoma de México, México City, 04510, Mexico
Lopez-Jacome, Luis Esau:
Laboratorio de Infectología, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Carlos-Martinez, Alberto:
Laboratorio de Microscopía Electrónica, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Reyes-Medina, Naxieli:
Laboratorio de Microscopía Electrónica, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Tamez-Pedroza, Luis:
Cirugía Plástica, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Martinez-Pardo, Maria Esther:
Banco de Tejidos Radioesterilizados, Instituto Nacional de Investigaciones Nucleares, Ocoyoacac, 52045, Mexico
Reyes-Frías M.D.L.:
Banco de Tejidos Radioesterilizados, Instituto Nacional de Investigaciones Nucleares, Ocoyoacac, 52045, Mexico
Lecona, Hugo:
Bioterio, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Baeza, Isabel:
Laboratorio de Biomembranas, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, 07738, Mexico
Martinez-Gutierrez, Fidel:
Laboratorio de Antimicrobianos, Biopelículas y Microbiota, Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, San Luis Potosí, 78210, Mexico
Centro de Investigación en Ciencias de la Salud y Biomedicina, Universidad Autonoma de San Luis Potosi, San Luis Potosi, 78210, Mexico
Marquez-Gutierrez, Erik:
Cirugía Plástica, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Martinez-Castanon, Gabriel:
Facultad de Estomatología, Universidad Autónoma de San Luis Potosí, San Luis Potosí, 78210, Mexico
Sanchez-Sanchez, Roberto:
Unidad de Ingeniería de Tejidos Terapia Celular y Medicina Regenerativa, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Mexico City, 14389, Mexico
Escuela de Ingeniería y Ciencias, Tecnológico de Monterrey, Mexico City, 64849, Mexico
gold, All Open Access; Gold
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