Isolation, expansion and differentiation of mesenchymal stromal cells from rabbits' bone marrow

Abstract: Tissue engineering has been a fundamental technique in the regenerative medicine field, once it permits to build tri-dimensional tissue constructs associating undifferentiated mesenchymal cells (or mesenchymal stromal cells - MSCs) and scaffolds in vitro. Therefore, many studies have been carried out using these cells from different animal species, and rabbits are often used as animal model for in vivo tissue repair studies. However, most of the information available about MSCs harvesting and characterization is about human and murine cells, which brings some doubts to researchers who desire to work with a rabbit model in tissue repair studies based on MSCs. In this context, this study aimed to add and improve the information available in the scientific literature providing a complete technique for isolation, expansion and differentiation of MSCs from rabbits. Bone marrow mononuclear cells (BMMCs) from humerus and femur of rabbits were obtained and to evaluate their proliferation rate, three different culture media were tested, here referred as DMEM-P, DMEM´S and α-MEM. The BMMCs were also cultured in osteogenic, chondrogenic and adipogenic induction media to prove their multipotentiality. It was concluded that the techniques suggested in this study can provide a guideline to harvest and isolate MSCs from bone marrow of rabbits in enough amount to allow their expansion and, based on the laboratory experience where the study was developed, it is also suggested a culture media formulation to provide a better cell proliferation rate with multipotentiality preservation

Pirfenidone Accelerates Wound Healing in Chronic Diabetic Foot Ulcers: A Randomized, Double-Blind Controlled Trial

Background. Diabetic foot ulcers are one disabling complication of diabetes mellitus. Pirfenidone (PFD) is a potent modulator of extracellular matrix. Modified diallyl disulfide oxide (M-DDO) is an antimicrobial and antiseptic agent. Aim. To evaluate efficacy of topical PFD + M-DDO in a randomized, double-blind trial versus ketanserin in the treatment of noninfected chronic DFU. Methods. Patients received PFD + M-DDO or ketanserin for 6 months. Relative ulcer volume (RUV) was measured every month; biopsies were taken at baseline and months 1 and 2 for histopathology and gene expression analysis for COL-1α, COL-4, KGF, VEGF, ACTA2 (α-SMA), elastin, fibronectin, TGF-β1, TGF-β3, HIF-1α, and HIF-1β. Results. Reduction of median RUV in the PFD + M-DDO group was 62%, 89.8%, and 99.7% at months 1–3 and 100% from months 4 to 6. Ketanserin reduced RUV in 38.4%, 56%, 60.8%, 94%, 94.8%, and 100% from the first to the sixth month, respectively. Healing score improved 4.5 points with PFD + M-DDO and 1.5 points with ketanserin compared to basal value. Histology analysis revealed few inflammatory cells and organized/ordered collagen fiber bundles in PFD + M-DDO. Expression of most genes was increased with PFD + M-DDO; 43.8% of ulcers were resolved using PFD + M-DDO and 23.5% with ketanserin. Conclusion. PFD + M-DDO was more effective than ketanserin in RUV reduction

Knowledge Evolution: Inert sciences to living science

Modern mentality tends to minimize what is real to a physical world that is accessible to its senses, instruments, reasoning and equations, ignoring other states of reality that, clearly throughout humanity’s history have been known. Modern human believes that he is capable of dispensing all knowledge from what he has been taught in the past by starting over again, trusting only their point of view and their own new prejudices. His attention increasingly focusing outwards prevents him from looking inwards, towards the center of consciousness, of being, which is, however, the first data that has been imposed on us and the basis on which necessarily everything else rests. A physical analysis of a piece of music or a painting, however scientific it might be, does not annul the meaning- so deeper and on another type of level-shows that the reality of a work of art is much more than its physical components. This objective work creates communication bonds interconnecting classical and modern science, relating different areas of knowledge. Like the invisible presence of microorganisms that participate in the evolution of nature, we intend to give a new approach to recovering the empirical knowledge long way forgotten by modern science in order to strengthen the reality of the parts that do not precede the whole, but when are born acquire sense together with the whole. Their role as “parts” is only a role in the cognitive process, not in the generative process.Fil: Castañeda Cataña, Mayra Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Amato, R.. Universidad de Buenos Aires. Facultad de Psicología; ArgentinaFil: Sepúlveda, Claudia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Carlucci, Maria Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentin

Características clínicas de la pancreatitis aguda en México

Antecedentes: La frecuencia y la etiología de la pancreatitis aguda (PA) varían de acuerdo a la región geográfica. Objetivo: Determinar la frecuencia, etiología y mortalidad de la PA en una población mexicana. Pacientes y métodos: En un periodo de seis años estudiamos prospectivamente a todos los pacientes con un primer episodio de PA. Resultados: Se incluyeron a 605 pacientes con edad promedio de 40 años. El 64,00% tenían sobrepeso y/o obesidad por índice de masa corporal (IMC). La etiología fue biliar en 66,60%, ingesta de alcohol en 15,90%, hipertrigliceridemia en 7,80%, poscolangiopancreatografía retrógrada endoscópica (CPRE) en 2,10% y de etiología no determinada en 7,20%. Se sospechó infección pancreática en 70 pacientes, los cuales fueron puncionados por tomografía axial computarizada (TAC) y en 32 se diagnosticó infección pancreática, siendo el Staphylococcus spp el microorganismo más común. La mortalidad global fue del 5,00% ( 70 años con 10,00%). La etiología alcohólica tuvo una mayor frecuencia de necrosis, infección pancreática y mortalidad. Conclusiones: La etiología de la PA más frecuente es la biliar, pero la causada por alcohol tiene más complicaciones. La mortalidad global fue del 5,00% y ésta se incremento en los pacientes de mayor edad

Systemic Sclerosis Pathogenesis and Emerging Therapies, beyond the Fibroblast

Systemic sclerosis (SSc) is a complex rheumatologic autoimmune disease in which inflammation, fibrosis, and vasculopathy share several pathogenic pathways that lead to skin and internal organ damage. Recent findings regarding the participation and interaction of the innate and acquired immune system have led to a better understanding of the pathogenesis of the disease and to the identification of new therapeutic targets, many of which have been tested in preclinical and clinical trials with varying results. In this manuscript, we review the state of the art of the pathogenesis of this disease and discuss the main therapeutic targets related to each pathogenic mechanism that have been discovered so far

Pirfenidone Accelerates Wound Healing in Chronic Diabetic Foot Ulcers: A Randomized, Double-Blind Controlled Trial

Background. Diabetic foot ulcers are one disabling complication of diabetes mellitus. Pirfenidone (PFD) is a potent modulator of extracellular matrix. Modified diallyl disulfide oxide (M-DDO) is an antimicrobial and antiseptic agent. Aim. To evaluate efficacy of topical PFD + M-DDO in a randomized, double-blind trial versus ketanserin in the treatment of noninfected chronic DFU. Methods. Patients received PFD + M-DDO or ketanserin for 6 months. Relative ulcer volume (RUV) was measured every month; biopsies were taken at baseline and months 1 and 2 for histopathology and gene expression analysis for COL-1α, COL-4, KGF, VEGF, ACTA2 (α-SMA), elastin, fibronectin, TGF-β1, TGF-β3, HIF-1α, and HIF-1β. Results. Reduction of median RUV in the PFD + M-DDO group was 62%, 89.8%, and 99.7% at months 1–3 and 100% from months 4 to 6. Ketanserin reduced RUV in 38.4%, 56%, 60.8%, 94%, 94.8%, and 100% from the first to the sixth month, respectively. Healing score improved 4.5 points with PFD + M-DDO and 1.5 points with ketanserin compared to basal value. Histology analysis revealed few inflammatory cells and organized/ordered collagen fiber bundles in PFD + M-DDO. Expression of most genes was increased with PFD + M-DDO; 43.8% of ulcers were resolved using PFD + M-DDO and 23.5% with ketanserin. Conclusion. PFD + M-DDO was more effective than ketanserin in RUV reduction