Applying the Cytocentric Principles to Regenerative Medicine for Reproducibility.

Purpose of Review: Cell and tissue products do not just reflect their present conditions; they are the culmination of all they have encountered over time. Currently, routine cell culture practices subject cell and tissue products to highly variable and non-physiologic conditions. This article defines five cytocentric principles that place the conditions for cells at the core of what we do for better reproducibility in Regenerative Medicine. Recent Findings: There is a rising awareness of the cell environment as a neglected, but critical variable. Recent publications have called for controlling culture conditions for better, more reproducible cell products. Summary: Every industry has basic quality principles for reproducibility. Cytocentric principles focus on the fundamental needs of cells: protection from contamination, physiologic simulation, and full-time conditions for cultures that are optimal, individualized, and dynamic. Here, we outline the physiologic needs, the technologies, the education, and the regulatory support for the cytocentric principles in regenerative medicine.

Marrow-isolated adult multilineage inducible cells embedded within a biologically-inspired construct promote recovery in a mouse model of peripheral vascular disease.

Peripheral vascular disease is one of the major vascular complications in individuals suffering from diabetes and in the elderly that is associated with significant burden in terms of morbidity and mortality. Stem cell therapy is being tested as an attractive alternative to traditional surgery to prevent and treat this disorder. The goal of this study was to enhance the protective and reparative potential of marrow-isolated adult multilineage inducible (MIAMI) cells by incorporating them within a bio-inspired construct (BIC) made of two layers of gelatin B electrospun nanofibers. We hypothesized that the BIC would enhance MIAMI cell survival and engraftment, ultimately leading to a better functional recovery of the injured limb in our mouse model of critical limb ischemia compared to MIAMI cells used alone. Our study demonstrated that MIAMI cell-seeded BIC resulted in a wide range of positive outcomes with an almost full recovery of blood flow in the injured limb, thereby limiting the extent of ischemia and necrosis. Functional recovery was also the greatest when MIAMI cells were combined with BICs, compared to MIAMI cells alone or BICs in the absence of cells. Histology was performed 28 days after grafting the animals to explore the mechanisms at the source of these positive outcomes. We observed that our critical limb ischemia model induces an extensive loss of muscular fibers that are replaced by intermuscular adipose tissue (IMAT), together with a highly disorganized vascular structure. The use of MIAMI cells-seeded BIC prevented IMAT infiltration with some clear evidence of muscular fibers regeneration.

Tissue engineering the retinal ganglion cell nerve fiber layer.

Retinal degenerative diseases, such as glaucoma and macular degeneration, affect millions of people worldwide and ultimately lead to retinal cell death and blindness. Cell transplantation therapies for photoreceptors demonstrate integration and restoration of function, but transplantation into the ganglion cell layer is more complex, requiring guidance of axons from transplanted cells to the optic nerve head in order to reach targets in the brain. Here we create a biodegradable electrospun (ES) scaffold designed to direct the growth of retinal ganglion cell (RGC) axons radially, mimicking axon orientation in the retina. Using this scaffold we observed an increase in RGC survival and no significant change in their electrophysiological properties. When analyzed for alignment, 81% of RGCs were observed to project axons radially along the scaffold fibers, with no difference in alignment compared to the nerve fiber layer of retinal explants. When transplanted onto retinal explants, RGCs on ES scaffolds followed the radial pattern of the host retinal nerve fibers, whereas RGCs transplanted directly grew axons in a random pattern. Thus, the use of this scaffold as a cell delivery device represents a significant step towards the use of cell transplant therapies for the treatment of glaucoma and other retinal degenerative diseases.

bFGF-containing electrospun gelatin scaffolds with controlled nano-architectural features for directed angiogenesis.

Current therapeutic angiogenesis strategies are focused on the development of biologically responsive scaffolds that can deliver multiple angiogenic cytokines and/or cells in ischemic regions. Herein, we report on a novel electrospinning approach to fabricate cytokine-containing nanofibrous scaffolds with tunable architecture to promote angiogenesis. Fiber diameter and uniformity were controlled by varying the concentration of the polymeric (i.e. gelatin) solution, the feed rate, needle to collector distance, and electric field potential between the collector plate and injection needle. Scaffold fiber orientation (random vs. aligned) was achieved by alternating the polarity of two parallel electrodes placed on the collector plate thus dictating fiber deposition patterns. Basic fibroblast growth factor (bFGF) was physically immobilized within the gelatin scaffolds at variable concentrations and human umbilical vein endothelial cells (HUVEC) were seeded on the top of the scaffolds. Cell proliferation and migration was assessed as a function of growth factor loading and scaffold architecture. HUVECs successfully adhered onto gelatin B scaffolds and cell proliferation was directly proportional to the loading concentrations of the growth factor (0-100 bFGF ng/mL). Fiber orientation had a pronounced effect on cell morphology and orientation. Cells were spread along the fibers of the electrospun scaffolds with the aligned orientation and developed a spindle-like morphology parallel to the scaffold's fibers. In contrast, cells seeded onto the scaffolds with random fiber orientation, did not demonstrate any directionality and appeared to have a rounder shape. Capillary formation (i.e. sprouts length and number of sprouts per bead), assessed in a 3-D in vitro angiogenesis assay, was a function of bFGF loading concentration (0 ng, 50 ng and 100 ng per scaffold) for both types of electrospun scaffolds (i.e. with aligned or random fiber orientation).

Condilomatosis vulvar grave / Severe vulvar condyloma

Se describe el caso clínico de una paciente que desde hacía 9 años presentaba numerosas verrugas en la vulva, además de prurito y dolor ocasional, más frecuentes al realizar determinadas actividades. Se efectuó vulvectomía simple y se comprobó la presencia de cambios típicos coilocíticos, de modo que no se trataba de lesiones malignas. Resultó imposible determinar el virus del papiloma humano, aunque por las manifestaciones clínicas podía corresponder a las cepas 6 y 11. La fémina fue egresada sin elementos de infección local, con tejido de granulación útil y compensada metabólicamente, pero con seguimiento por consulta externa hasta los 3 meses de operada(AU)

A case report of a patient who presented with numerous warts in her vulva for 9 years besides pruritus and occasional pain is described, more frequent when she was doing certain activities. Simple vulvectomy was performed and the presence of typical koilocytotic changes was confirmed, but there were not malignant lesions. It was impossible to determine the human papilloma virus, although by its clinical manifestations could be strains 6 and 11. The patient was discharged metabolically compensated with useful granulation tissue and without local infection, but she was followed for 3 months after surgery in the outpatient department(AU)

Condilomatosis vulvar grave / Severe vulvar condyloma

Se describe el caso clínico de una paciente que desde hacía 9 años presentaba numerosas verrugas en la vulva, además de prurito y dolor ocasional, más frecuentes al realizar determinadas actividades. Se efectuó vulvectomía simple y se comprobó la presencia de cambios típicos coilocíticos, de modo que no se trataba de lesiones malignas. Resultó imposible determinar el virus del papiloma humano, aunque por las manifestaciones clínicas podía corresponder a las cepas 6 y 11. La fémina fue egresada sin elementos de infección local, con tejido de granulación útil y compensada metabólicamente, pero con seguimiento por consulta externa hasta los 3 meses de operada.

A case report of a patient who presented with numerous warts in her vulva for 9 years besides pruritus and occasional pain is described, more frequent when she was doing certain activities. Simple vulvectomy was performed and the presence of typical koilocytotic changes was confirmed, but there were not malignant lesions. It was impossible to determine the human papilloma virus, although by its clinical manifestations could be strains 6 and 11. The patient was discharged metabolically compensated with useful granulation tissue and without local infection, but she was followed for 3 months after surgery in the outpatient department.

Papiloma cervical / Cervical Papilloma

Se presenta el caso de una paciente de 60 años de edad, sin pareja sexual en los últimos 6 años, con un tumor exofítico que ocupaba todo el tercio distal de la vagina, dependiente del labio anterior del cuello uterino, que sangraba fácilmente, tenía superficie irregular, medía aproximadamente 6 cm de diámetro y parecía ser maligno por su aspecto macroscópico; sin embargo, resultó ser un papiloma cervical benigno. La infrecuente aparición de formas tumorales exofíticas del virus del papiloma humano justificó el informe de este hallazgo clínico a la comunidad científica. Una vez más se acentúa la importancia de aplicar y cumplir el programa de detección precoz del cáncer cervicouterino, a fin de poder diagnosticar y tratar oportunamente cualquier lesión en el cuello del útero(AU)

The case of a 60 year-old patient is presented, who had not sexual partner in the last 6 years with an exophytic tumor that involved the whole distal third of the vagina and dependent on anterior lip of the cervix. The tumor bled easily and had irregular surface; its diameter was 6 cm approximately and it seemed to be malignant for its macroscopic aspect, but it was a benign cervical papilloma. The uncommon occurrence of tumoral exophytic forms of the human papillomavirus justified the report of this clinical finding to the scientific community. Once again the importance of applying and completing the early detection program of cervix cancer is emphasized in order to diagnose and treat any lesion appropriately in the cervix(AU)

Papiloma cervical / Cervical Papilloma

Se presenta el caso de una paciente de 60 años de edad, sin pareja sexual en los últimos 6 años, con un tumor exofítico que ocupaba todo el tercio distal de la vagina, dependiente del labio anterior del cuello uterino, que sangraba fácilmente, tenía superficie irregular, medía aproximadamente 6 cm de diámetro y parecía ser maligno por su aspecto macroscópico; sin embargo, resultó ser un papiloma cervical benigno. La infrecuente aparición de formas tumorales exofíticas del virus del papiloma humano justificó el informe de este hallazgo clínico a la comunidad científica. Una vez más se acentúa la importancia de aplicar y cumplir el programa de detección precoz del cáncer cervicouterino, a fin de poder diagnosticar y tratar oportunamente cualquier lesión en el cuello del útero.

The case of a 60 year-old patient is presented, who had not sexual partner in the last 6 years with an exophytic tumor that involved the whole distal third of the vagina and dependent on anterior lip of the cervix. The tumor bled easily and had irregular surface; its diameter was 6 cm approximately and it seemed to be malignant for its macroscopic aspect, but it was a benign cervical papilloma. The uncommon occurrence of tumoral exophytic forms of the human papillomavirus justified the report of this clinical finding to the scientific community. Once again the importance of applying and completing the early detection program of cervix cancer is emphasized in order to diagnose and treat any lesion appropriately in the cervix.

Effects of commonly used topical antimicrobial agents on Acinetobacter baumannii: an in vitro study.

Acinetobacter baumannii has recently emerged as an important pathogen among wounded soldiers in Iraq. Because of its ability to develop resistance to antimicrobial agents, wound infections with A. baumannii are difficult to treat and can lead to septicemia and even death. Use of appropriate topical antimicrobial agents in these circumstances could be one of the first steps in the prevention of A. baumannii wound infections. In this study, we present the in vitro effects of seven common topical antimicrobial creams and dressings on A. baumannii. A. baumannii was subjected to sensitivity tests with mupirocin, silver sulfadiazine, mafenide acetate, a double-antibiotic combination of polymyxin and bacitracin, a triple-antibiotic combination of neomycin, bacitracin, and polymyxin, and two silver-containing dressings. Zones of inhibition were measured after 24 hours of incubation. Of the evaluated antimicrobial agents, mafenide acetate was the most efficacious, followed by mupirocin and triple- and double-antibiotic combinations (in decreasing order). The silver-containing dressings yielded smaller zones of inhibition, compared to the previously mentioned agents, and no zone of inhibition was observed with silver sulfadiazine. Further in vivo studies on the effects of antimicrobial agents against A. baumannii are necessary to substantiate these findings and to determine the potential clinical relevance of these therapies.

Hamartoma esplénico (esplenadenoma) / Splenic hamartoma (splenadenoma)

presenta el caso de un paciente joven, que acudió a consulta por experimentar dolor en hemiabdomen izquierdo, con hepatomegalia de 3 cm a la palpación e imagen parenquimatosa de baja ecogenicidad cerca del hilio esplénico en el ultrasonido diagnóstico de abdomen, interpretada como un posible tumor intraparenquimatoso. Luego de realizadas todas las pruebas previstas fue intervenido quirúrgicamente, de donde se derivó que se trataba de un hamartoma esplénico. Dicho proceder resultó ser curativo(AU)

Correlación clínica, imagenológica y anatomopatológica de las muertes neonatales en 1998 / Clinical, imagenological and pathological correlation of neonatal Deaths in 1998

Se realizó un estudio descriptivo, transversal y retrospectivo de los 16 recién nacidos que fallecieron en el Hospital General Santiago durante 1998, a causa fundamentalmente de sepsis y malformaciones congénitas, corroboradas mediante una buena correlación entre el diagnóstico clínico, las imágenes radiográficas y el informe anatomopatológico de las defunciones(AU)