Development of secretome-based strategies to improve cell culture protocols in tissue engineering.

Advances in skin tissue engineering have promoted the development of artificial skin substitutes to treat large burns and other major skin loss conditions. However, one of the main drawbacks to bioengineered skin is the need to obtain a large amount of viable epithelial cells in short periods of time, making the skin biofabrication process challenging and slow. Enhancing skin epithelial cell cultures by using mesenchymal stem cells secretome can favor the scalability of manufacturing processes for bioengineered skin. The effects of three different types of secretome derived from human mesenchymal stem cells, e.g. hADSC-s (adipose cells), hDPSC-s (dental pulp) and hWJSC-s (umbilical cord), were evaluated on cultured skin epithelial cells during 24, 48, 72 and 120 h to determine the potential of this product to enhance cell proliferation and improve biofabrication strategies for tissue engineering. Then, secretomes were applied in vivo in preliminary analyses carried out on Wistar rats. Results showed that the use of secretomes derived from mesenchymal stem cells enhanced currently available cell culture protocols. Secretome was associated with increased viability, proliferation and migration of human skin epithelial cells, with hDPSC-s and hWJSC-s yielding greater inductive effects than hADSC-s. Animals treated with hWJSC-s and especially, hDPSC-s tended to show enhanced wound healing in vivo with no detectable side effects. Mesenchymal stem cells derived secretomes could be considered as a promising approach to cell-free therapy able to improve skin wound healing and regeneration.

La esquizofrenia y su concepto transcultural / Schizophrenia and it's cross-cultural concept

En la actualidad, la mente humana no es posible valorarla de forma directa, pues el campo del inconsciente todavía es un área muy desconocida, debido a la complejidad de los procesos cognitivos y subjetivos que alberga. Si la salud se concibe como el bienestar biopsicosocial de la persona, los profesionales de la salud mediante su trabajo deben hacer frente a las innumerables necesidades que pueden presentar las personas [Fragmento de texto] (AU)

La enfermera Genene Jones: El rol de ángel de la muerte en la obra Misery deStephen King / Enfermeira Genene Jones: O papel de anjo da morte na Miséry de Stephen King / Nurse Genene Jones: The Role of the Angel of Death in Stephen King's Misery

With the sublime emotional description that Stephen King achieves of the main characters inMisery, the writer manages to delve into the world of affections and feelings, exploring the art of care andits negative aspects. He manages to delve into the figure of the nurse Genene Jones, representative of therole of the angel of death, making visible theories that frame these typologies and their acts. Objective:to describe the behaviours, experiences, beliefs and values set out by Stephen King in the story "Misery"(1987). Methodology: Phenomenological-historical study with an exploratory approach in narrative andsecondary sources of the events that took place in the period between 1950-1990. Results: The piece is amasterful analysis of Genene Jones' nursing practice. It also reflects the transformation from the role ofan angel of death, carried out by a health professional, to become a serial killer. Conclusion: The studyof the horror literature allows the study of the nursing role to deepen and increase the knowledge of thenursing role, both for health professionals and nursing students and the general population. (AU)

Con la sublime descripción emocional que Stephen King logra de los protagonistas de Misery, elescritor consigue profundizar en el mundo de las afecciones y los sentimientos, explorando el arte delcuidado y sus aspectos negativos. Consigue ahondar en la figura de la enfermera Genene Jones,representante del rol de ángel de la muerte, visibilizando teorías que encuadran estas tipologías y susactos. Objetivo: describir las conductas, vivencias, creencias y valores expuestos por Stephen King en lanarración “Misery” (1987). Metodología: Estudio fenomenológico-histórico con abordaje exploratorio enfuentes narrativas y secundarias de los acontecimientos sucedidos en el periodo de tiempo comprendidoentre 1950-1990. Resultados: La obra es un análisis magistral del proceder enfermero de Genene Jones.Asimismo, refleja la transformación desde el rol de ángel de la muerte, llevado a cabo por un profesionalde la salud, hasta llegar a convertirse en un asesino en serie. Conclusión: El estudio de la literatura deterror permite profundizar e incrementar los conocimientos del rol enfermero, tanto a los profesionales dela salud como a los estudiantes de enfermería y la población en general. (AU)

Com a sublime descrição emocional que Stephen King consegue nos protagonistas da Miséria, oescritor mergulha no mundo dos afectos e sentimentos, explorando a arte do cuidado e os seus aspectosnegativos. A figura da enfermeira Genene Jones, um representante exemplar do papel do anjo da morte, éaprofundada, tornando visíveis as teorias que enquadram estas tipologias e os seus actos. Objectivo:descrever os comportamentos, experiências, crenças e valores expostos por Stephen King na história"Miséria" (1987). Metodologia: Estudo fenomenológico-histórico com uma abordagem exploratória emnarrativa, com recurso a fontes secundárias dos acontecimentos que tiveram lugar no período de 1950-1990. Resultados: O trabalho é uma análise do procedimento de enfermagem de Genene Jones e decomo o papel de anjo da morte, desempenhado por um profissional de saúde pode vir a personificar afigura de um assassino em série. Conclusão: O estudo da literatura de horror permite aprofundar, veroutras facetas, e aumentar o conhecimento do papel da enfermagem, tanto pelos profissionais de saúde eestudantes de enfermagem como pela população em geral. (AU)

Evaluation of myopic cornea lenticules. A histochemical and clinical correlation.

In this work, we have analyzed the main clinical and corneal histological parameters that may be associated to the spherical equivalent (SE), age and gender of individuals with myopic refractive errors. For this purpose, 108 cornea stroma lenticules were obtained from patients subjected to ReLEx-SMILE myopia correction. Histological analyses were carried out and histochemistry and immunohistochemistry were used to quantify key histological components of the cornea stroma, including mature collagen fibers, reticular and elastic fibers, glycoproteins, proteoglycans, type-V collagen and several crystallins. Clinical and histological data were analyzed to determine their association with SE, age and gender. Results showed a significant correlation between the age range of the patients and the expression of crystallins CRY-α-A, CRY-λ1 and type-V collagen and between CRY-λ1 and corneal thickness, spherical diopters (D) and SE, although correlation between CRY-λ1 and SE was non-significant when age was controlled. Comparison of cases with low myopia and high/moderate myopia found statistical differences for D and lenticule thickness and diameter. The binary logistic regression analysis allowed us to construct a model using two clinical parameters (D and lenticule thickness). Parameters showing significant correlation with the age were the corneal radius, keratometry reading (K), OZ, CRY-α-A and type-V collagen, whereas SE, lenticule thickness, OZ, CRY-λ1 and type-V collagen showed statistically significant differences between the youngest and the oldest patients. A binary logistic regression analysis model was generated including 3 variables (D, cornea radius and OZ). No gender differences were found. The specific clinical and histological modifications found to be associated to the SE and age could be useful for a better understanding of the mechanisms involved in the genesis or progression of myopia and could establish the basement for future therapeutic options.

Effective use of mesenchymal stem cells in human skin substitutes generated by tissue engineering.

Mesenchymal stem cells (MSCs) can differentiate toward epithelial cells and may be used as an alternative source for generation of heterotypical artificial human skin substitutes, thus, enhancing their development and translation potential to the clinic. The present study aimed at comparing four types of heterotypical human bioengineered skin generated using MSCs as an alternative epithelial cell source. Adipose-tissue-derived stem cells (ADSCs), dental pulp stem cells (DPSCs), Wharton's jelly stem cells (WJSCs) and bone marrow stem cells (BMSCs) were used for epidermal regeneration on top of dermal skin substitutes. Heterotypic human skin substitutes were evaluated before and after implantation in immune-deficient athymic mice for 30 d. Histological and genetic studies were performed to evaluate extracellular matrix synthesis, epidermal differentiation and human leukocyte antigen (HLA) molecule expression. The four cell types differentiated into keratinocytes, as shown by the expression of cytokeratin 10 and filaggrin 30 d post-grafting; also, they induced dermal fibroblasts responsible for the synthesis of extracellular fibrillar and non-fibrillar components, in a similar way among each other. WJSCs and BMSCs showed higher expression of cytokeratin 10 and filaggrin, suggesting these cells were more prone to epidermal regeneration. The absence of HLA molecules, even when the epithelial layer was differentiated, supports the future clinical use of these substitutes - especially ADSCs, DPSCs and WJSCs - with low rejection risk. MSCs allowed the generation of bioengineered human skin substitutes with potential clinical usefulness. According to their epidermal differentiation potential and lack of HLA antigens, WJSCs should preferentially be used.

Characterization of the human ridged and non-ridged skin: a comprehensive histological, histochemical and immunohistochemical analysis.

The structure of the human skin is directly dependent on its location and the mechanical forces to which it is subjected. In the present work, we have performed a comprehensive analysis of the human ridged and non-ridged skin to identify the differences and similarities between both skin types. For this purpose, human skin samples were obtained from dorsal hand skin (DHS), palmar hand skin (PHS), dorsal foot skin (DFS) and plantar foot skin (PFS) from the same cadaveric donors. Histological, histochemical and semiquantitative and quantitative immunohistochemical analyses were carried out to evaluate the epidermis, dermis and basement membrane. Results show that the epithelial layer of ridged skin had larger cell number and size than non-ridged skin for most strata. Melanocytes and Langerhans cells were more abundant in non-ridged skin, whereas Merkel cells were preferentially found in ridged skin. The expression pattern of CK5/6 was slightly differed between non-ridged and ridged skin. Involucrin expression was slightly more intense in non-ridged skin than in ridged skin. Collagen was more abundant in foot skin dermis than in hand skin, and in ridged skin as compared to non-ridged skin. Elastic fibers were more abundant in DHS. Biglycan was more abundant in foot skin than in hand skin. No differences were found for blood and lymphatic vessels. The basement membrane laminin was preferentially found in foot skin. These results revealed important differences at the epithelial, dermal and basement membrane levels that could contribute to a better knowledge of the human skin histology.

Wharton's jelly-derived mesenchymal cells as a new source for the generation of microtissues for tissue engineering applications.

Microtissues (MT) are currently considered as a promising alternative for the fabrication of natural, 3D biomimetic functional units for the construction of bio-artificial substitutes by tissue engineering (TE). The aim of this study was to evaluate the possibility of generating mesenchymal cell-based MT using human umbilical cord Wharton's jelly stromal cells (WJSC-MT). MT were generated using agarose microchips and evaluated ex vivo during 28 days. Fibroblasts MT (FIB-MT) were used as control. Morphometry, cell viability and metabolism, MT-formation process and ECM synthesis were assessed by phase-contrast microscopy, functional biochemical assays, and histological analyses. Morphometry revealed a time-course compaction process in both MT, but WJSC-MT resulted to be larger than FIB-MT in all days analyzed. Cell viability and functionality evaluation demonstrated that both MT were composed by viable and metabolically active cells, especially the WJSC during 4-21 days ex vivo. Histology showed that WJSC acquired a peripheral pattern and synthesized an extracellular matrix-rich core over the time, what differed from the homogeneous pattern observed in FIB-MT. This study demonstrates the possibility of using WJSC to create MT containing viable and functional cells and abundant extracellular matrix. We hypothesize that WJSC-MT could be a promising alternative in TE protocols. However, future cell differentiation and in vivo studies are still needed to demonstrate the potential usefulness of WJSC-MT in regenerative medicine.

Bioactive injectable aggregates with nanofibrous microspheres and human dental pulp stem cells: A translational strategy in dental endodontics.

Regeneration of the pulp-dentin complex with stem cells is a potential alternative to conventional root canal treatments. Human dental pulp stem cells (hDPSCs) have been extensively studied because of their ability to proliferate and differentiate into mineralized dental and non-dental tissues. Here we combined hDPSCs with two types of injectable poly-l-lactic acid (PLLA) microsphere with a nanofibrous or smooth surface to form bioactive injectable aggregates, and examined their ability to promote pulp regeneration in the root canal in an in vivo model. We investigated the biocompatibility, biosafety and odontogenic potential of fibrous (F-BIM) and smooth bioactive injectable microspheres (S-BIM) in vitro and in vivo. Our results demonstrated that PLLA microspheres and hDPSCs were able to form bioactive injectable aggregates that promoted dentin regeneration in both in vitro and in vivo models. Our results suggest that F-BIM and S-BIM may induce dentinogenesis upon in vivo grafting, and propose that the potential usefulness of the microsphere-hDPSC aggregates described here should be evaluated in clinical settings. Copyright © 2017 John Wiley & Sons, Ltd.

Membranes derived from human umbilical cord Wharton's jelly stem cells as novel bioengineered tissue-like constructs.

Cell-derived matrices were recently described as novel biomaterials generated by human cells allowed to grow and synthetize their own extracellular matrix in culture. In the present work, we generated and evaluated a novel tissue-like substitute (WDM) consisting of a membrane derived from cultured human Wharton's jelly stem cells. WDM were evaluated ex vivo and in vivo by histochemistry and immunohistochemistry for several mesenchymal cell markers and fibrillar and non-fibrillar extracellular matrix components. Results show that WDM were heterogeneous and consisted of dense cell-poor areas surrounded by cell-rich zones with abundant HWJSC. Histological analyses demonstrated that cell-poor areas were very rich in fibrillar and non-fibrillar extracellular matrix components such as collagen and proteoglycans, and cells in the WDM were highly viable and mostly PCNA-positive. HWJSC in the WDM expressed all markers of this cell type, including CD90, CD105, pan cytokeratin and CK8. In vivo analysis showed that the WDM was highly biocompatible and grafting this membrane in the muscle of laboratory rats was not associated to increased inflammation, necrosis, tumorigenesis or other side effects, while cells properly integrated at the damage site and showed high proliferation index. These results suggest that the structure and composition of the extracellular matrix of these novel WDM could reproduce the situation of native human tissues and that WDM implanted in vivo are highly biocompatible and rapidly integrate in the host tissues. For these reasons, we hypothesize that WDM could be used in regenerative medicine protocols.

Development of a multilayered palate substitute in rabbits: a histochemical ex vivo and in vivo analysis.

Current tissue engineering technology focuses on developing simple tissues, whereas multilayered structures comprising several tissue types have rarely been described. We developed a highly biomimetic multilayered palate substitute with bone and oral mucosa tissues using rabbit cells and biomaterials subjected to nanotechnological techniques based on plastic compression. This novel palate substitute was autologously grafted in vivo, and histological and histochemical analyses were used to evaluate biointegration, cell function, and cell differentiation in the multilayered palate substitute. The three-dimensional structure of the multilayered palate substitute was histologically similar to control tissues, but the ex vivo level of cell and tissue differentiation were low as determined by the absence of epithelial differentiation although cytokeratins 4 and 13 were expressed. In vivo grafting was associated with greater cell differentiation, epithelial stratification, and maturation, but the expression of cytokeratins 4, 13, 5, and 19 at did not reach control tissue levels. Histochemical analysis of the oral mucosa stroma and bone detected weak signals for proteoglycans, elastic and collagen fibers, mineralization deposits and osteocalcin in the multilayered palate substitute cultured ex vivo. However, in vivo grafting was able to induce cell and tissue differentiation, although the expression levels of these components were always significantly lower than those found in controls, except for collagen in the bone layer. These results suggest that generation of a full-thickness multilayered palate substitute is achievable and that tissues become partially differentiated upon in vivo grafting.

An immunohistochemical study of cytokeratins distribution of the human adult male and female urethra.

Surgical treatment of diseases affecting long urethral areas represents a challenge in urology. Recent developments of tissue-engineered urethral substitutes represent a hope for patients. However finding an ideal tissue source for urethral reconstruction first requires proper understanding of the native human urethra physiology and a deep knowledge of the histological and molecular features of the native human urethra. Here we present a comprehensive characterization of male and female urethra by histological, histochemical and immunohistochemical methods with a panel of 15 antibodies. The results demonstrated that the histology of the male and female urethra depend on the area where the sample is taken along its length. Proximal areas of male and female urethra have differential expression of the epithelial basal and suprabasal layer markers CK14 and CK10 which distinguished the prostatic/membranous and proximal female urethra from the bulbar/penile and distal female areas of the urethra. The distal male (penile) and female may be further divided by the distinct expression pattern of CK19. On the other hand, the expression of CK5/6 and CK19 also make a distinction of the proximal and distal female urethra. These results should facilitate a more informed selection of donor graft tissues for urethral replacement. Besides, novel bioengineered urethral tissue approaches should take into account the characterization of the different areas of the urethra presented in this work.

Relevant aspects in the surface properties in titanium dental implants for the cellular viability.

Roughness and topographical features are the most relevant of the surface properties for a dental implant for its osseointegration. For that reason, we studied the four surfaces more used in titanium dental implants: machined, sandblasted, acid etching and sandblasted plus acid etching. The roughness and wettability (contact angle and surface free energy) was studied by means 3D-interferometric microscope and sessile drop method. Normal human gingival fibroblasts (HGF) were obtained from small oral mucosa biopsies and were used for cell cultures. To analyze cell integrity, we first quantified the total amount of DNA and LDH released from dead cells to the culture medium. Then, LIVE/DEAD assay was used as a combined method assessing cell integrity and metabolism. All experiments were carried out on each cell type cultured on each Ti material for 24h, 48h and 72h. To evaluate the in vivo cell adhesion capability of each Ti surface, the four types of discs were grafted subcutaneously in 5 Wistar rats. Sandblasted surfaces were significantly rougher than acid etching and machined. Wettability and surface free energy decrease when the roughness increases in sand blasted samples. This fact favors the protein adsorption. The DNA released by cells cultured on the four Ti surfaces did not differ from that of positive control cells (p>0.05). The number of cells per area was significantly lower (p<0.05) in the sand-blasted surface than in the machined and surface for both cell types (7±2 cells for HGF and 10±5 cells for SAOS-2). The surface of the machined-type discs grafted in vivo had a very small area occupied by cells and/or connective tissue (3.5%), whereas 36.6% of the sandblasted plus acid etching surface, 75.9% of sandblasted discs and 59.6% of acid etching discs was covered with cells and connective tissue. Cells cultured on rougher surfaces tended to exhibit attributes of more differentiated osteoblasts than cells cultured on smoother surfaces. These surface properties justify that the sandblasted implants is able to significantly increase bone contact and bone growth with very good osseointegration results in vivo.

Ex vivo and in vivo modulatory effects of umbilical cord Wharton's jelly stem cells on human oral mucosa stroma substitutes.

Novel oral mucosa substitutes have been developed in the laboratory using human umbilical cord Wharton's jelly stem cells -HWJSC- as an alternative cell source. In the present work, we have generated human oral mucosa substitutes with oral mucosa keratinocytes and HWJSC to determine the influence of these cell sources on stromal differentiation. First, acellular and cellular stroma substitutes and bilayered oral mucosa substitutes with an epithelial layer consisting of oral mucosa keratinocytes -OM samples- or HWJSC -hOM- were generated. Then, tissues were analyzed by light and electron microscopy, histochemistry and immunohistochemistry to quantify all major extracellular matrix components after 1, 2 and 3 weeks of ex vivo development, and OM and hOM were also analyzed after in vivo grafting. The results showed that bioengineered oral mucosa stromas displayed an adequate fibrillar mesh. Synthesis of abundant collagen fibers was detected in OM and hOM after 3 weeks, and in vivo grafting resulted in an increased collagen synthesis. No elastic or reticular fibers were found. Glycoprotein synthesis was found at the epithelial-stromal layer when samples were grafted in vivo. Finally, proteoglycans, decorin, versican and aggrecan were strongly dependent on the in vivo environment and the presence of a well-structured epithelium on top. The use of HWJSC was associated to an increased synthesis of versican. These results confirm the usefulness of fibrin-agarose biomaterials for the generation of an efficient human oral mucosa stroma substitute and the importance of the in vivo environment and the epithelial-mesenchymal interaction for the adequate differentiation of the bioengineered stroma.

Human Dental Pulp Stem Cells. A promising epithelial-like cell source.

Several models of tissue-engineered human skin based on three-dimensional (3D) co-culture techniques have been proposed to the date. However, normal skin biopsies are not always available, especially in patients with a high percentage of skin affected by deep burning, and the generation of large amounts of cultured keratinocytes may take very long time, with an associated risk for the patients' survival. For those reasons, the search of alternative cell sources for tissue reconstruction is a clinical need. In this context, Human Dental Pulp Stem Cells (HDPSC) have the potential to differentiate into multiple cell lineages by the appropriate differentiation conditions, but skin epidermis differentiation has not been demonstrated so far. Here, we hypothesize that HDPSC may have pluripotent differentiation capability, and may be able to differentiate into skin epithelial keratinocytes in culture using organotypic 3D models based on the interaction with the subjacent dermal fibroblasts. By using HDPSC, the problems associated to the donor site availability and the proliferation capability of the epithelial cells could be solved. The rapid accessibility to these cells could be translated to a more immediate generation of a bioengineered human skin substitute for the future clinical treatment.

Sequential keratinocytic differentiation and maturation in a three-dimensional model of human artificial oral mucosa.

BACKGROUND AND OBJECTIVE: Oral mucosa shortage may limit or condition some clinical approaches in maxillofacial, periodontal and implant treatment. The availability of a human oral mucosa model generated by tissue engineering could help clinicians to address the lack of oral mucosa. In this work, we carried out a sequential maturation and differentiation study of the epithelial cell layer of an artificial human oral mucosa substitute based on fibrin-agarose biomaterials with fibroblasts and keratinocytes. MATERIAL AND METHODS: Histological, immunohistochemical and gene expression analyses were carried out in artificial human oral mucosa models developed and cultured for 1, 2 and 3 wk. RESULTS: Artificial oral mucosa models showed expression of tight junction proteins and cytokeratins from the first week of in vitro development. Mature samples of 3 wk of development subjected to air-liquid conditions showed signs of epithelial differentiation and expressed specific RNAs and proteins corresponding to adherent and gap junctions and basement lamina. Moreover, these mature samples overexpressed some desmosomal and tight junction transcripts, with gap junction components being downregulated. CONCLUSION: These results suggest that bioengineered human oral mucosa substitutes form a well-developed epithelial layer that was very similar to human native tissues. In consequence, the epithelial layer could be fully functional in these oral mucosa substitutes, thus implying that these tissues may have clinical usefulness.

Capacidad funcional, características del dolor y tratamiento farmacológico en pacientes con artrosis de rodilla / Functional capacity, pain characteristics and drug therapy in patients with knee osteoarthritis

Introducción. El objetivo de este estudio es valorar las características del dolor, la capacidad funcional y el tratamiento farmacológico de pacientes con gonartrosis que acuden a consultas de medicina familiar, así como los factores asociados con los mismos. Material y métodos. Se diseñó un estudio transversal. Por muestreo consecutivo fueron seleccionados pacientes mayores de 50 años que acudieron a consulta por dolor de rodilla y que cumplían criterios diagnósticos de gonartrosis (Sociedad Española de Reumatología), desde febrero a noviembre de 2011. A través de un protocolo específicamente diseñado se registraron variables sociodemográficas, clínicas, cuestionarios WOMAC y Lequesne, escala analógica visual de dolor, realización de radiografía de rodillas, y uso de fármacos. Resultados. Se incluyeron 86 pacientes, 70,9% mujeres, con rango de edad de 52-93 años (media: 73,7). El 96,5% tenía otras enfermedades crónicas. Tomaban analgésicos regularmente 83, con 87,9% (IC 95%:80,3-95,6) en el primer escalón analgésico de la Organización Mundial de la Salud (OMS). Los pacientes en los escalones 2 y 3 puntuaban significativamente más alto en la escala analógica visual que aquellos en el estadio 1 o que no tomaban analgésicos (media: 7,9 vs 5,9; p = 0,01). Con relación al cuestionario WOMAC existían diferencias significativas solo para 2 ítems: todos los que tomaban opioides tenían dificultad para levantarse de la cama (p = 0,001), y el 90% para levantarse del retrete (p = 0,005). El índice de Lequesne mostró un valor medio de 13,1 (DE: 4,1; rango: 3-21). Conclusiones. La gran mayoría de los pacientes con artrosis de rodilla que consultan en Atención Primaria tienen dolor fundamentalmente con el movimiento, lo que limita severamente su capacidad funcional. Los analgésicos se utilizan poco, por lo que queda un amplio margen terapéutico en estos pacientes (AU)

Introduction. Our objective was to assess the characteristics of pain, functional capacity, and pharmacological treatment in patients with knee osteoarthritis attending primary care centers and the factors associated with these characteristics. Material and methods. A cross-sectional study was performed in which patients over 50 years old consulting for knee pain and meeting diagnostic criteria for knee osteoarthritis (Spanish Rheumatology Society) were selected by consecutive sampling from February to November 2011. The following variables were recorded using a specifically designed protocol: sociodemographic and clinical variables, the WOMAC and Lequesne questionnaires, visual analog pain scale, radiographs of the knees, and drug use. Results. We included 86 patients, of whom 70.9% were women. Age ranged from 52 to 93 years (mean: 73.7) and 96.5% had other chronic diseases. Eighty three patients took analgesics regularly and 87.9% (95% CI: 80.3-95.6%) were in the first step of the World Health Organization (WHO) analgesic ladder. Patients in steps 2 and 3 scored significantly higher on the visual analog scale than those in stage 1 or not taking analgesics (mean: 7.9 vs 5.9, p = .01). The WOMAC scale showed significant differences in only two items: all those taking opioids had difficulty getting out of the bed (p = .001), and 90% had difficulty in rising from the toilet (p = .005). The Lequesne index showed a mean value of 13.1 (SD: 4.1; range 3-21). Conclusions. Most patients with knee osteoarthritis attending primary care essentially have pain on movement, severely limiting their functional capacity. Analgesics are used to a limited extent, leaving a wide therapeutic margin in these patients (AU)

Expression of epithelial markers by human umbilical cord stem cells. A topographical analysis.

INTRODUCTION: Human umbilical cord stem cells have inherent differentiation capabilities and potential usefulness in regenerative medicine. However, the epithelial differentiation capability and the heterogeneity of these cells have not been fully explored to the date. METHODS: We analyzed the expression of several undifferentiation and epithelial markers in cells located in situ in different zones of the umbilical cord -in situ analysis- and in primary ex vivo cell cultures of Wharton's jelly stem cells by microarray and immunofluorescence. RESULTS: Our results demonstrated that umbilical cord cells were heterogeneous and had intrinsic capability to express in situ stem cell markers, CD90 and CD105 and the epithelial markers cytokeratins 3, 4, 7, 8, 12, 13, 19, desmoplakin and zonula occludens 1 as determined by microarray and immunofluorescence, and most of these markers remained expressed after transferring the cells from the in situ to the ex vivo cell culture conditions. However, important differences were detected among some cell types in the umbilical cord, with subvascular zone cells showing less expression of stem cell markers and cells in Wharton's jelly and the amnioblastic zones showing the highest expression of stem cells and epithelial markers. CONCLUSIONS: These results suggest that umbilical cord mesenchymal cells have intrinsic potential to express relevant epithelial markers, and support the idea that they could be used as alternative cell sources for epithelial tissue engineering.

Retroperitoneal schwannoma. A complex surgical treatment of a tumor with uncertain behavior.

BACKGROUND: Retroperitoneal schwannoma is a rare nerve sheath tumor; the surgical removal of this tumor is sometimes compromised by its location. The aim of this study is to analyze our experience with the diagnosis and treatment of this type of tumor. METHOD: We present our experience between 1999 and 2011 in the diagnosis and treatment of retroperitoneal schwannoma. During that time, we diagnosed and treated five female patients (four adults and one infant) with the condition. The tumors appeared sporadically and were not associated with neurofibromatosis or other syndromes. Diagnosis was performed by computed tomography (CT) imaging in four cases and by magnetic resonance imaging (MRI) in one case. RESULTS: All patients underwent surgical treatment and complete resection of the lesion. An open resection was performed in four cases, and in the most recent case, the excision was conducted laparoscopically. In all of the cases, the histological diagnosis was retroperitoneal schwannoma, and in one case, there was a melanocytic variant that was not associated with Carney syndrome. At the time of this report, there has been no evidence of recurrence. CONCLUSION: Retroperitoneal schwannoma is a tumor that is difficult to diagnose with imaging techniques, and because of its localization, the tumor is difficult to remove surgically.

Medidas preventivas para evitar complicaciones de salud derivadas del uso de sondajes vesicales en pacientes lesionados medulares / Preventive measures to avoid health complications from the use of catheters in patients with spinal cord injury

Introducción: La lesión medular es uno de los sucesos más trágicos que puede acaecerle a una persona por las devastadoras consecuencias que conlleva la parálisis de los 2 ó 4 miembros en forma de paraplejía o tetraplejía, pérdida de las sensibilidades, disfunción vesical, intestinal, sexual y consecuentemente trascendentales secuelas psicológicas, sociales, laborales y económicas. Objetivos: El objetivo principal es determinar si ciertos tipos de sondas y técnicas de colocación de la sonda son mejores para la prevención de complicaciones a largo plazo en la incontinencia urinaria. Metodología: La estrategias de búsqueda para la identificación de los artículos de esta revisión los buscamos en bases de datos como Cinahl, Pubmed, Biblioteca Virtual de la Salud, Csic, Biblioteca Cochrane Plus. Las búsquedas se realizaron mediante el sistema de palabras clave. Se llevó a cabo la búsqueda mediante los descriptores DeCS /MeSH y además una búsqueda manual de artículos en revistas especializadas de Enfermería tanto españolas como extranjeras. Resultados: No hay estudios definitivos que revelen que la incidencia de la infección urinaria mejore con alguna técnica, estrategia o tipo de sonda; la dificultad de estos estudios consiste pues en que se necesita un periodo largo de tiempo y muchos pacientes abandonan el estudio. Conclusiones: Los datos disponibles sobre el sondaje intermitente no aportan pruebas convincentes de que la técnica especifica (estéril o limpia), tipo de sonda (recubierta o no recubierta); método (único uso o uso múltiple), personas (paciente u otro), o estrategia sea mejor que otra para todos los contextos clínicos. Las pruebas actuales son débiles y se recomiendan estudios bien diseñados. Valorar el impacto en la calidad de vida, supone la utilización de distintos métodos de evacuación urinaria en pacientes con lesión medular, utilizando un cuestionario específico y validado al español: King´s Health Questionnaire(AU)

Introduction: Spinal cord injury is one of the most tragic events that can befall a person for the devastating consequences associated with paralysis of 2 or 4 members in the form of paraplegia or quadriplegia, loss of sensitivity, bladder dysfunction, bowel, sexual and consequently transcendental psychologic social, labor and economical consequencies. Objectives: Main objective is determining if certain types of probes and catheterization techniques are the best for preventing long-term complications in urinary incontinence. Methodology: Search strategies for identifying articles of this review include databases such as Cinahl, Pubmed, Virtual Health Library, CSIC, and Cochrane Library. Searches were conducted using the keyword system: We carried out a search using Mesh descriptors / Mesh and a manual search of articles in Nursing journals from Spain and foreign countries. Results: There are no definitive studies that reveal that the incidence of UTI improves with some kind of technique, strategy or probe type. The main difficulty for these studies is the long duration; many patients leave the study. Conclusions: The available data on intermittent catheterization does not provide convincing evidence for the specific technique (sterile or clean), probe type (coated or uncoated), method (single use or multiple use), people (patient or other), or strategy is better than another for all clinical settings. The current evidence is uninformative and well-designed studies are recommended. Evaluating the impact on quality of life involves the use of different methods of evacuation in patients with spinal cord injury using a specific questionnaire validated in Spanish: King's Health Questionnaire(AU)

The effects of fibrin and fibrin-agarose on the extracellular matrix profile of bioengineered oral mucosa.

Several studies have developed efficient oral mucosa constructs using different types of scaffold. However, the changes in the morphology and gene and protein expression profile that could occur in these artificial constructs remain unknown. This study compared the histology and expression of several extracellular matrix molecules in human artificial oral mucosa developed using two different types of scaffolds: fibrin and fibrin-agarose. To that end, bioengineered oral mucosa stromas were constructed from biopsy samples of human oral mucosa and the substitute generated was analyzed at different periods of time in culture. Histological analysis was carried out by light and transmission electron microscopy and the expression of collagen types I, III, and VI, the proteoglycans decorin and biglycan, and the different chains of laminin, were assessed by immunoperoxidase technique. This study found that fibrin scaffolds accelerated fibroblast growth and remodeling of the scaffold, thus enhancing collagen fibrillogenesis. In the fibrin-agarose scaffold, the morphology and organization of the fibroblasts did not change during the culture period. All extracellular matrix proteins analyzed were expressed in both scaffolds. However, in fibrin scaffolds, these proteins were widely distributed and replaced the scaffold during the follow-up period. These results show that the substitutes generated showed histological and molecular similarities with native human oral mucosa stroma. In addition, it was observed that the nature of the biomaterial influenced the behaviour of the oral stromal fibroblasts, thereby modulating their growth, protein synthesis, and collagen fibrillogenesis.