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Introduction: Chronic venous insufficiency affects about 5% of the global adult population. Venous leg ulcers are one of the most frequent complications of this pathology, with a global prevalence of 2%. This disease affects both the quality of life of patients and, due to the high cost of the treatment, the health system. Compressive therapy and moist wound healing have been the gold standard treatment. However, when complications occur, they may not be effective. Case report: This is the case of a 66-year-old female patient with venous ulcers on her lower limbs and symptoms of fever and local pain that did not respond to conventional therapies. The patient was treated with a new dermal substitute made of an acellular type-I collagen membrane, which promotes the closure of the ulcer by stimulating the replacement of injured tissue with tissue similar to the healthy one. The condition of the patient improved at 16 weeks, and after 8 months of treatment there was no recurrence of the lesions. Conclusions: Acellular type-I collagen membrane developed by the Tissue Engineering Working Group of the Department of Pharmacy of the Universidad Nacional de Colombia is effective in treating venous ulcers of the lower limbs. Its low cost facilitates the access of the whole population to therapies based on its application. RESUMEN Introducción. La insuficiencia venosa crónica afecta alrededor del 5% de la población adulta en el mundo; una de sus mayores complicaciones son las úlceras en miembros inferiores, las cuales tienen una prevalencia mundial del 2%. Las úlceras afectan la calidad de vida de los pacientes e impactan al sistema de salud debido a los altos costos de atención que genera. El tratamiento de referencia es la terapia compresiva y la cura húmeda de las heridas, sin embargo estas intervenciones pueden no ser efectivas cuando las lesiones se complican. Presentación del caso. Paciente femenina de 66 años con úlceras venosas en miembros inferiores acompañadas de fiebre y dolor local que no respondían a las terapias convencionales. La paciente fue tratada con un nuevo sustituto dérmico basado en una membrana acelular de colágeno tipo I que contribuye al cierre de la úlcera al estimular el remplazo del tejido lesionado por tejido similar al sano, con lo cual tuvo mejoría a las 16 semanas; después de 8 meses de terminado el tratamiento no se presentó recurrencia de las lesiones. Conclusiones. La membrana acelular de colágeno tipo I desarrollada por el Grupo de Trabajo en Ingeniería de Tejidos del Departamento de Farmacia de la Universidad Nacional de Colombia es efectiva en el tratamiento de úlceras venosas en miembros inferiores y su bajo costo facilita el acceso de toda la población a terapias basadas en su aplicación.

Although many bone substitutes have been designed and produced, the development of bone tissue engineering products that mimic the microstructural characteristics of native bone remains challenging. It has been shown that pore orientation within collagen scaffolds influences bone matrix formation by the endochondral route. In addition, that the unidirectional orientation of the scaffolds can limit the growth of blood vessels. However, a comparison between the amount of bone that can be formed in scaffolds with different pore orientations in addition to analyzing the effect of loading osteogenic and proangiogenic factors is still required. In this work we fabricated uni- and multidirectional collagen sponges and evaluated their microstructural, physicochemical, mechanical and biological characteristics. Although the porosity and average pore size of the uni- and multidirectional scaffolds was similar (94.5% vs. 97.1% and 260 µm vs. 269 µm, respectively) the unidirectional sponges had a higher tensile strength, Young’s modulus and capacity to uptake liquids than the multidirectional ones (0.271 MPa vs. 0.478 MPa, 9.623 MPa vs. 3.426 MPa and 8000% mass gain vs. 4000%, respectively). Culturing of rat bone marrow mesenchymal stem cells demonstrated that these scaffolds support cell growth and osteoblastic differentiation in the presence of BMP-2 in vitro, although the pore orientation somehow affected cell attachment and differentiation. The evaluation of the ability of the scaffolds to support bone growth when loaded with BMP-2 or BMP-2 + VEGF in an ectopic rat model showed that they both supported bone formation. Histological analysis and quantification of mineralized matrix revealed that the pore orientation of the collagen scaffolds influenced the osteogenic process.

The role of periodontopathogens in inflammatory endothelial dysfunction is not known. This study characterizes a three-dimensional model with human coronary artery endothelial cells on three-dimensional (HCAEC-3D) type I collagen scaffolds to evaluate whether infection with Aggregatibacter actinomycetemcomitans induces a proinflammatory response associated with atherosclerosis. The HCAEC-3D culture was physicochemically characterized with regard to biocompatibility and barrier function. Then, the culture was infected with A. actinomycetemcomitans strain ATCC 29522 at multiplicities of infection (MOIs) of 1:1, 1:10, and 1:100. Cultures without infection and stimulated with A. actinomycetemcomitans lipopolysaccharide were used as controls. The secretion of soluble factors (IL-6, IL-1β, MCP-1, RANTES, MIP-1, IL-8, IL-1α, and TNF-α) was evaluated via flow cytometry; TGF-β1 was evaluated via enzyme-linked immunosorbent assay (ELISA). The adhesion and migration of fluorescent human THP-1 monocytes was evaluated. IL-8, MCP-1, and IL-6 secretion increased in a dose-dependent manner with A. actinomycetemcomitans infection and was significantly greater than that under control treatment. The concentration of TGF-β1 was significantly higher at MOI 1:100 than in controls. Treatment of the 3D cultures with A. actinomycetemcomitans at different MOIs induced significant differences in the adhesion of monocytes to the endothelium compared to the control without infection. Lastly, conditioned media from 3D cultures treated with A. actinomycetemcomitans induced monocyte migration. The effects of IL-8, MCP-1, IL-6, and TGF-β1 on the endothelium indicate the ability of A. actinomycetemcomitans to induce an inflammatory response through a mechanism of monocyte adhesion and migration and endothelial dysfunction.

Shortness of donor nerves has led to the development of nerve conduits that connect sectioned peripheral nerve stumps and help to prevent the formation of neuromas. Often, the standard diameters of these devices cannot be adapted at the time of surgery to the diameter of the nerve injured. In this work, scaffolds were developed to form filled nerve conduits with an inner matrix with unidirectional channels covered by a multidirectional pore zone. Collagen type I dispersions (5 mg/g and 8 mg/g) were sequentially frozen using different methods to obtain six laminar scaffolds (P1 to P5) formed by a unidirectional (U) pore/channel zone adjacent to a multidirectional (M) pore zone. The physicochemical and microstructural properties of the scaffolds were determined and compared, as well as their biodegradability, residual glutaraldehyde and cytocompatibility. Also, the Young's modulus of the conduits made by rolling up the bizonal scaffolds from the unidirectional to the multidirectional zone was determined. Based on these comparisons, the proliferation and differentiation of hASC were assessed only in the P3 scaffolds. The cells adhered, aligned in the same direction as the unidirectional porous fibers, proliferated, and differentiated into Schwann-like cells. Adjustable conduits made with the P3 scaffold were implanted in rats 10 mm sciatic nerve lesions to compare their performance with that of autologous sciatic nerve grafted lesions. The in vivo results demonstrated that the tested conduit can be adapted to the diameter of the nerve stumps to guide their growth and promote their regeneration. published

In mammals, long bones are formed by ossification of a cartilaginous mould during early stages of development, through the formation of structures called the primary ossification centre, the secondary ossification centres (SOCs) and the physeal cartilages (PCs). The PC is responsible for long bone growth. The morphology of the PC and the SOCs varies during different stages of femoral growth. In this respect, several details involving the process of murine femoral development are lacking. In the present study, a morphological characterization of femur development from the embryonic period to adulthood in mice was studied using micro-computed tomography (micro-CT). To achieve this aim, femora were collected at embryonic day (E) 14.5, E16.5 and E18.5 and at postnatal day (P)1, P7, P14, P35, P46 and P52. CT images were obtained using a micro-CT scanner (X-SkyScan 1172; Micro Photonics) and analysed using the micro-CT 3D visualization software Mimics (Materialise NV, Leuven, Belgium) and NRecon (Micro Photonics). The results of the present study revealed that the femur and its PCs and SOCs undergo morphological changes during different stages of development, including changes in their shape as well as position and thickness. These changes may be due to the response of the femur to mechanical loads imposed by muscle surrounding the bone during these stages of development. The result of the present study is important to improve our knowledge related to ossification and growth patterns of mouse femur during development.

Research on collagen type I scaffolds with Aloe vera is sparse. The aim of this work was to develop collagen type I scaffolds with gelatin-collagen microparticles and loaded with a dispersion of A. vera, to assess their performance as grafting material for healing of skin wounds. Scaffolds were evaluated in a Cavia porcellus model with full-thickness skin wound and compared with wounds healed by secondary intention (controls). Animals grafted with scaffolds without A. vera and their control wounds were also included in the study. Evaluation of enzymatic degradation and percentage of the scaffolds' free amino groups-as an indirect assessment of their cross-linking-were also carried out because A. vera contains compounds which affect their stability. We found that dispersions of lyophilized A. vera extract loaded on scaffolds do not have cytotoxic potential, and they decrease collagenase degradation of scaffolds in the range of 0.1 to 0.3% w/v in a dose-dependent manner. Only the A. vera dispersion with the highest concentration (0.3% w/v) decreased the percentage of free amino groups, which are the ones involved in the cross-link of collagen fibers. This finding suggests that cross-linking is not the mechanism by which the tested dispersions stabilize the scaffolds. Preclinical, histochemical, and histomorphometric analyses of repaired wound tissue indicate that loading collagen type I scaffolds, including microparticles of gelatin-collagen, with A. vera in the concentrations tested does not improve wound healing. Low biodegradability of the tested scaffolds caused by the inhibition of collagenase activity might account for these results.

Studies have been conducted on the pathogenicity of periodontopathogens in cultures of endothelial cells on two-dimensional (2D) polystyrene surfaces, where the monolayer formed is not exposed to proteins of the subendothelial matrix. In this work, we developed a culture system by seeding human coronary artery endothelial cells (HCAECs) onto three-dimensional (3D) scaffolds of collagen type I, a subendothelial protein. The inflammatory responses of the HCAEC monolayers, formed either on 3D scaffolds or directly on a 2D polystyrene plate, to lipopolysaccharide (LPS) from Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) were evaluated. The transcription of 3 genes, the secretion of 40 cytokines and 2 prostanoids, and the adhesion of monocytes to 2D and 3D cultures with or without exposure to lipopolysaccharides (control) were assessed. HCAECs exhibited differences in transcriptional and secretory profiles between the 3D and 2D models. In addition, the inflammatory responses of HCAEC to Aa-LPS and Pg-LPS differed between the two models. In 3D cultures treated with Aa-LPS, the levels of IL-8, RANTES, G-CSF, ICAM-1, IL-6, and TXA2 were significantly higher than those in the controls. In 2D cultures treated with Aa-LPS, IL-8, RANTES, G-CSF, ICAM-1, TNF-RI, PGI2, and TXA2 levels were significantly higher than those in their controls. In the presence of Aa-LPS, monocyte adhesion did not differ between treated and control 3D cultures but was significantly higher in treated 2D cultures than in the controls. In response to Pg-LPS, cytokine-prostaglandin secretion and monocyte adhesion did not differ between 3D and 2D cultures. These data indicate that HCAECs respond differently to these two types of LPS.

In the original article text presenting and discussing results shown in Fig. 6 omitted to mention that quantification of TGF-β2 and TGF-β3 was not included in Fig. 6a, c, e.

Previously, we have developed collagen type I scaffolds including microparticles of gelatin-collagen type I (SGC) that are able to control the release of a hydroglycolic extract of the Calendula officinalis flower. The main goal of the present work was to carry out the preclinical evaluation of SGC alone or loaded with the C. officinalis extract (SGC-E) in a lagomorph model of full-thickness skin wound. A total of 39 rabbits were distributed in three groups, of 13 animals each. The first group was used to compare wound healing by secondary intention (control) with wound healing observed when wounds were grafted with SGC alone. Comparison of control wounds with wounds grafted with SGC-E was performed in the second group, and comparison of wounds grafted with SGC with wounds grafted with SGC-E was performed in the third group. Clinical follow-ups were carried in all animals after surgery, and histological and histomorphometric analyses were performed on tissues taken from the healed area and healthy surrounding tissue. Histological and histomorphometric results indicate that grafting of SGC alone favors wound healing and brings a better clinical outcome than grafting SGC-E. In vitro collagenase digestion data suggested that the association of the C. officinalis extract to SGC increased the SGC-E cross-linking, making it difficult to degrade and affecting its biocompatibility.

Analysis of factors that play a role on the healing process in exudates from skin wounds might shed light on the effect that grafted artificial tissue has in wound regeneration and repair. The first objective of this work was to standardize an optic surface plasmon resonance method based on self-assembled monolayers to quantify healing mediator factors (angiopoietin-2, epidermal growth factor, tumour necrosis factor-α, transforming growth factor-β1, and vascular endothelial growth factor) in wound exudates. Optimal conditions for self-assembling of alkanethiol monolayers, immobilization of antibodies antifactors, and regeneration of sensor surfaces were established. A second objective was to compare healing of wounds grafted with artificial dermis with wounds left to heal by secondary intention (control) in a lagomorph model of full-thickness skin wound. Each animal included in this study had a control wound and an identical contralateral wound grafted with artificial dermis that was made by seeding autologous skin fibroblasts into unidirectional or multidirectional collagen type I scaffolds. Histological and histomorphometric analyses were carried out when animals were sacrificed, in addition to quantifying the factors in the exudates of wounds sampled 3 days after surgery. There were significant differences between the concentrations of evaluated factors in the exudates from grafted and control wounds. This finding coincides with differences observed in the histological and histomorphometric analyses of repaired tissue formed in treated and control wounds.

El cartílago articular es un tejido vulnerable a las lesiones de diferente etiología; siendo uno de los más afectados, el cartílago de la rodilla. Aunque la mayoría de los tratamientos convencionales reducen los síntomas, generalmente conducen a la formación de fibrocartílago; el cual, posee características diferentes a las del cartílago hialino de las articulaciones. Son pocas las aproximaciones terapéuticas que promueven el reemplazo del tejido dañado por cartílago hialino funcional; las más exitosas son las denominadas terapias avanzadas, que aplican células y productos de ingeniería de tejidos con el fin de estimular la regeneración del cartílago. La mayoría de ellas se basan en colocar soportes hechos con biomateriales de diferente origen, que sembrados o no con células exógenas o endógenas, reemplazan al cartílago dañado y promueven su regeneración. Este trabajo revisa algunas de las aproximaciones terapéuticas enfocadas en la regeneración del cartílago articular de rodilla; así como, los biomateriales más empleados en la elaboración de soportes para terapia celular e ingeniería de tejido cartilaginoso. The articular cartilage is prone to suffer lesions of different etiology, being the articular cartilage lesions of the knee the most common. Although most conventional treatments reduce symptoms they lead to the production of fibrocartilage, which has different characteristics than the hyaline cartilage of the joint. There are few therapeutic approaches that promote the replacement of damaged tissue by functional hyaline cartilage. Among them are the so-called advanced therapies, which use cells and tissue engineering products to promote cartilage regeneration. Most of them are based on scaffolds made of different biomaterials, which seeded or not with endogenous or exogenous cells, can be used as cartilage artificial replacement to improve joint function. This paper reviews some therapeutic approaches focused on the regeneration of articular cartilage of the knee and the biomaterials used to develop scaffolds for cell therapy and tissue engineering of cartilage.

Initial Enterococcus faecalis–endothelial cell molecular interactions which lead to enterococci associating in the host endothelial tissue, colonizing it and proliferating there can be assessed using in vitro models. Cultured human umbilical vein endothelial cells (HUVEC) have been used to study other Gram-positive bacteria–cell interactions; however, few studies have been aimed at establishing the relationship of E. faecalis with endothelial cells. The aggregation substance (AS) family of adhesins represents an E. faecalis virulence factor which has been implicated in endocarditis severity and bacterial persistence. The Asc10 protein (a member of this family) promotes bacterium–bacterium aggregation and bacterium–host cell binding. Evaluating Asc10 role in bacterial internalization by cultured enterocytes has shown that this adhesin facilitates E. faecalis endocytosis by HT-29 cells. A few eukaryotic cell structural components, such as cytoskeletal proteins, have been involved in E. faecalis entry into cell-lines; it is thus relevant to determine whether Asc10, as well as microtubules and actin microfilaments, play a role in E. faecalis internalization by cultured endothelial cells. The role of Asc10 and cytoskeleton proteins in E. faecalis ability to enter HUVEC was assessed in the present study, as well as cell apoptosis induction by enterococcal internalization by HUVEC; the data indicated increased cell apoptosis and that cytoskeleton components were partially involved in E. faecalis entry to endothelial cells, thereby suggesting that E. faecalis Asc10 protein would not be a critical factor for bacterial entry to cultured HUVEC.

This work evaluates the effect of processing variables on some physicochemical and mechanical properties of multi- and unidirectional laminar collagen type I scaffolds. The processing variables considered in this study included microstructure orientation (uni- and multidirectional fiber/pore controlled by freeze-drying methodology), cross-linking (chemical - using genipin and glutaraldehyde, and physical - using a dehydrothermal method), and collagen concentration (2, 5 and 8mg/ml). The biocompatibility of the scaffolds obtained in each of the evaluated manufacturing processes was also assessed. Despite previous research on collagen-based platforms, the effects that these processing variables have on the properties of collagen scaffolds are still not completely understood. Unidirectional scaffolds presented higher resistance to failure under stress than multidirectional ones. The cross-linking degree was found to decrease when the concentration of collagen increased whilst using chemical cross-linkers, and to increase with the concentration of collagen for the dehydrothermal cross-linked scaffolds. Pore orientation indexes of both unidirectional and multidirectional scaffolds were not influenced by collagen concentration. Cross-linked scaffolds were more hydrophobic than non-cross-linked ones, and presented water vapor permeability adequate for use in low-to-moderate exuding wounds. Pore size ranges were compatible with cell in-growth, independently of the employed cross-linking and freezing methodologies. Moreover, scaffolds cross-linked with glutaraldehyde presented higher in-growth of primary oral mucosa fibroblasts than those cross-linked with genipin or with the dehydrothermal treatment. This multi-factor analysis is expected to contribute to the design of collagen type I platforms, which are usable on several potential soft tissue-engineering applications.

Several studies have evaluated proteins secreted by fibroblasts comprising skin substitutes, finding that they are secreted in combinations and concentrations that promote wound healing. However, assessment of proteins secreted by oral fibroblasts forming a part of oral substitutes is scarce. In our previous work, collagen type-I scaffolds (CSs) and autologous artificial connective tissue (AACT) were produced and implanted in rabbit oral lesions, evidencing that AACT outperforms CS. The present work determined the secreted factor profile of AACT in the time of grafting as well as that of the AACT embedded in the clot. It also evaluated the proliferation and viability of AACT fibroblasts to establish the dwell time of these cells in the grafted area. Finally, it assessed whether CS, AACT, and clot-embedded AACT increase fibroblast recruitment induced by a fibrin clot, because the cell migratory response has been associated with the wound-healing outcome. We found that some of the factors secreted by AACT fibroblasts are significantly different from those secreted by clot-embedded AACT fibroblasts. Also, that the profile of proteins secreted by AACT fibroblasts and clot-embedded AACT fibroblasts is different from already reported protein secretion profiles of other engineered tissues used in treating oral mucosa wounds. It was also found that AACT fibroblasts are viable when grafted and remain in the treated area for almost 2 weeks, and that the migratory response of fibroblasts to tissue-substitute stimulus is significantly less than the migratory response induced by the clot alone. Overall, data suggest that AACT secretion of proteins is modulated by three-dimensionality and environment factors. This bioactivity and the fact that AACT does not increase fibroblast migration can be held accountable for AACT's good performance as a graft.

Previous studies have shown meconium-induced lung injury occurs throughout release of inflammatory cytokines. The exact mechanism of cytokine-induced apoptosis is not known. In this study we hypothesized that meconium-induced apoptosis in the lungs is mediated through the production of inducible nitric oxide (NO). We studied two groups of newborn rabbit pups: one group was instilled with meconium and other with normal saline. We measured precursors of NO in lung lavage from both groups of rabbits and NO levels were calculated accordingly. The levels of NO and NO-derivatives increased significantly in both groups. However NO expression in meconium group 2 h after meconium instillation was significantly higher than in saline-instilled group suggesting NO production plays a role in meconium-induced inflammation.

Res_eng:Research in developing artificial tissue has emerged during the two last decades around the world. In the same manner that surgical grafting of natural tissue, artificial tissue grafting aims to promote the replacement of lost tissue by one that has similar morphology and function. Artificial tissue does not completely prevent fibrotic scarring and wound contracture. However, in many cases its use has proved to result in better healing. Currently, in the Latin America region the advantages and disadvantages of skin and oral wound therapies based on grafting artificial tissue are only known by a reduced group of scientists working in the field and by the few clinicians willing to use them. For that reason, it is important to inform on skin and oral mucosa tissue engineered products and the advantages and disadvantages of applying them. This chapter surveys the origin of artificial tissue manufacturing, some biological bases of tissue engineering, skin and oral mucosa tissue substitutes approved for human use, and their clinical application.

Aiming to develop biological skin dresses with improved performance in the treatment of skin wounds, acellular collagen I scaffolds were modified with polymeric microparticles and the subsequent loading of a hydroglycolic extract of Calendula officinalis flowers. Microparticles made of gelatin-collagen were produced by a water-in-oil emulsion/cross-linking method. Thereafter, these microparticles were mixed with collagen suspensions at three increasing concentrations and the resulting mixtures lyophilized to make microparticle-loaded porous collagen scaffolds. Resistance to enzymatic degradation, ability to associate with the C. officinalis extract, and the extract release profile of the three gelatin-collagen microparticle-scaffold prototypes were assessed in vitro and compared to collagen scaffolds without microparticles used as control. Data indicated that the incorporation of gelatin-collagen microparticles increased the resistance of the scaffolds to in vitro enzymatic degradation, as well as their association with the C. officinalis flower extract. In addition, a sharp decrease in cytotoxicity, as well as more prolonged release of the extract, was attained. Overall results support the potential of these systems to develop innovative dermal substitutes with improved features. Furthermore, the gelatin-collagen mixture represents a low-cost and scalable alternative with high clinical transferability, especially appealing in developing countries.

Previously, we found that oral autologous artificial connective tissue (AACT) had a different protein secretion profile to that of clot-embedded AACT. Other oral mucosa substitutes, having different cell types and scaffolds, had dissimilar secretion profiles of proteins (including that for AACT) that influence healing outcome; thus, to ascertain the profiles of factors secreted by artificial tissue and whether they are influenced by their microstructure might help in understanding their bioactivity. An important component of tissue microstructure is the fiber orientation of the scaffold used for manufacturing it. This work developed a surface plasmon resonance (SPR) methodology to quantify factors secreted by oral artificial connective tissue (ACT) in culture medium, and a method to manufacture unidirectional laminar collagen I scaffolds. The SPR methodology was used for assessing differences in the protein secretion profile of ACT made with collagen scaffolds having different fiber orientation (unidirectional vs multidirectional). Oral fibroblasts seeded onto unidirectional scaffolds increased the secretion of six factors involved in modulating healing compared to those seeded onto multidirectional scaffolds. Histological analysis of uni- and multidirectional ACT showed that cells differ in their alignment and morphology. This SPR-methodology led to nanoscale detection of paracrine factors and might be useful to study biomarkers of three-dimensional cell growth, cell differentiation, and wound-healing progression.

Impaired wound healing is an important clinical problem in diabetes mellitus and results in failure to completely heal diabetic foot ulcers (DFUs), which may lead to lower extremity amputations. In the present study, collagen based dressings were prepared to be applied as support for the delivery of neurotensin (NT), a neuropeptide that acts as an inflammatory modulator in wound healing. The performance of NT alone and NT–loaded collagen matrices to treat wounds in streptozotocin (STZ) diabetic induced mice was evaluated. Results showed that the prepared dressings were not-cytotoxic up to 72h after contact with macrophages (Raw 264.7) and human keratinocyte (HaCaT) cell lines. Moreover, those cells were shown to adhere to the collagen matrices without noticeable change in their morphology. NT–loaded collagen dressings induced faster healing (17% wound area reduction) in the early phases of wound healing in diabetic wounded mice. In addition, they also significantly reduced inflammatory cytokine expression namely, TNF-α (p

Previous studies demonstrated that basic fibroblast growth factor (bFGF) decreases elastin gene transcription in pulmonary fibroblasts. In this study we pursue the identification of the element and the trans-acting factors responsible. Gel shift analyses show that bFGF increases protein binding to a sequence located at -564 to -558 base pairs (bp), which possesses homology to both AP1 and cAMP-response consensus elements yet displays a unique affinity for heterodimer binding. Site-directed mutation of the -564- to -558-bp sequence results in an increase in promoter activity and abrogates the effect of bFGF. Western blot analysis shows that bFGF induces a sustained increase in the steady-state levels of Fra 1, and co-transfection of a Fra 1 expression vector with an elastin promoter reporter construct results in an inhibition of elastin promoter activity. Overall the results suggest that bFGF represses elastin gene transcription by increasing the amount of the Fra 1 that subsequently binds to the -564- to -558-bp as a heterodimer with c-Jun to form an inhibitory complex. We propose that the identified bFGF response element can serve to down-regulate elastin transcription in elastogenic cells and, conversely, can serve to up-regulate elastogenesis in cells where endogenous bFGF signaling is attenuated or altered.

Currently, the treatment of articular cartilage defects results in tissue repair that leads to osteoarthritis [1]. Autologous chondrocyte implantation (ACI) is a technique based in obtaining autogenous chondrocytes from a cartilage biopsy. After culturing, the cells are seeded onto biodegradable scaffolds that provide the environment required for growing them. The grown chondrocytes replace and remodel the scaffold to produce artificial cartilage used for grafting. Indeed, surgical procedures based on artificial tissue have been already marketed in USA and Europe (Carticel®, BioCart). Unfortunately, the clinical implementation of ACI has been limited by its high cost: Colombian patients who need ACI must travel to USA, Italy or Israel where authorized cell processing facilities are located. Therefore, there is a need for the development of in-site production of tissue engineered cartilage to provide patients of all socioeconomic status with ACI at a reasonable cost. A commonly biomaterial used to manufacture scaffolds for engineering artificial cartilage is collagen type II, a major component of the cartilage extracellular matrix. In this work scaffolds were produced using pure collagen type II isolated from bovine trachea. The microstructure of them was also studied.

Previous studies have demonstrated that insulin-like growth factor-I (IGF-I) increases elastin gene transcription in aortic smooth muscle cells and that this up-regulation is accompanied by a loss of protein binding to the proximal promoter. Sp1 has been identified as one of the factors whose binding is lost, and in the present study we show that Sp3 binding is also abrogated by IGF-I, but in a selected manner. In functional analyses using Drosophila SL-2 cells, Sp1 expression can drive transcription from the elastin proximal promoter, while co-expression of Sp3 results in a repression of Sp1 activity. Footprint and gel shift analyses position the IGF-I responsive sequences to a putative retinoblastoma control element (RCE). Mutation of the putative RCE sequence as assessed by transient transfection of smooth muscle cells results in an increase in reporter activity equal in magnitude to that conferred by IGF-I on the wild type promoter. Together these results support the hypothesis that IGF-I-mediated increase in elastin transcription occurs via a mechanism of derepression involving the abrogation of a repressor that appears to be Sp3 binding to the RCE.

The holographic gratings on photopolymer films are studied by different spatial frequency and thickness to determine the evolution of diffraction efficiency during holographic recording. The experimental results are presented like a function of energy exposure, thickness and spatial frequency. The photopolymer emulsion is prepared with potassium dichromate and nickel (II) chloride hexahydrate in polyvinyl alcohol matrix. The maximum diffraction efficiency reached is at spatial frequency (f2) for sample A.

The study was carried out during real time holographic gratings formation when the coating emulsions age as a function from the time. The experiments are studied by influence of hologram parameters to get the diffraction efficiency at room conditions through changes of electrical potential application and pH dependence on coating solutions for hologram recording process.

This work assessed wound healing response in rabbit oral lesions grafted with autologous artificial connective tissue or acellular collagen scaffolds. Autologous artificial oral connective tissue (AACT) was produced using rabbit fibroblasts and collagen I scaffolds. Before implantation, AACT grafts were assayed to demonstrate the presence of fibroblasts and extracellular matrix components, as well as the expression of characteristic genes and secretion of chemokines, cytokines, and growth factors. AACT grafts were tested in the rabbits from which the fibroblasts were obtained, whereas acellular collagen type I scaffolds (CS) were evaluated in a separate group of rabbits. In both cases, contralateral wounds closed by secondary intention were used as controls. In a separate experiment, AACT-grafted wounds were directly compared with contralateral CS-grafted wounds in the same animals. Wound contraction and histological parameters were examined to evaluate closure differences between the treatments in the three animal experiments performed. Contraction of wounds grafted with AACT and CS was significantly lower than in their controls (p

Volume holography is a research topic that was generated considerably a big interest of the storage information during the last decades. The different systems of storage holographic are principally determined by the physic and chemical characteristics of the storage material. We use an organic conductive material based on polyvinyl alcohol (PVA) for the volume holographic storage. We presented some electro-optics results and physic - chemicals properties of the material used for holographic storage. The influences in the diffraction efficiency parameter as change different variables into material and the qualitative results of other electro-optical parameters examined.

The sugar matrix is used to record of phase holograms; it was modified with the purpose of obtaining a hydrophobic material to improve the stability of the registered image and to stimulate the photosensitivity of the sugar. The new material is formed by a sugar, pectin and vanillin dissolution. The diffraction efficiency parameter increases in comparison with only the sugar matrix, obtaining already of 10%.

Holographic properties of dichromated gelatin (DCG) colored with triphenyl dyes (TDCG), were studied. We described experimental techniques for its register of these holograms. This emulsion has excellent resolution and behavior. That increase the photo sensibility respect to conventional DCG and is easy to use.

Experimental techniques are described to register holograms in DC-PVA doped with organic colorants. This material has excellent resolution and behavior to increase the photo sensibility. We report some preliminary results.

The photopolymers are very attractive materials for applications and as holographic storage, for theirs high modulation and photosensitivity. We report the evolution of diffraction efficiency parameter from volume holographic gratings recorder in PVA doped with CuCl 2 (2H 2 O), which is the result of the physic-chemical changes between the light and the material. The resultant variation of the grating diffraction efficiency parameter is examined.

Polymer films mixed with metals salts are a novel organic material with applications in technology process for optical information storages. In this work we show quantitatively some optical and electrical characteristics of a composite organic conductive as holographic material. We show a material with the optical properties that depend of physics and chemical changes, which we register, the process and manipulate adequately parameters to obtain better results in the diffraction efficiency.

Characterization of the organic conductive materials is studied by voltage application during holographic gratings formation and when the exposure time of 900 seconds was reached. The behavior of the material is analyzed by diffracted intensity and exposure energy. Diffraction efficiency parameter of holographic gratings were studied in an organic conductive material like dichromated poly(vinyl alcohol) films doped with nickel(II) chloride hexahydrate. The study includes a comparative analysis between the experimental results obtained.

Experimental results to the saturation and diffraction efficiency from holographic gratings are presented in this investigation. The experiments were carried out during real time holographic gratings formation. Dichromated poly(vinyl alcohol) was doped with nickel(II) chloride hexahydrate and it is used like optical material. The influence of the hologram parameters to get the maximum diffraction efficiency is studied at room conditions. This study contributes to get more information about the behavior of this material for holographic gratings recording.

Experimental techniques are described for recording holograms in dichromate gelatin doped with an organic dye. This material has excellent resolution, and increase the photo sensibility. We present some basic studies about the possible form of to storage information, and reported results. This material is cheap, easy of to use, and it decrease the register time.

This work presents experimental results of intensity changes by polarization conditions at the resultant diffraction patters. The substrate used as retarder plate was a commercial transparency film for use with plain paper copier (3M-PP2900 TM ). The conductive material composition was introduce to dichromated poly(vinyl alcohol) by adding a metallic salt as nickel(II) chloride hexahydrate. Some electro-optical characteristics of organic conductive material that are used in the holographic gratings storage specifically when applied voltage.

The electro-optical changes of holographic gratings were analyzed when hologram formation was carried out in presence of applied voltage. Diffraction efficiency parameter of holographic gratings were studied in dichromated poly(vinyl alcohol) films and dichromated poly(vinyl alcohol) doped with nickel(II) chloride hexahydrate. The electrical changes of the materials were analyzed as a function of surface and volume resistivity.

We showing some properties using materials as dichromate polyvinyl alcohol (DC-PVA), it is doped with natural colorant, it were investigated and compared to the pure dichromated polyvinyl alcohol. The material increasing the sensibility, and present a good photo sensibility Preliminary studies shows, some properties with the ability to storage information, and present a technique to obtain holograms with these colored materials, and some experimental results. The best advantage of this DYE-DC-PVA holograms is the speed of recording.

Previous studies have demonstrated that insulin-like growth factor-I (IGF-I) increases elastin gene transcription in aortic smooth muscle cells and that this up-regulation is accompanied by a loss of protein binding to the proximal promoter. Sp1 has been identified as one of the factors whose binding is lost, and in the present study we show that Sp3 binding is also abrogated by IGF-I, but in a selected manner. In functional analyses using Drosophila SL-2 cells, Sp1 expression can drive transcription from the elastin proximal promoter, while co-expression of Sp3 results in a repression of Sp1 activity. Footprint and gel shift analyses position the IGF-I responsive sequences to a putative retinoblastoma control element (RCE). Mutation of the putative RCE sequence as assessed by transient transfection of smooth muscle cells results in an increase in reporter activity equal in magnitude to that conferred by IGF-I on the wild type promoter. Together these results support the hypothesis that IGF-I-mediated increase in elastin transcription occurs via a mechanism of derepression involving the abrogation of a repressor that appears to be Sp3 binding to the RCE.

The experimental data allow us to determine the imaging quality of holographic gratings with photosensitive film using organic material based on a polyvinyl alcohol matrix doped with potassium dichromate and nickel (II) chloride hexahydrate. The diffraction efficiency is estimated by different spatial frequencies, and the readout image quality is analyzed by the modulation transfer function. The experiment is carried out, with and without voltage application, at different spatial frequencies to obtain the image quality of photosensitive film.

The holographic gratings on photopolymer films are studied by three different thicknesses for samples A, B, and C. The photopolymer emulsion is prepared with potassium dichromate and nickel (II) chloride hexahydrate in polyvinyl alcohol matrix. The evolution of diffraction efficiency is evaluated during holographic recording with and without voltage as a function of energy exposure by changing the thickness. The curves of diffraction efficiency reach a peak when the films are continuously exposed to energy for a period of time. Sample B obtains the highest diffraction efficiency

PURPOSE. The pharmacologic activity of compounds isolated from Physalis peruviana has been demonstrated. The use of this fruit juice for treating pterygium has been reported in Colombian traditional medicine. However, studies demonstrating the fruit juice’s pharmacologic activity when used in this disease have not been published to date. In the present study the anti-inflammatory and cytostatic activities of P. peruviana fruit juice in a rabbit eye inflammatory model were investigated. METHODS. A novel rabbit eye inflammation model was developed for studying the juice’s anti-inflammatory activity (based on an adaptation of the Draize test). Cytostatic activity was evaluated by measuring and comparing growth rates of cultured fibroblasts exposed and not exposed to various fruit juice concentrations. RESULTS. P. peruviana fruit juice exhibited a mild anti-inflammatory activity compared with methylprednisolone, a known anti-inflammatory drug. An interesting dose-dependent cytostatic effect on cultured fibroblasts was also established. CONCLUSIONS. The data found suggest that the P. peruviana fruit juice anti-pterygium effect described in traditional medicine may be related to its inhibiting fibroblast growth. The present study contributes to the pharmacologic knowledge regarding a remedy commonly used in Colombian traditional medicine. (Invest Ophthalmol Vis Sci. 2008;49:3074 –3079)