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1.
J Adv Res ; 51: 149-160, 2023 09.
Article in English | MEDLINE | ID: mdl-36481476

ABSTRACT

BACKGROUND: The extracellular matrix (ECM) is a vital structure with a dynamic and complex organization that plays an essential role in tissue homeostasis. In the skin, the ECM is arranged into two types of compartments: interstitial dermal matrix and basement membrane (BM). All evidence in the literature supports the notion that direct dysregulation of the composition, abundance or structure of one of these types of ECM, or indirect modifications in proteins that interact with them is linked to a wide range of human skin pathologies, including hereditary, autoimmune, and neoplastic diseases. Even though the ECM's key role in these pathologies has been widely documented, its potential as a therapeutic target has been overlooked. AIM OF REVIEW: This review discusses the molecular mechanisms involved in three groups of skin ECM-related diseases - genetic, autoimmune, and neoplastic - and the recent therapeutic progress and opportunities targeting ECM. KEY SCIENTIFIC CONCEPTS OF REVIEW: This article describes the implications of alterations in ECM components and in BM-associated molecules that are determinant for guaranteeing its function in different skin disorders. Also, ongoing clinical trials on ECM-targeted therapies are discussed together with future opportunities that may open new avenues for treating ECM-associated skin diseases.


Subject(s)
Neoplasms , Skin Diseases , Humans , Extracellular Matrix/chemistry , Extracellular Matrix/metabolism , Extracellular Matrix Proteins/analysis , Extracellular Matrix Proteins/metabolism , Skin/metabolism , Neoplasms/metabolism , Skin Diseases/therapy , Skin Diseases/metabolism , Skin Diseases/pathology
2.
J Photochem Photobiol B ; 221: 112242, 2021 Aug.
Article in English | MEDLINE | ID: mdl-34214837

ABSTRACT

The aim of this work is to understand the inactivation efficiency of medium pressure mercury lamps, measured in terms of growth inhibition as well as cell death, damage and response, using three strains from three different Aspergillus species (A. fumigatus, A. niger and, A. terreus) spiked in filtered surface water. A complete characterization of the effect of the treatment on each strain of the fungal species was assessed considering spores' morphology, cell wall integrity and enzymatic activity, the formation of pyrimidine dimers in the DNA and proteome analysis. Results showed that, when subjected to medium pressure mercury lamps, A. niger is the most resistant to inactivation, that both A. fumigatus and A. niger suffer more morphological changes and present a higher number of damaged spores and A. terreus presented more dead spores. DNA damages detected in A. niger were able to be repaired to some extent, under both light and dark conditions. Finally, proteome analysis showed that the UV radiation treatment triggered different types of stress response, including cell wall reorganization and DNA repair in A. fumigatus and A. terreus, and oxidative stress responses like the increase in production of citric acid and itaconic acid in A. niger and A. terreus, respectively.


Subject(s)
Aspergillus/radiation effects , Light , Mercury/chemistry , Water Microbiology , Aspergillus/physiology , DNA Damage/radiation effects , Permeability/radiation effects , Proteome/radiation effects , Spores, Fungal/radiation effects
3.
Sci Rep ; 11(1): 8576, 2021 04 21.
Article in English | MEDLINE | ID: mdl-33883599

ABSTRACT

Mycorrhizas are known to have a positive impact on plant growth and ability to resist major biotic and abiotic stresses. However, the metabolic alterations underlying mycorrhizal symbiosis are still understudied. By using metabolomics and transcriptomics approaches, cork oak roots colonized by the ectomycorrhizal fungus Pisolithus tinctorius were compared with non-colonized roots. Results show that compounds putatively corresponding to carbohydrates, organic acids, tannins, long-chain fatty acids and monoacylglycerols, were depleted in ectomycorrhizal cork oak colonized roots. Conversely, non-proteogenic amino acids, such as gamma-aminobutyric acid (GABA), and several putative defense-related compounds, including oxylipin-family compounds, terpenoids and B6 vitamers were induced in mycorrhizal roots. Transcriptomic analysis suggests the involvement of GABA in ectomycorrhizal symbiosis through increased synthesis and inhibition of degradation in mycorrhizal roots. Results from this global metabolomics analysis suggest decreases in root metabolites which are common components of exudates, and in compounds related to root external protective layers which could facilitate plant-fungal contact and enhance symbiosis. Root metabolic pathways involved in defense against stress were induced in ectomycorrhizal roots that could be involved in a plant mechanism to avoid uncontrolled growth of the fungal symbiont in the root apoplast. Several of the identified symbiosis-specific metabolites, such as GABA, may help to understand how ectomycorrhizal fungi such as P. tinctorius benefit their host plants.


Subject(s)
Basidiomycota/metabolism , Plant Roots/microbiology , Quercus/microbiology , Gene Expression Profiling , Gene Expression Regulation, Plant , Metabolic Networks and Pathways , Metabolomics , Plant Roots/metabolism , Quercus/metabolism , Symbiosis , gamma-Aminobutyric Acid/biosynthesis
5.
Biochim Biophys Acta Mol Basis Dis ; 1866(6): 165752, 2020 06 01.
Article in English | MEDLINE | ID: mdl-32119897

ABSTRACT

Mood disorders like major depression and bipolar disorder (BD) are among the most prevalent forms of mental illness. Current knowledge of the neurobiology and pathophysiology of these disorders is still modest and clear biological markers are still missing. Thus, a better understanding of the underlying pathophysiological mechanisms to identify potential therapeutic targets is a prerequisite for the design of new drugs as well as to develop biomarkers that help in a more accurate and earlier diagnosis. Multiple pieces of evidence including genetic and neuro-imaging studies suggest that mood disorders are associated with abnormalities in endoplasmic-reticulum (ER)-related stress responses, mitochondrial function and calcium signalling. Furthermore, deregulation of the innate immune response has been described in patients diagnosed with mood disorders, including depression and BD. These disease-related events are associated with functions localized to a subdomain of the ER, known as Mitochondria-Associated Membranes (MAMs), which are lipid rafts-like domains that connect mitochondria and ER, both physically and biochemically. This review will outline the current understanding of the role of mitochondria and ER dysfunction under pathological brain conditions, particularly in major depressive disorder (MDD) and BD, that support the hypothesis that MAMs can act in these mood disorders as the link connecting ER-related stress response and mitochondrial impairment, as well as a mechanisms behind sterile inflammation arising from deregulation of innate immune responses. The role of MAMs in the pathophysiology of these pathologies and its potential relevance as a potential therapeutic target will be discussed.


Subject(s)
Endoplasmic Reticulum/metabolism , Mitochondria/metabolism , Mitochondrial Membranes/metabolism , Mood Disorders/immunology , Calcium Signaling/genetics , Endoplasmic Reticulum/pathology , Endoplasmic Reticulum Stress/genetics , Humans , Immunity, Innate/genetics , Membrane Microdomains/genetics , Membrane Microdomains/pathology , Mitochondria/pathology , Mitochondrial Membranes/pathology , Mood Disorders/genetics , Mood Disorders/metabolism , Mood Disorders/pathology
6.
Int J Endocrinol ; 2019: 4518742, 2019.
Article in English | MEDLINE | ID: mdl-31467527

ABSTRACT

BACKGROUND: The incidence of gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) has been increasing in the last five decades, but there is no large-scale data regarding these tumours in Portugal. We conducted a cross-sectional, multicentric study in main Portuguese centers to evaluate the clinical, pathological, and therapeutic profile of GEP-NENs. METHODS: From November, 2012, to July, 2014, data from 293 patients diagnosed with GEP-NENs from 15 centers in Portugal was collected and registered in an online electronic platform. RESULTS: Median age at diagnosis was 56.5 (range: 15-87) years with a preponderance of females (54.6%). The most frequent primary sites were the pancreas (31.1%), jejunum-ileum (24.2%), stomach (13.7%), and rectum (8.5%). Data regarding hormonal status was not available in most patients (82.3%). Stratified by the tumour grade (WHO 2010 classification), we observed 64.0% of NET G1, 24.7% of NET G2, and 11.3% of NEC. Poorly differentiated tumours occurred mainly in older patients (p = 0.017), were larger (p < 0.001), and presented more vascular (p = 0.004) and lymphatic (p = 0.001) invasion. At the time of diagnosis, 44.4% of GEP-NENs presented metastatic disease. Surgery (79.6%) and somatostatin analogues (30.7%) were the most frequently used therapies of GEP-NENs with reported grading. CONCLUSION: In general, Portuguese patients with GEP-NENs presented similar characteristics to other populations described in the literature. This cross-sectional study represents the first step to establish a national database of GEP-NENs that may aid in understanding the clinical and epidemiological features of these tumours in Portugal.

7.
Reumatismo ; 71(2): 68-74, 2019 Jul 09.
Article in English | MEDLINE | ID: mdl-31309776

ABSTRACT

The objective of our study was to compare pain and health-related quality of life among patients with knee osteoarthritis (OA), chronic nonspecific low back pain (CNLBP) and fibromyalgia (FM). This cross-sectional study included 87 subjects, who were divided into three groups according to their diagnosis: knee OA (n=29), CNLBP (n=29) and FM (n=29), between March 2013 and March 2014. Pain intensity was measured using the Numeric Pain Rating Scale (NPRS), quality of pain using the McGill Pain Questionnaire (MPQ) and health-related quality of life using the 36-item Short-Form Health Survey (SF-36). Painful body areas were marked on a pain map. No statistically significant differences between groups were found for NPRS. Regarding MPQ, the knee OA group presented a lower pain-rating index in contrast to the CNLBP and FM groups, and no significant differences were found between the CNLBP and FM groups. A greater number of characteristic pain descriptors and painful locations were found in the FM group than in the CNLBP or knee OA groups. Regarding SF-36, the FM group presented statistically significant lower values for bodily pain in contrast to the knee OA group. Even though the global pain intensity was similar between groups, the findings suggest that the FM group presented the worst pain experience and a lower health-related quality of life than the knee OA group in terms of bodily pain. They also suggest that the pain experience was worse for the CNLBP group than for the knee OA group but health-related quality of life was similar.


Subject(s)
Chronic Pain/diagnosis , Fibromyalgia/diagnosis , Low Back Pain/diagnosis , Osteoarthritis, Knee/diagnosis , Pain Measurement , Quality of Life , Adult , Aged , Aged, 80 and over , Cross-Sectional Studies , Female , Humans , Male , Middle Aged
8.
J Tissue Eng Regen Med ; 12(2): e1063-e1067, 2018 02.
Article in English | MEDLINE | ID: mdl-28342296

ABSTRACT

Design of tissue engineering strategies deals with the need to balance both biomaterials characteristics and techniques specificities, often resulting in cell-compromising processing conditions. One important factor often disregarded is the osmotic pressure to which cells are exposed. An in-house microfluidic system was used to prove that addition of an osmotic regulator significantly benefits the generation of viable cell-laden hydrogels under harsh processing conditions. Human adipose-derived stem cells were resuspended in 1.5% alginate and 1% gellan gum (GG; w/v) solutions containing different concentrations (0.12 m, 0.25 m and 1.5 m) of sucrose as osmotic regulator. GG (in water) and alginate (in water or phosphate-buffered saline) solutions were used to vary the conditions under which cells were kept prior processing. Independently of the polymer, addition of sucrose did not affect the processing conditions or the viscosity of the solutions, except at 1.5 m. The obtained results clearly demonstrate that inclusion of 0.25 m sucrose during processing of the cell-laden hydrogels allowed to keep cell viability around 80%, in opposition to the 20% observed in its absence, both for GG and alginate-derived hydrogels prepared in water. Impressively, the level of cell viability observed with the inclusion of 0.25 m sucrose, 76% for GG and 86% for alginate, was similar to that obtained with the standard alginate solution prepared in phosphate-buffered saline (82%). The beneficial effect of sucrose was observed within the first 5 min of processing and was maintained for prolonged experimental setups with viability values above 50%, even after a 2-h time-frame and independently of the material.


Subject(s)
Osmotic Pressure , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Adipose Tissue/cytology , Alginates/pharmacology , Cell Survival/drug effects , Humans , Hydrogels/pharmacology , Stem Cells/cytology , Stem Cells/drug effects , Stem Cells/metabolism , Sucrose/pharmacology
9.
Eur Spine J ; 24(3): 452-7, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25597041

ABSTRACT

PURPOSE: To investigate the association between symptom severity and physical activity participation in people with acute non-specific low back pain (LBP). METHODS: The sample included a total of 999 patients who presented to primary care with an acute episode of low back pain. Symptom severity, in terms of activity limitation and severity of pain; and physical activity participation before (habitual) and after pain onset were assessed using self-report questionnaires. All participants were interviewed within 14 days of pain onset. RESULTS: At interview most of the participants (87.5 %) reported having moderate to extreme activity limitation due to back pain. There was a significant decrease in physical activity participation after pain onset (mean difference: -176 min, 95 % CI 327-400; p < 0.0001) but no association between habitual or change in physical activity participation and symptom severity was observed (p > 0.21). CONCLUSION: Pain onset causes a significant and immediate decrease in physical activity participation, but this change does not seem to be associated with symptom severity.


Subject(s)
Low Back Pain/physiopathology , Motor Activity , Severity of Illness Index , Acute Disease , Adolescent , Adult , Aged , Female , Humans , Low Back Pain/psychology , Low Back Pain/therapy , Male , Middle Aged , Primary Health Care , Self Report , Young Adult
10.
Biomaterials ; 43: 23-31, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25591958

ABSTRACT

The detection, isolation and sorting of cells holds an important role in cell therapy and regenerative medicine. Also, injectable systems have been explored for tissue regeneration in vivo, because it allows repairing complex shaped tissue defects through minimally invasive surgical procedures. Here we report the development of chitosan microparticles with a size of 115.8 µm able to capture and expand a specific cell type that can also be regarded as an injectable biomaterial. Monoclonal antibodies against cell surface antigens specific to endothelial cells and stem cells were immobilized on the surface of the microparticles. Experimental results showed that particles bioconjugated with specific antibodies provide suitable surfaces to capture a target cell type and subsequent expansion of the captured cells. Primarily designed for an application in tissue engineering, three main challenges are accomplished with the herein presented microparticles: separation, scale-up expansion of specific cell type and successful use as an injectable system to form small tissue constructs in situ.


Subject(s)
Biocompatible Materials/chemistry , Chitosan/chemistry , Microspheres , Adipose Tissue/pathology , Antibodies, Monoclonal/chemistry , Biotin/chemistry , Biotinylation , DNA/chemistry , Flow Cytometry , Human Umbilical Vein Endothelial Cells , Humans , Materials Testing , Particle Size , Regeneration , Regenerative Medicine/methods , Tissue Engineering/instrumentation , Tissue Engineering/methods
11.
Eur J Phys Rehabil Med ; 51(2): 149-54, 2015 Apr.
Article in English | MEDLINE | ID: mdl-24755776

ABSTRACT

BACKGROUND: Fibromyalgia (FM) is a rheumatic disease characterized by chronic widespread pain and symptoms such as fatigue, sleep disturbances, cognitive difficulties, and depression. Postural instability is a debilitating disorder increasingly recognized as part of FM. OBJECTIVE: To assess and compare postural control and balance self-efficacy in women with and without FM and verify the association of these variables with pain, symptom severity, and strength. DESIGN: Case-control study SETTING: Physiotherapeutic Clinical Research and Electromyography Laboratory Department of Physical Therapy, Speech Therapy, and Occupational Therapy, School of Medicine, University of Sao Paulo, Sao Paulo, Brazil. POPULATION: Case-control study of 117 women ranging from age 35 to 60 years. Of these, 67 had FM. METHODS: Posture control was assessed with the modified clinical test of sensory interaction on balance with patients in forceplates, balance self-efficacy with the Activities-specific Balance Confidence Scale, pain severity with the Visual Analog Scale, tender point pain threshold with digital algometry, symptom severity with the fibromyalgia impact questionnaire, and lower limb strength with a dynamometer. RESULTS: Individuals with FM had impaired postural control showing increased speed of oscillation of the center of gravity (P=0.004) and decreased balance self-efficacy (P<0.001). They had moderate to excellent correlations of balance self-efficacy with pain (r=0.7, P<0.01), muscle strength (r=0.52, P<0.01), and symptom severity (r=0.78, P<0.10) compared with the control group. Correlation of postural control with the same variables was weak. CONCLUSIONS: Patients with FM have impaired postural control and low balance self-efficacy that are associated with pain, muscle strength, and symptom severity. CLINICAL REHABILITATION IMPACT: Postural control and balance self-efficacy needs to be assessed in patients with FM and the treatment goals should be the improvement of postural control and balance self-efficacy.


Subject(s)
Fibromyalgia/therapy , Muscle Weakness/etiology , Pain/etiology , Postural Balance/physiology , Sensation Disorders/etiology , Adult , Brazil , Case-Control Studies , Female , Fibromyalgia/complications , Fibromyalgia/psychology , Humans , Middle Aged , Muscle Weakness/psychology , Pain/psychology , Pain Measurement , Pain Threshold , Physical Endurance , Quadriceps Muscle/physiopathology , Self Efficacy , Sensation Disorders/psychology , Severity of Illness Index , Sickness Impact Profile
12.
Rev. bras. plantas med ; 17(4,supl.3): 1177-1181, 2015. tab
Article in Portuguese | LILACS | ID: lil-776595

ABSTRACT

RESUMO O presente trabalho teve como objetivo avaliar a propagação vegetativa do assa-peixe Vernonia polyanthes Less utilizando estacas apicais e diferentes substratos. O experimento foi conduzido em condições de casa de vegetação com irrigação controlada na Faculdade de Ciências Agronômicas da UNESP campus Botucatu- SP. O delineamento experimental utilizado foi o inteiramente casualizado, com sete tratamentos (Solo de textura argilosa; 50% Solo de textura argilosa + 50% Húmus; Solo de textura média; 50% Solo de textura média + 50% Húmus; Solo de textura arenosa; 50% Solo de textura arenosa + 50% Húmus; Húmus) e quatro repetições, sendo 10 estacas por repetição. As características avaliadas foram: comprimento da maior raiz, comprimento da parte aérea, número de brotações, porcentagem de enraizamento, volume radicular, nota para qualidade das estacas e massa seca da parte aérea e das raízes. Os dados foram submetidos à análise de variância pelo teste F e as médias comparadas pelo teste de Scott & Knott a 5% de probabilidade. Para a análise estatística, utilizou-se o programa SISVAR ®. A partir da análise, observou-se efeito altamente significativo dos tratamentos utilizados para todas as variáveis analisadas, com exceção da massa seca da parte aérea, que não apresentou diferença significativa. Para obtenção de melhor enraizamento da espécie em questão, o recomendado é a utilização do substrato: solo de textura argilosa ou média, acrescido de húmus na proporção de 1:1.


ABSTRAT The current study aimed to evaluate the vegetative propagation of assa-peixe using apical cuttings and different substrates. The trial was conducted under greenhouse conditions with controlled irrigation, in the Faculdade de Ciências Agronômicas of UNESP in Botucatu- SP campus. The experimental design was completely randomized, with seven treatments and four replications, with 10 cuttings per replicate. The characteristics were: longest root length, shoot length, number of shoots, rooting percentage, root volume, score of quality of cuttings and dry mass of shoots and roots. The data were subjected to analysis of variance by F test and the averages were compared by the Scott Knott test at 5% probability. For statistical analysis, we used the SISVAR ®. From the analysis, we observed highly relevant effect of the treatments used for all variables, with the exception of the dry weight of shoots, which did not present significant difference. In order to obtain a better rooting of the species concerned, the recommended method would be to use substrates such as clayey soil or medium plus humus in the ratio 1:1.


Subject(s)
/analysis , Vernonia/classification , Plants, Medicinal/classification , Analysis of Variance , Grassland
13.
Acta Biomater ; 10(12): 4928-4938, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25192821

ABSTRACT

We have developed a bilayered dermal-epidermal scaffold for application in the treatment of full-thickness skin defects. The dermal component gels in situ and adapts to the lesion shape, delivering human dermal fibroblasts in a matrix of fibrin and cross-linked hyaluronic acid modified with a cell adhesion-promoting peptide. Fibroblasts were able to form a tridimensional matrix due to material features such as tailored mechanical properties, presence of protease-degradable elements and cell-binding ligands. The epidermal component is a robust membrane containing cross-linked hyaluronic acid and poly-l-lysine, on which keratinocytes were able to attach and to form a monolayer. Amine-aldehyde bonding at the interface between the two components allows the formation of a tightly bound composite scaffold. Both parts of the scaffold were designed to provide cell-type-specific cues to allow for cell proliferation and form a construct that mimics the skin environment.


Subject(s)
Fibroblasts/physiology , Fibroblasts/transplantation , Keratinocytes/physiology , Keratinocytes/transplantation , Skin Transplantation/instrumentation , Skin, Artificial , Tissue Scaffolds , Biomimetic Materials , Cell Adhesion/physiology , Cell Proliferation/physiology , Cell Survival/physiology , Cells, Cultured , Equipment Design , Equipment Failure Analysis , Fibroblasts/cytology , Humans , Keratinocytes/cytology
14.
Acta Biomater ; 10(7): 3145-55, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24650971

ABSTRACT

Skin regeneration remains a challenge, requiring a well-orchestrated interplay of cell-cell and cell-matrix signalling. Cell sheet (CS) engineering, which has the major advantage of allowing the retrieval of the intact cell layers along with their naturally organized extracellular matrix (ECM), has been poorly explored for the purpose of creating skin substitutes and skin regeneration. This work proposes the use of CS technology to engineer cellular constructs based on human keratinocytes (hKC), key players in wound re-epithelialization, dermal fibroblasts (hDFb), responsible for ECM remodelling, and dermal microvascular endothelial cells (hDMEC), part of the dermal vascular network and modulators of angiogenesis. Homotypic and heterotypic three-dimensional (3-D) CS-based constructs were developed simultaneously to target wound re-vascularization and re-epithelialization. After implantation of the constructs in murine full-thickness wounds, human cells were engrafted into the host wound bed and were present in the neotissue formed up to 14 days post-implantation. Different outcomes were obtained by varying the composition and organization of the 3-D constructs. Both hKC and hDMEC significantly contributed to re-epithelialization by promoting rapid wound closure and early epithelial coverage. Moreover, a significant increase in the density of vessels at day 7 and the incorporation of hDMEC in the neoformed vasculature confirmed its role over neotissue vacularization. As a whole, the obtained results confirmed that the proposed 3-D CS-based constructs provided the necessary cell machinery, when in a specific microenvironment, guiding both re-vascularization and re-epithelialization. Although dependent on the nature of the constructs, the results obtained sustain the hypothesis that different CS-based constructs lead to improved skin healing.


Subject(s)
Neovascularization, Physiologic , Re-Epithelialization , Skin/cytology , Animals , Cells, Cultured , Humans , In Situ Hybridization , Male , Mice , Skin Transplantation
15.
Biomed Microdevices ; 16(1): 69-78, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24122322

ABSTRACT

The use of bottom-up approaches in tissue engineering applications is advantageous since they enable the combination of various layers that could be made from different materials and/or incorporate different biochemical cues. Regarding the complex structure and the vascular system of the bone tissue, the aim of this study was to develop an innovative bottom-up approach that allows the construction of 3D biodegradable scaffolds from 2D microfabricated membranes with precise shape, pore size and porosity. For that purpose, poly (caprolactone) (PCL) and starch ­ poly (caprolactone) (SPCL (30 % starch)) blended sheets were used as substrates to produce the microfabricated membranes using micro hotembossing. The use of this micro fabrication process allowed accurately imprinting micropillars and microholes in reproducible way. The assembling of the microfabricated membranes was performed using an easy, highly reproducible and inexpensive approach based on its successive stacking. Additionaly, the suitability of the microfabricated membranes to support the attachment and the cytoskeletal organization of human bone marrow stem cells (hBMSCs), macrovascular endothelial cells and osteoblasts derived from hBMSCs was demonstrated. Furthermore, hBMSCs proliferated and maintained the expression of the stromal progenitor marker STRO-1 when cultured on both PCL and SPCL microfabricated membranes. The proposed methodology constitutes a promising alternative to the traditional processing methods used to prepare tissue engineering scaffolds.


Subject(s)
Bone and Bones/chemistry , Microtechnology/methods , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Cell Adhesion , Cell Differentiation , Cells, Cultured , Human Umbilical Vein Endothelial Cells , Humans , Immunohistochemistry , Mesenchymal Stem Cells/chemistry , Microscopy, Electron, Scanning , Osteoblasts/chemistry , Polymers/chemistry , Porosity
16.
Biomacromolecules ; 14(11): 3997-4008, 2013 Nov 11.
Article in English | MEDLINE | ID: mdl-24093541

ABSTRACT

Among the wide range of strategies to target skin repair/regeneration, tissue engineering (TE) with stem cells at the forefront, remains as the most promising route. Cell sheet (CS) engineering is herein proposed, taking advantage of particular cell-cell and cell-extracellular matrix (ECM) interactions and subsequent cellular milieu, to create 3D TE constructs to promote full-thickness skin wound regeneration. Human adipose derived stem cells (hASCs) CS were obtained within five days using both thermoresponsive and standard cell culture surfaces. hASCs-based constructs were then built by superimposing three CS and transplanted into full-thickness excisional mice skin wounds with delayed healing. Constructs obtained using thermoresponsive surfaces were more stable than the ones from standard cell culture surfaces due to the natural adhesive character of the respective CS. Both CS-generating strategies lead to prolonged hASCs engraftment, although no transdifferentiation phenomena were observed. Moreover, our findings suggest that the transplanted hASCs might be promoting neotissue vascularization and extensively influencing epidermal morphogenesis, mainly through paracrine actions with the resident cells. The thicker epidermis, with a higher degree of maturation characterized by the presence of rete ridges-like structures, as well as a significant number of hair follicles observed after transplantation of the constructs combining the CS obtained from the thermoresponsive surfaces, reinforced the assumptions of the influence of the transplanted hASCs and the importance of the higher stability of these constructs promoted by cohesive cell-cell and cell-ECM interactions. Overall, this study confirmed the potential of hASCs CS-based constructs to treat full-thickness excisional skin wounds and that their fabrication conditions impact different aspects of skin regeneration, such as neovascularisation, but mainly epidermal morphogenesis.


Subject(s)
Adipose Tissue/cytology , Epidermal Cells , Morphogenesis , Stem Cells/cytology , Tissue Engineering , Wound Healing , Adipose Tissue/chemistry , Animals , Cells, Cultured , Extracellular Matrix/chemistry , Humans , Male , Mice , Mice, Inbred BALB C , Stem Cells/chemistry
17.
J Environ Manage ; 121: 87-95, 2013 May 30.
Article in English | MEDLINE | ID: mdl-23524400

ABSTRACT

Selected bacterial strains were immobilised on the surface of hydroxyapatite (Ca10(PO4)6(OH)2 - HAp) of natural origin (fish bones). The capacity of the material, alone and in combination with the bacterial strains to act as heavy metal removers from aqueous streams was assessed. Pseudomonas fluorescens (S3X), Microbacterium oxydans (EC29) and Cupriavidus sp. (1C2) were chosen based on their resistance to heavy metals and capacity of adsorbing the metals. These systems were tested using solutions of Zn(II), Cd(II) and in solutions containing both metals. A synergistic effect between the strains and HAp, which is effective in removing the target heavy metals on its own, was observed, as the combination of HAp with the bacterial strains led to higher adsorption capacity for both elements. For the solutions containing only one metal the synergistic effect was greater for higher metal concentrations; 1C2 and EC29 were the most effective strains for Zn(II) and Cd(II) respectively, while S3X was less effective. Overall, an almost four-fold increase was observed for the maximum adsorption capacity for Zn(II) when 1C2 was employed - 0.433 mmol/g in comparison of 0.121 mmol/g for the unmodified HAp. For Cd(II), on the other hand, an almost three-fold increase was registered with EC29 bacterial strain - 0.090 vs 0.036 mmol/g for the unmodified HAp. When the solutions containing both metals were tested, the effect was more marked for lower concentrations.


Subject(s)
Biofilms , Cupriavidus/metabolism , Durapatite , Metals, Heavy/isolation & purification , Pseudomonas fluorescens/metabolism , Water Pollutants, Chemical/isolation & purification , Adsorption , Animals , Bone and Bones , Gadus morhua , Metals, Heavy/metabolism , Water Pollutants, Chemical/metabolism
18.
Eur J Phys Rehabil Med ; 49(2): 197-204, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23486303

ABSTRACT

BACKGROUND: Fibromyalgia is a chronic pain syndrome associated with sleep disorders, fatigue and psychological symptoms. Combinations therapies, such as electrotherapy and therapeutic exercises have been used in the clinical practice. AIM: To assess the efficacy of high-frequency transcutaneous electrical nerve stimulation (TENS) as an adjuvant therapy to aerobic and stretching exercises, for the treatment of fibromyalgia. DESIGN: Controlled clinical trial. SETTING: Unit of rehabilitation of a public hospital. POPULATION: Twenty-eight women aged 52.4±7.5 years, with fibromyalgia. METHODS: A visual analogue scale measured pain intensity; tender points pain threshold, by dolorimetry; and quality of life, by the Fibromyalgia Impact Questionnaire. All subjects participated in an eight-week program consisting of aerobic exercises, followed by static stretching of muscle chains. In TENS group, high-frequency (150 Hz) was applied on bilateral tender points of trapezium and supraspinatus. RESULTS: TENS group had a greater pain reduction (mean change score=-2.0±2.9 cm) compared to Without TENS group (-0.7±3.7 cm). There was a difference between mean change scores of each group for pain threshold (right trapezium: 0.2±1 kg/cm² in TENS group and -0.2±1.2 kg/cm² in Without TENS group). In the evaluation of clinically important changes, patients receiving TENS had relevant improvement of pain, work performance, fatigue, stiffness, anxiety and depression compared to those not receiving TENS. CONCLUSION: It has suggested that high-frequency TENS as an adjuvant therapy is effective in relieving pain, anxiety, fatigue, stiffness, and in improving ability to work of patients with fibromyalgia. CLINICAL REHABILITATION IMPACT: High-frequency TENS may be used as a short-term complementary treatment of fibromyalgia.


Subject(s)
Fibromyalgia/therapy , Transcutaneous Electric Nerve Stimulation , Combined Modality Therapy , Disability Evaluation , Exercise Therapy , Female , Fibromyalgia/psychology , Humans , Middle Aged , Pain Management , Pain Measurement , Quality of Life , Surveys and Questionnaires , Treatment Outcome
19.
Acta Biomater ; 9(6): 6790-7, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23462554

ABSTRACT

A significant number of therapeutics derived from natural polymers and plants have been developed to replace or to be used in conjunction with existing dressing products. The use of the therapeutic properties of aloe vera could be very useful in the creation of active wound dressing materials. The present work was undertaken to examine issues concerning structural features, topography, enzymatic degradation behavior, antibacterial activity and cellular response of chitosan/aloe vera-based membranes. The chitosan/aloe vera-based membranes that were developed displayed satisfactory degradation, roughness, wettability and mechanical properties. A higher antibacterial potency was displayed by the blended membranes. Moreover, in vitro assays demonstrated that these blended membranes have good cell compatibility with primary human dermal fibroblasts. The chitosan/aloe vera-based membranes might be promising wound dressing materials.


Subject(s)
Aloe/chemistry , Chitosan/chemistry , Fibroblasts/cytology , Fibroblasts/physiology , Guided Tissue Regeneration/instrumentation , Membranes, Artificial , Tissue Scaffolds , Biocompatible Materials/chemical synthesis , Cells, Cultured , Equipment Design , Equipment Failure Analysis , Feasibility Studies , Guided Tissue Regeneration/methods , Humans , Materials Testing , Tissue Engineering/instrumentation
20.
Acta Biomater ; 9(2): 5234-42, 2013 Feb.
Article in English | MEDLINE | ID: mdl-22995408

ABSTRACT

Vascularization is the most pressing issue in tissue engineering (TE) since ensuring that engineered constructs are adequately perfused after in vivo transplantation is essential for the construct's survival. The combination of endothelial cells with current TE strategies seems the most promising approach but doubts persist as to which type of endothelial cells to use. Umbilical cord blood (UCB) cells have been suggested as a possible source of endothelial progenitors. Osteoblasts obtained from human adipose-derived stem cells (hASCs) were co-cultured with the mononuclear fraction of human UCB for 7 and 21 days on carrageenan membranes. The expression of vWF and CD31, and the DiI-AcLDL uptake ability allowed detection of the presence of endothelial and monocytic lineages cells in the co-culture for all culture times. In addition, the molecular expression of CD31 and VE-cadherin increased after 21 days of co-culture. The functionality of the system was assessed after transplantation in nude mice. Although an inflammatory response developed, blood vessels with cells positive for human CD31 were detected around the membranes. Furthermore, the number of blood vessels in the vicinity of the implants increased when cells from the mononuclear fraction of UCB were present in the transplants compared to transplants with only hASC-derived osteoblasts. These results show how endothelial progenitors present in the mononuclear fraction of UCB can be sustained by hASC-derived osteoblast co-culture and contribute to angiogenesis even in an in vivo setting of inflammatory response.


Subject(s)
Adipose Tissue/cytology , Endothelial Cells/cytology , Fetal Blood/cytology , Leukocytes, Mononuclear/cytology , Osteoblasts/cytology , Stem Cells/cytology , Animals , Blood Vessels/drug effects , Carrageenan/pharmacology , Cell Separation , Coculture Techniques , Endothelial Cells/drug effects , Female , Flow Cytometry , Gene Expression Regulation/drug effects , Humans , Immunohistochemistry , Implants, Experimental , Leukocytes, Mononuclear/drug effects , Mice , Mice, Inbred BALB C , Mice, Nude , Osteoblasts/drug effects , Osteoblasts/transplantation , Phenotype , Stem Cells/drug effects
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