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1.
J Vis Exp ; (205)2024 Mar 22.
Artículo en Inglés | MEDLINE | ID: mdl-38587382

RESUMEN

The myocardium is composed of cardiomyocytes and an even greater number of fibroblasts, the latter being responsible for extracellular matrix production. From the early stages of heart development throughout the lifetime, in both normal and pathological conditions, the composition of the extracellular matrix changes and influences myocardium structure and function. The purpose of the method described here is to obtain the substrate for the culture of cardiac cells in vitro (termed cardiac ECM), mimicking the myocardial extracellular matrix in vivo. To this end, fibroblasts isolated from the adult human heart were cultured to confluence on gelatin-coated dishes to produce the myocardium-specific extracellular matrix. The subsequent removal of cardiac fibroblasts, while preserving the deposited cardiac ECM, produced the substrate for studying the influence of the myocardium-specific extracellular matrix on other cells. Importantly, the composition of the fibroblast-derived coating of the culture dish changes according to the in vivo activity of the fibroblasts isolated from the heart, allowing subsequent studies of cell-matrix interactions in different normal and pathological conditions.


Asunto(s)
Matriz Extracelular , Miocardio , Adulto , Humanos , Células Cultivadas , Miocitos Cardíacos , Fibroblastos
2.
Methods Mol Biol ; 2803: 3-12, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38676881

RESUMEN

The extracellular matrix (ECM) forms most of the tissue microenvironment and is in a constant and dynamic equilibrium with cells. The decellularization process employs physical or chemical methods, or a combination of them, to remove the cellular components of tissues and organs while preserving the architecture and composition of the ECM. Depending on the methodology used, the decellularized ECM (dECM) is then suitable for research or clinical applications. Here, we describe an optimized protocol for the efficient decellularization of the human myocardium to generate 3D scaffolds of well-preserved cardiac extracellular matrix that can be used for in vitro or in vivo studies.


Asunto(s)
Matriz Extracelular Descelularizada , Miocardio , Ingeniería de Tejidos , Andamios del Tejido , Humanos , Andamios del Tejido/química , Miocardio/citología , Miocardio/metabolismo , Ingeniería de Tejidos/métodos , Matriz Extracelular Descelularizada/química , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Microambiente Celular
3.
PLoS One ; 17(10): e0276224, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36260645

RESUMEN

Extracellular matrix (ECM) is a fundamental component of the heart, guiding vital cellular processes during organ homeostasis. Most cardiovascular diseases lead to a remarkable remodeling of the ECM, accompanied by the formation of a fibrotic tissue that heavily compromises the heart function. Effective therapies for managing fibrosis and promoting physiological ECM repair are not yet available. The production of a decellularized extracellular matrix (d-ECM) serving as a three-dimensional and bioactive scaffold able to modulate cellular behavior and activities is considered crucial to achieve a successful regeneration. The protocol represents a step-by-step method to obtain a decellularized cardiac matrix through the combination of sodium dodecyl sulphate (SDS) and Triton X-100. Briefly, cardiac samples obtained from left ventricles of explanted, pathological human hearts were dissected and washed to remove residual body fluids. Samples were then snap-frozen and sliced by a cryostat into 350 µm thick sections. The sections obtained were decellularized using a solution containing 1% Triton X-100 and 1% SDS in combination, for 24 hours, until observing the color change from brownish-red to translucent-white. As a result, the protocol shows efficiency in preserving ECM architecture and protein composition during the whole process, suggesting that it is worthwhile, highly reproducible and produces a well- preserved decellularized extracellular matrix from cardiac samples. Notwithstanding, some limitations need to be addressed, such as the risk for microbial contamination and the unpredictable trend of the protocol when applied to decellularize samples other than myocardium, vessels, or skin. These issues require antibiotics mixture supplement during the procedure followed by UV sterilization, and appropriate adjustments for a tissue-specific utilization, respectively. The protocol is intended to produce a cardiac d-ECM for cell settlement, representing the ideal scaffold for tissue engineering purposes.


Asunto(s)
Matriz Extracelular , Ingeniería de Tejidos , Humanos , Octoxinol/farmacología , Dodecil Sulfato de Sodio/farmacología , Matriz Extracelular/metabolismo , Ingeniería de Tejidos/métodos , Regeneración , Antibacterianos/metabolismo , Andamios del Tejido
4.
Diagnostics (Basel) ; 12(8)2022 Aug 21.
Artículo en Inglés | MEDLINE | ID: mdl-36010371

RESUMEN

The increasing incidence of periprosthetic joint infections (PJIs) has led to a growing interest in developing strategies to prevent and treat this severe complication. The surgical site's application of antiseptic solutions to eliminate contaminating bacteria and eradicate the bacterial biofilm has been increasing over time. Even though it has been proven that combining antimicrobials could enhance their activities and help overcome acquired microbial resistance related to the topical use of antibiotics, the toxicity of integrated solutions is not well described. This study aimed to evaluate the cytotoxicity of solutions containing povidone-iodine (PI) and hydrogen peroxide (H2O2), alone or in combination, after 1.3 and 5 min of exposure. Chondrocytes, tenocytes, and fibroblast-like synoviocytes were used for cytotoxicity analysis. Trypan blue stain (0.4% in PBS) was applied to evaluate the dead cells. All solutions tested showed a progressive increase in toxicity as exposure time increased except for PI at 0.3%, which exhibited the lowest toxicity. The combined solutions reported a reduced cellular killing at 3 and 5 min than H2O2 at equal concentrations, similar results to PI solutions.

5.
Front Nutr ; 9: 888485, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35719154

RESUMEN

Poor nutritional status is common (estimated prevalence 5-69%) in acute coronavirus disease-2019 (COVID-19), and has been associated with hospitalization, the need for intensive care, and mortality. Body composition (BC) and muscle function have also been related in such patients to poor disease outcomes. As the evidence in the literature is limited, a cross-sectional study was carried out to determine the frequency of malnutrition in a cohort of post-acute COVID-19 patients referred to a rehabilitation center after hospital discharge. BC and muscle strength were assessed and the differences between bedridden and not bedridden patients were specifically evaluated. The study sample was composed of 144 post-acute COVID-19 patients (mean age 64.8 years; males = 95), 37% of whom were bedridden (males = 60%). Nutritional status was evaluated with Mini-Nutritional Assessment (MNA) and Controlling Nutritional status (CONUT). Fat-free mass (FFM) and skeletal muscle mass (SM) were estimated using bioelectrical impedance analysis (BIA). Raw BIA variables (phase angle = PhA and impedance ratios = IRs) were also determined and handgrip strength (HGS) was measured. Dynapenia was identified according to the 2019 EWGSOP criteria. According to MNA, 18% (n. 26) of patients were malnourished and 62% (n. 89) were at risk of malnutrition. As for CONUT, 21% (n. 31) of cases had moderate-severe malnutrition and 58% (n. 83) had light malnutrition. Abnormalities of raw BIA variables (low PhA and high IRs) and low HGS were more common in bedridden patients, in those who were malnourished, or had low FFM or SM. Dynapenic patients were 65% men and 47% women. In conclusion, malnutrition, BC alterations, and low HGS occur in post-acute COVID-19 patients and are more common in bedridden patients. Further studies are needed to identify reliable algorithms for assessing nutritional status in post-acute COVID-19 patients undergoing rehabilitation.

6.
Front Physiol ; 13: 879046, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35669580

RESUMEN

Although human Cardiac Progenitor Cells (hCPCs) are not retained by host myocardium they still improve cardiac function when injected into ischemic heart. Emerging evidence supports the hypothesis that hCPC beneficial effects are induced by paracrine action on resident cells. Extracellular vesicles (EVs) are an intriguing mechanism of cell communication based on the transport and transfer of peptides, lipids, and nucleic acids that have the potential to modulate signaling pathways, cell growth, migration, and proliferation of recipient cells. We hypothesize that EVs are involved in the paracrine effects elicited by hCPCs and held accountable for the response of the infarcted myocardium to hCPC-based cell therapy. To test this theory, we collected EVs released by hCPCs isolated from healthy myocardium and evaluated the effects they elicited when administered to resident hCPC and cardiac fibroblasts (CFs) isolated from patients with post-ischemic end-stage heart failure. Evidence emerging from our study indicated that hCPC-derived EVs impacted upon proliferation and survival of hCPCs residing in the ischemic heart and regulated the synthesis and deposition of extracellular-matrix by CFs. These findings suggest that beneficial effects exerted by hCPC injection are, at least to some extent, ascribable to the delivery of signals conveyed by EVs.

7.
Artículo en Inglés | MEDLINE | ID: mdl-34886363

RESUMEN

Few data are available on the body composition of pole dancers. Bioelectrical impedance analysis (BIA) is a method that is used to estimate fat-free mass (FFM) and fat mass (FM), while raw BIA variables, such as the impedance ratio (IR) and phase angle (PhA), are markers of body cell mass and the ratio between extracellular and total body water. The aim of this study was to evaluate the body composition of pole dancers compared to controls, in particular, those raw BIA variables that are considered as markers of muscle composition. Forty female pole dancers and 59 controls participated in the study. BIA was performed on the whole body and upper and lower limbs, separately, at 5, 50, 100 and 250 kHz. The FFM, FFM index, FM and body fat percentage (BF%) were predicted. The bioelectrical impedance indexes IR and PhA were also considered. Pole dancers exhibited higher FFMI and BI indexes and lower BF%. PhA was greater and IRs were smaller in pole dancers than in controls for the whole body and upper limbs. Considering the training level, FFM, whole-body IR and PhA were higher in the professionals than non-professionals. Raw BIA variables significantly differed between the pole dancers and controls, suggesting a higher BCM; furthermore, practicing pole dancing was associated with a greater FFM and lower FM.


Asunto(s)
Composición Corporal , Baile , Impedancia Eléctrica , Femenino , Humanos
8.
Nutrition ; 91-92: 111445, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34628279

RESUMEN

OBJECTIVES: Musculoskeletal fitness and body composition are major components of health-related physical fitness that are expected to be linked to each other. The aim of this study was to explore the association of musculoskeletal fitness (expressed as hand grip strength [HGS]) and raw bioelectrical impedance analysis (BIA) variables and other predictors in the second and third decades of life. METHODS: Four groups of healthy normal-weight and overweight participants were studied: 130 male adolescents (age 16.0 ± 1.1 y), 143 female adolescents (16.0 ± 0.9 y), 141 young men (25.5 ± 2.0 y), and 141 young women (25.2 ± 1.9 y). Raw BIA variables (impedance [Z] and phase angle [PhA]) for the whole body were measured at four frequencies (5, 50, 100, and 250 kHz), and HGS was measured in both upper limbs. RESULTS: HGS was higher in males than females and higher in young men than male adolescents. A fairly strong correlation emerged in both genders between HGS and raw BIA variables-that is, bioimpedance index (height2/Z), PhA, and impedance ratios (Z at high frequency/Z at low frequency). In multiple regression analysis, bioimpedance index, PhA, and impedance ratio were more effective predictors of HGS than BIA-derived estimates of body composition or anthropometric variables such as height, weight, and body mass index. In both genders, HGS was significantly higher in young adults than in adolescents for the same PhA or impedance ratio. CONCLUSIONS: This study provides some insight into the use of raw BIA variables and HGS in the first decades of life, suggesting a new approach for a reliable assessment of muscle quality in terms of both muscle structure and strength.


Asunto(s)
Composición Corporal , Fuerza de la Mano , Adolescente , Índice de Masa Corporal , Impedancia Eléctrica , Ejercicio Físico , Femenino , Humanos , Masculino , Adulto Joven
9.
J Vis Exp ; (175)2021 09 08.
Artículo en Inglés | MEDLINE | ID: mdl-34570106

RESUMEN

Extracellular matrix (ECM) provides biophysical and biochemical stimuli to support self-renewal, proliferation, survival, and differentiation of surrounding cells due to its content of diverse bioactive molecules. Due to these characteristics, the ECM has been recently considered a promising candidate for the creation of biological scaffolds to boost tissue regeneration. Emerging studies have demonstrated that decellularized human tissues could resemble the native ECM in their structural and biochemical profiles, preserving the three-dimensional (3D) architecture and the content of fundamental biological molecules. Hence, decellularized ECM can be employed to promote tissue remodeling, repair, and functional reconstruction of many organs. Selecting the appropriate decellularization procedure is crucial to obtain acellular tissues that retain the characteristics of the ideal microenvironment for cells. The protocol described here provides a detailed step-by-step description of the decellularization method to obtain a reproducible and effective cell-free biological ECM. Skin fragments from patients undergoing plastic surgery were scaled down and decellularized using a combination of sodium dodecylsulfate (SDS), Triton X-100, and antibiotics. To promote the regular and homogeneous transport of the solution through the samples, they were enclosed in embedding cassettes to ensure protection from mechanical insults. After the decellularization procedure, the snow-white color of skin fragments indicated complete and successful decellularization. Additionally, decellularized samples showed an intact and well-preserved architecture. The results suggest that the proposed decellularization method was effective, fast, and reproducible and protected samples from architectural damages.


Asunto(s)
Matriz Extracelular , Medicina Regenerativa , Diferenciación Celular , Humanos , Octoxinol , Ingeniería de Tejidos , Andamios del Tejido
10.
Clin Nutr ; 40(9): 5238-5248, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34474193

RESUMEN

BACKGROUND & AIMS: Bioimpedance analysis-derived phase angle (PhA), as marker of body cell mass and cell integrity, might be altered in obesity, a condition which is characterized by alterations in muscle structure and function. The aim of this systematic review was to evaluate whether and to which extent PhA varies in individuals/patients with excess body weight focusing on: a) changes in PhA due to obesity; b) changes in PhA after bariatric interventions or training programs. METHODS: According to PRISMA criteria, a systematic literature search until February 2021 using PubMed, Embase, Scopus, and Web of Science was performed. Selection criteria included studies on patients with obesity without comorbidities other than metabolic diseases. RESULTS: A total of 278 articles were first identified. After removing duplicates and excluding studies that did not fulfil the inclusion criteria, the full text of the remaining 80 potentially relevant studies was examined to finally retrieve 11 cross-sectional and 10 longitudinal studies. Few studies have shown that PhA is lower in individuals/patients with obesity than in controls. The only study on the matter showed a decrease with age after the fourth decade of life. Four out of five studies reported consistently greater mean values in men than in women. In two studies PhA was lower in patients with severe obesity; in addition, Patients with low PhA had higher BMI and PhA was inversely correlated with fat mass. Longitudinal studies showed that PhA markedly decreased after bariatric surgery and slightly increased after training programs. CONCLUSIONS: Thus, a relatively low number of studies have evaluated PhA in individuals/patients with obesity with sometimes contradictory and preliminary results. PhA might be useful to evaluate muscle quality in individuals/patients with obesity but further studies are needed to more accurately associate this variable with changes in muscle structure and strength, as well as in metabolic functions.


Asunto(s)
Índice de Masa Corporal , Impedancia Eléctrica , Evaluación Nutricional , Obesidad/fisiopatología , Sarcopenia/diagnóstico , Adulto , Cirugía Bariátrica , Estudios Transversales , Femenino , Humanos , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Fuerza Muscular , Obesidad/complicaciones , Obesidad/cirugía , Periodo Posoperatorio , Periodo Preoperatorio , Sarcopenia/etiología , Factores Sexuales
11.
J Funct Morphol Kinesiol ; 6(3)2021 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-34287311

RESUMEN

Official tests are used to assess the fitness status of soccer referees, and their results correlate with match performance. However, FIFA-approved tests expose the referees to high physical demands and are difficult to implement during the sportive year. The aim of our study was to evaluate the correlation between the 6 × 40-m sprint and Yo-Yo Intermittent Recovery Level 1 (IR1) official tests and other field-based tests that require no or little equipment, are not time-consuming, and impose low physical demands. All tests were performed by male referees from the Regional Section of the Italian Referee Association (n = 30). We observed a strong correlation between 6 × 40-m sprint and Illinois agility tests (r = 0.63, p = 0.001) and a moderate correlation between Yo-Yo IR1 and hand-grip strength in the dominant (r = 0.45, p = 0.014) and non-dominant hand (r = 0.41, p = 0.031). Interestingly, only a moderate correlation (r = -0.42, p = 0.025) was observed between the FIFA official tests, 6 × 40-m sprint and Yo-Yo IR1. These results suggest that Illinois agility and hand-grip tests could represent simple and low-physical-impact tools for repeated assessment and monitoring of referee fitness throughout the sportive season.

12.
Int J Mol Sci ; 21(21)2020 Oct 24.
Artículo en Inglés | MEDLINE | ID: mdl-33114386

RESUMEN

Cardiac adverse remodeling is characterized by biological changes that affect the composition and architecture of the extracellular matrix (ECM). The consequently disrupted signaling can interfere with the balance between cardiogenic and pro-fibrotic phenotype of resident cardiac stromal primitive cells (CPCs). The latter are important players in cardiac homeostasis and can be exploited as therapeutic cells in regenerative medicine. Our aim was to compare the effects of human decellularized native ECM from normal (dECM-NH) or failing hearts (dECM-PH) on human CPCs. CPCs were cultured on dECM sections and characterized for gene expression, immunofluorescence, and paracrine profiles. When cultured on dECM-NH, CPCs significantly upregulated cardiac commitment markers (CX43, NKX2.5), cardioprotective cytokines (bFGF, HGF), and the angiogenesis mediator, NO. When seeded on dECM-PH, instead, CPCs upregulated pro-remodeling cytokines (IGF-2, PDGF-AA, TGF-ß) and the oxidative stress molecule H2O2. Interestingly, culture on dECM-PH was associated with impaired paracrine support to angiogenesis, and increased expression of the vascular endothelial growth factor (VEGF)-sequestering decoy isoform of the KDR/VEGFR2 receptor. Our results suggest that resident CPCs exposed to the pathological microenvironment of remodeling ECM partially lose their paracrine angiogenic properties and release more pro-fibrotic cytokines. These observations shed novel insights on the crosstalk between ECM and stromal CPCs, suggesting also a cautious use of non-healthy decellularized myocardium for cardiac tissue engineering approaches.


Asunto(s)
Matriz Extracelular/metabolismo , Insuficiencia Cardíaca/patología , Células Madre Mesenquimatosas/citología , Adulto , Anciano , Animales , Supervivencia Celular , Células Cultivadas , Técnicas de Cocultivo , Citocinas/genética , Citocinas/metabolismo , Matriz Extracelular/genética , Femenino , Fibrosis , Insuficiencia Cardíaca/genética , Insuficiencia Cardíaca/metabolismo , Humanos , Peróxido de Hidrógeno/metabolismo , Masculino , Células Madre Mesenquimatosas/metabolismo , Persona de Mediana Edad
13.
Med Eng Phys ; 85: 7-15, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-33081966

RESUMEN

Decellularized extracellular matrix is one of the most promising biological scaffold supporting in vitro tissue growth and in vivo tissue regeneration in both preclinical research and clinical practice. In case of thick tissues or even organs, conventional static decellularization methods based on chemical or enzymatic treatments are not effective in removing the native cellular material without affecting the extracellular matrix. To overcome this limitation, dynamic decellularization methods, mostly based on perfusion and agitation, have been proposed. In this study, we developed a low-cost scalable 3D-printed sample-holder for agitation-based decellularization purposes, designed for treating multiple specimens simultaneously and for improving efficiency, homogeneity and reproducibility of the decellularization treatment with respect to conventional agitation-based approaches. In detail, the proposed sample-holder is able to house up to four specimens and, immersed in the decellularizing solution within a beaker placed on a magnetic stirrer, to expose them to convective flow, enhancing the solution transport through the specimens while protecting them. Computational fluid dynamics analyses were performed to investigate the fluid phenomena establishing within the beaker and to support the sample-holder design. Exploratory biological tests performed on human skin specimens demonstrated that the sample-holder reduces process duration and increases treatment homogeneity and reproducibility.


Asunto(s)
Matriz Extracelular , Ingeniería de Tejidos , Humanos , Perfusión , Impresión Tridimensional , Reproducibilidad de los Resultados , Andamios del Tejido
14.
Med Eng Phys ; 84: 1-9, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32977905

RESUMEN

Physical stimuli are crucial for the structural and functional maturation of tissues both in vivo and in vitro. In tissue engineering applications, bioreactors have become fundamental and effective tools for providing biomimetic culture conditions that recapitulate the native physical stimuli. In addition, bioreactors play a key role in assuring strict control, automation, and standardization in the production process of cell-based products for future clinical application. In this study, a compact, easy-to-use, tunable stretch bioreactor is proposed. Based on customizable and low-cost technological solutions, the bioreactor was designed for providing tunable mechanical stretch for biomimetic dynamic culture of different engineered tissues. In-house validation tests demonstrated the accuracy and repeatability of the imposed mechanical stimulation. Proof of concepts biological tests performed on engineered cardiac constructs, based on decellularized human skin scaffolds seeded with human cardiac progenitor cells, confirmed the bioreactor Good Laboratory Practice compliance and ease of use, and the effectiveness of the delivered cyclic stretch stimulation on the cardiac construct maturation.


Asunto(s)
Reactores Biológicos , Ingeniería de Tejidos , Humanos , Andamios del Tejido
15.
Artículo en Inglés | MEDLINE | ID: mdl-32512767

RESUMEN

National and international healthcare organizations propose guidelines for physical activity worldwide, defining its characteristics. These guidelines' practical applications are difficult to estimate, since they are not fully followed. The aim of the present cross-sectional observational study was to assess awareness about guidelines for physical activity and to evaluate their practical applications in a sample of the Italian population. In total, 310 participants completed an online survey (mean age 29.10 ± 4.44), assessing the habits, beliefs and health effects of physical activity. In total, 39.35% of respondents were inactive. In total, 6.91% of active respondents did not perform a warm-up phase at the beginning of each training session and 77.14% did not check their own heart rate during the training session. Approximately half of respondents reported erroneous beliefs about the type, frequency and volume of physical activity, compared to data proposed by the guidelines. The preventive effect of physical activity was clearly perceived for cardiovascular diseases, diabetes, metabolic syndrome and depression. Several subjects misinterpreted the preventive role of physical activity in colon and breast cancers, and in femur and vertebral fractures. Habits and beliefs about physical activity in the general population are far from the guidelines and recommendations. Therefore, it is necessary to strengthen the conscious practice of physical activity further.


Asunto(s)
Enfermedades Cardiovasculares , Ejercicio Físico , Objetivos , Conocimientos, Actitudes y Práctica en Salud , Adulto , Actitud hacia los Computadores , Enfermedades Cardiovasculares/prevención & control , Estudios Transversales , Femenino , Humanos , Masculino , Encuestas y Cuestionarios , Adulto Joven
16.
Artículo en Inglés | MEDLINE | ID: mdl-32266249

RESUMEN

The complex and highly organized environment in which cells reside consists primarily of the extracellular matrix (ECM) that delivers biological signals and physical stimuli to resident cells. In the native myocardium, the ECM contributes to both heart compliance and cardiomyocyte maturation and function. Thus, myocardium regeneration cannot be accomplished if cardiac ECM is not restored. We hypothesize that decellularized human skin might make an easily accessible and viable alternate biological scaffold for cardiac tissue engineering (CTE). To test our hypothesis, we decellularized specimens of both human skin and human myocardium and analyzed and compared their composition by histological methods and quantitative assays. Decellularized dermal matrix was then cut into 600-µm-thick sections and either tested by uniaxial tensile stretching to characterize its mechanical behavior or used as three-dimensional scaffold to assess its capability to support regeneration by resident cardiac progenitor cells (hCPCs) in vitro. Histological and quantitative analyses of the dermal matrix provided evidence of both effective decellularization with preserved tissue architecture and retention of ECM proteins and growth factors typical of cardiac matrix. Further, the elastic modulus of the dermal matrix resulted comparable with that reported in literature for the human myocardium and, when tested in vitro, dermal matrix resulted a comfortable and protective substrate promoting and supporting hCPC engraftment, survival and cardiomyogenic potential. Our study provides compelling evidence that dermal matrix holds promise as a fully autologous and cost-effective biological scaffold for CTE.

17.
J Vis Exp ; (155)2020 01 19.
Artículo en Inglés | MEDLINE | ID: mdl-32009646

RESUMEN

Induced pluripotent stem cells (iPSCs) could be considered, to date, a promising source of pluripotent cells for the management of currently untreatable diseases, for the reconstitution and regeneration of injured tissues and for the development of new drugs. Despite all the advantages related to the use of iPSCs, such as the low risk of rejection, the lessened ethical issues, and the possibility to obtain them from both young and old patients without any difference in their reprogramming potential, problems to overcome are still numerous. In fact, cell reprogramming conducted with viral and integrating viruses can cause infections and the introduction of required genes can induce a genomic instability of the recipient cell, impairing their use in clinic. In particular, there are many concerns about the use of c-Myc gene, well-known from several studies for its mutation-inducing activity. Fibroblasts have emerged as the suitable cell population for cellular reprogramming as they are easy to isolate and culture and are harvested by a minimally invasive skin punch biopsy. The protocol described here provides a detailed step-by-step description of the whole procedure, from sample processing to obtain cell cultures, choice of reagents and supplies, cleaning and preparation, to cell reprogramming by the means of a commercial non-modified RNAs (NM-RNAs)-based reprogramming kit. The chosen reprogramming kit allows an effective reprogramming of human dermal fibroblast to iPSCs and small colonies can be seen as early as 24 h after the first transfection, even with modifications with the respect to the standard datasheet. The reprogramming procedure used in this protocol offers the advantage of a safe reprogramming, without the risk of infections caused by viral vector-based methods, reduces the cellular defense mechanisms, and allows the generation of xeno-free iPSCs, all critical features that are mandatory for further clinical applications.


Asunto(s)
Abdomen/anatomía & histología , Separación Celular/métodos , Dermis/citología , Fibroblastos/citología , Células Madre Pluripotentes Inducidas/citología , Piel/citología , Adulto , Diferenciación Celular , Reprogramación Celular , Fibroblastos/microbiología , Humanos
18.
J Funct Morphol Kinesiol ; 5(2)2020 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-33467238

RESUMEN

The brachial plexus represents a complex anatomical structure in the upper limb. This "network" of peripheral nerves permits the rearrangement of motor efferent fibers, coming from different spinal nerves, in several terminal branches directed to upper limb muscles. Moreover, afferent information coming from different cutaneous regions in upper limb are sorted in different spinal nerves through the brachial plexus. Severe brachial plexus injuries are a rare clinical condition in the general population and in sport medicine, but with dramatic consequences on the motor and sensory functions of the upper limb. In some sports, like martial arts, milder injuries of the brachial plexus can occur, with transient symptoms and with a full recovery. Clinical evaluation represents the cornerstone in the assessment of the athletes with brachial plexus injuries. Electrodiagnostic studies and imaging techniques, like magnetic resonance and high-frequency ultrasound, could be useful to localize the lesion and to define an appropriate treatment and a functional prognosis. Several conservative and surgical techniques could be applied, and multidisciplinary rehabilitative programs could be performed to guide the athlete toward the recovery of the highest functional level, according to the type of injury.

19.
J Funct Morphol Kinesiol ; 5(4)2020 Dec 05.
Artículo en Inglés | MEDLINE | ID: mdl-33467304

RESUMEN

Neuromotor training can improve motor performance in athletes and patients. However, few data are available about their effect on reaction time (RT). We investigated the influence of video observation/motor imagery (VO/MI) on simple RT to visual and auditory stimuli. The experimental group comprised 21 cadets who performed VO/MI training over 4 weeks. Nineteen cadets completed a sham intervention as control. The main outcome measure was RT to auditory and visual stimuli for the upper and lower limbs. The RT to auditory stimuli improved significantly post-intervention in both groups (control vs. experimental mean change for upper limbs: -40 ms vs. -40 ms, p = 0.0008; for lower limbs: -50 ms vs. -30 ms, p = 0.0174). A trend towards reduced RT to visual stimuli was observed (for upper limbs: -30 ms vs. -20 ms, p = 0.0876; for lower limbs: -30 ms vs. -20 ms, p = 0.0675). The interaction term was not significant. Only the specific VO/MI training produced a linear correlation between the improvement in the RT to auditory and visual stimuli for the upper (r = 0.703) and lower limbs (r = 0.473). In conclusion, VO/MI training does not improve RT when compared to control, but it may be useful in individuals who need to simultaneously develop a fast response to different types of stimuli.

20.
Medicina (Kaunas) ; 55(8)2019 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-31394838

RESUMEN

Tendinopathies are very common in general population and a huge number of tendon-related procedures take place annually worldwide, with significant socio-economic repercussions. Numerous treatment options are commonly used for tendon disorders. Besides pharmacological and physical therapy, nutrition could represent an additional tool for preventing and treating this complex pathology that deserve a multidisciplinary approach. In recent years, nutraceutical products are growing up in popularity since these seem to favor the prevention and the healing processes of tendon injuries. This narrative literature review aims to summarize current understanding and the areas of ongoing research about the management of tendinopathies with the help of oral supplementation.


Asunto(s)
Suplementos Dietéticos/normas , Tendinopatía/tratamiento farmacológico , Tendinopatía/fisiopatología , Tendinopatía/terapia , Suplementos Dietéticos/efectos adversos , Humanos , Tendones/efectos de los fármacos , Tendones/fisiopatología
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