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BACKGROUND: Dissection of the axillary lymph nodes during surgery for breast cancer with lymph node involvement is burdened by a complication: lymphedema. Approximately half of women undergoing axillary dissection suffer from it, with a notable impact in terms of perceived discomfort, presented quality of life, and alteration of body image. There is also no shortage of problems in the patient's social and professional life. METHODS: The present review aims to select Randomized Controlled Trials (RCTs) present in the literature regarding the effects of yoga as an alternative therapy in patients with breast cancer-related lymphedema. A search of four databases was undertaken: Cochrane, Pubmed, Scopus, and Web of Science. The searches were conducted on 19 May 2024, and updated to 30 June 2024 without date limits. RCTs without language limitations, in any context, and with any yoga variant were considered. RESULTS: The postulated search strings highlighted a total of 69 potentially eligible studies. The study selection system consisted of two levels of screening, (1) abstract selection and (2) full-text selection, for a total of three studies included in the review. The three RCTs included involved mixed treatment sessions in an outpatient setting with a yoga teacher and at home using a DVD. In the various studies, the outcome measures concerned quality of life, ROM, spinal mobility, limb volume, and tissue induration. CONCLUSIONS: According to the analysis of the data obtained, yoga as an alternative therapy could be useful if combined with the usual care routine in women with lymphedema related to sensory cancer, in terms of improving physical, professional, and emotional quality of life and reducing symptoms such as fatigue, pain, and insomnia. Furthermore, yoga could bring about a reduction in tissue induration of the limb, greater spinal mobility evaluated in terms of improvement of the pelvic and kyphotic angle, and greater strength in shoulder abduction.
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Single-limb stance (SLS) is a demanding postural task, widely used for balance assessment in both research and clinical practice. Despite extensive data on elderly and clinical populations, less is known about younger and healthier adults. Our aim in this study was to assess balance during a SLS task among a cohort of healthy adults to determine whether there are age or sex group or testing condition differences in performances. In this cross-sectional study, we involved 120 participants aged 30-65 years and divided them into four age sub-groups with equal numbers of males and females in each. We assessed balance during a 45-s SLS task on a] the Delos Postural Proprioceptive System for both lower limbs in two conditions - open eyes (OE) and closed eyes (CE). We calculated stability (SI) and autonomy (AU) indices and used analysis of variance to determine that there was no significant effect of limb dominance or sex on balance parameters. However, there was a significant interaction effect between age group and testing condition for both SI and AU (p < .001 for both), with balance worsening as age increased only in the CE condition. These results highlight a pattern of balance decline with age when vision is eliminated from balance performance, underscoring the critical relationship between sensory input and postural control as people age.
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Background: Parkinson's disease (PD) is a neurodegenerative disorder characterized by motor symptoms such as tremors, rigidity, and bradykinesia. Rehabilitation utilizing mirror neurons leverages the brain's capacity for action observation (AO) and motor imagery (MI) to enhance motor function. This approach involves patients imitating movements observed in therapists or videos, aiming to improve gait, coordination, and overall quality of life. Mirror neuron activation facilitates motor learning and may decelerate disease progression, thus enhancing patient mobility and independence. Methods: This scoping review aimed to map current evidence on PD therapies employing mirror neuron-based rehabilitation. Databases searched included PubMed, PEDro, and Cochrane. The review included randomized controlled trials (RCTs) and systematic reviews that examined the effects of AO and MI in PD rehabilitation. Results: Five studies met the inclusion criteria, encompassing various rehabilitation techniques focusing on AO and MI. These studies consistently demonstrated positive outcomes, such as reduced disease severity and improved quality of life, gait, and balance in PD patients. The activation of mirror neurons through AO and MI was shown to facilitate motor learning and contribute to improved functional mobility. Conclusions: Although the included studies support the beneficial impact of AO and MI techniques in PD rehabilitation, numerous questions remain unresolved. Further research is necessary to evaluate the potential integration of these techniques into standard physiotherapy routines for PD patients. This review highlights the promise of AO and MI in enhancing motor rehabilitation for PD, suggesting the need for more comprehensive studies to validate and refine these therapeutic approaches.
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BACKGROUND: Among non-pharmacological interventions, Multiwave Locked System (MLS) Laser therapy has been used in patients with several musculoskeletal pathologies and in combination with other therapeutical interventions. The effects of sole MLS therapy on pain and function in patients with chronic non-specific low-back pain are unknown. OBJECTIVE: The objective of this study was to investigate the effects of MLS Laser therapy on pain, function, and disability in patients with chronic non-specific low back pain in comparison to a placebo treatment group. METHODS: Forty-five patients were randomized into two groups: the MLS Laser group and the Sham Laser group, undergoing 8 sessions of either a MLS Laser therapy or a Sham Laser therapy, respectively. At the beginning of the therapy (T0), at the end of the therapy (T1), and 1 month after the end of therapy (T2) patients were assessed for low back pain (by means of a VAS scale), function (by means of kinematic and electromyographic assessment of a forward bending movement) and self-reported disability (by means of the Roland-Morris and Oswestry Disability questionnaires). RESULTS: There was a significant reduction of pain and disability in both groups at T1 and T2 in comparison with T0. At T2 patients in the MLS group showed a significantly lower pain in comparison with patients in the Sham group (VAS = 2.2 ± 2 vs. 3.6 ± 2.4; p< 0.05). No differences between the two groups were found for function and disability. CONCLUSION: Both MLS Laser and Sham Laser therapies lead to a significant and comparable reduction in pain and disability in patients with chronic non-specific low back pain. However, one month after treatment, MLS Laser therapy has been found to be significantly more effective in reducing pain as compared to sham treatment.
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Dolor Crónico , Evaluación de la Discapacidad , Dolor de la Región Lumbar , Dimensión del Dolor , Humanos , Dolor de la Región Lumbar/terapia , Dolor de la Región Lumbar/rehabilitación , Dolor de la Región Lumbar/fisiopatología , Masculino , Femenino , Método Doble Ciego , Persona de Mediana Edad , Adulto , Dolor Crónico/terapia , Dolor Crónico/rehabilitación , Resultado del Tratamiento , Terapia por Luz de Baja Intensidad/métodosAsunto(s)
Neuropatía Radial , Humanos , Neuropatía Radial/cirugía , Aparatos Ortopédicos , Diseño de EquipoRESUMEN
OBJECTIVE: To provide a comprehensive overview of rehabilitation treatment strategies for focal hand dystonia (FHD) in musicians, examining their evolution and effectiveness. DATA SOURCES: A systematic search of five databases, PubMed, PEDro, Cochrane Library, Trip, and Google Scholar, to identify relevant articles on FHD rehabilitation. The last search was performed on 20 December 2023. METHODS: Inclusion criteria were applied to 190 initially identified articles, resulting in 17 articles for review. Exclusions were made for duplicates, irrelevant titles, abstracts, and non-rehabilitation interventions. RESULTS: Ten different rehabilitation approaches were identified over 20 years. While no definitive intervention protocol exists, a multimodal approach is commonly recommended. CONCLUSIONS: This scoping review underscores the diversity of rehabilitation strategies for FHD. It suggests the potential of multimodal approaches, emphasizing the need for further large-scale clinical efficacy studies.
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Trastornos Distónicos , Humanos , Trastornos Distónicos/rehabilitación , Trastornos Distónicos/etiología , Mano/fisiopatología , Música , Enfermedades Profesionales/rehabilitaciónRESUMEN
INTRODUCTION: The management of pain and functional recovery following a radial capitellum fracture poses a significant clinical challenge, especially in individuals whose professions, such as physiotherapy, demand optimal joint functionality. Transcranial Direct Current Stimulation (tDCS) emerges as a potential non-pharmacological intervention for pain management, necessitating exploration in the context of orthopedic injuries. CASE PRESENTATION: A 41-year-old male physiotherapist presented with a MASON 2 radial capitellum fracture following a fall, experiencing notable pain (NPRS 6/7) and functional impairment (DASH 45/100, PRTEE 43/100). Conservative management, involving immobilization and potential surgical consideration, was employed, followed by tDCS for pain management. Post-tDCS, significant improvements were observed in pain and functional scores (NPRS to 0, DASH to 14.2, PRTEE to 7), alongside enhancements in range of motion and muscle strength. CLINICAL DISCUSSION: The application of tDCS showcased notable efficacy in pain reduction and functional improvement, highlighting its potential in augmenting pain management strategies post-fracture. However, the variability in responses and lack of standardized application protocols necessitate further research to optimize its clinical utility. The balance between immobilization for fracture healing and mobilization for preventing stiffness and facilitating recovery was pivotal in managing the fracture and ensuring functional improvement. CONCLUSIONS: This case underscores the potential of tDCS in managing pain and facilitating functional recovery in radial capitellum fractures, warranting further exploration and standardization of its application in clinical practice. The integrated, patient-centric approach, involving interdisciplinary collaboration and personalized care, was crucial in ensuring positive outcomes and provides a framework for managing similar orthopedic cases.
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Rotationplasty (RP) is a special surgical technique for bone tumors of the lower limb and is the chosen procedure for children under 6 with bone sarcoma in the distal femur. Leg reconstruction results in an unusual aspect of the limb potentially giving life-long emotional outcomes, especially considering the young age of most RP patients. Although the high level of the quality of life of these patients has been previously reported, aspects related to long-term psychological well-being, self-esteem and life satisfaction, particularly regarding the gender, procreation and parenting, have never been explored. The aim of this study was to assess the general degree of psychological well-being of RP patients, with specific reference to gender, procreation and parenting. Twenty long-term RP survivors of high-grade bone sarcoma participated in the study. They were administered the following validated questionnaires: HADS for psychological well-being (degree of anxiety and depression), Temperament and Character Inventory (TCI), RSES for self-esteem, SF-36 for quality of life, SWLS extended to life satisfaction, and ABIS for body image integration. Data on education, marriage, employment and parenthood were gathered. All the scores obtained were very close to normal references. The only gender difference was found for the TCI Cooperativeness scale, which was higher in women than in men. A satisfactory psychological well-being in terms of both self-esteem and integration of the prosthetic joint limb into one's body image, with relatively limited amount of anxiety/depression, good quality of life, and good temperament and character traits, was found. No major gender differences were reported.
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During osteoarthritis development, chondrocytes are subjected to a functional derangement. This increases their susceptibility to stressful conditions such as oxidative stress, a characteristic of the aging tissue, which can further provoke extrinsic senescence by DNA damage responses. It was previously observed that IκB kinase α knockdown increases the replicative potential of primary human OA chondrocytes cultured in monolayer and the survival of the same cells undergoing hypertrophic-like differentiation in 3-D. In this paper we investigated whether IKKα knockdown could modulate oxidative stress-induced senescence of OA chondrocytes undergoing a DDR and particularly the involvement in this process of the DNA mismatch repair system, the principal mechanism for repair of replicative and recombinational errors, devoted to genomic stability maintenance in actively replicating cells. This repair system is also implicated in oxidative stress-mediated DNA damage repair. We analyzed microsatellite instability and expression of the mismatch repair components in human osteoarthritis chondrocytes after IKKα knockdown and H2O2 exposure. Only low MSI levels and incidence were detected and exclusively in IKKα proficient cells. Moreover, we found that IKKα proficient and deficient chondrocytes differently regulated MMR proteins after oxidative stress, both at mRNA and protein level, suggesting a reduced susceptibility of IKKα deficient cells. Our data suggest an involvement of the MMR system in the response to oxidative stress that tends to be more efficient in IKKαKD cells. This argues for a partial contribution of the MMR system to the better ability to recover DNA damage already observed in these cells.
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Condrocitos , Osteoartritis , Condrocitos/metabolismo , Daño del ADN , Reparación de la Incompatibilidad de ADN/genética , Reparación del ADN/genética , Humanos , Peróxido de Hidrógeno/farmacología , Quinasa I-kappa B/genética , Quinasa I-kappa B/metabolismo , Osteoartritis/genética , Estrés Oxidativo/genéticaRESUMEN
According to previous research, natural polyamines exert a role in regulating cell committment and differentiation from stemness during skeletal development. In order to assess whether distinct polyamine patterns are associated with different skeletal cell types, primary cultures of stem cells, chondrocytes or osteoblasts were dedicated for HPLC analysis of intracellular polyamines. Spermine (SPM) and Spermidine (SPD) levels were higher in adipose derived stem cells (ASC) compared to mature skeletal cells, i.e. chondrocytes and osteoblasts, confirming the connection of polyamine content with stemness. To establish whether polyamines can protect ASC against oxidative DNA damage in a 3-D differentiation model, the level of γH2AX was measured by western blot, and found to correlate with age and BMI of patients. Addition of either polyamine to ASC was able to hinder DNA damage in the low micromolecular range, with marked reduction of γH2AX level at 10 µM SPM and 5 µM SPD. Molecular analysis of the mechanisms that might underlie the protective effect of polyamine supplementation evidences a possible involvement of autophagy. Altogether, these results support the idea that polyamines are able to manage both stem cell differentiation and cell oxidative damage, and therefore represent appealing tools for regenerative and cell based applications.
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Daño del ADN , Células Madre Mesenquimatosas/metabolismo , Espermidina/metabolismo , Espermina/metabolismo , Adulto , Anciano , Células Cultivadas , Daño del ADN/efectos de los fármacos , Histonas/análisis , Histonas/metabolismo , Humanos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Persona de Mediana Edad , Espermidina/farmacología , Espermina/farmacologíaRESUMEN
INTRODUCTION: Recent evidence suggests that GSK3 activity is chondroprotective in osteoarthritis (OA), but at the same time, its inactivation has been proposed as an anti-inflammatory therapeutic option. Here we evaluated the extent of GSK3ß inactivation in vivo in OA knee cartilage and the molecular events downstream GSK3ß inactivation in vitro to assess their contribution to cell senescence and hypertrophy. METHODS: In vivo level of phosphorylated GSK3ß was analyzed in cartilage and oxidative damage was assessed by 8-oxo-deoxyguanosine staining. The in vitro effects of GSK3ß inactivation (using either LiCl or SB216763) were evaluated on proliferating primary human chondrocytes by combined confocal microscopy analysis of Mitotracker staining and reactive oxygen species (ROS) production (2',7'-dichlorofluorescin diacetate staining). Downstream effects on DNA damage and senescence were investigated by western blot (γH2AX, GADD45ß and p21), flow cytometric analysis of cell cycle and light scattering properties, quantitative assessment of senescence associated ß galactosidase activity, and PAS staining. RESULTS: In vivo chondrocytes from obese OA patients showed higher levels of phosphorylated GSK3ß, oxidative damage and expression of GADD45ß and p21, in comparison with chondrocytes of nonobese OA patients. LiCl mediated GSK3ß inactivation in vitro resulted in increased mitochondrial ROS production, responsible for reduced cell proliferation, S phase transient arrest, and increase in cell senescence, size and granularity. Collectively, western blot data supported the occurrence of a DNA damage response leading to cellular senescence with increase in γH2AX, GADD45ß and p21. Moreover, LiCl boosted 8-oxo-dG staining, expression of IKKα and MMP-10. CONCLUSIONS: In articular chondrocytes, GSK3ß activity is required for the maintenance of proliferative potential and phenotype. Conversely, GSK3ß inactivation, although preserving chondrocyte survival, results in functional impairment via induction of hypertrophy and senescence. Indeed, GSK3ß inactivation is responsible for ROS production, triggering oxidative stress and DNA damage response.
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Condrocitos/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/metabolismo , Cloruro de Litio/farmacología , Obesidad/patología , Osteoartritis de la Rodilla/patología , Proliferación Celular/efectos de los fármacos , Tamaño de la Célula/efectos de los fármacos , Células Cultivadas , Senescencia Celular/efectos de los fármacos , Condrocitos/citología , Condrocitos/patología , Daño del ADN , Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Glucógeno Sintasa Quinasa 3 beta , Humanos , Indoles/farmacología , Maleimidas/farmacología , Obesidad/enzimología , Osteoartritis de la Rodilla/enzimología , Estrés Oxidativo/efectos de los fármacos , Fosforilación/efectos de los fármacosRESUMEN
BACKGROUND: Healthcare-Associated Infections (HAIs) are one of the most frequent complications occurring in healthcare facilities. Contaminated environmental surfaces provide an important potential source for transmission of many healthcare-associated pathogens, thus indicating the need for new and sustainable strategies. AIM: This study aims to evaluate the effect of a novel cleaning procedure based on the mechanism of biocontrol, on the presence and survival of several microorganisms responsible for HAIs (i.e. coliforms, Staphyloccus aureus, Clostridium difficile, and Candida albicans) on hard surfaces in a hospital setting. METHODS: The effect of microbial cleaning, containing spores of food grade Bacillus subtilis, Bacillus pumilus and Bacillus megaterium, in comparison with conventional cleaning protocols, was evaluated for 24 weeks in three independent hospitals (one in Belgium and two in Italy) and approximately 20000 microbial surface samples were collected. RESULTS: Microbial cleaning, as part of the daily cleaning protocol, resulted in a reduction of HAI-related pathogens by 50 to 89%. This effect was achieved after 3-4 weeks and the reduction in the pathogen load was stable over time. Moreover, by using microbial or conventional cleaning alternatively, we found that this effect was directly related to the new procedure, as indicated by the raise in CFU/m2 when microbial cleaning was replaced by the conventional procedure. Although many questions remain regarding the actual mechanisms involved, this study demonstrates that microbial cleaning is a more effective and sustainable alternative to chemical cleaning and non-specific disinfection in healthcare facilities. CONCLUSIONS: This study indicates microbial cleaning as an effective strategy in continuously lowering the number of HAI-related microorganisms on surfaces. The first indications on the actual level of HAIs in the trial hospitals monitored on a continuous basis are very promising, and may pave the way for a novel and cost-effective strategy to counteract or (bio)control healthcare-associated pathogens.
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Infección Hospitalaria/prevención & control , Desinfección/métodos , Hospitales , Recuento de Colonia MicrobianaRESUMEN
BACKGROUND: In osteoarthritis (OA), an inflammatory environment is responsible for the imbalance between the anabolic and catabolic activity of chondrocytes and, thus, for articular cartilage derangement. This study was aimed at providing further insight into the impairment of the anabolic cytokine IL-4 and its receptors in human OA cartilage, as well as the potential ability of IL-4 to antagonize the catabolic phenotype induced by IL-1ß. METHODOLOGY/PRINCIPAL FINDINGS: The in vivo expression of IL-4 and IL-4 receptor subunits (IL-4R, IL-2Rγ, IL-13Rα1) was investigated on full thickness OA or normal knee cartilage. IL-4 expression was found to be significantly lower in OA, both in terms of the percentage of positive cells and the amount of signal per cell. IL-4 receptor type I and II were mostly expressed in mid-deep cartilage layers. No significant difference for each IL-4 receptor subunit was noted. IL-4 anti-inflammatory and anti-catabolic activity was assessed in vitro in the presence of IL-1ß and/or IL-4 for 24 hours using differentiated high density primary OA chondrocyte also exhibiting the three IL-4 R subunits found in vivo. Chemokines, extracellular matrix degrading enzymes and their inhibitors were evaluated at mRNA (real time PCR) and protein (ELISA or western blot) levels. IL-4 did not affect IL-1ß-induced mRNA expression of GRO-α/CXCL1, IL-8/CXCL8, ADAMTS-5, TIMP-1 or TIMP-3. Conversely, IL-4 significantly inhibited RANTES/CCL5, MIP-1α/CCL3, MIP-1ß/CCL4, MMP-13 and ADAMTS-4. These results were confirmed at protein level for RANTES/CCL5 and MMP-13. CONCLUSIONS/SIGNIFICANCE: Our results indicate for the first time that OA cartilage has a significantly lower expression of IL-4. Furthermore, we found differences in the spectrum of biological effects of IL-4. The findings that IL-4 has the ability to hamper the IL-1ß-induced release of both MMP-13 and CCL5/RANTES, both markers of OA chondrocytes, strongly indicates IL-4 as a pivotal anabolic cytokine in cartilage whose impairment impacts on OA pathogenesis.
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Cartílago Articular/metabolismo , Citocinas/metabolismo , Interleucina-1beta/farmacología , Interleucina-4/metabolismo , Osteoartritis/metabolismo , Adolescente , Adulto , Anciano , Quimiocina CCL5/metabolismo , Niño , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Femenino , Humanos , Masculino , Metaloproteinasa 13 de la Matriz/metabolismo , Persona de Mediana Edad , Adulto JovenRESUMEN
INTRODUCTION: Recent evidence suggests that tissue accumulation of senescent p16INK4a-positive cells during the life span would be deleterious for tissue functions and could be the consequence of inherent age-associated disorders. Osteoarthritis (OA) is characterized by the accumulation of chondrocytes expressing p16INK4a and markers of the senescence-associated secretory phenotype (SASP), including the matrix remodeling metalloproteases MMP1/MMP13 and pro-inflammatory cytokines interleukin-8 (IL-8) and IL-6. Here, we evaluated the role of p16INK4a in the OA-induced SASP and its regulation by microRNAs (miRs). METHODS: We used IL-1-beta-treated primary OA chondrocytes cultured in three-dimensional setting or mesenchymal stem cells differentiated into chondrocyte to follow p16INK4a expression. By transient transfection experiments and the use of knockout mice, we validate p16INK4a function in chondrocytes and its regulation by one miR identified by means of a genome-wide miR-array analysis. RESULTS: p16INK4a is induced upon IL-1-beta treatment and also during in vitro chondrogenesis. In the mouse model, Ink4a locus favors in vivo the proportion of terminally differentiated chondrocytes. When overexpressed in chondrocytes, p16INK4a is sufficient to induce the production of the two matrix remodeling enzymes, MMP1 and MMP13, thus linking senescence with OA pathogenesis and bone development. We identified miR-24 as a negative regulator of p16INK4a. Accordingly, p16INK4a expression increased while miR-24 level was repressed upon IL-1-beta addition, in OA cartilage and during in vitro terminal chondrogenesis. CONCLUSIONS: We disclosed herein a new role of the senescence marker p16INK4a and its regulation by miR-24 during OA and terminal chondrogenesis.
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Condrocitos/patología , Condrogénesis/fisiología , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , MicroARNs/metabolismo , Osteoartritis/metabolismo , Animales , Artritis Experimental/metabolismo , Artritis Experimental/patología , Western Blotting , Diferenciación Celular/fisiología , Senescencia Celular/fisiología , Condrocitos/metabolismo , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunohistoquímica , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 13 de la Matriz/metabolismo , Células Madre Mesenquimatosas/citología , Ratones , Ratones Noqueados , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , Osteoartritis/patología , Reacción en Cadena en Tiempo Real de la Polimerasa , TransfecciónRESUMEN
The first step in skeleton development is the condensation of mesenchymal precursors followed by any of two different types of ossification, depending on the type of bone segment: in intramembranous ossification, the bone is deposed directly in the mesenchymal anlagen, whereas in endochondral ossification, the bone is deposed onto a template of cartilage that is subsequently substituted by bone. Polyamines and polyamine-related enzymes have been implicated in bone development as global regulators of the transcriptional and translational activity of stem cells and pivotal transcription factors. Therefore, it is tempting to investigate their use as a tool to improve regenerative medicine strategies in orthopedics. Growing evidence in vitro suggests a role for polyamines in enhancing differentiation in both adult stem cells and differentiated chondrocytes. Adipose-derived stem cells have recently proved to be a convenient alternative to bone marrow stromal cells, due to their easy accessibility and the high frequency of stem cell precursors per volume unit. State-of-the-art "prolotherapy" approaches for skeleton regeneration include the use of adipose-derived stem cells and platelet concentrates, such as platelet-rich plasma (PRP). Besides several growth factors, PRP also contains polyamines in the micromolar range, which may also exert an anti-apoptotic effect, thus helping to explain the efficacy of PRP in enhancing osteogenesis in vitro and in vivo. On the other hand, spermidine and spermine are both able to enhance hypertrophy and terminal differentiation of chondrocytes and therefore appear to be inducers of endochondral ossification. Finally, the peculiar activity of spermidine as an inducer of autophagy suggests the possibility of exploiting its use to enhance this cytoprotective mechanism to counteract the degenerative changes underlying either the aging or degenerative diseases that affect bone or cartilage.
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Sistemas de Liberación de Medicamentos , Músculo Esquelético/efectos de los fármacos , Poliaminas/farmacología , Células Madre/efectos de los fármacos , Ingeniería de Tejidos , Animales , Diferenciación Celular/efectos de los fármacos , Humanos , Músculo Esquelético/citología , Músculo Esquelético/metabolismo , Poliaminas/metabolismo , Células Madre/metabolismoRESUMEN
BACKGROUND: The non-canonical NF-κB activating kinase IKKα, encoded by CHUK (conserved-helix-loop-helix-ubiquitous-kinase), has been reported to modulate pro- or anti- inflammatory responses, cellular survival and cellular differentiation. Here, we have investigated the mechanism of action of IKKα as a novel effector of human and murine chondrocyte extracellular matrix (ECM) homeostasis and differentiation towards hypertrophy. METHODOLOGY/PRINCIPAL FINDINGS: IKKα expression was ablated in primary human osteoarthritic (OA) chondrocytes and in immature murine articular chondrocytes (iMACs) derived from IKKα(f/f):CreERT2 mice by retroviral-mediated stable shRNA transduction and Cre recombinase-dependent Lox P site recombination, respectively. MMP-10 was identified as a major target of IKKα in chondrocytes by mRNA profiling, quantitative RT-PCR analysis, immunohistochemistry and immunoblotting. ECM integrity, as assessed by type II collagen (COL2) deposition and the lack of MMP-dependent COL2 degradation products, was enhanced by IKKα ablation in mice. MMP-13 and total collagenase activities were significantly reduced, while TIMP-3 (tissue inhibitor of metalloproteinase-3) protein levels were enhanced in IKKα-deficient chondrocytes. IKKα deficiency suppressed chondrocyte differentiation, as shown by the quantitative inhibition of.Alizarin red staining and the reduced expression of multiple chondrocyte differentiation effectors, including Runx2, Col10a1 and Vegfa,. Importantly, the differentiation of IKKα-deficient chondrocytes was rescued by a kinase-dead IKKα protein mutant. CONCLUSIONS/SIGNIFICANCE: IKKα acts independent of its kinase activity to help drive chondrocyte differentiation towards a hypertrophic-like state. IKKα positively modulates ECM remodeling via multiple downstream targets (including MMP-10 and TIMP-3 at the mRNA and post-transcriptional levels, respectively) to maintain maximal MMP-13 activity, which is required for ECM remodeling leading to chondrocyte differentiation. Chondrocytes are the unique cell component in articular cartilage, which are quiescent and maintain ECM integrity during tissue homeostasis. In OA, chondrocytes reacquire the capacity to proliferate and differentiate and their activation results in pronounced cartilage degeneration. Τηυσ, our findings are also of potential relevance for defining the onset and/or progression of OA disease.
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Condrocitos/citología , Condrocitos/metabolismo , Matriz Extracelular/metabolismo , Quinasa I-kappa B/metabolismo , Animales , Diferenciación Celular/genética , Diferenciación Celular/fisiología , Células Cultivadas , Humanos , Quinasa I-kappa B/genética , Immunoblotting , Inmunohistoquímica , Metaloproteinasa 10 de la Matriz/metabolismo , Metaloproteinasa 13 de la Matriz/metabolismo , Ratones , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
A newly formed memory is temporarily fragile and becomes stable through a process known as consolidation. Stable memories may again become fragile if retrieved or reactivated, and undergo a process of reconsolidation to persist and strengthen. Both consolidation and reconsolidation require an initial phase of transcription and translation that lasts for several hours. The identification of the critical players of this gene expression is key for understanding long-term memory formation and persistence. In rats, the consolidation of inhibitory avoidance (IA) memory requires gene expression in both the hippocampus and amygdala, two brain regions that process contextual/spatial and emotional information, respectively; IA reconsolidation requires de novo gene expression in the amygdala. Here we report that, after IA learning, the levels of the transcription factor CCAAT enhancer binding protein δ (C/EBPδ) are significantly increased in both the hippocampus and amygdala. These increases are essential for long-term memory consolidation, as their blockade via antisense oligodeoxynucleotide-mediated knockdown leads to memory impairment. Furthermore, C/EBPδ is upregulated and required in the amygdala for IA memory reconsolidation. C/EBPδ is found in nuclear, somatic, and dendritic compartments, and a dendritic localization of C/EBPδ mRNA in hippocampal neuronal cultures suggests that this transcription factor may be translated at synapses. Finally, the induction of long-term potentiation at CA3-CA1 synapses by tetanic stimuli in acute slices, a cellular model of long-term memory, leads to an accumulation of C/EBPδ in the nucleus. We conclude that the transcription factor C/EBPδ plays a critical role in memory consolidation and reconsolidation.
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Proteína delta de Unión al Potenciador CCAAT/fisiología , Memoria/fisiología , Amígdala del Cerebelo/metabolismo , Animales , Femenino , Hipocampo/metabolismo , Masculino , Inhibición Neural/fisiología , Técnicas de Cultivo de Órganos , Embarazo , Ratas , Ratas Long-Evans , Tiempo de Reacción/fisiologíaRESUMEN
The molecular mechanisms underlying spermine osteo-inductive activity on human adipose-derived stem cells (ASCs) grown in 3-dimensional (3D) cultures were investigated. Spermine belongs to the polyamine family, naturally occurring, positively charged polycations that are able to control several cellular processes. Spermine was used at a concentration close to that found in platelet-rich plasma (PRP), an autologous blood product containing growth and differentiation factors, which has recently become popular in in vitro and in vivo bone healing and engineering. Adipose tissue was surgically obtained from the hypodermis of patients undergoing hip arthroplasty. Patient age negatively affected both ASC yield and ASC ability to form 3D constructs. ASC 3D cultures were seeded in either non differentiating or chondrogenic conditions, with or without the addition of 5 µM spermine to evaluate its osteogenic potential across 1, 2, and 3 weeks of maturation. Osteogenic medium was used as a reference. The effects of the addition of spermine on molecular markers of osteogenesis, at both gene and protein level, and mineralization were evaluated. The effects of spermine were temporally defined and responsible for the progression from the early to the mature osteoblast differentiation phases. Spermine initially promoted gene and protein expression of Runx-2, an early marker of the osteoblast lineage; then, it increased ß-catenin expression and activation, which led to the induction of Osterix gene expression, the mature osteoblast commitment factor. The finding that spermine induces ASC to differentiate toward mature osteoblasts supports the use of these easily accessible mesenchymal stem cells coupled with PRP for orthopedic applications.
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Tejido Adiposo/citología , Osteoblastos/citología , Osteoblastos/metabolismo , Osteogénesis/efectos de los fármacos , Espermina/farmacología , Células Madre/citología , beta Catenina/metabolismo , Adulto , Anciano , Apoptosis/efectos de los fármacos , Agregación Celular/efectos de los fármacos , Recuento de Células , Diferenciación Celular/efectos de los fármacos , Linaje de la Célula/efectos de los fármacos , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Condrogénesis/efectos de los fármacos , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Femenino , Citometría de Flujo , Humanos , Cinética , Masculino , Persona de Mediana Edad , Osteoblastos/efectos de los fármacos , Plasma Rico en Plaquetas/metabolismo , Transporte de Proteínas/efectos de los fármacos , Células Madre/efectos de los fármacos , Células Madre/metabolismo , Factores de Transcripción/metabolismoRESUMEN
Aging is associated with a gradual decline in cognitive functions, and more dramatic cognitive impairments occur in patients affected by Alzheimer's disease (AD). Electrophysiological and molecular studies performed in aged animals and in animal models of AD have shown that cognitive decline is associated with significant modifications in synaptic plasticity (i.e., activity-dependent changes in synaptic strength) and have elucidated some of the cellular mechanisms underlying this process. Morphological studies have revealed a correlation between the quality of memory performance and the extent of structural changes of synaptic contacts occurring during memory consolidation. We briefly review recent experimental evidence here.