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Chronic diabetic wounds present formidable challenges, marked by uncontrolled bacterial infections, prolonged inflammation, and impaired angiogenesis. The evolving landscape of photo-responsive antibacterial therapy holds great promise in addressing these multifaceted issues, with a particular focus on leveraging the distinctive properties of 2D heterojunction materials. In this investigation, we engineered composite sprayed hydrogels, seamlessly integrating Bi/MoS2 nano-heterojunctions. Capitalizing on the synergistic interplay between photocatalytic antibacterial and photothermal antibacterial mechanisms, the Bi/MoS2 heterojunction, guided by its localized surface plasmon resonance, demonstrated outstanding antibacterial efficacy within a mere 10-minute exposure to 808â¯nm near-infrared light. This accelerated sterilization both in vitro and in vivo, consequently expediting wound healing. The sprayed composite gel not only furnishes protective shielding for skin tissues but also fosters endothelial cell proliferation, vascularization, and angiogenesis. This safe and ultrafast sterilizing hydrogel presents immense potential for application in antimicrobial dressings, thereby offering a promising avenue for diabetic wound healing.
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Carcinoid syndrome is a rare condition resulting from neuroendocrine tumors (NETs) that secrete vasoactive substances like serotonin. This report describes the case of a 61-year-old man with a history of chronic obstructive pulmonary disease (COPD) and hypertension who presented with new-onset angioedema, loss of consciousness, and a fall. He had been treated for COPD exacerbations during ER visits without improvement and was unaware of a prior mesenteric carcinoid tumor diagnosis from 2012. The next emergency evaluation revealed significant airway and facial edema necessitating intubation. Imaging and biopsy identified a well-differentiated grade 1 NET with extensive liver metastases. Laboratory tests showed elevated levels of serum serotonin, chromogranin A, and 24-hour urine 5-hydroxyindoleacetic acid (5-HIAA). Post-discharge, a PET scan confirmed metastatic lesions primarily in the liver and small bowel, with an unresectable mesenteric mass. The patient was treated with lanreotide and became symptom-free. This case underscores the need to consider carcinoid syndrome in patients with COPD presenting with unexplained respiratory symptoms, as timely diagnosis and treatment can significantly enhance patient outcomes.
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Chronic diabetic wounds pose significant challenges due to uncontrolled bacterial infections, prolonged inflammation, and impaired angiogenesis. The rapid advancement of photo-responsive antibacterial therapy showed promise in addressing these complex issues, particularly utilizing 2D heterojunction materials, which offer unique properties. Herein, we designed an in situ sprayed Bi/BiOCl 0D/2D heterojunction composite fibrin gel with the characteristics of rapid formation and effective near-infrared activation for the treatment of non-healing diabetes-infected wounds. The sprayed composite gel can provide protective shielding for skin tissues and promote endothelial cell proliferation, vascularization, and angiogenesis. The Bi/BiOCl 0D/2D heterojunction, with its localized surface plasmon resonance (LSPR), can overcome the wide bandgap limitation of BiOCl, enhancing the generation of local heat and reactive oxygen species under near-infrared irradiation. This facilitated bacterial elimination and reduced inflammation, supporting the accelerated healing of diabetes-infected wounds. Our study underscores the potential of LSPR-enhanced heterojunctions as advanced wound therapies for chronic diabetic wounds. This article is protected by copyright. All rights reserved.
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Protocatechuic acid (PCA) is a natural component with multiple biological activities. However, the underlying mechanisms of the effects of PCA on anti-ulcerative colitis (UC) are unclear. A UC mouse model was established by allowing the mice to freely drink a dextran sulfate sodium solution. The mice were administered PCA intragastrically for 7 days. Histological pathology, intestinal flora, and ferroptosis regulators were determined in vivo. Additionally, ferroptotic Caco-2 cells were modeled to investigate the role of PCA in ferroptosis. Our results showed that PCA reduced the levels of the disease activity index, inflammatory factors, and histological damage in UC mice. We also found that the regulation of intestinal flora, especially Bacteroidetes, was one of the potential mechanisms underlying the protective effects of PCA anti-UC. Moreover, PCA downregulated the level of ferroptosis in the colon tissue, as evidenced by a reduced iron overload, decreased glutathione depletion, and a lower level of malondialdehyde production compared with the model group. Similar effects of PCA on ferroptosis were observed in Erastin-treated Caco-2 cells. The results obtained using reactive oxygen species assays and the changes in mitochondrial structure observed via scanning electron microscopy also support these results. Our findings suggested that PCA protected against UC by regulating intestinal flora and ferroptosis.
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Colitis Ulcerosa , Colitis , Ferroptosis , Microbioma Gastrointestinal , Humanos , Ratones , Animales , Dextranos/farmacología , Células CACO-2 , Colitis Ulcerosa/inducido químicamente , Colon , Sulfatos/farmacología , Sodio , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Colitis/patología , Ratones Endogámicos C57BLRESUMEN
The ability to control stem cell function is the key to stem cell-based therapy and living tissue regeneration. In natural conditions, histone deacetylases (HDAC) are regarded as the important defining epigenetic reprogramming for stem cell differentiation. To date, human adipose-derived stem cells (hADSCs) have been widely utilised for bone tissue engineering applications. The present study aimed to examine the effect of a novel HDAC2&3-selective inhibitor, MI192, on hADSCs epigenetic reprogramming for regulating its osteogenic potential in vitro. The results confirmed that MI192 treatment reduced the hADSCs viability in a time and dose-dependent manner. The optimal concentration and pre-treatment time of MI192 for hADSCs osteogenic induction was 30 µM and 2 days representatively. A quantitative biochemical assay confirmed that the pre-treatment with MI192 (30 µM) for 2 days significantly enhanced hADSCs alkaline phosphatase (ALP) specific activity (P<0.05) compared with that of the valproic acid (VPA) pre-treatment group. Real-time PCR analysis revealed that MI192 pre-treatment up-regulated hADSCs gene expressions of osteogenic markers (e.g., Runx2, Col1, and OCN) under the osteogenic induction. DNA flow cytometric analysis indicated that two days' pre-treatment with MI192 (30 µM) resulted in G2/M arrest in hADSCs and this G2/M arrest was reversible. Our results suggest that MI192 is capable of epigenetic reprogramming of hADSCs via HDAC inhibition for controlling the cell cycle, resulting in enhancing hADSCs osteogenic differentiation, which indicates the potential of using MI192 for promoting bone tissue regeneration.
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Inhibidores de Histona Desacetilasas , Osteogénesis , Humanos , Inhibidores de Histona Desacetilasas/farmacología , Tejido Adiposo/metabolismo , Apoptosis , Células Cultivadas , Línea Celular Tumoral , Puntos de Control de la Fase G2 del Ciclo Celular , Diferenciación Celular , Células Madre/metabolismo , Epigénesis GenéticaRESUMEN
To date, few studies have explored the role of central obesity on the association between diet quality, measured by the health eating index (HEI), inflammatory eating index (DII), and low-grade inflammation-related serum inflammatory markers. In this paper, we use the data from the 2015-2018 National Health and Nutrition Examination Survey (NHANES) to explore this. Dietary intakes were measured during two 24-h dietary recall interviews and using USDA Food Pattern Equivalence Database (FPED) dietary data. Serum inflammatory markers were obtained from NHANES Laboratory Data. Generalized structural equation models (GSEMs) were used to explore the mediating relationship. Central obesity plays a significant mediating role in the association between HEI-2015 and high-sensitivity C-reactive protein (hs-CRP), mediating 26.87% of the associations between the two; it also mediates 15.24% of the associations between DII and hs-CRP. Central obesity plays a mediating role in 13.98% of the associations between HEI-2015 and white blood cells (WBC); it also mediates 10.83% of the associations between DII and WBC. Our study suggests that central obesity plays a mediating role in the association of dietary quality with low-grade inflammation-related serum inflammatory markers (hs-CRP and WBC).
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Proteína C-Reactiva , Obesidad Abdominal , Adulto , Humanos , Proteína C-Reactiva/metabolismo , Encuestas Nutricionales , Estudios Transversales , Dieta , Inflamación , Biomarcadores , ObesidadRESUMEN
The significant rise of human immunodeficiency virus (HIV) epidemic among young students has become a new feature of acquired immune deficiency syndrome in China, and unprotected anal intercourse (UAI) among male students who have sex with men (SMSM) played a central role in this process. The aim of this study was to investigate the prevalence of UAI, as well as analyze the factors associated with UAI among SMSM in Qingdao, China. From May 2021 to April 2022, men who aged 15 to 30, studied in a high school or college in Qingdao and had anal sex with men in the past 6 months were recruited through snowball method relying on the nongovernmental organization. An anonymous electronic questionnaire was administrated to collect information on socio-demographic characteristics, sexual behaviors, substance use before sex, HIV prevention services and self-esteem. Univariate and multivariate logistic regressions were used to assess factors related to UAI. Among the 341 SMSM included in the study, 40.5% engaged in UAI in the past 6 months. Being migrants from other provinces [odds ratios (OR) = 2.04, 95% confidence intervals (CI): 1.10-3.78], failing to use condoms for the first anal intercourse (OR = 3.38, 95% CI: 1.85-6.18), drinking alcohol before sex (OR = 2.31, 95% CI: 1.25-4.28) and being low level self-esteem (OR = 1.77, 95% CI: 1.09-2.87) were positively associated with UAI. SMSM who had homosexual intercourse more than once a week (OR = 1.76, 95% CI: 1.03-3.00) or had multiple male sex partners (OR = 1.99, 95% CI: 1.20-3.30) were more likely to engage in UAI. Received peer education in the past 12 months (OR = 0.48, 95% CI: 0.27-0.86) was associated with lower odds of UAI. The situation of UAI among SMSM in Qingdao was a public health issue of concern. It is necessary to take targeted measures, such as focusing on first-time sex, advancing sexual health education, expanding peer education, implementing alcohol use screening and maintaining the self-esteem of SMSM, so as to reduce the occurrence of high-risk behaviors among SMSM and curb the spread of HIV on campus.
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Síndrome de Inmunodeficiencia Adquirida , Conducta Sexual , Masculino , Humanos , Estudios Transversales , Prevalencia , China/epidemiologíaRESUMEN
In order to explore the relationship between the Healthy Eating Index (HEI-2015) and cardiovascular disease (CVD), and the mediating role of obesity and depressive symptoms, we used the data from the 2011-2018 National Health and Nutrition Examination Survey (NHANES) for further study. A total of 12,644 participants were included in the study. The HEI was derived using NHANES personal food data and USDA Food Pattern Equivalence Database (FPED) dietary data. The risk of cardiovascular disease was determined using the Framingham Heart Study's multifactorial calculation tool. The weighted multiple logistic regression model was used to explore the association between the HEI-2015 and CVD, and the generalized structural equation was used to explore the mediating effects of obesity and depression, respectively and jointly. Higher HEI-2015 scores were associated with a lower risk of CVD compared to lower quartiles. Obesity, depressive symptoms, and their chain effects all played significant mediating roles in the association between the HEI-2015 and CVD, with proportional mediations of 9.03%, 2.23% and 0.25%, respectively. Our results suggest that higher dietary quality is associated with a lower risk of CVD, mediated by obesity, depressive symptoms, and the chain effect of obesity and depressive symptoms.
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Enfermedades Cardiovasculares , Depresión , Humanos , Encuestas Nutricionales , Depresión/epidemiología , Enfermedades Cardiovasculares/epidemiología , Enfermedades Cardiovasculares/etiología , Dieta/efectos adversos , Obesidad/epidemiologíaRESUMEN
Woody plant encroachment has been long observed in the southern Great Plains (SGP) of the United States. However, our understanding of its spatiotemporal variability, which is the basis for informed and targeted management strategy, is still poor. This study investigates the encroachment of evergreen forest, which is the most important encroachment component in the SGP. A validated evergreen forest map of the SGP (30 m resolution, for the time period 2015 to 2017) from our previous study was utilized (referred to as evergreen_base). Sample plots of evergreen forest (as of 2017) were collected across the study area, based on which a threshold of winter season (January and February) mean normalized difference vegetation index (NDVIwinter) was derived for each of the 5 sub-regions, using Landsat 7 surface reflectance data from 2015 to 2017. Then a NDVIwinter layer was created for each year within the four time periods of 1985-1989, 1995-1999, 2005-2009, and 2015-2017, with winter season surface reflectance data from Landsat 4, 5, and 7. By applying the sub-region specific NDVIwinter thresholds to the annual NDVIwinter layers and the evergreen_base, a SGP evergreen forest map was generated for each of those years. The annual evergreen forest maps within each time period were composited into one. According to the resulting four composite evergreen forest maps, mean annual encroachment rate (km2/year) was calculated at sub-region and ecoregion scales, over each of the three temporal stages 1990-1999, 2000-2009, and 2010-2017, respectively. To understand the spatiotemporal variability of the encroachment, the encroachment rate at each temporal stage was related to the corresponding initial evergreen forest area, mean annual precipitation (MAP), and mean annual burned area (MABA) through linear regression and pairwise comparison. Results suggest that most of the ecoregions have seen a slowing trend of evergreen forest encroachment since 1990. The temporal trend of encroachment rate tends to be consistent with that of MAP, but opposite to that of MABA. The spatial variability of the encroachment rate among ecoregions can be largely (>68%) explained by initial evergreen forest area but shows no significant relationship with MAP or MABA. These findings provide pertinent guidance for the combat of woody plant encroachment in the SGP under the context of climate change.
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Bosques , Árboles , Plantas , Cambio Climático , Modelos LinealesRESUMEN
Abnormal ovarian function is the main manifestation of female reproductive toxicity. Granulosa cells (GCs) play an important role in determining the fate of follicles and are the main effector cells of the female reproductive system. Excessive apoptosis of GCs leads to pathological folliculogenesis and further reproductive damage. However, drugs available for treatment of female reproductive toxicity are limited. Recent studies have confirmed that various natural products and bioactive ingredients of traditional Chinese medicine (TCM) can inhibit apoptosis of GCs and protect ovarian function. In this review, the mechanisms underlying the proapoptotic and antiapoptotic effects of natural products and bioactive ingredients of TCM on the proliferation, function, and apoptosis of GCs are summarized based on the findings of reports published over the past 10 years as reference for the treatment of female reproductive toxicity.
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Medicina Tradicional China , Folículo Ovárico , Femenino , Humanos , Células de la Granulosa , ApoptosisRESUMEN
[This corrects the article DOI: 10.3389/fimmu.2022.903796.].
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Heterogeneity of bone marrow mesenchymal stromal cells (MSCs, frequently referred to as "mesenchymal stem cells") clouds biological understanding and hampers their clinical development. In MSC cultures most commonly used in research and therapy, we have identified an MSC subtype characterized by CD317 expression (CD317pos (29.77 ± 3.00% of the total MSC population), comprising CD317dim (28.10 ± 4.60%) and CD317bright (1.67 ± 0.58%) MSCs) and a constitutive interferon signature linked to human disease. We demonstrate that CD317pos MSCs induced cutaneous tissue damage when applied a skin explant model of inflammation, whereas CD317neg MSCs had no effect. Only CD317neg MSCs were able to suppress proliferative cycles of activated human T cells in vitro, whilst CD317pos MSCs increased polarization towards pro-inflammatory Th1 cells and CD317neg cell lines did not. Using an in vivo peritonitis model, we found that CD317neg and CD317pos MSCs suppressed leukocyte recruitment but only CD317neg MSCs suppressed macrophage numbers. Using MSC-loaded scaffolds implanted subcutaneously in immunocompromised mice we were able to observe tissue generation and blood vessel formation with CD317neg MSC lines, but not CD317pos MSC lines. Our evidence is consistent with the identification of an immune stromal cell, which is likely to contribute to specific physiological and pathological functions and influence clinical outcome of therapeutic MSCs.
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Células Madre Mesenquimatosas , Animales , Humanos , Células Madre Mesenquimatosas/metabolismo , Ratones , Transducción de Señal , Células del Estroma , Células TH1RESUMEN
Epigenetic approaches using the histone deacetylase 2 and 3 inhibitor-MI192 have been reported to accelerate stem cells to form mineralised tissues. Gelatine methacryloyl (GelMA) hydrogels provide a favourable microenvironment to facilitate cell delivery and support tissue formation. However, their application for bone repair is limited due to their low mechanical strength. This study aimed to investigate a GelMA hydrogel reinforced with a 3D printed scaffold to support MI192-induced human bone marrow stromal cells (hBMSCs) for bone formation. Cell culture: The GelMA (5 wt%) hydrogel supported the proliferation of MI192-pre-treated hBMSCs. MI192-pre-treated hBMSCs within the GelMA in osteogenic culture significantly increased alkaline phosphatase activity (p ≤ 0.001) compared to control. Histology: The MI192-pre-treated group enhanced osteoblast-related extracellular matrix deposition and mineralisation (p ≤ 0.001) compared to control. Mechanical testing: GelMA hydrogels reinforced with 3D printed poly(ethylene glycol)-terephthalate/poly(butylene terephthalate) (PEGT/PBT) scaffolds exhibited a 1000-fold increase in the compressive modulus compared to the GelMA alone. MI192-pre-treated hBMSCs within the GelMA-PEGT/PBT constructs significantly enhanced extracellular matrix collagen production and mineralisation compared to control (p ≤ 0.001). These findings demonstrate that the GelMA-PEGT/PBT construct provides enhanced mechanical strength and facilitates the delivery of epigenetically-activated MSCs for bone augmentation strategies.
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Epigenetics plays a critical role in regulating mesenchymal stem cells' (MSCs) fate for tissue repair and regeneration. There is increasing evidence that the inhibition of histone deacetylase (HDAC) isoform 3 can enhance MSC osteogenesis. This study investigated the potential of using a selective HDAC2 and 3 inhibitor, MI192, to promote human dental pulp stromal cells (hDPSCs) bone-like tissue formation in vitro and in vivo within porous Bombyx Mori silk scaffolds. Both 2 and 5 wt% silk scaffolds were fabricated and characterised. The 5 wt% scaffolds possess thicker internal lamellae, reduced scaffold swelling and degradation rates, whilst increased compressive modulus in comparison to the 2 wt% silk scaffold. MI192 pre-treatment of hDPSCs on 5 wt% silk scaffold significantly enhanced hDPSCs alkaline phosphatase activity (ALP). The expression of osteoblast-related genes (RUNX2, ALP, Col1a, OCN) was significantly upregulated in the MI192 pre-treated cells. Histological analysis confirmed that the MI192 pre-treated hDPSCs-silk scaffold constructs promoted bone extracellular matrix (ALP, Col1a, OCN) deposition and mineralisation compared to the untreated group. Following 6 weeks of subcutaneous implantation in nude mice, the MI192 pre-treated hDPSCs-silk scaffold constructs enhanced the vascularisation and extracellular matrix mineralisation compared to untreated control. In conclusion, these findings demonstrate the potential of using epigenetic reprogramming and silk scaffolds to promote hDPSCs bone formation efficacy, which provides evidence for clinical translation of this technology for bone augmentation.
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Inhibidores de Histona Desacetilasas , Ingeniería de Tejidos , Animales , Benzamidas , Células Cultivadas , Pulpa Dental/metabolismo , Epigénesis Genética , Inhibidores de Histona Desacetilasas/farmacología , Humanos , Isoquinolinas , Ratones , Ratones Desnudos , Osteogénesis/genética , Seda/farmacología , Células del Estroma/metabolismo , Andamios del TejidoRESUMEN
CONTEXT: The osteogenic potential of the human dental pulp stromal cells (hDPSCs) was reduced in the state of oxidative stress. Resveratrol (RSV) possesses numerous biological properties, including osteogenic potential, growth-promoting and antioxidant activities. OBJECTIVE: This study investigates the osteogenic potential of RSV by activating the Sirt1/Nrf2 pathway on oxidatively stressed hDPSCs and old mice. MATERIALS AND METHODS: The hDPSCs were subjected to reactive oxygen species (ROS) fluorescence staining, cell proliferation assay, ROS activity assay, superoxide dismutase (SOD) enzyme activity, the glutathione (GSH) concentration assay, alkaline phosphatase staining, real-time polymerase chain reaction (RT-PCR) and Sirt1 immunofluorescence labelling to assess the antioxidant stress and osteogenic ability of RSV. Forty female Kunming mice were divided into Old, Old-RSV, Young and Young-RSV groups to assess the repair of calvarial defects of 0.2 mL RSV of 20 mg/kg/d for seven days by injecting intraperitoneally at 4 weeks after surgery using micro-computed tomography, nonlinear optical microscope and immunohistochemical analysis. RESULTS: RSV abates oxidative stress by alleviating the proliferation, mitigating the ROS activity, increasing the SOD enzyme activity and ameliorating the GSH concentration (RSV IC50 in hDPSCs is 67.65 ± 9.86). The antioxidative stress and osteogenic capabilities of RSV were confirmed by the up-regulated gene expression of SOD1, xCT, RUNX2 and OCN, as well as Sirt1/Nrf2. The collagen, bone matrix formation and Sirt1 expression, are significantly increased after RSV treatment in mice. DISCUSSION AND CONCLUSIONS: For elderly or patients with oxidative stress physiological states such as hypertension, heart disease, diabetes, etc., RSV may potentially improve bone augmentation surgery in regenerative medicine.
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Osteogénesis/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Resveratrol/farmacología , Células del Estroma/efectos de los fármacos , Factores de Edad , Animales , Animales no Consanguíneos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Pulpa Dental/citología , Femenino , Humanos , Ratones , Factor 2 Relacionado con NF-E2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Sirtuina 1/metabolismo , Células del Estroma/citología , Superóxido Dismutasa/metabolismoRESUMEN
The fast degradation of collagen-based membranes in the biological environment remains a critical challenge, resulting in underperforming Guided Bone Regeneration (GBR) therapy leading to compromised clinical results. Photoactive atelocollagen (AC) systems functionalised with ethylenically unsaturated monomers, such as 4-vinylbenzyl chloride (4VBC), have been shown to generate mechanically competent materials for wound healing, inflammation control and drug delivery, whereby control of the molecular architecture of the AC network is key. Building on this platform, the sequential functionalisation with 4VBC and methacrylic anhydride (MA) was hypothesised to generate UV-cured AC hydrogels with reduced swelling ratio, increased proteolytic stability and barrier functionality for GBR therapy. The sequentially functionalised atelocollagen precursor (SAP) was characterised via TNBS and ninhydrin colourimetric assays, circular dichroism and UV-curing rheometry, which confirmed nearly complete consumption of collagen's primary amino groups, preserved triple helices and fast (< 180 s) gelation kinetics, respectively. Hydrogel's swelling ratio and compression modulus were adjusted depending on the aqueous environment used for UV-curing, whilst the sequential functionalisation of AC successfully generated hydrogels with superior proteolytic stability in vitro compared to both 4VBC-functionalised control and the commercial dental membrane Bio-Gide®. These in vitro results were confirmed in vivo via both subcutaneous implantation and a proof-of-concept study in a GBR calvarial model, indicating integrity of the hydrogel and barrier defect, as well as tissue formation following 1-month implantation in rats. STATEMENT OF SIGNIFICANCE: Collagen-based membranes remain a key component in Guided Bone Regeneration (GBR) therapy, but their properties, e.g. proteolytic stability and soft tissue barrier functionality, are still far from optimal. This is largely attributed to the complex molecular configuration of collagen, which makes chemical accessibility and structure-function relations challenging. Here, we fabricated a UV-cured hydrogel network of atelocollagen, whereby triple helices were sequentially functionalised with two distinct ethylenically unsaturated monomers. The effects of the sequential functionalisation and UV-curing on the macroscopic properties, degradation behaviour and GBR capability were investigated in vitro and in vivo. The results highlight the key role of the sequential functionalisation and provide important insights for the design of future, longer-lasting resorbable membranes for GBR therapy.
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Regeneración Tisular Dirigida , Membranas Artificiales , Animales , Regeneración Ósea , Colágeno/farmacología , Ratas , Cicatrización de HeridasRESUMEN
Human bone marrow stromal cells (hBMSCs) have been extensively utilised for bone tissue engineering applications. However, they are associated with limitations that hinder their clinical utility for bone regeneration. Cell fate can be modulated via altering their epigenetic functionality. Inhibiting histone deacetylase (HDAC) enzymes have been reported to promote osteogenic differentiation, with HDAC3 activity shown to be causatively associated with osteogenesis. Therefore, this study aimed to investigate the potential of using an HDAC2 & 3 selective inhibitor - MI192 to induce epigenetic reprogramming of hBMSCs and enhance its therapeutic efficacy for bone formation. Treatment with MI192 caused a time-dose dependant reduction in hBMSCs viability. MI192 was also found to substantially alter hBMSCs epigenetic function through reduced HDAC activity and increased histone acetylation. hBMSCs were pre-treated with MI192 (50 µM) for 48 h prior to osteogenic induction. MI192 pre-treatment significantly upregulated osteoblast-related gene/protein expression (Runx2, ALP, Col1a and OCN) and enhanced alkaline phosphatase specific activity (ALPSA) (1.43-fold) (P ≤ 0.001). Moreover, MI192 substantially increased hBMSCs extracellular matrix calcium deposition (1.4-fold) (P ≤ 0.001) and mineralisation when compared to the untreated control. In 3D microtissue culture, MI192 significantly promoted hBMSCs osteoblast-related gene expression and ALPSA (> 2.41-fold) (P ≤ 0.001). Importantly, MI192 substantially enhanced extracellular matrix deposition (ALP, Col1a, OCN) and mineralisation (1.67-fold) (P ≤ 0.001) within the bioassembled-microtissue (BMT) construct. Following 8-week intraperitoneal implantation within nude mice, MI192 treated hBMSCs exhibited enhanced extracellular matrix deposition and mineralisation (2.39-fold) (P ≤ 0.001) within the BMT when compared to the untreated BMT construct. Taken together, these results demonstrate that MI192 effectively altered hBMSCs epigenetic functionality and is capable of promoting hBMSCs osteogenic differentiation in vitro and in vivo, indicating the potential of using epigenetic reprogramming to enhance the therapeutic efficacy of hBMSCs for bone augmentation strategies.
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Células Madre Mesenquimatosas , Animales , Médula Ósea , Células de la Médula Ósea , Regeneración Ósea , Diferenciación Celular , Células Cultivadas , Epigénesis Genética , Humanos , Ratones , Ratones Desnudos , Osteogénesis/genéticaRESUMEN
The use of human dental pulp stromal cells (hDPSCs) has gained increasing attention as an alternative stem cell source for bone tissue engineering. The modification of the cells' epigenetics has been found to play an important role in regulating differentiation, with the inhibition of histone deacetylases 3 (HDAC3) being linked to increased osteogenic differentiation. This study aimed to induce epigenetic reprogramming using the HDAC2 and 3 selective inhibitor, MI192 to promote hDPSCs osteogenic capacity for bone regeneration. MI192 treatment caused a time-dose-dependent change in hDPSC morphology and reduction in viability. Additionally, MI192 successfully augmented hDPSC epigenetic functionality, which resulted in increased histone acetylation and cell cycle arrest at the G2/M phase. MI192 pre-treatment exhibited a dose-dependent effect on hDPSCs alkaline phosphatase activity. Quantitative PCR and In-Cell Western further demonstrated that MI192 pre-treatment significantly upregulated hDPSCs osteoblast-related gene and protein expression (alkaline phosphatase, bone morphogenic protein 2, type I collagen and osteocalcin) during osteogenic differentiation. Importantly, MI192 pre-treatment significantly increased hDPSCs extracellular matrix collagen production and mineralisation. As such, for the first time, our findings show that epigenetic reprogramming with the HDAC2 and 3 selective inhibitor MI192 accelerates the osteogenic differentiation of hDPSCs, demonstrating the considerable utility of this MSCs engineering approach for bone augmentation strategies.
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Benzamidas/farmacología , Pulpa Dental/citología , Inhibidores de Histona Desacetilasas/farmacología , Isoquinolinas/farmacología , Osteogénesis/efectos de los fármacos , Acetilación/efectos de los fármacos , Fosfatasa Alcalina/metabolismo , Benzamidas/administración & dosificación , Ciclo Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Inhibidores de Histona Desacetilasas/administración & dosificación , Histonas/metabolismo , Humanos , Isoquinolinas/administración & dosificación , Tercer Molar/citología , Osteogénesis/fisiología , Células del Estroma/metabolismoRESUMEN
It is well-known that extracellular ATP acts as an autocrine/paracrine signal to regulate cell functions by inducing intracellular Ca2+ signalling through its cognate receptors, namely, the ligand-gated ion channel P2X receptors that mediate Ca2+ influx and/or the Gq/11-coupled P2Y receptors that link to Ca2+ release from the ER. The reduction in ER Ca2+ can trigger further extracellular Ca2+ entry by activating the store-operated Ca2+ (SOC) channel. Mesenchymal stem cells (MSC) play an important role in the homeostasis of residing tissues and have promising applications in regenerative medicines. MSC can release ATP spontaneously or in response to diverse stimuli, and express multiple P2X and Gq/11-coupled P2Y receptors that participate in ATP-induced Ca2+ signalling and regulate cell function. There is increasing evidence to show the contribution of the SOC channel in ATP-induced Ca2+ signalling in MSC. In this mini-review, we discuss the current understanding of the expression of the SOC channel in MSC and its potential role in mediating ATP-induced Ca2+ signalling and regulation of MSC differentiation, proliferation and migration.
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Células Madre Mesenquimatosas , Receptores Purinérgicos P2 , Adenosina Trifosfato , Calcio/metabolismo , Señalización del Calcio , Células Madre Mesenquimatosas/metabolismo , Receptores Purinérgicos P2/metabolismo , Transducción de SeñalRESUMEN
In this study, the complete mitogenome of an entomopathogenic fungus Akanthomyces lecanii is assembled and annotated. This circular mitogenome is 24,643 bp in length and consists of 2 rRNA genes (rnl and rns), 26 tRNA genes and 14 standard protein-coding genes of the oxidative phosphorylation system. Only one intron (group IA) is identified, which invades rnl and carries an ORF coding for ribosomal protein S3. Phylogenetic analysis based on concatenated mitochondrial nucleotide sequences confirms A. lecanii in Cordycipitaceae, and A. lecanii clusters together with Akanthomyces muscarius.