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1.
Int J Biol Macromol ; 244: 125355, 2023 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-37327940

RESUMO

Composite films of Bacillus megaterium H16 derived PHB with 1%Poly-L-lactic acid (PLLA), 1%Poly-ε-caprolactone (PCL), and 0.3 % graphene nanoplatelets (GNP) were produced by solvent cast method. The composite films were characterized by SEM, DSC-TGA, XRD, and ATR-FTIR. The ultrastructure of PHB and its composites depicted an irregular surface morphology with pores after the evaporation of chloroform. The GNPs were seen to be integrated inside the pores. The B. megaterium H16 derived-PHB and its composites demonstrated good biocompatibility which was evaluated in vitro on HaCaT and L929 cells by MTT assay. The cell viability was best for PHB followed by PHB/PLLA/PCL > PHB/PLLA/GNP > PHB/PLLA. PHB and its composites were highly hemocompatible as it resulted in <1 % hemolysis. The PHB/PLLA/PCL and PHB/PLLA/GNP composites can serve as ideal biomaterials for skin tissue engineering.


Assuntos
Bacillus megaterium , Materiais Biocompatíveis , Materiais Biocompatíveis/farmacologia , Materiais Biocompatíveis/química , Engenharia Tecidual/métodos , Poliésteres/química , Pele , Hidroxibutiratos/química
2.
J Nanosci Nanotechnol ; 18(12): 8142-8154, 2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30189931

RESUMO

Silica (SiO2) is the inevitable form of silicon owing to its high affinity for oxygen, existing as a geogenic element perpetrating multifarious health problems when bioavailable via anthropogenic activities. The hydrated form of silica viz. orthosilicic acid (H4SiO4) excessively displays grave toxicity, attributed to prolonged exposure and incessant H+ ions generating capacity inflicting pulmonary toxicity and renal toxicity silica. The diverse deleterious potency of silica highlights the desirability of selective and sensitive detection of toxic species (mainly orthosilicic acid) bioaccumulation in affected living human cells. In this paper we have reported, the design of water-dispersible turn-on fluorimetric sensing material for the detection of orthosilicic acid in the aqueous phase and in live cells. The sensing material was prepared by adsorbing a suitable rhodamine derivative (i.e., Rhodamine B hydrazide (Rh1)) on water dispersible TiO2 nanoparticles. The function of the sensing system, which is composed of Rh1 and TiO2 (Rh1@TiO2), is accredited to H+ ion (from orthosilicic acid) induced spirolactam ring-opening of the rhodamine derivative generating orange fluorescence and bright pink colouration. The sensing system was efficiently utilized for fluorimetric detection and imaging of orthosilicic acid accumulation in-vitro in human kidney cells (HK cells). To the best of our knowledge, this is the first time this sensing system (Rh1@TiO2) is reported for detection of toxic silica species accumulation in-vitro in human kidney cells. The advantages, such as good water dispersibility, the absence of organic solvents during fluorimetric studies, quick turn-on type signal transduction, low-level imaging, which are offered by the synthesized sensing material (Rh1@TiO2), make it a potential candidate to fabricate medical tool for early identification of silicainduced nephrotoxicity, which can help to reduce the burden and risk of chronic kidney disease development.


Assuntos
Hidrazinas , Nanopartículas , Rodaminas , Titânio , Humanos , Rim/diagnóstico por imagem , Rim/efeitos dos fármacos , Nanopartículas/toxicidade , Rodaminas/toxicidade , Dióxido de Silício/toxicidade , Água
3.
Sci Total Environ ; 645: 297-317, 2018 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-30029111

RESUMO

Silica is a trace-geogenic compound with limited-bioavailability. It inflicts health-perils like pulmonary-silicosis and chronic kidney disease (CKD), when available via anthropogenic-disturbances. Amidst silica-imposed pathologies, pulmonary toxicological-mechanisms are well-described, ignoring the renal-pathophysiological mechanisms. Hence, the present-study aimed to elucidate cellular-cum-molecular toxicological-mechanisms underlying silica-induced renal-pathology in-vitro. Various toxicity-assessments were used to study effects of silica on the physiological-functions of HK-cells (human-kidney proximal-tubular cells - the toxin's prime target) on chronic (1-7 days) sub-toxic (80 mg/L) and toxic (100-120 mg/L) dosing. Results depicted that silica triggered dose-cum-time dependent cytotoxicity/cell-death (MTT-assay) that significantly increased on long-term dosing with ≥100 mg/L silica; establishing the nephrotoxic-potential of this dose. Contrarily, insignificant cell-death on sub-toxic (80 mg/L) dosing was attributed to rapid intracellular toxin-clearance at lower-doses preventing toxic-effects. The proximal-tubular (HK-cells) cytotoxicity was found to be primarily mediated by silica-triggered incessant oxidative-stress (elevated ROS).·This enhanced ROS inflicted severe inflammation and subsequent fibrosis, evident from increased pro-inflammatory-cum-fibrogenic cytokines generation (IL-1ß, IL-2, IL-6, TNF-α and TGF-ß). Simultaneously, ROS induced persistent DNA-damage (Comet-assay) that stimulated G2/M arrest for p53-mediated damage-repair, aided by checkpoint-promoter (Chk1) activation and mitotic-inducers (i.e. Cdc-25, Cdk1, cyclinB1) inhibition. However, DNA-injuries surpassed the cellular-repair, which provoked the p53-gene to induce mitochondrial-mediated apoptotic cell-death via activation of Bax, cytochrome-c and caspase-cascade (9/3). This persistent apoptotic cell-death and simultaneous incessant inflammation culminated in the development of tubular-atrophy and fibrosis, the major pathological-manifestations of CKD. These findings provided novel-insights into the pathological-mechanisms (cellular and molecular) of silica-induced CKD, inflicted on chronic toxic-dosing (≥100 mg/L).Thereby, encouraging the development of therapeutic-strategies (e.g. anti-oxidant treatment) for specific molecular-targets (e.g. ROS) to retard silica-induced CKD-progression, for reduction in the global-CKD burden.


Assuntos
Substâncias Perigosas/toxicidade , Dióxido de Silício/toxicidade , Apoptose , Humanos , Rim/efeitos dos fármacos , Mitocôndrias , Estresse Oxidativo , Espécies Reativas de Oxigênio , Oligoelementos
4.
Rev Diabet Stud ; 14(4): 334-353, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29590227

RESUMO

In recent years, there has been an alarming increase in the incidence of diabetes, with one in every eleven individuals worldwide suffering from this debilitating disease. As the available treatment options fail to reduce disease progression, novel avenues such as the bioartificial pancreas are being given serious consideration. In the past decade, the research focus has shifted towards the field of tissue engineering, which helps to design biological substitutes for repair and replacement of non-functional or damaged organs. Scaffolds constitute an integral part of tissue engineering; they have been shown to mimic the native extracellular matrix, thereby supporting cell viability and proliferation. This review offers a novel compilation of the recent advances in polymeric scaffolds, which are used for pancreatic tissue engineering. Furthermore, in this article, the design strategies for bioartificial pancreatic constructs and their future applications in cell-based therapy are discussed.


Assuntos
Diabetes Mellitus Tipo 1/terapia , Pâncreas , Engenharia Tecidual , Alicerces Teciduais , Matriz Extracelular , Humanos
5.
Artif Cells Nanomed Biotechnol ; 46(3): 637-649, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-28639839

RESUMO

With increasing gap in the demand and supply of vital organs for transplantation there is a pressing need to bridge the gap with substitutes. One way to make substitutes is by tissue engineering which involves combining several types of synthetic or biomaterials, cells and growth factors cross-linked together to synthesize a functional scaffold for repair or replacement of non-functional organs. Nanoparticle based composites are gaining importance in tissue engineering due to their ability to enhance cell attachment and proliferation. The current study focuses on synthesizing agarose composites embedded with chitosan-coated silver nanoparticles using glutaraldehyde as the cross-linker. The synthesis of chitosan coated silver nanoparticles within the scaffold was confirmed with UV-visible spectroscopy. Physical and chemical characterization of the synthesized nanoparticles were done by XRD, FTIR, TGA and SEM. DMA showed higher mechanical strength of the scaffolds. The scaffolds showed degradation of ∼37% within a span of four weeks. The higher physical support provided by the synthesized scaffolds was shown by in-vitro cell viability assay. Broad spectrum anti-bacterial activity and superior hemocompatibility further showed the advantage it offered for growing cells. Thus a biopolymer based nanocomposite was synthesized, with intended widespread use as scaffold for engineering of soft tissues due to its enhanced biocompatibility and greater surface area for cell growth.


Assuntos
Quitosana , Materiais Revestidos Biocompatíveis , Teste de Materiais , Nanopartículas/química , Sefarose , Engenharia Tecidual , Quitosana/química , Quitosana/farmacologia , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Células HeLa , Humanos , Sefarose/química , Sefarose/farmacologia , Prata/química , Prata/farmacologia
6.
Anal Chem ; 89(23): 12698-12704, 2017 12 05.
Artigo em Inglês | MEDLINE | ID: mdl-29115126

RESUMO

Methylmercury (CH3Hg+) is the common form of organic mercury and is more toxic than its inorganic or elemental forms. Mercury is emanated in the course of various natural events and human activities and converts to methylmercury by anaerobic organisms. CH3Hg+ are ingested by fish and subsequently bioaccumulated in their tissue and, eventually, enter the human diet, causing serious health issues. Therefore, selective and sensitive detection of bioaccumulated CH3Hg+ in fish samples is essential. Herein, the development of a simple, highly sensitive and selective aggregation-induced emission (AIE)-based turn-on probe for both inorganic mercury ions and organicmercury species is reported. The probe's function is based on mercury ion-promoted transmetalation reaction of aryl boronic acid. The probe, a tetraphenylethylene (TPE)-monoboronic acid (1), was successfully utilized for AIE-based fluorescence imaging study on methylmercury-contaminated live cells and zebrafish for the first time. Both Hg(II) and CH3Hg+ ensued a fast transmetalation of TPE-boronic acid causing drastic reduction in the solubility of the resulting product (TPE-HgCl/TPE-HgMe) in the working solvent system. At the dispersed phase, the aggregated form of TPE-mercury ions recovers planarity because of restricted rotational freedom promoting aggregation-induced emission. Simple design, cost-effective synthesis, high selectivity, inexpensive instrumentation, fast signal transduction, and low limit of detection (0.12 ppm) are some of the key merits of this analytical tool.


Assuntos
Mercúrio/análise , Compostos de Metilmercúrio/análise , Microscopia de Fluorescência/métodos , Animais , Ácidos Borônicos/síntese química , Ácidos Borônicos/química , Ácidos Borônicos/toxicidade , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/química , Corantes Fluorescentes/toxicidade , Células HEK293 , Humanos , Limite de Detecção , Peixe-Zebra
7.
Chemosphere ; 177: 239-249, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28292724

RESUMO

Chronic-Kidney-Disease of Unknown-etiology (CKDu) has been reported in developing-countries like Sri-Lanka, India and Central-America without sparing the Indian sub-district (namely Canacona) located in south-Goa. The disease etiology is unlinked to common causes of diabetes and hypertension and assumed to be environmentally induced due to its asymptomatic-nature and occurrence in groundwater relying communities. This study aimed to understand environmental risk-factors underlying CKDu-etiology using Indian sub-district (Canacona) as case-study. Biochemical-analysis of CKDu-affected and non-affected individual's blood and detailed hydro-geochemical analyses of CKDu-affected and non-affected region's groundwater (drinking-water)were conducted. Trace geogenic-element-silica was highly dominant in affected-region's groundwater, thus its nephrotoxic-potential was analysed via in-vitro cytotoxicity-assays on human-kidney-cell-lines. All CKDu-affected-subjects showed increased-levels of serum-urea (52.85 mM),creatinine (941.5 µM),uric-acid (1384.5 µM), normal blood-glucose (4.65 mM), being distinct biomarkers of environmentally-induced CKD-'chronic-tubulo-interstitial-nephritis'. Affected-subjects reported high blood-lead levels (1.48 µM)suggesting direct-nephrotoxicity resulting in impaired blood-clearance and also exhibits indirect-nephrotoxicity by disrupting calcium-homeostasis causing skeletal-disorders and prolonged-consumption of NSAID's (pain-alleviation), indirectly causing renal-damage. Affected-region's groundwater was acidic (pH-5.6), resulting in borderline-lead (9.98 µgL-1) and high-silica (115.5 mgL-1)contamination. Silica's bio-availability (determining its nephrotoxicity) was enhanced at groundwater's acidic-pH and Ca-Mg-deficient-composition (since these cations complex with silica reducing bioavailability). Silica exhibited renal-proximal-tubular-cytotoxicity on long-term exposure comparable with affected-region's groundwater silica-levels, by apoptosis-mediated-cell-death resulting in tubular-atrophy, interstitial-fibrosis and irreversible renal-damage (CKD). Thus this study provides novel-insights into nephrotoxic-potential of trace-geogenic-element-silica in CKDu causation. It highlights direct-indirect nephrotoxicity exhibited by lead at low-levels due to its bio-accumulative-capacity. Silica's nephrotoxic-potential can be considered when deciphering etiology of CKDu-problem in developing-countries (relying on groundwater).


Assuntos
Água Potável/análise , Chumbo/análise , Insuficiência Renal Crônica/induzido quimicamente , Dióxido de Silício/análise , Oligoelementos/análise , Adulto , Antropometria , Anti-Inflamatórios não Esteroides/química , Apoptose , Biomarcadores/sangue , Glicemia/análise , Ciclo Celular , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células , Creatinina/sangue , Feminino , Água Subterrânea/análise , Células HEK293 , Humanos , Concentração de Íons de Hidrogênio , Índia , Chumbo/sangue , Masculino , Metais/química , Insuficiência Renal Crônica/epidemiologia , Fatores de Risco , Ureia/sangue
8.
BMC Dev Biol ; 16: 8, 2016 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-27068127

RESUMO

BACKGROUND: It has been suggested that the ectopic expression of PDX1, a dominant pancreatic transcription factor, plays a critical role in the developmental programming of the pancreas even from cells of unrelated tissues such as keratinocytes and amniotic fluid stem cells. In this study we have chosen to drive pancreatic development in human amnion epithelial cells by inducing endogenous PDX1 expression. Further, we have investigated the role of Epidermal Growth Factor (EGF) and Poly-L-Ornithine (PLO) on this differentiation process. RESULTS: Human amnion epithelial cells expressed high levels of endogenous PDX1 upon transduction with an adenoviral vector expressing murine Pdx1. Other markers of various stages of pancreatic differentiation such as NKX6.1, SOX17, RFX6, FOXA2, CFTR, NEUROD1, PAX4 and PPY were also expressed upon Pdx1 transduction. Although initial expression of pancreatic progenitor markers was higher in culture conditions lacking EGF, for a sustained and increased expression EGF was required. Culture on PLO further increased the positive impact of EGF. CONCLUSION: Pancreatic marker expression subsequent to mPdx1 transduction suggests that this approach may facilitate the in vitro differentiation of hAECs into cells of the endocrine pancreas. This result may have important implications in diabetes therapy.


Assuntos
Âmnio/citologia , Diferenciação Celular/efeitos dos fármacos , Fator de Crescimento Epidérmico/farmacologia , Células Epiteliais/citologia , Proteínas de Homeodomínio/metabolismo , Peptídeos/farmacologia , Transativadores/metabolismo , Animais , Biomarcadores/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Camundongos , Transdução Genética
9.
Mar Pollut Bull ; 88(1-2): 148-54, 2014 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-25277552

RESUMO

Aliphatic hydrocarbons are one of the major environmental pollutants with reduced bioavailability. The present study focuses on the effect of hydroxy cucurbit[6]uril on the bioavailability of hydrocarbons. A bacterial consortium was used for biodegradation studies under saline and non-saline conditions. Based on denaturing gradient gel electrophoresis results it was found that the consortium under saline conditions had two different strains. The experiment was conducted in microcosms with tetradecane, hexadecane, octadecane and mixture of the mentioned hydrocarbons as the sole carbon source. The residual hydrocarbon was quantified using gas chromatography every 24h. It was found that biodegradation of tetradecane and hexadecane, as individual carbon source increased in the presence of hydroxy CB[6], probably due to the increase in their bioavailability. In case of octadecane this did not happen. Bioavailability of all three aliphatic hydrocarbons was increased when provided as a mixture to the consortium under saline conditions.


Assuntos
Hidrocarbonetos Aromáticos com Pontes/metabolismo , Hidrocarbonetos/metabolismo , Imidazóis/metabolismo , Consórcios Microbianos , Alcanos/metabolismo , Alcanos/farmacocinética , Sequência de Bases , Biodegradação Ambiental , Disponibilidade Biológica , Hidrocarbonetos Aromáticos com Pontes/síntese química , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Cromatografia Gasosa , Eletroforese em Gel de Gradiente Desnaturante , Células HeLa/efeitos dos fármacos , Humanos , Hidrocarbonetos/farmacocinética , Imidazóis/síntese química , Imidazóis/farmacologia , Espectroscopia de Ressonância Magnética , Consórcios Microbianos/genética , Dados de Sequência Molecular , Espectrometria de Massas por Ionização por Electrospray
10.
Carbohydr Res ; 397: 37-45, 2014 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-25199581

RESUMO

Four new D-glucose derived m-s-m type gemini surfactants with variable spacer and tail length have been synthesized by a simple and efficient synthetic methodology utilizing the free C-3 hydroxy group of diisopropylidene glucose. The synthetic route to these gemini surfactants with a quaternary ammonium group as polar head group involves a sequence of simple reactions including alkylation, imine formation, quaternization of amine etc. The surface properties of the new geminis were evaluated by surface tension and conductivity measurements. These gemini surfactants showed low cytotoxicity by MTT assay on HeLa cell line. The DNA binding capabilities of these surfactants were determined by agarose gel electrophoresis, fluorescence titration, and DLS experiments. The preliminary studies by agarose gel electrophoresis indicated chain length dependent DNA binding abilities, further supported by ethidium bromide exclusion experiments. Two of the D-glucose derived gemini surfactants showed effective binding with pET-28a plasmid DNA (pDNA) at relatively low N/P ratio (i.e., cationic nitrogen/DNA phosphate molar ratio).


Assuntos
DNA/metabolismo , Tensoativos/química , Tensoativos/metabolismo , Tensoativos/farmacologia , Calcitriol/análogos & derivados , Calcitriol/química , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Química Sintética , Eletroforese em Gel de Ágar , Glucose , Células HeLa/efeitos dos fármacos , Humanos , Micelas , Tamanho da Partícula , Relação Estrutura-Atividade , Propriedades de Superfície , Tensão Superficial , Tensoativos/síntese química
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