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1.
Eur J Med Res ; 29(1): 82, 2024 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-38287418

RESUMO

BACKGROUND: Cadmium (Cd) exposure has been found to have detrimental effects on the development of the central nervous system and cognitive ability in children. However, there is ongoing debate regarding the impact of maternal Cd exposure on the cognitive ability of offspring. In this study, we aimed to investigate the mechanisms underlying the influence of maternal Cd exposure on the cognitive ability of offspring rats. METHODS: Here, we constructed a model of cadmium poisoning in first-generation rats through gavage. The cognitive and memory abilities of its offspring were evaluated by water maze experiment. Then, we used the gene chip to find out the key genes, and we performed qRT-PCR detection of these genes. Subsequently, enrichment analysis was employed to identify pathways. Finally, we constructed a co-expression network consisting of LncRNAs and mRNAs to elucidate the biological functions and regulatory mechanisms of LncRNAs. RESULTS: The results of the water maze trial demonstrated that the offspring of rats exposed to cadmium in the first generation had reduced cognitive and memory abilities. Through an analysis of gene expression in the hippocampus of the cadmium-treated rats' offspring and the control group, we identified a correlation between the islet secretion pathway and the cognitive impairment observed in the offspring. Utilizing various algorithms, we identified Cpa1 and Prss1 as potential key genes associated with the cognitive impairment caused by cadmium. The results of qRT-PCR demonstrated a decrease in the expression levels of these genes in the hippocampus of the cadmium-treated rats' offspring. In addition, in the co-expression network, we observed that Cpa1 was co-expressed with 11 LncRNAs, while Prss1 was associated with 4 unexplored LncRNAs. Furthermore, we conducted an analysis to examine the relationship between Cpa1, Prss1-related transcription factors, and LncRNAs. CONCLUSION: Overall, this study provides novel insights into the molecular effects of first generation Cd exposure on the cognitive ability of offspring. The target genes and signaling pathways investigated in this study could serve as potential targets for improving neurodevelopment and cognitive ability in children.


Assuntos
Deficiências da Aprendizagem , RNA Longo não Codificante , Humanos , Criança , Ratos , Animais , Cádmio/toxicidade , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos
2.
J Microencapsul ; 40(3): 157-170, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36825613

RESUMO

AIMS: The aim of this study is to develop a liposome that could exert unparalleled antioxidant effects. In the present study, a vitamin C (VC)/vitamin E (VE)-co-loaded multivesicular liposome (VCVE-MVL) was constructed. METHODS: Vitamins were encapsulated in soybean phosphatidylcholine (SPC) and cholesterol (CHO) by multi-emulsification method. The concentration of VC was determined by Fast Blue method. The concentration of VE was determined by high performance liquid chromatography (HPLC). Vitamin antioxidant capacity in vitro and in vivo was determined using ß-carotene bleaching. RESULTS: VCVE-MVL with particle diameter of 848.55 ± 0.29 nm and SPAN of 0.16 ± 0.11 were obtained. The encapsulation efficiency of VC reached 48.51% (w/w)±0.15. Compared with VC/VE solution, VCVE-MVL had a higher permeation efficiency. In addition, the in vitro and ex-vivo antioxidant tests verified the adequate antioxidant activity of VCVE-MVL. CONCLUSIONS: In conclusion, the antioxidant synergistic system we constructed and demonstrated its potential applications in the cosmetics industry.


Assuntos
Antioxidantes , Vitamina E , Antioxidantes/farmacologia , Antioxidantes/química , Ácido Ascórbico , Lipossomos/química , Vitaminas
3.
J Food Sci ; 87(6): 2766-2774, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35502685

RESUMO

Rice is a recognized source of cadmium (Cd) exposure. In this study, a total of 102 rice samples were obtained from main producing rice regions of China, and the bioavailability of Cd in rice were assessed by using an in vitro digestion method combined with a Caco-2 cell monolayer model. The results revealed that the average (range of) concentration of Cd in the rice samples was 0.2840 (0.0123-2.4710) mg/kg, and the average (range of) bioavailability of Cd in the rice samples was 23.94% (7.47-39.73%). We analyzed the recent rice consumption data and calculated the target hazard quotients (THQs) to evaluate the noncarcinogenic health risk of Cd in rice consumption for adults and children, respectively. We suggested that the most of investigated regions showed low risks of Cd in rice for general population, and the regions of Cd in rice near the limit deserves might be attracted more attention after taking bioavailability into account. PRACTICAL APPLICATION: The cadmium content of rice itself will affect its corresponding bioavailability to a certain extent. By studying the in vitro bioavailability of cadmium rice with different pollution levels, it is helpful for the dose selection of animal experiments or toxicity tests, and finally, we may provide some references for the establishment of appropriate treatment scheme of rice Cd poisoning.


Assuntos
Oryza , Poluentes do Solo , Disponibilidade Biológica , Células CACO-2 , Cádmio/análise , China , Humanos , Medição de Risco , Solo , Poluentes do Solo/análise
4.
J Food Sci ; 86(8): 3730-3742, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34309019

RESUMO

Cadmium (Cd) is a widespread heavy metal pollutant in the environment that damages human health. In this study, the bioaccessibility and bioavailability of Cd in different Cd-contaminated rice (low pollution level cadmium rice (Rice-L, 0.111 mg/kg), medium pollution level cadmium rice (Rice-M, 0.400 mg/kg), and high pollution level cadmium rice (Rice-H, 0.655 mg/kg)) were estimated and determined by an in vitro digestion model Rijksinstituut voor volksgezondheiden milieu (RIVM), Caco-2 cell model, and mouse model. The results indicated that Cd in the oral cavity (15.65-28.28%) displayed the lowest bioaccessibility comparing with small intestine (90.04-94.73%) and the stomach (99.30-100.70%) in vitro after cooking. In addition, the results showed that the bioaccessibility of Cd in CdCl2 , CdCl2 +normal rice (Rice-N), Rice-H, Rice-M, Rice-L group were 99.29%, 92.57%, 90.04%, 94.73%, and 91.11%, respectively; the in vitro bioavailability of Cd in CdCl2 , CdCl2 +Rice-N, Rice-H, Rice-M, and Rice-L group were 27.50%, 20.78%, 21.90%, 26.90%, 36.46%, respectively, we found that the group of CdCl2 is significantly higher than CdCl2 +Rice-N and Rice-H (p < 0.05), while the targets hazard quotient (THQ) value of rice ingested without considering the in vitro bioavailability is 2.7-4.6 times than the THQ value with considered and the relative bioavailability (RBA) of Cd in Rice-L, Rice-M, Rice-H are 80.25%, 64.32%, and 60.91%, respectively. These results indicate that the rice substrate has impact on the bioaccessibility and bioavailability of Cd, and might overestimate the health risks of Cd if bioavailability was not considered. PRACTICAL APPLICATION: Studying the bioaccessibility and bioavailability of cadmium in rice is a promising strategy to obtain a more accurate human health risk assessment of cadmium exposure in rice, as well as provide a theoretical basis for the formulation of cadmium limit standard in grain, which was also conducive to the rational and full utilization of rice resources in China.


Assuntos
Oryza , Poluentes do Solo , Animais , Disponibilidade Biológica , Células CACO-2 , Cádmio/análise , Contaminação de Alimentos/análise , Humanos , Camundongos , Solo
5.
Spectrochim Acta A Mol Biomol Spectrosc ; 246: 118973, 2021 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-33017793

RESUMO

Geographical discrimination and adulteration analysis play significant roles in edible oil analysis. A novel method for discrimination and adulteration analysis of edible oils were proposed in this study. The two-dimensional correlation spectra of edible oils were obtained by solvents perturbation and the convolutional neural networks (CNNs) were constructed to analyze the synchronous and asynchronous correlation spectra of the edible oils. The differences for geographical origins of oils or oil types could be amplificated through the networks. For different networks, the layer sequences and the filter number of convolutional layers may affect the analysis results. A group of sesame oils from different geographical origins and a group of olive oils adulterated by other vegetable oils were adopted to evaluate the proposed method. The results show that the proposed method may provide an alternative method for edible oil discrimination and adulteration analysis in practical applications. For the two datasets, the prediction accuracy could be 97.3% and 88.5%, respectively.


Assuntos
Contaminação de Alimentos , Óleos de Plantas , Contaminação de Alimentos/análise , Redes Neurais de Computação , Azeite de Oliva/análise , Óleos de Plantas/análise , Análise Espectral
6.
Food Funct ; 11(10): 8987-8995, 2020 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-33001081

RESUMO

Obesity is an increasingly severe global health problem, leading to chronic inflammation and metabolic disorders in both peripheral tissues and the central nervous system. Matcha is a powdered green tea, and it is very popular in recent years as a beverage and food additive. Matcha green tea has been reported to have outstanding potential in regulating obesity-related metabolic syndrome. However, there are few studies on the regulation mechanism of matcha green tea on the central nervous system. In this study, we established a high-fat diet-induced obese mouse model. The results showed that dietary supplementation with matcha could effectively inhibit the weight gain, fat accumulation, glycemia and lipidemia increase, and excessive activation of microglia in the arcuate nucleus of the hypothalamus. Furthermore, we used different concentrations (100%, 80%, 60%, 40%, and 20%, v/v) of ethanol solution to prepare matcha ethanol extracts, and investigated their effects on palmitic acid-induced inflammation of microglial BV-2 cells. The results showed that matcha ethanol extracts could significantly reduce the release of inflammatory cytokines and the expression and phosphorylation of JAK2 and STAT3.


Assuntos
Camellia sinensis/química , Hipotálamo/efeitos dos fármacos , Janus Quinase 2/imunologia , Obesidade/tratamento farmacológico , Obesidade/imunologia , Extratos Vegetais/administração & dosagem , Fator de Transcrição STAT3/imunologia , Animais , Antioxidantes/administração & dosagem , Humanos , Hipotálamo/imunologia , Janus Quinase 2/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Fator de Transcrição STAT3/genética , Transdução de Sinais/efeitos dos fármacos , Chá/química
7.
Toxins (Basel) ; 12(3)2020 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-32235760

RESUMO

Deoxynivalenol (DON), along with 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON), occur in grains and cereal products and is often hazardous to humans and livestock. In this study, 579 wheat samples and 606 maize samples intended for consumption were collected from China in 2017 and analyzed to determine the co-occurrence of type-B trichothecenes (DON, 3-ADON, and 15-ADON). All the wheat samples tested positive for DON, while 99.83% of the maize samples were DON-positive with mean DON concentrations of 165.87 and 175.30 µg/kg, respectively. Per the Chinese standard limits for DON, 3.63% of wheat and 2.97% of the maize samples were above the maximum limit of 1000 µg/kg. The DON derivatives (3-ADON and 15-ADON) were less frequently found and were present at lower levels than DON in wheat. 3-ADON and 15-ADON had incidences of 13.53% and 76.40%, respectively, in maize. By analyzing the distribution ratio of DON and its derivatives in wheat and maize, DON (95.51%) was the predominant toxin detected in wheat samples, followed by 3.97% for the combination of DON + 3-ADON, while DON + 3-ADON + 15-ADON and DON + 15-ADON were only found in 0.17% and 0.35% of wheat samples, respectively. Additionally, a large amount of the maize samples were contaminated with DON + 15-ADON (64.19%) and DON (22.11%). The samples with a combination of DON + 3-ADON and DON + 3-ADON + 15-ADON accounted for 1.32% and 12.21%, respectively. Only one maize sample did not contain all three mycotoxins. Our study shows the necessity of raising awareness of the co-occurrence of mycotoxin contamination in grains from China to protect consumers from the risk of exposure to DON and its derivatives.


Assuntos
Grão Comestível/química , Contaminação de Alimentos/análise , Tricotecenos/análise , Triticum/química , Zea mays/química , Acetilação , Cromatografia Líquida de Alta Pressão , Limite de Detecção , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem , Tricotecenos/química , Triticum/microbiologia , Zea mays/microbiologia
8.
Nat Prod Res ; 34(22): 3253-3256, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30676779

RESUMO

ß-glucans are a heterogeneous group of natural polysaccharides. They are ubiquitously found in bacterial or fungal cell walls, cereals, seaweed, and mushrooms. The beneficial role of ß-glucan in tumor, insulin resistance, dyslipidemia, hypertension, and obesity is being continuously documented. Ample evidence showed that ß-glucan could act on several receptors, such as Dectin, complement receptor (CR3), TLR-2, 4, 6 and scavenger. Based on the above, we wanted to explore whether agaricus bisporus-derived ß-glucan acted on these receptors on Raw 264.7 macrophages and 3T3-L1 adipocytes.


Assuntos
Adipócitos/efeitos dos fármacos , Agaricus/química , Antígenos CD36/metabolismo , Macrófagos/efeitos dos fármacos , beta-Glucanas/farmacologia , Células 3T3-L1 , Adipócitos/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Citometria de Fluxo/instrumentação , Fluoresceína-5-Isotiocianato/farmacocinética , Polissacarídeos Fúngicos/farmacocinética , Polissacarídeos Fúngicos/farmacologia , Macrófagos/metabolismo , Camundongos , Células RAW 264.7 , beta-Glucanas/farmacocinética
9.
Int J Mol Sci ; 20(20)2019 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-31614951

RESUMO

Obesity is closely associated with neuroinflammation in the hypothalamus, which is characterized by over-activated microglia and excessive production of pro-inflammatory cytokines. The present study was aimed at elucidating the effects of (-)-epigallocatechin gallate (EGCG) on palmitic acid-stimulated BV-2 microglia and high-fat-diet-induced obese mice. The results indicated the suppressive effect of EGCG on lipid accumulation, pro-inflammatory cytokines (TNF-α, IL-6, and IL-1ß) release, and microglial activation in both cellular and high-fat-diet rodent models. These results were associated with lower phosphorylated levels of the janus kinase 2/signal transducers and activators of transcription 3 (JAK2/STAT3) signaling pathway. In conclusion, EGCG can attenuate high-fat-induced hypothalamic inflammation via inhibiting the JAK2/STAT3 signaling pathways in microglia.


Assuntos
Fármacos Antiobesidade/farmacologia , Catequina/análogos & derivados , Microglia/efeitos dos fármacos , Obesidade/tratamento farmacológico , Animais , Fármacos Antiobesidade/uso terapêutico , Catequina/farmacologia , Catequina/uso terapêutico , Linhagem Celular , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Hipotálamo/efeitos dos fármacos , Hipotálamo/imunologia , Hipotálamo/metabolismo , Inflamação/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Janus Quinase 2/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Microglia/metabolismo , Obesidade/metabolismo , Ácido Palmítico/metabolismo , Ácido Palmítico/farmacologia , Polifenóis/farmacologia , Fator de Transcrição STAT3/metabolismo , Chá/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
10.
J Mater Sci Mater Med ; 21(5): 1473-8, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20162322

RESUMO

Dentine carious lesions can be remineralized under optimal conditions, while the surface characteristics of the caries-attacked area may play an important role in the remineralization process. To understand such a surface mechanism, we examined the microstructures of the remineralized area pretreated with different methods. It was found that dentinal tubules on the remineralized surface orientated differently from intrinsic dentine tubules, with the specific alignment angle determined by different surface treatments. Various surface treatments included in this study were 37% phosphoric acid treatment (the etched group), 37% phosphoric acid etching followed by the application of 10% sodium hypochlorite treatment (the deproteinized group), and untreatment (the control group). These findings are helpful for understanding the non-restorative repair of dentine lesions and the remineralization process of the caries-affected dentin surface.


Assuntos
Cárie Dentária/patologia , Cárie Dentária/terapia , Dentina/patologia , Ácidos Fosfóricos/química , Humanos , Hipoclorito de Sódio/farmacologia
11.
Brain Res ; 1325: 183-90, 2010 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-20167210

RESUMO

The neuroprotective effects of N-stearoyl-L-tyrosine (NSTyr) on cognitive function and neuronal plasticity during chronic cerebral hypoperfusion (CCH) in rats were investigated. After induction of CCH, NSTyr was administered daily for 3 months intraperitoneally. Cognitive functions were evaluated by Morris water maze and hippocampal long-term potentiation (LTP). Neuropathological changes were examined using light micrograph and Fluoro-Jade B staining. Neuronal plasticity was assessed by measuring the expression of MAP-2, GAP-43 and synaptophysin on hippocampal regions of rats with immunohistochemistry and western blotting. CCH resulted in significant spatial memory impairment and inhibition of LTP, and led to neurodegeneration in the CA1 region of the hippocampus in the model rats compared with the sham-operated rats. In the model rats treated with NSTyr, cognitive function improved. The expression levels of MAP-2 and synaptophysin protein in hippocampal areas in the model rats were less than those in the sham-operated rats, and increased in the model rats treated with NSTyr. However, no statistical significance of GAP-43 expression among the sham, model and NSTyr groups was observed. These data indicate that NSTyr exerts protective effects on cognitive function of rats after CCH, which may be related to the changes of neurodegeneration and neuronal plasticity in the hippocampal area of rats.


Assuntos
Transtornos Cerebrovasculares/tratamento farmacológico , Cognição/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Plasticidade Neuronal/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Tirosina/análogos & derivados , Animais , Transtornos Cerebrovasculares/patologia , Transtornos Cerebrovasculares/fisiopatologia , Doença Crônica , Modelos Animais de Doenças , Proteína GAP-43/metabolismo , Hipocampo/patologia , Hipocampo/fisiopatologia , Potenciação de Longa Duração/efeitos dos fármacos , Aprendizagem em Labirinto/efeitos dos fármacos , Transtornos da Memória/tratamento farmacológico , Transtornos da Memória/patologia , Transtornos da Memória/fisiopatologia , Proteínas Associadas aos Microtúbulos/metabolismo , Ratos , Ratos Sprague-Dawley , Percepção Espacial/efeitos dos fármacos , Sinaptofisina/metabolismo , Tirosina/farmacologia
12.
Molecules ; 14(10): 4051-64, 2009 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-19924046

RESUMO

A series of lipoamino acids were synthesized and their neuroprotective effect against brain ischemia induced by oxygen-glucose deprivation (OGD) on rat cerebral slices was evaluated. Among these compounds, N-stearoyl-L-tyrosine (4), N-stearoyl-L-serine (5) and N-stearoyl-L-threonine (6) exhibited good neuroprotective activity. We found that the neuroprotective activity of lipoamino acids depended on the acyl group, the presence of a free carboxylic function and a free hydroxyl group at the branched chain of the amino acids. The results also showed that 5 was the most active compound, protecting rat brain slices against OGD as well as hydrogen peroxide (H(2)O(2)) insult at the range of 1-10 M.


Assuntos
Isquemia Encefálica/prevenção & controle , Encéfalo/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Serina/análogos & derivados , Treonina/análogos & derivados , Tirosina/análogos & derivados , Anaerobiose , Animais , Encéfalo/irrigação sanguínea , Peróxido de Hidrogênio/toxicidade , Fármacos Neuroprotetores/síntese química , Fármacos Neuroprotetores/química , Ratos , Serina/síntese química , Serina/química , Serina/farmacologia , Treonina/síntese química , Treonina/química , Treonina/farmacologia , Tirosina/síntese química , Tirosina/química , Tirosina/farmacologia
13.
J Huazhong Univ Sci Technolog Med Sci ; 28(4): 483-6, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18704318

RESUMO

In this study, calcium phosphate cements (CPC) were prepared by mixing cement powders of tetracalcium phosphate (TTCP) with a cement liquid of phosphate acid saline solution. Tetracycline (TTC)-CPC, chitosan-CPC and chitosan-TTC-CPC were investigated with different premixed schedule. It was demonstrate that both TTC and chitosan worked on the phase transition and crystal characteristics. TTCP mixed with phosphate acid saline solution had similar features of Fourier transform-infrared spectrometry (FT-IR) no matter it was mixed with chitosan or TTC or both. TTC premixed with cement liquid or powder had significant different features of FT-IR and 876 cm(-1) seemed to be a special peak for TTC when TTC was premixed with cement liquid. This was also supported by XRD analysis, which showed that TTC premixed with cement liquid improved phase transition of TTCP to OCP. Chitosan, as organic additive, regulates the regular crystal formation and inhibits the phase transition of TTCP to OCP, except when it is mingled with cement liquid premixed with TTC in field scanning electron microscope. It was concluded that the premixed schedule influences the crystal formation and phase transition, which may be associated with its biocompatibility and bioactivities in vivo.


Assuntos
Materiais Biocompatíveis/química , Cimentos Ósseos/química , Fosfatos de Cálcio/química , Quitosana/química , Tetraciclina/química , Cimentos Dentários/química , Fosfatos/química , Difração de Raios X
14.
Shanghai Kou Qiang Yi Xue ; 17(2): 187-90, 2008 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-18470426

RESUMO

PURPOSE: To investigate the effect of premix schedule on crystal formation of self-hardening calcium phosphate cements (CPC). METHODS: CPC were prepared by mixing cement powders of tetracalcium phosphate(TTCP) with a cement liquid of phosphate acid saline solution. TTC-CPC, chitosan-CPC and chitosan-TTC-CPC were investigated with different premix schedules. The crystal formation was investigated by X-ray diffraction(XRD) and field scanning electron microscope(FSEM). RESULTS: TTC and chitosan both affected the phase transition and crystal character in CPC. Chitosan, as organic additive, regulated the regular crystal formation and inhibited the phase transition of TTCP into OCP. CONCLUSIONS: The premix schedule affects the crystal formation and phase transition, which may affect its biocompatibility and bioactivities in vivo.


Assuntos
Fosfatos de Cálcio/química , Cimentos Dentários , Cimentos Ósseos , Quitosana , Difração de Raios X
15.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-260129

RESUMO

In this study, calcium phosphate cements (CPC) were prepared by mixing cement powders of tetracalcium phosphate (TTCP) with a cement liquid of phosphate acid saline solution. Tetracycline (TTC)-CPC, chitosan-CPC and chitosan-TTC-CPC were investigated with different premixed schedule. It was demonstrate that both TTC and chitosan worked on the phase transition and crystal characteristics. TTCP mixed with phosphate acid saline solution had similar features of Fourier transform-infrared spectrometry (FT-IR) no matter it was mixed with chitosan or TTC or both. TTC premixed with cement liquid or powder had significant different features of FT-IR and 876 cm-1seemed to be a special peak for TTC when TTC was premixed with cement liquid. This was also supported by XRD analysis, which showed that TTC premixed with cement liquid improved phase transition of TTCP to OCP. Chitosan, as organic additive, regulates the regular crystal formation and inhibits the phase transition of TTCP to OCP, except when it is mingled with cement liquid premixed with TTC in field scanning electron microscope. It was concluded that the premixed schedule influences the crystal formation and phase transition, which may be associated with its biocompatibility and bioactivities in vivo.

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