Improvement of the Adhesion and Proliferation of Human Osteoblastic-Like Cells with Biomimetic Nanoapatite Coatings

2007 ◽  
Vol 330-332 ◽  
pp. 385-388 ◽  
Author(s):  
Zhi Jun Pan ◽  
Bing Gang Guan ◽  
Di Sheng Yang ◽  
Jie Feng ◽  
Wei Qi Yan
Keyword(s):  
Cell Adhesion ◽  
Cell Morphology ◽  
Laser Scanning ◽  
Cell Attachment ◽  
Confocal Laser Scanning Microscope ◽  
Biological Behavior ◽  
Confocal Laser ◽  
Analysis System ◽  
Cell Adhesion And Proliferation

Biomimetic nanoapatite coatings was developed by functionally modified methods with a combination of topographic, chemical and biomimetic treatments on the surface of titanium (Ti) substrate. The biological behavior and bioactivity of functionally modified SLA implants with chemical and biomimetic treatments (SCB-treated Ti) using body like solution were investigated to compare with untreated Ti and SLA Ti plates as controls. The cell attachment, proliferation, alkaline phosphotatse (AKP) activity, cell morphology and differentiation were evaluated by using MTT, RT-PCR, scanning electron microscopy (SEM) and confocal laser-scanning microscope (CLSM) analysis system. The results showed that the cell adhesion and proliferation was enhanced on functionalized titanium surface with nano-scale apatite compared to the controls. SEM micrographs also revealed that the osteoblast-like cells spreadly grew along the surface. Cell morphology and differentiation could be further observed distinctly by CLSM graphs. Moreover, mRNA expression of alkaline phosphotatse in nucleus on the SCB-treated Ti increased obviously on the third day compared with the controls. The in vitro results demonstrated the remarkable improvement on cell adhesion and proliferation of the biomimetic nanoapatite on SCB-treated Ti, which could be used for orthopaedic/dental implants.

scholarly journals Grape Seed Extract as a Potential Remineralizing Agent: A Comparative in vitro Study

2012 ◽  
Vol 13 (4) ◽  
pp. 425-430 ◽  
Author(s):  
Shiny Benjamin ◽  
Roshni LNU ◽  
Sabeena Susan Thomas ◽  
Mohan Thomas Nainan
Keyword(s):  
Laser Scanning ◽  
Grape Seed Extract ◽  
Grape Seed ◽  
Confocal Laser Scanning Microscope ◽  
Seed Extract ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Caries Lesions ◽  
Calcium Glycerophosphate

ABSTRACT Objective Remineralization is an effective treatment that may stop or reverse early tooth decay. Grape seed extract (GSE) is the potential remineralizing agent under investigation. Materials and methods Sound human tooth sections were obtained from the cervical portion of the root and stored in demineralizing solution at 37°C for 96 hours to induce artificial root caries lesions. The sections were divided into four treatment groups including 6.5% grape seed extract, sodium monofluorophosphate (220 ppm) with 0.05% calcium glycerophosphate, 0.5% calcium glycerophosphate and control (no treatment). An in vitro pH cycling model was used to cycle the demineralized specimens through treatment solutions, acidic buffer and neutral buffer for 8 days at 6 cycles per day. Subsequently, they were evaluated using confocal laser scanning microscope. Data were analyzed using analysis of variance (p < 0.05). Results GSE revealed less demineralization and more remineralization compared with other groups. Conclusion GSE promotes remineralization of artificial root caries lesions. Clinical significance The search for the perfect remineralizing agent continues to this day. GSE could be a welcome addition to the remineralization armamentarium. Abbreviations and acronyms GSE: Grape seed extract; ppm: Parts per million; CaGP: Calcium glycerophosphate; CLSM: Confocal laser scanning microscope; ANOVA: Analysis of variance; PA: Proanthocyanidin; CEJ: Cementoenamel junction; mM: Millimole; CaCl2.2H2O: Calcium chloride dihydrate; KH2PO4: Potassium dehydrate phosphate; K2HPO4: Dipotassium phosphate; dH2O: Deionized water; w/v: Weight by volume; ROD: Relative optical density; nm: Nanometer; SD: Standard deviation. How to cite this article Benjamin S, Roshni, Thomas SS, Nainan MT. Grape Seed Extract as a Potential Remineralizing Agent: A Comparative in vitro Study. J Contemp Dent Pract 2012;13(4):425-430.

Synthesis, singlet oxygen generation, photocytotoxicity and subcellular localization of azobisporphyrins as potentially photodynamic therapeutic agents in vitro cell study

2017 ◽  
Vol 21 (02) ◽  
pp. 122-127 ◽  
Author(s):  
Yunman Zheng ◽  
Sizhe Zhu ◽  
Lijun Jiang ◽  
Fengshou Wu ◽  
Chi Huang ◽  
...  
Keyword(s):  
Singlet Oxygen ◽  
Hela Cells ◽  
Cellular Localization ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Oxygen Generation ◽  
Scanning Confocal Microscopy

Three azobisporphyrins (Por1, Por2 and Por3) were synthesized by coupling two molecules of (4-nitrophenyl/pyridyl) porphyrins in the presence of KOH/butanol. The structures of porphyrins were confirmed by UV, IR, NMR and mass spectra and elemental analysis. With tetraphenylporphyrin (H2TPP) as a control, the singlet oxygen (1O[Formula: see text] generation of porphyrins was evaluated through 1,3-diphenylisobenzofuran (DPBF) method. The order of ability to generate 1O2 for three azobisporphyrins was Por 1 [Formula: see text]Por 2 > Por 3[Formula: see text] H2TPP. The photocytotoxicity and sub-cellular localization of azobisporphyrins over Hela cells were studied through MTT analysis and confocal laser scanning microscope, respectively. The results indicated Por 1 and Por 2 displayed the low dark-cytotoxicity, while Por 3 induced a concentration-dependent cytotoxicity to Hela cells with the concentration of porphyrins ranging from 1 to 100 [Formula: see text] M. With the light dose at 4 J/cm2, Por 3 killed more than 60% Hela cells at 2 [Formula: see text] M, indicating a high photocytoxicity. As seen from the laser scanning confocal microscopy images, Por 3 was mainly localized in cell membrane, while Por 1 and Por 2 do not displayed significant fluorescent emission in Hela cells. These results suggest the synthesized cationic azobisporphyrin could be used as a potential therapeutic agent for photodynamic therapy of cancers.

A Facile Synthesis of Acid-Activatable Nanoparticles for Efficient Intracellular Doxorubicin Release

2017 ◽  
Vol 46 ◽  
pp. 20-30 ◽  
Author(s):  
Cao Ming ◽  
Xiao Wan Song ◽  
Yu Jiao Zhang ◽  
Chang Zhi Xu ◽  
Peng Chen ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Hela Cells ◽  
Laser Scanning ◽  
Human Cancer ◽  
Polymeric Nanoparticles ◽  
Confocal Laser Scanning Microscope ◽  
Ph Sensitive ◽  
Confocal Laser ◽  
Cell Nuclei

pH responsive polymeric nanoparticles have emerged as a promising technology platform for targeted and controlled drug delivery in recent years. In this paper, endosomal pH-activatable doxorubicin (DOX) and core-crosslinked polymeric nanoparticles (DCNPs) were prepared and investigated for potent growth inhibition of human cancer cells in vitro. In vitro drug release studies, DOX conjugated nanoparticles with hydrazone bond showed a pH sensitive release phenomenon, that is, the releasing is significantly faster at mildly acidic condition with pH of 5.5 than that at physiological condition. Confocal laser scanning microscope (CLSM) observations revealed that DOX conjugated nanoparticles delivered and released DOX into the cytosols as well as cell nuclei of Hela cells following 6 h incubation. MTT assays demonstrated that these pH-sensitive DOX nanoparticles exhibited high antitumor effect to HeLa cells. The conjugated DOX polymeric nanoparticles may be a promising candidate as a nanoscale and pH-sensitive drug delivery vehicle for cancer therapy.

scholarly journals Mitochondrial function in vitrified versus slow-frozen murine embryos

2019 ◽  
Vol 15 (2) ◽  
pp. 150-152
Author(s):  
Razif Dasiman ◽  
Mimi-Sophia Sarbandi ◽  
Nor-Shahida Abdul Rahman ◽  
Salina Othman ◽  
Mastura Malek ◽  
...  
Keyword(s):  
Mitochondrial Function ◽  
Laser Scanning ◽  
Developmental Stages ◽  
Confocal Laser Scanning Microscope ◽  
Slow Freezing ◽  
Confocal Laser ◽  
Mitochondrial Functions ◽  
Murine Embryos ◽  
Cryopreserved Embryos

The effects of vitrification and slow-freezing on mitochondrial functions of in vitro produced murine embryos at various developmental stages were investigated using the Confocal Laser Scanning Microscope (CLSM). Oocytes were obtained from superovulated females, fertilized with sperm and cultured. Resulting 2-, 4- and 8-cell embryos were collected and cryopreserved by vitrification and slow-freezing. Mitochondria were stained with MitoTracker Red (CMXRos). Images were viewed by CLSM and analyzed using QWin SoftwareV.3. Fluorescent intensities were used to indicate viability. Results showed that mitochondrial fluorescence intensities of cryopreserved embryos were significantly lower as compared to non-cryopreserved embryos (p<0.01). Vitrification was found to be superior to slow-freezing at all developmental stages, based on mitochondrial function.

Microtubule and microfilament organization in immature, in vitro matured and in vitro fertilized prepubertal goat oocytes

Zygote ◽  
2005 ◽  
Vol 13 (2) ◽  
pp. 155-165 ◽  
Author(s):  
Esther Velilla ◽  
Elisabet Rodríguez-Gonzalez ◽  
Francesca Vidal ◽  
Maria-Teresa Paramio
Keyword(s):  
Laser Scanning ◽  
Polar Body ◽  
Germinal Vesicle ◽  
Fluorescein Isothiocyanate ◽  
Confocal Laser Scanning Microscope ◽  
Meiotic Spindle ◽  
Confocal Laser ◽  
Cortical Region ◽  
Microtubule Networks

The aim of our study was to analyse the cytoskeletal organization of prepubertal goat oocytes. Microtubule and microfilament organization during in vitro maturation of prepubertal and adult goat oocytes and presumptive zygotes of in vitro matured–in vitro fertilized (IVM-IVF) prepubertal goat oocytes were analysed. Oocytes were matured in M-199 with hormones and serum and inseminated with frozen-thawed spermatozoa. Oocytes and presumptive zygotes were treated with anti-α-tubulin antibody and fluorescein isothiocyanate (FITC)-labelled goat anti-mouse antibody to stain the microtubules. Microfilaments were localized by means of phalloidin 5 μg/ml conjugated with fluorescein isothiocyanate (FITC-phalloidin). DNA was stained with propidium iodide. Stained oocytes were observed under a confocal laser scanning microscope. At the germinal vesicle nuclear stage, microfilaments were distributed at the cortex of the oocytes. After in vitro maturation, 91.7% of metaphase II (MII) oocytes from adult goats displayed microfilaments in the cortex and within the polar body and were characterized by the presence of a microfilament thickening at the cortical region over the meiotic spindle. In prepubertal goat MII oocytes only 5.7% of oocytes displayed microfilaments at the cortex and within the polar body. After insemination, most of the zygotes displayed microfilaments distributed at the cortex. An undefined microtubular network was observed in adult and prepubertal goat oocytes at the germinal vesicle stage. After in vitro maturation, 100% of MII oocytes from adult goats displayed microtubules on the meiotic spindle and within the polar body. This pattern of distribution was observed in 71.6% of prepubertal goat oocytes. Undefined microtubule networks were present in most of the zygotes analysed. In conclusion, cytoskeletal differences were found between prepubertal and adult goat MII oocytes. Furthermore, most of the zygotes from IVM-IVF prepubertal goat oocytes displayed cytoskeletal anomalies.

scholarly journals Biofilms: Survival Mechanisms of Clinically Relevant Microorganisms

2002 ◽  
Vol 15 (2) ◽  
pp. 167-193 ◽  
Author(s):  
Rodney M. Donlan ◽  
J. William Costerton
Keyword(s):  
Medical Devices ◽  
Laser Scanning ◽  
Antimicrobial Agents ◽  
Cell Attachment ◽  
Disease Process ◽  
Open Water ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Host Immune System ◽  
Native Valve

SUMMARY Though biofilms were first described by Antonie van Leeuwenhoek, the theory describing the biofilm process was not developed until 1978. We now understand that biofilms are universal, occurring in aquatic and industrial water systems as well as a large number of environments and medical devices relevant for public health. Using tools such as the scanning electron microscope and, more recently, the confocal laser scanning microscope, biofilm researchers now understand that biofilms are not unstructured, homogeneous deposits of cells and accumulated slime, but complex communities of surface-associated cells enclosed in a polymer matrix containing open water channels. Further studies have shown that the biofilm phenotype can be described in terms of the genes expressed by biofilm-associated cells. Microorganisms growing in a biofilm are highly resistant to antimicrobial agents by one or more mechanisms. Biofilm-associated microorganisms have been shown to be associated with several human diseases, such as native valve endocarditis and cystic fibrosis, and to colonize a wide variety of medical devices. Though epidemiologic evidence points to biofilms as a source of several infectious diseases, the exact mechanisms by which biofilm-associated microorganisms elicit disease are poorly understood. Detachment of cells or cell aggregates, production of endotoxin, increased resistance to the host immune system, and provision of a niche for the generation of resistant organisms are all biofilm processes which could initiate the disease process. Effective strategies to prevent or control biofilms on medical devices must take into consideration the unique and tenacious nature of biofilms. Current intervention strategies are designed to prevent initial device colonization, minimize microbial cell attachment to the device, penetrate the biofilm matrix and kill the associated cells, or remove the device from the patient. In the future, treatments may be based on inhibition of genes involved in cell attachment and biofilm formation.

scholarly journals Evaluation of the anti-biofilm effect of poloxamer-based thermoreversible gel of silver nanoparticles as a potential medication for root canal therapy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ting Liu ◽  
Aerdake Aman ◽  
Muniremu Ainiwaer ◽  
Liang Ding ◽  
Fei Zhang ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Root Canal ◽  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Elemental Content ◽  
Periodontal Ligament Fibroblasts ◽  
Strong Activity ◽  
Thermoreversible Gel

AbstractThe purpose of this study was to design silver nanoparticles (AgNPs) poloxamer thermoreversible gel (AgNPs-PL) and investigate whether this gel could provide sustained antibacterial activity against Enterococcus faecalis (E. faecalis) in the root canal. The gels fabricated were characterized in terms of gelatin temperature, particle size, in-vitro Ag+ release, and elemental content. Cytotoxicity of AgNPs-PL on primary human periodontal ligament fibroblasts (HPDLFs) was examined by CCK-8 assay. Characterization of AgNPs-PL gel revealed that it contained particles existing as large clumps/fused aggregates of different shapes, with a mean diameter of 21.624 ± 14.689 nm, exhibited sustained release of Ag+ for 9 days, and non-toxic to HPDLFs at a low dose (4–32 μg/mL) through 24, 48, and 72 h exposures. The antibacterial effect of 16 and 32 μg/mL concentrations of AgNPs-PL was compared with blank poloxamer gel (PL) and calcium hydroxide (CH) using three methods: (I) agar counting plate, (II) scanning electron microscope (SEM) observations, and (III) confocal laser scanning microscope (CLSM) analysis. AgNPs-PL at the two doses above was more effective than PL and CH in removing E. faecalis biofilm at 1, 3, 9 days. Thus, AgNPs-PL exhibits strong activity against E. faecalis and is easy to produce, with a continuous release profile of Ag+. AgNPs-PL gel may be a candidate for a new root canal disinfection.

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