Genome to phenome tools: In vivo and in vitro transfection of Crassostrea virginica hemocytes

2020 ◽  
Vol 103 ◽  
pp. 438-441
Author(s):  
Adrienne N. Tracy ◽  
Raghavendra Yadavalli ◽  
Kiara S. Reed ◽  
Rahul Parnaik ◽  
Nicole J. Poulton ◽  
...  
Keyword(s):  
Crassostrea Virginica ◽  
In Vitro Transfection

scholarly journals Chitosan-miRNA functionalized microporous titanium oxide surfaces via a layer-by-layer approach with a sustained release profile for enhanced osteogenic activity

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Kaimin Wu ◽  
Mengyuan Liu ◽  
Nan Li ◽  
Li Zhang ◽  
Fanhui Meng ◽  
...  
Keyword(s):  
Sustained Release ◽  
Osteogenic Differentiation ◽  
Sodium Hyaluronate ◽  
Release Profile ◽  
Titanium Implants ◽  
Layer By Layer ◽  
Matrix Mineralization ◽  
In Vitro Transfection

Abstract Background The biofunctionalization of titanium implants for high osteogenic ability is a promising approach for the development of advanced implants to promote osseointegration, especially in compromised bone conditions. In this study, polyelectrolyte multilayers (PEMs) were fabricated using the layer-by-layer approach with a chitosan-miRNA (CS-miRNA) complex and sodium hyaluronate (HA) as the positively and negatively charged polyelectrolytes on microarc-oxidized (MAO) Ti surfaces via silane-glutaraldehyde coupling. Methods Dynamic contact angle and scanning electron microscopy measurements were conducted to monitor the layer accumulation. RiboGreen was used to quantify the miRNA loading and release profile in phosphate-buffered saline. The in vitro transfection efficiency and the cytotoxicity were investigated after seeding mesenchymal stem cells (MSCs) on the CS-antimiR-138/HA PEM-functionalized microporous Ti surface. The in vitro osteogenic differentiation of the MSCs and the in vivo osseointegration were also evaluated. Results The surface wettability alternately changed during the formation of PEMs. The CS-miRNA nanoparticles were distributed evenly across the MAO surface. The miRNA loading increased with increasing bilayer number. More importantly, a sustained miRNA release was obtained over a timeframe of approximately 2 weeks. In vitro transfection revealed that the CS-antimiR-138 nanoparticles were taken up efficiently by the cells and caused significant knockdown of miR-138 without showing significant cytotoxicity. The CS-antimiR-138/HA PEM surface enhanced the osteogenic differentiation of MSCs in terms of enhanced alkaline phosphatase, collagen production and extracellular matrix mineralization. Substantially enhanced in vivo osseointegration was observed in the rat model. Conclusions The findings demonstrated that the novel CS-antimiR-138/HA PEM-functionalized microporous Ti implant exhibited sustained release of CS-antimiR-138, and notably enhanced the in vitro osteogenic differentiation of MSCs and in vivo osseointegration. This novel miRNA-functionalized Ti implant may be used in the clinical setting to allow for more effective and robust osseointegration.

scholarly journals Exploitation of De Novo Helper-Lipids for Effective Gene Delivery.

2009 ◽  
Vol 11 (4) ◽  
pp. 56 ◽  
Author(s):  
Tomoaki Kurosaki ◽  
Takashi Kitahara ◽  
Mugen Teshima ◽  
Koyo Nishida ◽  
Junzo Nakamura ◽  
...  
Keyword(s):  
Gene Delivery ◽  
De Novo ◽  
Transfection Efficiency ◽  
The Other ◽  
Penetration Enhancers ◽  
Cell Toxicity ◽  
In Vivo Transfection ◽  
In Vitro Transfection

Purpose: In gene delivery, a fusogenic lipid such as dioleyl phosphatidylethanolamine (DOPE) which is a component of cationic liposomal vector is important factor for effective transfection efficiency. We investigated the effect of penetration enhancers as alternative helper-lipids to DOPE. Methods: Transdermal penetraion enhancers such as N-lauroylsarcosine (LS), (R)-(+)-limonene (LM), vitamin E (VE), and phosphatidyl choline from eggs (EggPC) were used in this experiments as helper-lipids with N-[1-(2, 3-dioleyloxy) propyl]-N, N, N-trimethlylammonium chloride (DOTMA) and cholesterol (CHOL). We examined in vitro transfection efficiency, cytotoxicity, hematotoxicity, and in vivo transfection efficiency of plasmid DNA/cationic liposomes complexes. Results: In transfection experiments in vitro, the cationic lipoplexes containing LS had highest transfection efficiency among the other lipoplexes independently of FBS. Furthermore, the lipoplexes containing LS had lowest cell toxicity among the other lipoplexes in the presence of FBS. As the results of erythrocytes interaction experiment, DOTMA/LS/CHOL, DOTMA/VE/CHOL, and DOTMA/EggPC/CHOL lipoplexes showed extremely lower hematotoxicity. On the basis of these results, the in vivo transfection efficiencies of the lipoplexes were examined. The lipoplexes containing LS had the highest transfection activity among the other lipoplexes. Conclusion: In conclusion, several transdermal penetration enhancers are available for alternative helper-lipids to DOPE in cationic liposomal vectors. Among them, DOTMA/LS/CHOL lipoplexes showed superior characteristics in in vitro transfection efficiency, cell toxicity, hematotoxicity, and in vivo transfection efficiency.

scholarly journals Type X collagen gene regulation by Runx2 contributes directly to its hypertrophic chondrocyte–specific expression in vivo

2003 ◽  
Vol 162 (5) ◽  
pp. 833-842 ◽  
Author(s):  
Qiping Zheng ◽  
Guang Zhou ◽  
Roy Morello ◽  
Yuqing Chen ◽  
Xavier Garcia-Rojas ◽  
...  
Keyword(s):  
Binding Sites ◽  
Genomic Analysis ◽  
Comparative Genomic ◽  
Collagen Gene ◽  
Runt Domain ◽  
Specific Expression ◽  
Hypertrophic Chondrocyte ◽  
In Vitro Transfection

The α1(X) collagen gene (Col10a1) is the only known hypertrophic chondrocyte–specific molecular marker. Until recently, few transcriptional factors specifying its tissue-specific expression have been identified. We show here that a 4-kb murine Col10a1 promoter can drive β-galactosidase expression in lower hypertrophic chondrocytes in transgenic mice. Comparative genomic analysis revealed multiple Runx2 (Runt domain transcription factor) binding sites within the proximal human, mouse, and chick Col10a1 promoters. In vitro transfection studies and chromatin immunoprecipitation analysis using hypertrophic MCT cells showed that Runx2 contributes to the transactivation of this promoter via its conserved Runx2 binding sites. When the 4-kb Col10a1 promoter transgene was bred onto a Runx2+/− background, the reporter was expressed at lower levels. Moreover, decreased Col10a1 expression and altered chondrocyte hypertrophy was also observed in Runx2 heterozygote mice, whereas Col10a1 was barely detectable in Runx2-null mice. Together, these data suggest that Col10a1 is a direct transcriptional target of Runx2 during chondrogenesis.

scholarly journals Effect of Cationic Lipid Type in Folate-PEG-Modified Cationic Liposomes on Folate Receptor-Mediated siRNA Transfection in Tumor Cells

Pharmaceutics ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 181 ◽  
Author(s):  
Yoshiyuki Hattori ◽  
Satono Shimizu ◽  
Kei-ichi Ozaki ◽  
Hiraku Onishi
Keyword(s):  
Gene Silencing ◽  
Tumor Cells ◽  
Folate Receptor ◽  
Cationic Lipid ◽  
Intratumoral Injection ◽  
Cationic Liposomes ◽  
Kb Cells ◽  
In Vitro Transfection

In this study, we examined the effect of cationic lipid type in folate (FA)-polyethylene glycol (PEG)-modified cationic liposomes on gene-silencing effects in tumor cells using cationic liposomes/siRNA complexes (siRNA lipoplexes). We used three types of cationic cholesterol derivatives, cholesteryl (3-((2-hydroxyethyl)amino)propyl)carbamate hydroiodide (HAPC-Chol), N-(2-(2-hydroxyethylamino)ethyl)cholesteryl-3-carboxamide (OH-Chol), and cholesteryl (2-((2-hydroxyethyl)amino)ethyl)carbamate (OH-C-Chol), and we prepared three types of FA-PEG-modified siRNA lipoplexes. The modification of cationic liposomes with 1–2 mol % PEG-lipid abolished the gene-silencing effect in human nasopharyngeal tumor KB cells, which overexpress the FA receptor (FR). In contrast, FA-PEG-modification of cationic liposomes restored gene-silencing activity regardless of the cationic lipid type in cationic liposomes. However, the optimal amount of PEG-lipid and FA-PEG-lipid in cationic liposomes for selective gene silencing and cellular uptake were different among the three types of cationic liposomes. Furthermore, in vitro transfection of polo-like kinase 1 (PLK1) siRNA by FA-PEG-modified liposomes exhibited strong cytotoxicity in KB cells, compared with PEG-modified liposomes; however, in in vivo therapy, intratumoral injection of PEG-modified PLK1 siRNA lipoplexes inhibited tumor growth of KB xenografts, as well as that of FA-PEG-modified PLK1 siRNA lipoplexes. From these results, the optimal formulation of PEG- and FA-PEG-modified liposomes for FR-selective gene silencing might be different between in vitro and in vivo transfection.

scholarly journals Hypoglycemic and Hypolipidemic Effects of a Novel Pan-Peroxisome Proliferator-Activated Receptor Agonist, MBT1805, in db/db Mices

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Wang C ◽  
◽  
Jin X ◽  
Jin Q ◽  
Shi Y ◽  
...  
Keyword(s):  
Weight Gain ◽  
Peroxisome Proliferator ◽  
The Novel ◽  
Peroxisome Proliferator Activated Receptor ◽  
Ppar Agonist ◽  
Hepatocyte Damage ◽  
In Vitro Transfection

Background: MBT1805 is a novel pan-Peroxisome Proliferator-Activated Receptor (PPAR) agonist. Materials and Methods: In vitro, transfection and luciferase assays tested EC50 values of MBT1805. In vivo, hypoglycemic and hypolipidemic effects of MBT1805 were observed in db/db mice compared with Rosiglitazone. Results: In vitro, MBT1805 activates human PPARα, PPARγ and PPARδ with EC50 values of 8.46μM, 11.94μM, 11.15μM, respectively. Results showed that the bodyweight of db/db mice treated with MBT1805 was not changed. By contrast, Rosiglitazone-treated mice showed significant weight gain (p<0.05). MTB1805 decreased blood glucose level without causing noticeable hepatocytes damage. Conclusion: The novel balanced pan-PPAR agonist, MBT1805 has moderate hypoglycemic and hypolipidemic effects, and does not cause weight gain, hepatocyte damage and hepatic lipid deposition. These experimental results indicate that MBT1805 is safe in the treatment of type 2 diabetes.

Lipoplexes Formulation and Optimisation: In Vitro Transfection Studies Reveal No Correlation with In Vivo Vaccination Studies

2010 ◽  
Vol 7 (2) ◽  
pp. 175-187 ◽  
Author(s):  
Sarah E. McNeil ◽  
Anil Vangala ◽  
Vincent W. Bramwell ◽  
Peter J. Hanson ◽  
Yvonne Perrie
Keyword(s):  
In Vitro Transfection

Induction and Differentiation of Dental Epithelium in Vivo and in Vitro

1982 ◽  
Vol 40 ◽  
pp. 142-145
Author(s):  
E. J. Kollar
Keyword(s):  
Connective Tissue ◽  
Oral Mucosa ◽  
Basal Lamina ◽  
Functional Derivatives ◽  
Specific Source ◽  
Dental Epithelium ◽  
Connective Tissue Stroma

The differentiation and maintenance of many specialized epithelial structures are dependent on the underlying connective tissue stroma and on an intact basal lamina. These requirements are especially stringent in the development and maintenance of the skin and oral mucosa. The keratinization patterns of thin or thick cornified layers as well as the appearance of specialized functional derivatives such as hair and teeth can be correlated with the specific source of stroma which supports these differentiated expressions.

Ultrastructural Correlations between Clonal Human Tumor Colonies and in Vivo Neoplasms

1982 ◽  
Vol 40 ◽  
pp. 210-211
Author(s):  
M.J. Murphy ◽  
R.R. Price ◽  
J.C. Sloman
Keyword(s):  
Cultured Cells ◽  
Human Tumor ◽  
Cell Type ◽  
Tumor Mass ◽  
Initial Cell ◽  
Cloning Assay ◽  
Human Tumor Cloning ◽  
The Relationship

The in vitro human tumor cloning assay originally described by Salmon and Hamburger has been applied recently to the investigation of differential anti-tumor drug sensitivities over a broad range of human neoplasms. A major problem in the acceptance of this technique has been the question of the relationship between the cultured cells and the original patient tumor, i.e., whether the colonies that develop derive from the neoplasm or from some other cell type within the initial cell population. A study of the ultrastructural morphology of the cultured cells vs. patient tumor has therefore been undertaken to resolve this question. Direct correlation was assured by division of a common tumor mass at surgical resection, one biopsy being fixed for TEM studies, the second being rapidly transported to the laboratory for culture.

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