Human Hair Keratin Composite Scaffold: Characterisation and Biocompatibility Study on NIH 3T3 Fibroblast Cells

The aim of this study was to transform human hair keratin waste into a scaffold for soft tissue engineering to heal wounds. The keratin was extracted using the Shindai method. Keratin and polyvinyl alcohol (PVA) was cross-linked with alginate dialdehyde and converted into a scaffold by the freeze-drying method using gentamycin sulphate (GS) as a model drug. The scaffold was subjected to Fourier transform infrared spectra (FTIR), swelling index, porosity, water absorption, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), X-ray diffraction (XRD), drug release, and cell viability (MTT) analysis. The scaffold was tested for keratinocyte growth using the murine fibroblast cell line (NIH 3T3 cells). The outcome from the keratin had a molecular weight band between 52–38 kDa in SDS-PAGE (Sodium dodecylsulfate-Polyacrylamide gel electrophoresis). A porous scaffold was capable of water absorption (73.64 ± 14.29%), swelling ability (68.93 ± 1.33%), and the release of GS shown as 97.45 ± 4.57 and 93.86 ± 5.22 of 1:4 and 1:3 scaffolds at 16 h. The physicochemical evaluation revealed that the prepared scaffold exhibits the proper structural integrity: partially crystalline with a strong thermal property. The scaffold demonstrated better cell viability against the murine fibroblast cell line (NIH 3T3 cells). In conclusion, we found that the prepared composite scaffold (1:4) can be used for wound healing applications.

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Comparative study of a new composite biomaterial fluor-hydroxyapatite on fibroblast cell line NIH-3T3 by direct test

Biologia ◽

10.2478/s11756-008-0043-x ◽

2008 ◽

Vol 63 (2) ◽

Cited By ~ 6

Author(s):

Marica Theiszová ◽

Soňa Jantová ◽

Silvia Letašiová ◽

Ľuboš Valík ◽

Martin Palou

Keyword(s):

Comparative Study ◽

Cell Line ◽

Dna Content ◽

Fibroblast Cell ◽

Cell Number ◽

Fibroblast Cell Line ◽

3T3 Cells ◽

End Points ◽

Basal Cytotoxicity ◽

Nih 3T3

AbstractThe number of biomaterials used in biomedical applications has rapidly increased in the past two decades. Fluorapatite (FA) is one of the inorganic constituents of bone or teeth used for hard tissue repairs and replacements. Fluor-hydroxyapatite (FHA) is a new synthetically prepared composite that in its structure contains the same molecular concentration of OH− groups and F− ions. The aim of this experimental investigation was to use the embryonal mouse fibroblast cell line NIH-3T3 for comparative study of basal cytotoxicity of fluoridated biomaterials FHA and FA discs. Hydroxyapatite (HA) disc, high-density polyethylene as negative control and polyvinyl chloride (PVC) containing organotin stabilizer as positive control were used as standard biomaterials. The appropriateness of the use of NIH-3T3 cells and their sensitivity for tested biomaterials were evaluated on the basis of five cytotoxic end points: cell proliferation, cell morphology, lactate dehydrogenase (LDH) released, protein and DNA cell content. The basal cytotoxicity of FHA, FA and HA discs was measured by direct contact method. FHA composite, FA and HA demonstrated in cell line NIH-3T3 nearly similar basal cytotoxicity increasing with the time of treatment. After 72 h of biomaterials treatment, about 25% inhibition of cell number, unchanged morphology of dividing cells, 6.31–0.16% increase of released LDH, about 10% inhibition of cell protein content and about 20% inhibition of DNA content was found. On the other hand, from the growth rates it resulted that NIH-3T3 cells, affected by tested biomaterials, divided about 20% slowlier than the control (untreated cells). Using the linear regression analysis we found out that deviations in measurements of cytotoxicity by four methods were as follows: less than 10% for cell number, protein and DNA content methods and 12.4% for released LDH method. Based on a good correlation of the cytotoxicity of biomaterials obtained from all end points we could conclude that fibroblast NIH-3T3 cell line was appropriate for measuring the basal cytoxicity of tested biomaterials.

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MicroRNA-27a-3p promotes proliferation and migration by up-regulation of perlecan in murine fibroblast cell line NIH/3T3

Archives of Medical Science ◽

10.5114/aoms/97204 ◽

2021 ◽

Author(s):

Jin Wang ◽

Qingyue Ma ◽

Wenquan Pang ◽

Shangzhi Li ◽

Yunwen Zou ◽

...

Keyword(s):

Cell Viability ◽

Fibroblast Cell ◽

3T3 Cells ◽

Expression Levels ◽

Gene Assay ◽

And Migration ◽

Skin Scar ◽

Nih 3T3 ◽

Proliferation And Migration ◽

Nih 3T3 Cells

IntroductionSkin scar is a common cutaneous complication, the outcome of which is unpleasant. Several microRNAs (miRs) participate in the process of skin scar formation. We aimed to explore the role of miR-27a-3p in NIH/3T3 mouse fibroblasts as well as the downstream protein and signaling cascades.Material and methodsmiR-27a-3p was aberrantly expressed in NIH/3T3 cells, followed by measurements of cell viability, migration and expressions of proteins related to proliferation and migration. Perlecan expression in cells aberrantly expressing miR-27a-3p was examined by Western blot analysis. Reporter gene assay was conducted to assess the relationship between miR-27a-3p and perlecan. Then, whether miR-27a-3p affected NIH/3T3 cells through regulating perlecan was ascertained. The effects of aberrantly expressed miR-27a-3p and perlecan on expression levels of VEGF, bFGF and key kinases in the MAPK/ERK and the PI3K/AKT pathways were detected.ResultsCell viability and migration were enhanced and protein expression levels of Cyclin D1, MMP-2 and MMP-9 were up-regulated by miR-27a-3p overexpression in NIH/3T3 cells. Then, we found that perlecan was positively correlated with miR-27a-3p expression, and its knockdown abrogated the effects of miR-27a-3p overexpression on NIH/3T3 cells. Finally, we found that the expression levels of VEGF and bFGF as well as phosphorylated levels of MAPK, ERK, PI3K and AKT were increased by miR-27a-3p overexpression, and those increases were reversed by perlecan knockdown.ConclusionsmiR-27a-3p promotes proliferation and migration of NIH/3T3 cells through up-regulating perlecan expression. Meanwhile, miR-27a-3p up-regulates expression levels of VEGF and bFGF, and activates MAPK/ERK and PI3K/AKT pathways through up-regulating perlecan expression.

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Effect of heat treatment on cytotoxicity of self-adhesive resin cements: Cell viability analysis

European Journal of Dentistry ◽

10.4103/ejd.ejd_34_18 ◽

2018 ◽

Vol 12 (02) ◽

pp. 281-286 ◽

Cited By ~ 3

Author(s):

Celso Afonso Klein-Júnior ◽

Roberto Zimmer ◽

Guilherme Scotta Hentschke ◽

Denise Cantarelli Machado ◽

Rubem Beraldo dos Santos ◽

...

Keyword(s):

Heat Treatment ◽

Cell Viability ◽

Fibroblast Cell ◽

Resin Cements ◽

Adhesive Resin ◽

Viability Analysis ◽

3T3 Fibroblasts ◽

Mixing Control ◽

Nih 3T3

ABSTRACT Objective: The aim of the study was to assess, in vitro, the influence on cytotoxicity of heat treatment applied before photopolymerization, while mixing three self-adhesive resin cements, in an NIH/3T3 fibroblast cell culture, based on cell viability measures. Methods: Samples were divided into three groups: (1) no heat treatment while mixing (control), (2) 37°C, and (3) 60°C heat treatment while mixing. Cements were light-cured immediately after mixing and immersed in Dulbecco's Modified Eagle Media for the extraction of possibly uncured products after 24 h and 7 days. Cultures contained 0.5 mL of NIH/3T3 fibroblasts per well at a concentration of 0.4 × 105 cells/mL and specific extracts for each sample. Statistical Analysis Used: Data were statistically analyzed with ANOVA and post hoc Student–Newman–Keuls (significance of 5%). Results: Cement cytotoxicity increased with time, as shown by the higher values observed at 7 days. There was a slight difference in intragroup cytotoxicity levels between 24 h and 7 days. Heat treatment at 60°C was associated with a major decrease in cytotoxicity levels in all three groups, both at 24 h and at 7 days, with no differences among the cements. Conclusions: Heat treatment at 60°C should be considered as a strategy to reduce cytotoxicity of self-adhesive resin cements, as evidenced by the results observed at 24 h and 7 days of analysis.

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Reactivity to TGF-β is one step towards transformation in a murine fibroblast cell line

International Journal of Cancer ◽

10.1002/ijc.2910510124 ◽

1992 ◽

Vol 51 (1) ◽

pp. 139-143

Author(s):

Michaela Götschl ◽

Petra Höfler ◽

Georg Bauer

Keyword(s):

Cell Line ◽

Fibroblast Cell ◽

Fibroblast Cell Line ◽

Murine Fibroblast ◽

One Step

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Chemical constituents of Eremomastax speciosa (Hochst.) Cufod leaves and its cytotoxic potential on NIH-3T3 cells

Bulletin of the Chemical Society of Ethiopia ◽

10.4314/bcse.v34i3.18 ◽

2021 ◽

Vol 34 (3) ◽

pp. 633-640

Author(s):

M. O. Eve ◽

T. N. Alfred ◽

I. I. Akripo ◽

E. E. Ubana ◽

I. M. Choudhary

Keyword(s):

Fatty Acid ◽

Chemical Constituents ◽

Acid Ethyl Ester ◽

Fibroblast Cell ◽

Fatty Acid Esters ◽

Gas Chromatography Mass Spectrometry ◽

3T3 Cells ◽

Nih 3T3 ◽

Acid Esters ◽

Nih 3T3 Cells

This study aimed at assessing the cytotoxicity of Eremomastax speciosa crude extract on NIH-3T3 fibroblast cell lines and reporting the chemical constituents in the extract. The MTT assay on NIH-3T3 cells showed a significantly lower (p < 0.05) inhibition from E. speciosa (IC50 > 30 µg/mL) compared to cyclohexamide (IC50 > 0.8 µg/mL). This result validates the non-toxicity observed with the use of E. speciosa on normal cells at low to moderate doses. Four compounds were isolated and identified from their EIMS as well as 1D and 2D NMR spectroscopic data namely hydroxyandrographolide (1), stigmasterol glucoside (2), (Z)-4-coumaric acid 4-O-β-D-apiofuranosyl-(1’’→2’)-O-β-D-glucopyranoside (3) and 5-methoxy-4,4′-di-O-methyl- secolariciresinol-9′-monoacetate (4). These compounds are isolated from this species for the first time. Thirteen volatile constituents were detected in the extract using gas chromatography mass spectrometry (GC-MS). Besides 6,10,14-trimethy-2-pentadecanone (12.63%), mostly fatty acid esters were detected in high amounts notably ethyl hexadecanoate (16.00%), ethyl-9,12,15-octadecatrienoate (11.51%) and 9,12-octadecadienoic acid ethyl ester (8.05%). This study revealed many unsaturated fatty acid esters in E. speciosa and is noteworthy that ω-3 and ω-6 fatty acid esters were predominant, hence an added nutritional value to this plant. KEY WORDS: Eremomastax speciosa, Secondary metabolites, NIH-3T3 cytotoxicity, NMR, GC-MS Bull. Chem. Soc. Ethiop. 2020, 34(3), 633-640. DOI: https://dx.doi.org/10.4314/bcse.v34i3.18

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