Impedance testing of porous Si3N4 scaffolds for skeletal implant applications

Abstract Si3N4 ceramics show excellent characteristics of mechanical and chemical resistance in combination with good biocompatibility, antibacterial property and radiolucency. Therefore, they are intensively studied as structural materials in skeletal implant applications. Despite their attractive properties, there are limited data in the field about in vitro studies of cellular growth on ceramic implant materials. In this study, the growth of bone cells was investigated on porous silicon nitride (Si3N4) ceramic implant by using electrochemical impedance spectroscopy (EIS). Partial sintering was performed at 1700 °C with limited amount of sintering additive for the production of porous Si3N4 scaffolds. All samples were then sterilized by using ethylene oxide followed by culturing MG-63 osteosarcoma cells on the substrates for in vitro assays. At 20 and 36 h, EIS was performed and results demonstrated that magnitude of the impedance as a result of the changes in the culture medium increased after incubation with osteosarcoma cells. The changes are attributed to the cellular uptake of charged molecules from the medium. Si3N4 samples appear to show large impedance magnitude changes, especially between 100 and 1 Hz. Impedance changes were also correlated with WST-1 measurements (36 h) and DAPI results.

Download Full-text

Targeting Clusterin Induces Apoptosis, Reduces Growth Ability and Invasion and Mediates Sensitivity to Chemotherapy in Human Osteosarcoma Cells

Current Pharmaceutical Biotechnology ◽

10.2174/1389201020666190821151120 ◽

2020 ◽

Vol 21 (2) ◽

pp. 131-139 ◽

Cited By ~ 3

Author(s):

Xiaohui Wang ◽

Ying Yu ◽

Lingna Zang ◽

Peng Zhang ◽

Jinfeng Ma ◽

...

Keyword(s):

Clinicopathological Features ◽

Cellular Growth ◽

Chemotherapeutic Drug ◽

Osteosarcoma Cells ◽

Human Osteosarcoma ◽

Response To Chemotherapy ◽

Human Osteosarcoma Cells ◽

U2os Cells ◽

High Grade Osteosarcoma

Objective: The aim of the study was to investigate the expression of sCLU in relation to the clinicopathological features and prognosis of patients with untreated High-Grade Osteosarcoma (HGOS) and to evaluate sCLU as a target for osteosarcoma (OS) therapies. Methods: The expression of sCLU in 98 patients of HGOS enrolled from April 2005 to March 2015 at the affiliated hospital of Qingdao University was evaluated by immunohistochemistry. The sCLU expression, clinical data and survival were compared. siRNA-mediated sCLU gene silencing on cell apoptosis, viability, invasion and chemosensitivity to doxorubicin in U2OS cells in vitro was evaluated. Results: sCLU expression was found in 59 (60%) of the 98 patients. A positive correlation was observed between sCLU expression and metastatic disease (P = 0.036) and a negative correlation between sCLU expression and response to chemotherapy (P = 0.002). Targeting sCLU expression in U2OS cells induced significant reduction in cellular growth and higher rates of spontaneous endogenous apoptosis. In addition, targeting sCLU expression inhibited the invasion of U2OS cells. Furthermore, targeting sCLU expression significantly sensitized to chemotherapeutic drug, doxorubicin. Conclusions: The overexpression of sCLU was significantly correlated with metastasis and chemosensitivity in patients with HGOS. sCLU may be a promising therapeutic or chemopreventive target for human OS treatment.

Download Full-text

Thyroid hormone, vitamin D and retinoid receptor expression and signalling in primary cultures of rat osteoblastic and immortalised osteosarcoma cells

Journal of Endocrinology ◽

10.1677/joe.0.1540063 ◽

1997 ◽

Vol 154 (1) ◽

pp. 63-74 ◽

Cited By ~ 24

Author(s):

R Bland ◽

R L Sammons ◽

M C Sheppard ◽

G R Williams

Keyword(s):

Vitamin D ◽

Retinoic Acid ◽

Bone Cells ◽

Primary Cultures ◽

Receptor Expression ◽

Receptor Protein ◽

Osteosarcoma Cell ◽

Osteosarcoma Cells ◽

Retinoid Receptor

Abstract 3,5,3′-Tri-iodothyronine (T3), 1α,25(OH)2-vitamin D3 (D3) and retinoids activate related nuclear receptors which interact by heterodimerisation to regulate gene expression. Actions of each hormone are discrete and may be specified by changes in the relative concentrations of their receptors (T3R, vitamin D receptor (VDR), retinoic acid receptor (RAR), retinoid X receptor (RXR)). T3, D3 and retinoids are essential for skeletal development and maintenance and we have previously shown complex interactions amongst their signalling pathways in osteosarcoma cells. In these studies we demonstrate that similar T3R, VDR, RAR and RXR proteins are co-expressed in both osteoblast lineage cell primary cultures and osteosarcoma cells by Western blotting. We investigated whether hormone interactions in bone result from changes in receptor stoichiometry. Cells were treated with combinations of T3, D3, 9-cis retinoic acid (9-cis RA) and all-trans retinoic acid (RA) that are known from previous studies to produce complex cell specific responses. No alteration in expression of any receptor protein was seen in response to any hormone combination in three phenotypically distinct osteosarcoma cell lines. Thus, in contrast to studies of overexpressed receptors in vitro, changes in the physiological concentrations of endogenous T3R, VDR, RAR and RXR do not specify discrete hormone actions in osteoblastic cells. Other unidentified factors are likely to modulate hormone action in these bone cells. Journal of Endocrinology (1997) 154, 63–74

Download Full-text

A Practical Approach for the In Vitro Safety and Efficacy Assessment of an Anti-Ageing Cosmetic Cream Enriched with Functional Compounds

Molecules ◽

10.3390/molecules26247592 ◽

2021 ◽

Vol 26 (24) ◽

pp. 7592

Author(s):

Nicola Zerbinati ◽

Sabrina Sommatis ◽

Cristina Maccario ◽

Serena Di Francesco ◽

Maria Chiara Capillo ◽

...

Keyword(s):

3D Models ◽

Dermal Fibroblasts ◽

In Vitro Assays ◽

In Vitro Tests ◽

Efficacy Assessment ◽

Normal Human ◽

Good Biocompatibility ◽

Anti Inflammatory ◽

Inflammatory Effect

(1) Background: Cosmeceuticals are topical products applied to human skin to prevent skin ageing and maintain a healthy skin appearance. Their effectiveness is closely linked to the compounds present in a final formulation. In this article, we propose a panel of in vitro tests to support the efficacy assessment of an anti-ageing cream enriched with functional compounds. (2) Methods: biocompatibility and the irritant effect were evaluated on reconstructed human epidermis (RHE) and corneal epithelium (HCE) 3D models. After a preliminary MTT assay, normal human dermal fibroblasts (NHDF) and keratinocytes (HaCaT) were used to evaluate the extracellular matrix (ECM) protein synthesis, and interleukin-6 (IL-6) and metalloproteinase-1 (MMP-1) production. (3) Results: data collected showed good biocompatibility and demonstrated the absence of the irritant effect in both 3D models. Therefore, we demonstrated a statistical increase in collagen and elastin productions in NHDF cells. In HaCaT cells, we highlighted an anti-inflammatory effect through a reduction in IL-6 levels in inflammatory stimulated conditions. Moreover, the reduction of MMP-1 production after UV-B radiation was demonstrated, showing significant photo-protection. (4) Conclusion: a multiple in vitro assays approach is proposed for the valid and practical assessment of the anti-ageing protection, anti-inflammatory and biocompatible claims that can be assigned to a cosmetic product containing functional compounds.

Download Full-text

Poly(3-hydroxybutyrate): Promising biomaterial for bone tissue engineering

Acta Pharmaceutica ◽

10.2478/acph-2020-0007 ◽

2020 ◽

Vol 70 (1) ◽

pp. 1-15 ◽

Cited By ~ 3

Author(s):

Barbara Dariš ◽

Željko Knez

Keyword(s):

Tissue Engineering ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Biomedical Applications ◽

Bone Cells ◽

Drug Carriers ◽

Physico Chemical ◽

Good Biocompatibility

AbstractPoly(3-hydroxybutyrate) is a natural polymer, produced by different bacteria, with good biocompatibility and biodegradability. Cardiovascular patches, scaffolds in tissue engineering and drug carriers are some of the possible biomedical applications of poly(3-hydroxybutyrate). In the past decade, many researchers examined the different physico-chemical modifications of poly(3-hydroxybutyrate) in order to improve its properties for use in the field of bone tissue engineering. Poly(3-hydroxybutyrate) composites with hydroxyapatite and bioglass are intensively tested with animal and human osteoblasts in vitro to provide information about their biocompatibility, biodegradability and osteoinductivity. Good bone regeneration was proven when poly(3-hydroxy-butyrate) patches were implanted in vivo in bone tissue of cats, minipigs and rats. This review summarizes the recent reports of in vitro and in vivo studies of pure poly(3-hydroxy-butyrate) and poly(3-hydroxybutyrate) composites with the emphasis on their bioactivity and biocompatibility with bone cells.

Download Full-text

In vitro - studies with antler bone cells: Structure forming capacity, osteocalcin production and influence of sex steroids

Osteologie/Osteology ◽

10.1024/1019-1291.15.4.245 ◽

2006 ◽

Vol 15 (04) ◽

pp. 245-257 ◽

Cited By ~ 3

Author(s):

H. J. Rolf ◽

K. G. Wiese ◽

H. Siggelkow ◽

H. Schliephake ◽

G. A. Bubernik

Keyword(s):

Sex Steroids ◽

Bone Cells ◽

In Vitro Studies ◽

Osteocalcin Production

Download Full-text

A in Vivo Assay for Standardizing Heparin Preparations

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646810 ◽

1979 ◽

Vol 41 (03) ◽

pp. 576-582

Author(s):

A R Pomeroy

Keyword(s):

In Vitro Assays ◽

British Pharmacopoeia ◽

In Vitro Method ◽

Standard Preparation ◽

Reliable Estimation ◽

Assay Procedure ◽

Accuracy And Precision ◽

Heparin Preparation

SummaryThe limitations of currently used in vitro assays of heparin have demonstrated the need for an in vivo method suitable for routine use.The in vivo method which is described in this paper uses, for each heparin preparation, four groups of five mice which are injected intravenously with heparin according to a “2 and 2 dose assay” procedure. The method is relatively rapid, requiring 3 to 4 hours to test five heparin preparations against a standard preparation of heparin. Levels of accuracy and precision acceptable for the requirements of the British Pharmacopoeia are obtained by combining the results of 3 to 4 assays of a heparin preparation.The similarity of results obtained the in vivo method and the in vitro method of the British Pharmacopoeia for heparin preparations of lung and mucosal origin validates this in vivo method and, conversely, demonstrates that the in vitro method of the British Pharmacopoeia gives a reliable estimation of the in vivo activity of heparin.

Download Full-text

Potentially Thrombogenic Materials in Factor IX Concentrates

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647857 ◽

1975 ◽

Vol 33 (03) ◽

pp. 617-631 ◽

Cited By ~ 47

Author(s):

H. S Kingdon ◽

R. L Lundblad ◽

J. J Veltkamp ◽

D. L Aronson

Keyword(s):

Human Plasma ◽

Antithrombin Iii ◽

Factor Ix ◽

In Vitro Assays ◽

Substantial Agreement ◽

Coefficient Of Correlation

SummaryFactor IX concentrates manufactured from human plasma and intended for therapeutic infusion in man have been suspected for some time of being potentially thrombogenic. In the current studies, assays were carried out in vitro and in vivo for potentially thrombogenic materials. It was possible to rank the various materials tested according to the amount of thrombogenic material detected. For concentrates not containing heparin, there was substantial agreement between the in vivo and in vitro assays, with a coefficient of correlation of 0.77. There was no correlation between the assays for thrombogenicity and the antithrombin III content. We conclude that many presently available concentrates of Factor IX contain substantial amounts of potentially thrombogenic enzymes, and that this fact must be considered in arriving at the decision whether or not to use them therapeutically.

Download Full-text

New Platinum (II) Ternary Complexes of Formamidine and Pyrophosphate: Synthesis, Characterization and DFT Calculations and In vitro Cytotoxicity

Combinatorial Chemistry & High Throughput Screening ◽

10.2174/1386207323666200218115700 ◽

2020 ◽

Vol 23 (7) ◽

pp. 611-623

Author(s):

Ahmed A. Soliman ◽

Fawzy A. Attaby ◽

Othman I. Alajrawy ◽

Azza A.A. Abou-hussein ◽

Wolfgang Linert

Keyword(s):

Cell Line ◽

Cancer Cell ◽

Theoretical Calculations ◽

Thermal Analyses ◽

Cancer Cell Line ◽

Cellular Growth ◽

Planar Geometry ◽

Square Planar ◽

Vis Spectroscopy

Aim and Objective: Platinum (II) and platinum (IV) of pyrophosphate complexes have been prepared and characterized to discover their potential as antitumor drugs. This study was conducted to prepare and characterize new ternary platinum (II) complexes with formamidine and pyrophosphate as an antitumor candidate. Materials and Methods: The complexes have been characterized by mass, infrared, UV-Vis. spectroscopy, elemental analysis, magnetic susceptibility, thermal analyses, and theoretical calculations. They have been tested for their cytotoxicity, which was carried out using the fastcolorimetric assay for cellular growth and survival against MCF-7 (breast cancer cell line), HCT- 116 (colon carcinoma cell line), and HepG-2 (hepatocellular cancer cell line). Results: All complexes are diamagnetic, and the electronic spectral data displayed the bands due to square planar Pt(II) complexes. The optimized complexes structures (1-4) indicated a distorted square planar geometry where O-Pt-O and N-Pt-N bond angles were 82.04°-96.44°, respectively. Conclusion: The complexes showed noticeable cytotoxicity and are considered as promising antitumor candidates for further applications.

Download Full-text

In-silico Studies of Isolated Phytoalkaloid Against Lipoxygenase: Study Based on Possible Correlation

Combinatorial Chemistry & High Throughput Screening ◽

10.2174/1386207321666180220125406 ◽

2018 ◽

Vol 21 (3) ◽

pp. 215-221

Author(s):

Haroon Khan ◽

Muhammad Zafar ◽

Helena Den-Haan ◽

Horacio Perez-Sanchez ◽

Mohammad Amjad Kamal

Keyword(s):

In Silico ◽

Docking Studies ◽

In Vivo Studies ◽

In Vitro Assays ◽

Chronic Obstructive ◽

Lipoxygenase Inhibitors ◽

Lipoxygenase Inhibition ◽

In Silico Studies

Aim and Objective: Lipoxygenase (LOX) enzymes play an important role in the pathophysiology of several inflammatory and allergic diseases including bronchial asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis, rheumatoid arthritis and chronic obstructive pulmonary disease. Inhibitors of the LOX are believed to be an ideal approach in the treatment of diseases caused by its over-expression. In this regard, several synthetic and natural agents are under investigation worldwide. Alkaloids are the most thoroughly investigated class of natural compounds with outstanding past in clinically useful drugs. In this article, we have discussed various alkaloids of plant origin that have already shown lipoxygenase inhibition in-vitro with possible correlation in in silico studies. Materials and Methods: Molecular docking studies were performed using MOE (Molecular Operating Environment) software. Among the ten reported LOX alkaloids inhibitors, derived from plant, compounds 4, 2, 3 and 1 showed excellent docking scores and receptor sensitivity. Result and Conclusion: These compounds already exhibited in vitro lipoxygenase inhibition and the MOE results strongly correlated with the experimental results. On the basis of these in vitro assays and computer aided results, we suggest that these compounds need further detail in vivo studies and clinical trial for the discovery of new more effective and safe lipoxygenase inhibitors. In conclusion, these results might be useful in the design of new and potential lipoxygenase (LOX) inhibitors.

Download Full-text

Repurposing of auranofin against bacterial infections: An In silico and In vitro study

Current Computer - Aided Drug Design ◽

10.2174/1386207323666200717155640 ◽

2020 ◽

Vol 16 ◽

Author(s):

Nidhi Sharma ◽

Arti Singh ◽

Ruchika Sharma ◽

Anoop Kumar

Keyword(s):

Broad Spectrum ◽

In Silico ◽

Antibacterial Agent ◽

Bacterial Infections ◽

In Vitro Assays ◽

Infection Model ◽

Synergistic Activity ◽

Bacterial Strains ◽

Molecular Docking Study

Aim: The aim of the study was to find out the role of auranofin as a promising broad spectrum antibacterial agent. Methods: In-vitro assays (Percentage growth retardation, Bacterial growth kinetics, Biofilm formation assay) and In-silico study (Molegro virtual docker (MVD) version 6.0 and Molecular operating environment (MOE) version 2008.10 software). Results: The in vitro assays have shown that auranofin has good antibacterial activity against Gram positive and Gram negative bacterial strains. Further, auranofin has shown synergistic activity in combination with ampicillin against S. aureus and B. subtilis whereas in combination with neomycin has just shown additive effect against E. coli, P. aeruginosa and B. pumilus. In vivo results have revealed that auranofin alone and in combination with standard drugs significantly decreased the bioburden in zebrafish infection model as compared to control. The molecular docking study have shown good interaction of auranofin with penicillin binding protein (2Y2M), topoisomerase (3TTZ), UDP-3-O-[3- hydroxymyristoyl] N-acetylglucosaminedeacetylase (3UHM), cell adhesion protein (4QRK), β-lactamase (5CTN) and arylsulphatase (1HDH) enzyme as that of reference ligand which indicate multimodal mechanism of action of auranofin. Finally, MTT assay has shown non-cytotoxic effect of auranofin. Conclusion: In conclusion, auranofin in combination with existing antibiotics could be developed as a broad spectrum antibacterial agent; however, further studies are required to confirm its safety and efficacy. This study provides possibility of use of auranofin apart from its established therapeutic indication in combination with existing antibiotics to tackle the problem of resistance.

Download Full-text