scholarly journals Use of Impedance Spectroscopy for the Characterization of In-Vitro Osteoblast Cell Response in Porous Titanium Bone Implants

Metals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1077
Author(s):  
Mercè Giner ◽  
Alberto Olmo ◽  
Miguel Hernández ◽  
Paloma Trueba ◽  
Ernesto Chicardi ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Impedance Spectroscopy ◽  
Cell Viability ◽  
Cellular Response ◽  
Stress Shielding ◽  
Porous Titanium ◽  
Titanium Implants ◽  
Size And Morphology ◽  
Porous Substrates

The use of titanium implants with adequate porosity (content, size and morphology) could solve the stress shielding limitations that occur in conventional titanium implants. Experiments to assess the cellular response (adhesion, proliferation and differentiation of osteoblasts) on implants are expensive, time-consuming and delicate. In this work, we propose the use of impedance spectroscopy to evaluate the growth of osteoblasts on porous titanium implants. Osteoblasts cells were cultured on fully-dense and 40 vol.% porous discs with two ranges of pore size (100–200 μm and 355–500 μm) to study cell viability, proliferation, differentiation (Alkaline phosphatase activity) and cell morphology. The porous substrates 40 vol.% (100–200 µm) showed improved osseointegration response as achieved more than 80% of cell viability and higher levels of Cell Differentiation by Alkaline Phosphatase (ALP) at 21 days. This cell behavior was further evaluated observing an increase in the impedance modulus for all study conditions when cells were attached. However, impedance levels were higher on fully-dense due to its surface properties (flat surface) than porous substrates (flat and pore walls). Surface parameters play an important role on the global measured impedance. Impedance is useful for characterizing cell cultures in different sample types.

scholarly journals Rapid Fabrication of Microfluidic Devices for Biological Mimicking: A Survey of Materials and Biocompatibility

Micromachines ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 346
Author(s):  
Hui Ling Ma ◽  
Ana Carolina Urbaczek ◽  
Fayene Zeferino Ribeiro de Souza ◽  
Paulo Augusto Gomes Garrido Carneiro Leão ◽  
Janice Rodrigues Perussi ◽  
...  
Keyword(s):  
Shear Stress ◽  
Cell Viability ◽  
Cellular Response ◽  
Fluid Shear Stress ◽  
Umbilical Vein ◽  
Microfluidic Devices ◽  
In Vitro Assays ◽  
Stress Effect

Microfluidics is an essential technique used in the development of in vitro models for mimicking complex biological systems. The microchip with microfluidic flows offers the precise control of the microenvironment where the cells can grow and structure inside channels to resemble in vivo conditions allowing a proper cellular response investigation. Hence, this study aimed to develop low-cost, simple microchips to simulate the shear stress effect on the human umbilical vein endothelial cells (HUVEC). Differentially from other biological microfluidic devices described in the literature, we used readily available tools like heat-lamination, toner printer, laser cutter and biocompatible double-sided adhesive tapes to bind different layers of materials together, forming a designed composite with a microchannel. In addition, we screened alternative substrates, including polyester-toner, polyester-vinyl, glass, Permanox® and polystyrene to compose the microchips for optimizing cell adhesion, then enabling these microdevices when coupled to a syringe pump, the cells can withstand the fluid shear stress range from 1 to 4 dyne cm2. The cell viability was monitored by acridine orange/ethidium bromide (AO/EB) staining to detect live and dead cells. As a result, our fabrication processes were cost-effective and straightforward. The materials investigated in the assembling of the microchips exhibited good cell viability and biocompatibility, providing a dynamic microenvironment for cell proliferation. Therefore, we suggest that these microchips could be available everywhere, allowing in vitro assays for daily laboratory experiments and further developing the organ-on-a-chip concept.

scholarly journals Biological characterization of implant surfaces - in vitro study

2015 ◽  
Vol 44 (4) ◽  
pp. 195-199 ◽  
Author(s):  
Priscilla Barbosa Ferreira Soares ◽  
Camilla Christian Gomes Moura ◽  
Huberth Alexandre da Rocha Júnior ◽  
Paula Dechichi ◽  
Darceny Zanetta-Barbosa
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Viability ◽  
Total Protein ◽  
Cell Attachment ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Serum Total Protein ◽  
Trypan Blue Exclusion ◽  
Experimental Surface

<title>Abstract</title><sec><title>Objective</title><p>Evaluate the biological performance of titanium alloys grade IV under different surface treatments: sandblasting and double etching (Experimental surface 1; Exp1, NEODENT); surface with wettability increase (Experimental surface 2; Exp2, NEODENT) on response of preliminary differentiation and cell maturation.</p></sec><sec><title>Material and method</title><p>Immortalized osteoblast cells were plated on Exp1 and Exp2 titanium discs. The polystyrene plate surface without disc was used as control group (C). Cell viability was assessed by measuring mitochondrial activity (MTT) at 4 and 24 h (n = 5), cell attachment was performed using trypan blue exclusion within 4 hours (n = 5), serum total protein and alkaline phosphatase normalization was performed at 4, 7 and 14 days (n = 5). Data were analyzed using one-way ANOVA and Tukey test.</p></sec><sec><title>Result</title><p>The values of cell viability were: 4h: C– 0.32±0.01<sup>A</sup>; Exp1– 0.34±0.08<sup>A</sup>; Exp2– 0.29±0.03<sup>A</sup>. 24h: C– 0.43±0.02<sup>A</sup>; Exp1– 0.39±0.01<sup>A</sup>; Exp2– 0.37±0.03<sup>A</sup>. The cell adhesion counting was: C– 85±10<sup>A</sup>; Exp1- 35±5<sup>B</sup>; Exp2– 20±2<sup>B</sup>. The amounts of serum total protein were 4d: C– 40±2<sup>B</sup>; Exp1– 120±10<sup>A</sup>; Exp2– 130±20<sup>A</sup>. 7d: C– 38±2<sup>B</sup>; Exp1– 75±4<sup>A</sup>; Exp2– 70±6<sup>A</sup>. 14 d: C– 100±3<sup>A</sup>; Exp1– 130±5<sup>A</sup>; Exp2– 137±9<sup>A</sup>. The values of alkaline phosphatase normalization were: 4d: C– 2.0±0.1<sup>C</sup>; Exp1– 5.1±0.8<sup>B</sup>; Exp2– 9.8±2.0<sup>A</sup>. 7d: C– 1.0±0.01<sup>C</sup>; Exp1– 5.3±0.5<sup>A</sup>; Exp2– 3.0±0.3<sup>B</sup>. 14 d: C– 4.1±0.3<sup>A</sup>; Exp1– 4.4±0.8<sup>A</sup>; Exp2– 2.2±0.2<sup>B</sup>. Different letters related to statistical differences.</p></sec><sec><title>Conclusion</title><p>The surfaces tested exhibit different behavior at dosage of alkaline phosphatase normalization showing that the Exp2 is more associated with induction of cell differentiation process and that Exp1 is more related to the mineralization process.</p></sec>

Fabrication and Bioactivity of Porous Titanium Implant

2007 ◽  
Vol 342-343 ◽  
pp. 613-616
Author(s):  
Xia Lu ◽  
Li Ang Xing ◽  
Pei Zhi Wang ◽  
Jun Fu
Keyword(s):  
Alkaline Phosphatase ◽  
Titanium Implant ◽  
Study Groups ◽  
Porous Titanium ◽  
Bone Sialoprotein ◽  
Control Group ◽  
Ha Coating ◽  
Titanium Material ◽  
Polymerase Chain

The aim of this study is to fabricate an implant framework for tissue engineering by sintering titanium beads and coating with hydroxyapatite and test its biocompatibility and bioactivity in vitro. The porous titanium with and without hydroxyapatite coating were involved in study groups. Osteoblastic proliferation, activity of alkaline phosphatase, mRNA of osteocalcin and bone sialoprotein were detected by MTT-assay, ALP test and real-time polymerase chain reaction respectively. The results indicated that the porous titanium material with/without HA coating could promote osteoblastic proliferation significantly contrast to the control group. However, only porous titanium with HA coating increased alkaline phosphatase, osteocalcin and bone sialoprotein gene expression apparently and had statistically differences with the other two groups. Abstract no. is TE-Po-044

Controlled implant/soft tissue interaction by nanoscale surface modifications of 3D porous titanium implants

Nanoscale ◽  
2015 ◽  
Vol 7 (21) ◽  
pp. 9908-9918 ◽  
Author(s):  
Elisabeth Rieger ◽  
Agnès Dupret-Bories ◽  
Laetitia Salou ◽  
Marie-Helene Metz-Boutigue ◽  
Pierre Layrolle ◽  
...  
Keyword(s):  
Surface Modification ◽  
Soft Tissue ◽  
Surface Modifications ◽  
Porous Titanium ◽  
Titanium Implants ◽  
Tissue Response ◽  
Soft Tissue Response ◽  
Modification Of Titanium

Nanoscale surface modification of titanium microbeads can control the soft tissue response in vitro and in vivo.

scholarly journals Improved Corrosion Behavior and Biocompatibility of Porous Titanium Samples Coated with Bioactive Chitosan-Based Nanocomposites

Materials ◽  
2021 ◽  
Vol 14 (21) ◽  
pp. 6322
Author(s):  
Cristina García-Cabezón ◽  
Vanda Godinho ◽  
Coral Salvo-Comino ◽  
Yadir Torres ◽  
Fernando Martín-Pedrosa
Keyword(s):  
Corrosion Resistance ◽  
Stress Shielding ◽  
Porous Titanium ◽  
Substrate Material ◽  
Titanium Implants ◽  
Potential Candidate ◽  
Protection Efficiency ◽  
Cellular Behavior ◽  
One Step ◽  
Current Densities

Porous titanium implants can be a good solution to solve the stress shielding phenomenon. However, the presence of pores compromises mechanical and corrosion resistance. In this work, porous titanium samples obtained using a space-holder technique are coated with Chitosan, Chitosan/AgNPs and Chitosan/Hydroxyapatite using only one step and an economic electrodeposition method. The coatings’ topography, homogeneity and chemical composition were analyzed. A study of the effect of the porosity and type of coating on corrosion resistance and cellular behavior was carried out. The electrochemical studies reveal that porous samples show high current densities and an unstable oxide film; therefore, there is a need for surface treatments to improve corrosion resistance. The Chitosan coatings provide a significant improvement in the corrosion resistance, but the Chitosan/AgNPs and Chitosan/HA coatings showed the highest protection efficiency, especially for the more porous samples. Furthermore, these coatings have better adherence than the chitosan coatings, and the higher surface roughness obtained favors cell adhesion and proliferation. Finally, a combination of coating and porous substrate material with the best biomechanical balance and biofunctional behavior is proposed as a potential candidate for the replacement of small, damaged bone tissues.

scholarly journals Combination therapy of cold atmospheric plasma (CAP) with temozolomide in the treatment of U87MG glioblastoma cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Eda Gjika ◽  
Sonali Pal-Ghosh ◽  
Megan E. Kirschner ◽  
Li Lin ◽  
Jonathan H. Sherman ◽  
...  
Keyword(s):  
Combination Therapy ◽  
Cell Viability ◽  
Cellular Response ◽  
Atmospheric Plasma ◽  
Treatment Modalities ◽  
Glioblastoma Cells ◽  
M Cell ◽  
Cell Viability Assay ◽  
Cold Atmospheric Plasma

Abstract Cold atmospheric plasma (CAP) technology, a relatively novel technique mainly investigated as a stand-alone cancer treatment method in vivo and in vitro, is being proposed for application in conjunction with chemotherapy. In this study, we explore whether CAP, an ionized gas produced in laboratory settings and that operates at near room temperature, can enhance Temozolomide (TMZ) cytotoxicity on a glioblastoma cell line (U87MG). Temozolomide is the first line of treatment for glioblastoma, one of the most aggressive brain tumors that remains incurable despite advancements with treatment modalities. The cellular response to a single CAP treatment followed by three treatments with TMZ was monitored with a cell viability assay. According to the cell viability results, CAP treatment successfully augmented the effect of a cytotoxic TMZ dose (50 μM) and further restored the effect of a non-cytotoxic TMZ dose (10 μM). Application of CAP in conjunction TMZ increased DNA damage measured by the phosphorylation of H2AX and induced G2/M cell cycle arrest. These findings were supported by additional data indicating reduced cell migration and increased αvβ3 and αvβ5 cell surface integrin expression as a result of combined CAP–TMZ treatment. The data presented in this study serve as evidence that CAP technology can be a suitable candidate for combination therapy with existing chemotherapeutic drugs. CAP can also be investigated in future studies for sensitizing glioblastoma cells to TMZ and other drugs available in the market.

Osteogenic differentiation of equine cord blood multipotent mesenchymal stromal cells within coralline hydroxyapatite scaffolds in vitro

2011 ◽  
Vol 24 (05) ◽  
pp. 354-362 ◽  
Author(s):  
R. J. Figueroa ◽  
T. G. Koch ◽  
D. H. Betts
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Viability ◽  
Osteogenic Differentiation ◽  
Alkaline Phosphatase Activity ◽  
Stromal Cells ◽  
Culture Medium ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Coralline Hydroxyapatite ◽  
Osteogenic Induction

SummaryObjective: To investigate the osteogenic differentiation potential of equine umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) within coralline hydro-xyapatite scaffolds cultured in osteogenic induction culture medium.Methods: Scaffolds seeded with equine CBMSC were cultured in cell expansion culture medium (control) or osteogenic induction medium (treatment). Cell viability and distribution were confirmed by the MTT cell viability assay and DAPI nuclear fluorescence staining, respectively. Osteogenic differentiation was evaluated after 10 days using reverse transcription polymerase chain reaction, alkaline phosphatase activity, and secreted osteocalcin concentration. Cell morphology and matrix deposition were assessed by scanning electron microscopy (SEM) after 14 days in culture.Results: Cells showed viability and adequate distribution within the scaffold. Successful osteogenic differentiation within the scaffolds was demonstrated by the increased expression of osteogenic markers such as Runx2, osteopontin, osteonectin, collagen IA increased levels of alkaline phosphatase activity increased osteocalcin protein secretion and bone-like matrix presence in the scaffold pores upon SEM evaluation.Clinical significance: These results demonstrate that equine CB-MSC maintain viability and exhibit osteogenic potential in coralline hydroxyapatite scaffolds when induced in vitro. Equine CB-MSC scaffold constructs deserve further investigation for their potential role as biologically active fillers to enhance bone-gap repair in the horse.

scholarly journals Combined Fluorescence-Based in Vitro Assay for the Simultaneous Detection of Cell Viability and Alkaline Phosphatase Activity during Osteogenic Differentiation of Osteoblast Precursor Cells

2020 ◽  
Vol 3 (2) ◽  
pp. 30 ◽  
Author(s):  
Sebastian Wilkesmann ◽  
Fabian Westhauser ◽  
Joerg Fellenberg
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Viability ◽  
Osteogenic Differentiation ◽  
Simultaneous Detection ◽  
Cell Number ◽  
Precursor Cells ◽  
Vitro Assay ◽  
Alp Activity ◽  
Bone Substitute Materials

Novel bone substitute materials need to be evaluated in terms of their osteogenic differentiation capacity and possible unwanted cytotoxic effects in order to identify promising candidates for the therapy of bone defects. The activity of alkaline phosphatase (ALP) is frequently quantified as an osteogenic marker, while various colorimetric assays, like MTT assay, are used to monitor cell viability. In addition, the DNA or protein content of the samples needs to be quantified for normalization purposes. As this approach is time consuming and often requires the analysis of multiple samples, we aimed to simplify this process and established a protocol for the combined fluorescence-based quantification of ALP activity and cell viability within one single measurement. We demonstrate that the fluorogenic substrate 4-methylumbelliferone-phosphate (4-MUP) and the commonly used para-nitrophenylphosphate (p-NPP) produce comparable and highly correlating results. We further show that fluorescein–diacetate (FDA) can be used to quantify both cell viability and cell number without interfering with the quantification of ALP activity. The measurement of additional normalization parameters is, therefore, unnecessary. Therefore, the presented assay allows for a time-efficient, simple and reliable analysis of both ALP activity and cell viability from one sample and might facilitate experiments evaluating the osteogenic differentiation of osteoblast precursor cells.

In vitro study of antibacterial and osteogenic activity of titanium metal releasing strontium and silver ions

2020 ◽  
Vol 35 (6) ◽  
pp. 670-680
Author(s):  
Yaichiro Okuzu ◽  
Shunsuke Fujibayashi ◽  
Seiji Yamaguchi ◽  
Kazutaka Masamoto ◽  
Bungo Otsuki ◽  
...  
Keyword(s):  
Surface Treatment ◽  
Cellular Response ◽  
Low Cost ◽  
In Vitro Study ◽  
Silver Ions ◽  
Titanium Implants ◽  
Dental Surgery ◽  
Osteogenic Response

Peri-prosthetic infection and loosening of implants are major problems in orthopaedic and dental surgery. To address these issues, surface treatment methods for titanium implants have been improved by modifying the alkali and heat treatment. We have previously fabricated calcium-treated Ti metal that releases Sr ions (CaSr-Ti), which resulted in a higher in vitro osteogenic response and early in vivo bone bonding. Further, we developed a Ti metal that released both Sr and Ag ions (CaSrAg-Ti). In this study, we evaluated the antibacterial ability and osteogenic cellular response of CaSrAg-Ti and CaSr-Ti in vitro using rat bone marrow stromal cells (BMSCs) cultured on implant samples and extract mediums (EMs) made by immersing the implant samples in the medium. CaSrAg-Ti did not show cytotoxicity and was associated with a slightly higher osteogenic response when compared to CaSr-Ti, without inhibiting the effect of Sr. The osteogenic response was also observed in the cells cultured with the CaSrAg-Ti EM; however, the response was not as high as that of the cells on the CaSrAg-Ti implant sample. Significantly higher antibacterial activity was observed along with an antibacterial efficacy of more than 95% against methicillin-susceptible Staphylococcus aureus and Escherichia coli. The main advantages of our surface treatment are its simplicity and low cost. Therefore, our treatment is promising for clinical applications in orthopaedic or dental Ti-based implants with antibacterial and early bone-bonding abilities.

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