Response Of MG63 Osteoblast-like Cells Research Articles

Modulation of MG-63 Osteogenic Response on Mechano-Bactericidal Micronanostructured Titanium Surfaces.

Despite recent advances in the development of orthopedic devices, implant-related failures that occur as a result of poor osseointegration and nosocomial infection are frequent. In this study, we developed a multiscale titanium (Ti) surface topography that promotes both osteogenic and mechano-bactericidal activity using a simple two-step fabrication approach. The response of MG-63 osteoblast-like cells and antibacterial activity toward Pseudomonas aeruginosa and Staphylococcus aureus bacteria was compared for two distinct micronanoarchitectures of differing surface roughness created by acid etching, using either hydrochloric acid (HCl) or sulfuric acid (H2SO4), followed by hydrothermal treatment, henceforth referred to as either MN-HCl or MN-H2SO4. The MN-HCl surfaces were characterized by an average surface microroughness (Sa) of 0.8 ± 0.1 μm covered by blade-like nanosheets of 10 ± 2.1 nm thickness, whereas the MN-H2SO4 surfaces exhibited a greater Sa value of 5.8 ± 0.6 μm, with a network of nanosheets of 20 ± 2.6 nm thickness. Both micronanostructured surfaces promoted enhanced MG-63 attachment and differentiation; however, cell proliferation was only significantly increased on MN-HCl surfaces. In addition, the MN-HCl surface exhibited increased levels of bactericidal activity, with only 0.6% of the P. aeruginosa cells and approximately 5% S. aureus cells remaining viable after 24 h when compared to control surfaces. Thus, we propose the modulation of surface roughness and architecture on the micro- and nanoscale to achieve efficient manipulation of osteogenic cell response combined with mechanical antibacterial activity. The outcomes of this study provide significant insight into the further development of advanced multifunctional orthopedic implant surfaces.

Effect of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys

Various types of titanium base alloys are utilized as implant materials and as minis crews in orthodontics. The purpose of the current study is to enhancing titanium-based alloys with the additions of Cobalt in different percentage (5%, 10%, and 15%) to enhance primary osteoblast attachment and implant tissue incorporation. the reaction between human MG63 osteoblast-like cells and titanium – cobalt alloy face is substantial in biomaterials and tissue engineering field. The major target was to estimate the proteomic response of MG63 osteoblast-like cells in various cobalt percentages. The samples with and without cobalt addition was produced by using powder technology in order to get an acceptable initial surface roughness. Also sample characterizations includes (scanning electron microscopy (SEM), X-ray diffraction (XRD), vitro test (Biocompatibility MTT assay with MG-63 Cells). Results obtained from XRD and microstructure observations, sample without cobalt addition refer to an alloy with single I± phase, while after the additions of cobalt in different percentage (5%, 10%, 15%) the samples consist of two I±-Ti and intermetallic compound Ti2Co and the amount of Ti2Co phase slightly increase with increasing cobalt content. From MTT assays Proliferation and cells attachment of MG63 were cultured after 2 days Cells grown for the pure titanium alloys possess finer and circular confines compared with the grown cells at (titanium – cobalt) alloys with all cobalt percentages. Rising cobalt percentage makes the MG63 cell sharper and enlarges which means cobalt aid to enhancing both cell proliferation and growth. Cytotoxicity Cell Viability results was kept, exhibition no kind of attack caused by titanium or cobalt materials, The feature, of cobalt additions, in contrast to commercial pure titanium alloys, is the presence of active and non- cytotoxic compound (Ti2Co) which presents several attractive features, make the surface more attractive to the bone . How to Cite this Article Pubmed Style Fatimah J. Al-Hasani. of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys. SRP. 2020; 11(5): 804-813. doi:10.31838/srp.2020.5.119 Web Style Fatimah J. Al-Hasani. of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys. http://www.sysrevpharm.org/?mno=131488 [Access: March 30, 2021]. doi:10.31838/srp.2020.5.119 AMA (American Medical Association) Style Fatimah J. Al-Hasani. of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys. SRP. 2020; 11(5): 804-813. doi:10.31838/srp.2020.5.119 Vancouver/ICMJE Style Fatimah J. Al-Hasani. of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys. SRP. (2020), [cited March 30, 2021]; 11(5): 804-813. doi:10.31838/srp.2020.5.119 Harvard Style Fatimah J. Al-Hasani (2020) of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys. SRP, 11 (5), 804-813. doi:10.31838/srp.2020.5.119 Turabian Style Fatimah J. Al-Hasani. 2020. of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Reviews in Pharmacy, 11 (5), 804-813. doi:10.31838/srp.2020.5.119 Chicago Style Fatimah J. Al-Hasani. Effect of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys. Systematic Reviews in Pharmacy 11 (2020), 804-813. doi:10.31838/srp.2020.5.119 MLA (The Modern Language Association) Style Fatimah J. Al-Hasani. Effect of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Alloys. Systematic Reviews in Pharmacy 11.5 (2020), 804-813. Print. doi:10.31838/srp.2020.5.119 APA (American Psychological Association) Style Fatimah J. Al-Hasani (2020) of Cobalt Addition on the Cytotoxicity and Cell Attachment of Titanium Reviews in Pharmacy, 11 (5), 804-813. doi:10.31838/srp.2020.5.119

Electrospun Polyhydroxybutyrate/Poly(ε-caprolactone)/58S Sol-Gel Bioactive Glass Hybrid Scaffolds with Highly Improved Osteogenic Potential for Bone Tissue Engineering.

Electrospinning of biopolymer and inorganic substances is one of the efficient ways to combine various advantageous properties in one single fibrous structure with potential for tissue engineering applications. In the present study, to integrate the high stiffness of polyhydroxybutyrate (PHB), the flexibility of poly(ε-caprolactone) (PCL) and the bioactivity of 58S bioactive glass, PHB/PCL/58S sol-gel bioactive glass hybrid scaffolds were fabricated using combined electrospinning and sol-gel method. Physical features such as fiber diameter distribution, mechanical strength and Young's modulus were characterized thoroughly. FTIR analysis demonstrated the successful incorporation of 58S bioactive glass into the blend polymers, which greatly improved the hydrophilicity of PHB/PCL fibermats. The primary biological response of MG-63 osteoblast-like cells on the prepared fibrous scaffolds was evaluated, proving that the 58S glass sol containing hybrid scaffold were not only favorable to MG-63 cell adhesion but also slightly enhanced cell viability and significantly increased alkaline phosphate activity .

Response of MG63 osteoblast-like cells to ordered nanotopographies fabricated using colloidal self-assembly and glancing angle deposition.

Ordered surface nanostructures have attracted much attention in different fields including biomedical engineering because of their potential to study the size effect on cellular response and modulation of cell fate. However, the ability to fabricate large-area ordered nanostructures is typically limited due to high costs and low speed of fabrication. Herein, highly ordered nanostructures with large surface areas (>1.5 × 1.5 cm(2)) were fabricated using a combination of facile techniques including colloidal self-assembly, colloidal lithography, and glancing angle deposition (GLAD). An ordered tantalum (Ta) pattern with 60-nm-height was generated using colloidal lithography. A monolayer of colloidal crystal, i.e., hexagonal close packed 720 nm polystyrene particles, was self-assembled and used as a mask. Ta patterns were subsequently generated by evaporation of Ta through the mask. The feature size was further increased by 100 or 200 nm using GLAD, resulting in the fabrication of four different surfaces (FLAT, Ta60, GLAD100, and GLAD200). Cell adhesion, proliferation, and mineralization of MG63 osteoblast-like cells were investigated on these ordered nanostructures over a 1 week period. Our results showed that cell adhesion, spreading, focal adhesion formation, and filopodia formation of the MG63 osteoblast-like cells were inhibited on the GLAD surfaces, especially the initial (24 h) attachment, resulting in a lower cell density on the GLAD surfaces. After 1 week culture, alkaline phosphatase activity and the amount of Ca was higher on the GLAD surfaces compared with Ta60 and FLAT controls, suggesting that the GLAD surfaces facilitate differentiation of osteoblasts. This study demonstrates that ordered Ta nanotopographies synthesized by combining colloidal lithography with GLAD can improve the mineralization of osteoblast-like cells providing a new platform for biomaterials and bone tissue engineering.

Proteomic analysis of the biological response of MG63 osteoblast-like cells to titanium implants

Understanding of the interaction between human MG63 osteoblast-like cells and surfaces is necessary in the field of tissue engineering and biomaterials. Various titanium surfaces are widely used as not only implant materials, but also as miniscrews in orthodontics. Our goal was to assess the proteomic response of MG63 osteoblast-like cells to different titanium surfaces. MG63 osteoblast-like cells were cultured on three different titanium surfaces: a smooth surface (S), a sandblasted with large grit and acid-etched surface (SLA), and a surface coated with a thin layer of hydroxyapatite (HA). Cells grown on the rougher surfaces (SLA and HA) exhibited downregulated cell proliferation and morphological changes. In the proteomic analysis, cells grown on the SLA surface showed upregulated expression of protocadherin-β3 precursor, kinase insert domain receptor, fibroblast growth factor receptor-3, and insulin-like growth factor I, while the expression levels of cell adhesion kinase, collagen α-1(I) chain precursor, collagen type XI α2, and cadherin-11 were upregulated in cells grown on the HA surface. These proteins are known to be involved in osteoblast adhesion, growth, and differentiation. Thus, the surface properties of dental materials can influence the expression of proteins involved in osseointegration-related processes. Proteomic analysis may reveal changes in novel proteins that explain why osseointegration varies depending on surface properties.

Effect of blasting treatment and Fn coating on MG63 adhesion and differentiation on titanium: a gene expression study using real-time RT-PCR

Biomaterial surface properties, via alterations in the adsorbed protein layer, and the presence of specific functional groups can influence integrin binding specificity, thereby modulating cell adhesion and differentiation processes. The adsorption of fibronectin, a protein directly involved in osteoblast adhesion to the extracellular matrix, has been related to different physical and chemical properties of biomaterial surfaces. This study used blasting particles of different sizes and chemical compositions to evaluate the response of MG63 osteoblast-like cells on smooth and blasted titanium surfaces, with and without fibronectin coatings, by means of real-time reverse transcription-polymerase chain reaction (qRT-PCR) assays. This response included (a) expression of the α(5), α(v) and α(3) integrin subunits, which can bind to fibronectin through the RGD binding site, and (b) expression of alkaline phosphatase (ALP) and osteocalcin (OC) as cell-differentiation markers. ALP activity and synthesis of OC were also tested. Cells on SiC-blasted Ti surfaces expressed higher amounts of the α(5) mRNA gene than cells on Al(2)O(3)-blasted Ti surfaces. This may be related to the fact that SiC-blasted surfaces adsorbed higher amounts of fibronectin due to their higher surface free energy and therefore provided a higher number of specific cell-binding sites. Fn-coated Ti surfaces decreased α(5) mRNA gene expression, by favoring the formation of other integrins involved in adhesion over α(5)β(1). The changes in α(5) mRNA expression induced by the presence of fibronectin coatings may moreover influence the osteoblast differentiation pathway, as fibronectin coatings on Ti surfaces also decreased both ALP mRNA expression and ALP activity after 14 and 21 days of cell culture.

Influence of surface topography on osteoblast response to fibronectin coated calcium phosphate thin films

The ability to engineer biomaterial surfaces that are capable of a dynamic interaction with cells and tissues is central to the development of medical implants with improved functionality. An important consideration in this regard is the role played by the extracellular proteins that bind to an implant surface in vivo. Deliberate use of an ad-layer of such proteins on an implant surface has been observed to guide and direct cell response. However, the role that changes in surface topography might play in determining the nature of this cell–protein–surface interaction has not been investigated in detail. In this study, calcium phosphate (CaP) thin films have been deposited onto substrates with varying topography such that this is reflected in the (conformal) CaP surface features. A fibronectin (FN) ad-layer was then deposited from solution onto each surface and the response of MG63 osteoblast-like cells investigated. The results revealed that in all cases, the presence of the adsorbed FN layer on the CaP thin films improved MG63 cell adhesion, proliferation and promoted early onset differentiation. Moreover, the nature and scale of the response were shown to be influenced by the underlying CaP surface topography. Specifically, MG63 cell on FN-coated CaP thin films with regular topographical features in the nanometer range showed statistically significant differences in focal adhesion assembly, osteocalcin expression and alkaline phosphase activity compared to CaP thin films that lacked these topographical features. As such, these data indicate that surface topography can be used to further influence cell adhesion and downstream differentiation by enhancing the effects of a surface adsorbed FN layer.

Oxidized NiTi surfaces enhance differentiation of osteoblast‐like cells

A new oxidation treatment (OT) on NiTi shape memory alloys was developed in a previous work. This OT treatment significantly decreases Ni ion release into the exterior medium, and therefore is thought to be beneficial for NiTi cytocompatibility. As to confirm this expectation, the in vitro response of MG63 osteoblast-like cells cultured on untreated and oxidized NiTi surfaces was studied. An adhesion test at 1, 4, and 8 h of incubation was performed. Statistical differences were evidenced at 1 h of adhesion depending on the surface treatment and chemical composition of the substrate. However, at larger times of study, there were no statistically significant differences between untreated and oxidized surfaces. The proliferation test (until 9 days) showed that untreated and oxidized NiTi surfaces are not cytotoxic for MG63 cells. The differences of adhesion at short times did not affect the proliferation of MG63 cells. However, after 48 h of stimulation with ascorbic acid and dexamethasone, the MG63 cells cultured on oxidized surfaces showed higher alkaline phosphatase activity and osteocalcin levels. The improvement of osteoblast differentiation due to OT treatment could accelerate bone formation, and, therefore, could allow earlier loading of NiTi devices used in dental and orthopedic applications.

Response of MG63 osteoblast-like cells onto polycarbonate membrane surfaces with different micropore sizes

Response of different types of cells on materials is important for the applications of tissue engineering and regenerative medicine. It is recognized that the behavior of the cell adhesion, proliferation, and differentiation on materials depends largely on surface characteristics such as wettability, chemistry, charge, rigidity, and roughness. In this study, we examined the behavior of MG63 osteoblast-like cells cultured on a polycarbonate (PC) membrane surfaces with different micropore sizes (0.2–8.0 μm in diameter). Cell adhesion and proliferation to the PC membrane surfaces were determined by cell counting and MTT assay. The effect of surface micropore on the MG63 cells was evaluated by cell morphology, protein content, and alkaline phosphatase (ALP) specific activity. It seems that the cell adhesion and proliferation were progressively inhibited as the PC membranes had micropores with increasing size, probably due to surface discontinuities produced by track-etched pores. Increasing micropore size of the PC membrane results in improved protein synthesis and ALP specific activity in isolated cells. There was a statistically significant difference ( P<0.05) between different micropore sizes. The MG63 cells also maintained their phenotype under conditions that support a round cell shape. RT-PCR analysis further confirmed the osteogenic phenotype of the MG63 cells onto the PC membranes with different micropore sizes. In results, as micropore size is getting larger, cell number is reduced and cell differentiation and matrix production is increased. This study demonstrated that the surface topography plays an important role for pheonotypic expression of the MG63 osteoblast-like cells.

Effect of polymer molecular weight and addition of calcium stearate on response of MG63 osteoblast-like cells to UHMWPE particles

Periprosthetic osteolysis and implant loosening is associated with the presence of ultrahigh molecular weight polyethylene (UHMWPE) wear debris particles. Osteoblast phenotypic expression in vitro is affected by UHMWPE particles, suggesting that bone formation may also be affected by wear debris. Here we tested the hypothesis that the response of osteoblasts to UHMWPE can be modified by changes in UHMWPE particle chemistry. We used four different commercially available preparations of GUR UHMWPE particles to determine if chemical composition (±Ca-stearate) or polymer molecular weight (3.1–4.2 million or 5.4–6.5 million g/mol) modulates osteoblast response. Particles were characterized by size distribution, morphology, and number of particles added to the culture medium. They had an average equivalent circle diameter ranging from 0.46–1.26 μm. MG63 cell response was assessed by measuring cell number, cellular and cell layer alkaline phosphatase, and prostaglandin E 2 ( PGE 2) production. There were dose-dependent effects of the particles on cell response. Cell number and PGE 2 production were increased, while alkaline phosphatase specific activity was decreased. In addition, there was a marked difference between cultures treated with particles containing Ca-stearate and as a function of polymer molecular weight. Particles of higher molecular weight caused a greater stimulation of proliferation and inhibition of alkaline phosphatase than particles of lower molecular weight. The presence of Ca-stearate exerted a more pronounced depression of osteoblast phenotype as well as a significantly greater increase in PGE 2 release by the cells. The present study shows that chemical composition and polymer molecular weight of UHMWPE are capable of modulating osteoblast response to particles. The results suggest that osteoblast differentiation is inhibited by UHMWPE particles, whereas cell proliferation and PGE 2 production are stimulated. This may have direct effects on osteoblasts and bone formation, but also paracrine effects on cells of the monocytic lineage inducing bone resorption and promoting inflammation which may lead to aseptic loosening. The present results suggest that the cellular events in aseptic loosening may be modulated or even accelerated by changes in the composition of the UHMWPE used to fabricate implants.

Surface roughness modulates the response of MG63 osteoblast-like cells to 1,25-(OH)(2)D(3) through regulation of phospholipase A(2) activity and activation of protein kinase A.

Implant surface roughness influences osteoblast proliferation, differentiation, and local factor production. Moreover, the responsiveness of osteoblasts to systemic hormones such as 1, 25-(OH)(2)D(3) is altered by the effects of surface roughness; on the roughest Ti surfaces the effects of roughness and 1, 25-(OH)(2)D(3) are synergistic. Prostaglandin E(2) (PGE(2)) appears to be involved in mediating the effects of surface roughness on the cells, as well as in the response to 1,25-(OH)(2)D(3). However, it is not yet known through which signaling pathways surface roughness exerts its effects on the response of osteoblasts to 1, 25-(OH)(2)D(3). The present study examined the potential role of protein kinase A (PKA), phospholipase A(2)(PLA(2)), and protein kinase C (PKC) in this process. MG63 osteoblast-like human osteosarcoma cells were cultured on cpTi disks with R(a) values of 0. 54 microm (PT), 4.14 microm (SLA), or 4.92 microm (TPS). PKA was inhibited by adding H8 to the cultures; similarly, PLA(2) was inhibited with quinacrine or activated with melittin, and PKC was inhibited with chelerythrine. Inhibitors or activators were included in the culture media through the entire culture period or for the last 24 h of culture. In addition, cultures were treated for 24 h with inhibitors or activators in the presence of 1,25-(OH)(2)D(3). The effects on cell number and alkaline phosphatase specific activity were determined after 24 h; PKC activity was determined after 9 min and at 24 h. Cell number was reduced on rough surfaces, and alkaline phosphatase activity was increased. 1,25-(OH)(2)D(3) had a synergistic effect with surface roughness on alkaline phosphatase. However, neither surface roughness nor 1,25-(OH)(2)D(3) had an effect on PKC. H8 treatment for 24 h inhibited cell number and alkaline phosphatase on all surfaces; however, when it was present throughout the culture period, the PKA inhibitor had no effect on cell number, but decreased alkaline phosphatase-specific activity. H8 reduced the 1,25-(OH)(2)D(3)-mediated effect on cell number and alkaline phosphatase. Quinacrine inhibited cell proliferation and alkaline phosphatase on all surfaces and further reduced the 1,25-(OH)(2)D(3)-dependent decreases in both parameters. Melittin had no effect when applied for 24 h and did not modify the 1,25-(OH)(2)D(3) effect; however, when present throughout the culture period, it caused a decrease in proliferation and an increase in enzyme activity. Chelerythrine, the PKC inhibitor, only inhibited cell proliferation when it was present throughout the entire culture period. However, it decreased alkaline phosphatase in cultures treated for 24 h, but increased enzyme activity when it was present for the entire culture period. The results indicate that surface roughness and 1,25-(OH)(2)D(3) both mediate their effects through PLA(2) which catalyzes the rate-limiting step in PGE(2) production. Further downstream, PGE(2) activates PKA. Surface roughness-dependent effects are also mediated through PKC, but only after the cells have reached confluence and are undergoing phenotypic maturation. The effect of surface roughness on responsiveness to 1,25-(OH)(2)D(3) is mediated through PLA(2)/PKA and not through PKC.

Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition

The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness and composition are considered to influence the properties of adherent cells. The aim of this study was to determine the effect of chemical composition and surface roughness of commercially pure titanium (Ti) and Ti–6Al–4V alloy (Ti–A) on MG63 osteoblast-like cells. Unalloyed and alloyed Ti disks were machined and either fine-polished or wet-ground, resulting in smooth (S) and rough (R) finishes, respectively. Standard tissue culture plastic was used as a control. Surface topography and profile were evaluated by cold field emission scanning electron microscopy and profilometry, while chemical composition was determined using Auger electron spectroscopy and Fourier transform infrared spectroscopy. The effect on the cells was evaluated 24 h postconfluence by measuring cell number, [ 3H]-thymidine incorporation into DNA, cell and cell layer alkaline phosphatase specific activity (ALPase), osteocalcin and collagen production, [ 35S]-sulfate incorporation into proteoglycan, and prostaglandin E 2 (PGE 2) and transforming growth factor- β (TGF- β) production. When compared to plastic, the number of cells was reduced on the pure Ti surfaces, while it was equivalent on the Ti-A surfaces; [ 3H]-thymidine incorporation was reduced on all surfaces. The stimulatory effect of surface roughness on ALPase in isolated cells and the cell layer was more pronounced on the rougher surfaces, with enzyme activity on Ti-R being greater than on Ti-A-R. Osteocalcin production was increased only on the Ti-R surface. Collagen production was decreased on Ti surfaces except Ti-R; [ 35S]-sulfate incorporation was reduced on all surfaces. Surface roughness affected local factor production (TGF- β, PGE 2). The stimulatory effect of the rougher surfaces on PGE 2 and TGF- β was greater on Ti than Ti-A. In summary, cell proliferation, differentiation, protein synthesis and local factor production were affected by surface roughness and composition. Enhanced differentiation of cells grown on rough vs. smooth surfaces for both Ti and Ti-A surfaces was indicated by decreased proliferation and increased ALPase and osteocalcin production. Local factor production was also enhanced on rough surfaces, supporting the contention that these cells are more differentiated. Surface composition also played a role in cell differentiation, since cells cultured on Ti-R surfaces produced more ALPase than those cultured on Ti-A-R. While it is still unknown which material properties induce which cellular responses, this study suggests that surface roughness and composition may play a major role and that the best design for an orthopaedic implant is a pure titanium surface with a rough microtopography.