Increased Cell Attachment Research Articles

Arachidonic acid enhances TPA-induced differentiation in human leukemia HL-60 cells via reactive oxygen species-dependent ERK activation

The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), is a potent stimulator of differentiation in human leukemia cells; however, the effects of arachidonic acid (AA) on TPA-induced differentiation are still unclear. In the present study, we investigated the contribution of AA to TPA-induced differentiation of human leukemia HL-60 cells. We found that treatment of HL-60 cells with TPA resulted in increases in cell attachment and nitroblue tetrazolium (NBT)-positive cells, which were significantly enhanced by the addition of AA. Stimulation of TPA-induced intracellular reactive oxygen species (ROS) production by AA was detected in HL-60 cells via a DCHF-DA analysis, and the addition of the antioxidant, N-acetyl-cysteine (NAC), was able to reduce TPA+AA-induced differentiation in accordance with suppression of intracellular peroxide elevation by TPA+AA. Furthermore, activation of extracellular-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) by TPA+AA was identified in HL-60 cells, and the ERK inhibitor, PD98059, but not the JNK inhibitor, SP600125, inhibited TPA+AA-induced NBT-positive cells. Suppression of TPA+AA-induced ERK protein phosphorylation by PD98059 and NAC was detected, and AA enhanced ERK protein phosphorylation by TPA was in HL-60 cells. AA clearly increased TPA-induced HL-60 cell differentiation, as evidenced by a marked increase in CD11b expression, which was inhibited by NAC and PD98059 addition. Eicosapentaenoic acid (EPA) as well as AA showed increased intracellular peroxide production and differentiation of HL-60 cells elicited by TPA. Evidence of AA potentiation of differentiation by TPA in human leukemia cells HL-60 via activation of ROS-dependent ERK protein phosphorylation was first demonstrated herein.

Comparison of efficacies of different bone substitutes adhered to osteoblasts with and without extracellular matrix proteins

Background/purposeRegeneration of bony tissue is a major goal in periodontal and implant surgery. The adhesion efficacy of osteogenic cells to bone substitutes is a determinant for osteogenesis. The adsorbed proteins have a significant impact on the behavior of cells and the biological activity of the grafting materials. The aim of the present study was to compare the adhesion ability of different grafting materials with and without extracellular matrix (ECM) protein coating. Materials and methodsMG63 human osteosarcoma cells were cocultured with the same volume of different bone substitutes (Bio-Oss, Cerasorb, MBCP, and BoneCeramic). The nonadhesion cells in the suspension were counted. The medium pH values were measured. The viability of attached cells was measured by the dye WST-1 (water-soluble tetrazolium salts-1). The same procedures for cell viability were repeated for each bone substitute coated with ECM proteins (fibronectin, vitronectin, type I collagen) separately. ResultsAdsorption of ECM proteins increased cell adhesion to the bone substitutes. Vitronectin and fibronectin increased cell attachment to Cerasorb or BoneCeramic significantly more than type I collagen did. Type I collagen was better in increasing cell attachment to Bio-Oss, which had the lowest attached cell count both on dry material and with protein coating. ConclusionThe results indicated that ECM proteins increased cell attachment to bone substitutes in vitro. The preferential affinity of different bone substitutes to certain ECM proteins was evident. Cerasorb and BoneCeramic had better MG63 human osteosarcoma cell adhesion ability than Bio-Oss and MBCP.

Mg ion implantation on SLA-treated titanium surface and its effects on the behavior of mesenchymal stem cell

Magnesium (Mg) is one of the most important ions associated with bone osseointegration. The aim of this study was to evaluate the cellular effects of Mg implantation in titanium (Ti) surfaces treated with sand blast using large grit and acid etching (SLA). Mg ions were implanted into the surface via vacuum arc source ion implantation. The surface morphology, chemical properties, and the amount of Mg ion release were evaluated by scanning electron microscopy (SEM), Auger electron spectroscopy (AES), Rutherford backscattering spectroscopy (RBS), and inductively coupled plasma-optical emission spectrometer (ICP-OES). Human mesenchymal stem cells (hMSCs) were used to evaluate cellular parameters such as proliferation, cytotoxicity, and adhesion morphology by MTS assay, live/dead assay, and SEM. Furthermore, osteoblast differentiation was determined on the basis of alkaline phosphatase (ALP) activity and the degree of calcium accumulation. In the Mg ion-implanted disk, 2.3×1016ions/cm2 was retained. However, after Mg ion implantation, the surface morphology did not change. Implanted Mg ions were rapidly released during the first 7days in vitro. The MTS assay, live/dead assay, and SEM demonstrated increased cell attachment and growth on the Mg ion-implanted surface. In particular, Mg ion implantation increased the initial cell adhesion, and in an osteoblast differentiation assay, ALP activity and calcium accumulation. These findings suggest that Mg ion implantation using the plasma source ion implantation (PSII) technique may be useful for SLA-treated Ti dental implants to improve their osseointegration capacity.

Evaluation of in vitro bioactivity and MG63 Oesteoblast cell response for TiO2 coated magnesium alloys

The present investigation reports TiO2 coating on magnesium alloy AZ31 by sol–gel method via dip coating technique. TiO2 coated surface was characterized by thin film X-ray diffraction (TF-XRD), Fourier transform infrared red (FT-IR) spectroscopy, scanning electron microscopy (SEM) with energy-dispersive X-ray (EDX) spectroscopy, atomic force microscopy (AFM) and transmission electron microscopy (TEM) techniques. From TF-XRD results, the peaks at 2θ values of 25.14, 32.12, 68.73 and 70.11 confirm the presence of TiO2. The TiO2 is crystalline in nature and the crystallite size is about 32.4 nm. SEM-EDX, TEM and AFM show that the coated surface is uniform and nanoporous. FT-IR analysis shows that the peak in the range of 692 cm−1 is assigned to Ti–O–Ti stretching vibration. Contact angle measurements show that the coating is hydrophilic in nature. Bioactivity of the coating in simulated body fluid (SBF) was also examined, the hydroxyl functionalized surface greatly enhances the hydroxyapatite growth. The potentiodynamic polarization studies prove that the corrosion resistance of the TiO2 coated surface after immersion in SBF for 7 days is improved dramatically. Cell adhesion studies confirm the increased cell attachment on TiO2 coated surface when compared to uncoated alloy, due to less amount of Mg ion release from the substrate in the culture medium.

Effects of Tepary Bean (Phaseolus acutifolius) Protease Inhibitor and Semipure Lectin Fractions on Cancer Cells

Some natural and synthetic protease inhibitors (PI), such as the Bowman-Birk PI from soybean, have anticancer effects. We previously purified and characterized a Bowman-Birk-type PI from Tepary bean (Phaseolus acutifolius) seeds (TBPI). A semipure protein fraction containing this inhibitor, when tested its in vitro effect on transformed cells, showed a differential cytotoxic effect, as well as an increase in cell attachment to culture dishes. In this article we report that lectins were responsible for the cytotoxic effect previously observed, exhibiting a differential, antiproliferative effect on nontransformed cells and on different lineages of cancer cells. Although the purified TBPI lacked cytotoxicity, it was found to be responsible for the increase in cell adhesion, decreasing culture dishes’ extracellular matrix degradation, leading to a decrease of the in vitro cell invasion capacity. This effect coincided with the suppression of Matrix Metalloproteinase-9 activity. These results indicate that Tepary bean seeds contain at least 2 different groups of bioactive proteins with distinct effects on cancer cells.

RhoD regulates cytoskeletal dynamics via the actin nucleation–promoting factor WASp homologue associated with actin Golgi membranes and microtubules

The Rho GTPases have mainly been studied in association with their roles in the regulation of actin filament organization. These studies have shown that the Rho GTPases are essential for basic cellular processes, such as cell migration, contraction, and division. In this paper, we report that RhoD has a role in the organization of actin dynamics that is distinct from the roles of the better-studied Rho members Cdc42, RhoA, and Rac1. We found that RhoD binds the actin nucleation-promoting factor WASp homologue associated with actin Golgi membranes and microtubules (WHAMM), as well as the related filamin A-binding protein FILIP1. Of these two RhoD-binding proteins, WHAMM was found to bind to the Arp2/3 complex, while FILIP1 bound filamin A. WHAMM was found to act downstream of RhoD in regulating cytoskeletal dynamics. In addition, cells treated with small interfering RNAs for RhoD and WHAMM showed increased cell attachment and decreased cell migration. These major effects on cytoskeletal dynamics indicate that RhoD and its effectors control vital cytoskeleton-driven cellular processes. In agreement with this notion, our data suggest that RhoD coordinates Arp2/3-dependent and FLNa-dependent mechanisms to control the actin filament system, cell adhesion, and cell migration.

Immobilization of cell‐binding peptides on poly‐ε‐caprolactone film surface to biomimic the peripheral nervous system

Cell-material interactions are crucial for cell adhesion and proliferation on biomaterial surfaces. Immobilization of biomolecules leads to the formation of biomimetic substrates, improving cell response. We introduced RGD (Arg-Gly-Asp) sequences on poly-ε-caprolactone (PCL) film surfaces using thiol chemistry to enhance Schwann cell (SC) response. XPS elemental analysis indicated an estimate of 2-3% peptide functionalization on the PCL surface, comparable with carbodiimide chemistry. Contact angle was not remarkably reduced; hence, cell response was only affected by chemical cues on the film surface. Adhesion and proliferation of Schwann cells were enhanced after PCL modification. Particularly, RGD immobilization increased cell attachment up to 40% after 6 h of culture. It was demonstrated that SC morphology changed from round to very elongated shape when surface modification was carried out, with an increase in the length of cellular processes up to 50% after 5 days of culture. Finally RGD immobilization triggered the formation of focal adhesion related to higher cell spreading. In summary, this study provides a method for immobilization of biomolecules on PCL films to be used in peripheral nerve repair, as demonstrated by the enhanced response of Schwann cells.

Fabrication and Characterization of Hybrid Nanofibers from Poly(Vinyl Alcohol), Milk Protein and Metal Carbonates

Porous three dimensional nanofibrous membranes were fabricated from poly(vinyl alcohol) (PVA), milk protein and inorganic salts such as calcium carbonate (CaCO3) or magnesium carbonate (MgCO3). Microscopic investigations showed that the fibers have smooth morphology with an average diameter of 300-500 nm and a surface area of 5.29 m2g(-1). Thermal analysis of the composite nanofibers showed a decrease in glass transition temperature as compared to PVA nanofiber. Incorporation of CaCO3 and MgCO3 into the nanofiber matrix was confirmed by energy dispersive spectroscopy and X-ray diffraction analysis. The cytocompatibility of electrospun composite nanofiber sheets was evaluated using human lung fibroblasts (IMR-90). There was an increase in cell attachment and cell density on milk protein incorporated to PVA-CaCO3 and PVA-MgCO3 fibers within a week of cell seeding. The cytocompatibility and increase in cell adhesion property of the hybrid nanofiber may provide significant advantages for such materials in biomedical applications.

Directionality and Bipolarity of Olfactory Ensheathing Cells on Electrospun Nanofibers

As a preliminary to the construction of olfactory ensheathing cells (OECs) bearing scaffold for bridging larger lesions in the spinal cord, we have investigated the response of purified cultured OECs to nanoscale fibers of varying diameter using US FDA-approved, biodegradable poly(lactic-co-glycolic-acid). Conventional electrospinning produced fibers of approximately 700 nm diameter (nano-700) while nanocomposite electrospinning with quantum dots produced significantly more uniform fibers of a reduced diameter to approximately 237 nm (nano-250). OECs from adult rat were FACS purified, cultured at low density on either a flat surface or a meshwork of randomly orientated nano-700 and nano-250 fibers, and assessed using cytomorphometric analysis of immunofluorescent confocal images and by scanning electron microscopy. Compared with a flat surface, culture on a nano-700 mesh increases cell attachment. Cells change from rounded to stellate forms in random orientation. Further size reduction to the nano-250 favors bipolarity in cells with unidirectional orientation as observed in the case when transplanted OECs were used to bridge areas of damage in rat spinal cords.

IL-1β enhances cell adhesion to degraded fibronectin.

IL-1β is a prominent proinflammatory cytokine that mediates degradation of extracellular matrix proteins through increased expression of matrix metalloproteinases, which involves a signaling pathway in adherent cells that is restricted by focal adhesions. Currently, the mechanism by which IL-1β affects cell adhesion to matrix proteins is not defined, and it is not known whether degraded matrix proteins affect IL-1β signaling. We examined adhesion-related IL-1β signaling in fibroblasts attaching to native or MMP3-degraded fibronectin. IL-1β increased cell attachment, resistance to shear force and the numbers of focal adhesions containing activated β(1) integrins. IL-1β-enhanced attachment required FAK, kindlins 1/2, and talin. MMP3-degraded fibronectin-inhibited IL-1β-enhanced cell adhesion and promoted spontaneous ERK activation that was independent of IL-1β treatment. We conclude that IL-1β enhances the adhesion of anchorage-dependent cells to MMP3-degraded fibronectin, which, in turn, is associated with deregulated cellular responses to IL-1β. These data point to a novel role of IL-1β as a proadhesive signaling molecule in inflammation that employs kindlins and talin to regulate adhesion.

The effect of adherens junction components on keratinocyte adhesionin vitro: Potential implications for sealing the skin‐implant interface of intraosseous transcutaneous amputation prostheses

Amputation places a significant burden on healthcare systems worldwide as patients suffer life-long complications associated with the stump-socket interface. Skin penetrating, osseointegrated implants like intraosseous transcutaneous amputation prostheses, could overcome this, however, they rely on the formation and maintenance of an infection-free seal at the skin-implant interface. Epithelial cell migration around transcutaneous implants creates downgrowth, which leads to infection and implant failure. Epithelial cells form cell-cell attachments via adherens junctions and desmosomes that prevent cell migration via contact inhibition. If epithelial cells formed cell-cell attachments with an implant surface, it could facilitate stronger cell attachment and prevent downgrowth. In adherens junctions, E-cadherin is essential in homotypic cell attachment. In this study, we have demonstrated that cell-cell adherens junctions can be formed on substrates adsorbed with E-cadherin. We have assessed the effects of two E-cadherin peptides and determined an optimal concentration for increasing cell attachment via adherens junctions. We have demonstrated that adsorption of 15 μg/mL of the full extracellular domain of E-cadherin to titanium alloy significantly increases metabolic activity, cell area, and attachment of murine keratinocytes in vitro, with a fourfold increase in attachment via adherens junctions at 24, 48, and 72 h.

Platelet‐rich fibrin increases cell attachment, proliferation and collagen‐related protein expression of human osteoblasts

Platelet-rich fibrin (PRF) prepared by Choukroun's technique is derived from an autogenous preparation of concentrated platelets without any manipulation. PRF was found to increase osteoblast growth and proliferation. However, the underlying mechanisms are not yet completely understood. This study aimed to determine the effects of PRF on cell attachment, proliferation, phosphorylated Akt, heat shock protein 47 (HSP47) and lysyl oxidase (LOX) expression on human osteoblasts. Blood collection was carried out from 10 healthy volunteers. Cell attachment and proliferation were measured by colorimetric assay with WST-1 and alamar blue in human osteoblast cell line U2OS cells, respectively. Western blot was employed to evaluate the expression of p-Akt, HSP47 and LOX. PRF alone was found to stimulate U2OS cell attachment compared with untreated controls (p < 0.05). PRF was found to increase osteoblast proliferation during a 5-day incubation period (p < 0.05). PRF was found to increase Akt phosphorylation in a time-dependent manner (p < 0.05). Collagen-related proteins HSP47 and LOX were significantly elevated by stimulation with PRF compared with untreated controls (p < 0.05). It is suggested that PRF is capable of increasing osteoblast attachment, proliferation and simultaneously upregulating collagen-related protein production. These actions in combination would effectively promote bone regeneration.

Titania‐polymeric powder coatings with nano‐topography support enhanced human mesenchymal cell responses

Titanium implant osseointegration is dependent on the cellular response to surface modifications and coatings. Titania-enriched nanocomposite polymeric resin coatings were prepared through the application of advanced ultrafine powder coating technology. Their surfaces were readily modified to create nano-rough (<100 nm) surface nano-topographies that supported human embryonic palatal mesenchymal cell responses. Energy dispersive x-ray spectroscopy confirmed continuous and homogenous coatings with a similar composition and even distribution of titanium. Scanning electron microscopy (SEM) showed complex micro-topographies, and atomic force microscopy revealed intricate nanofeatures and surface roughness. Cell counts, mitochondrial enzyme activity reduction of yellow 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to dark purple, SEM, and inverted fluorescence microscopy showed a marked increase in cell attachment, spreading, proliferation, and metabolic activity on the nanostructured surfaces. Reverse Transcription- Polymerase Chain Reaction (RT-PCR) analysis showed that type I collagen and Runx2 expression were induced, and Alizarin red staining showed that mineral deposits were abundant in the cell cultures grown on nanosurfaces. This enhancement in human mesenchymal cell attachment, growth, and osteogenesis were attributed to the nanosized surface topographies, roughness, and moderate wetting characteristics of the coatings. Their dimensional similarity to naturally occurring matrix proteins and crystals, coupled with their increased surface area for protein adsorption, may have facilitated the response. Therefore, this application of ultrafine powder coating technology affords highly biocompatible surfaces that can be readily modified to accentuate the cellular response.

Covalent attachment of P15 peptide to titanium surfaces enhances cell attachment, spreading, and osteogenic gene expression

P15, a synthetic 15 amino acid peptide, mimics the cell-binding domain within the alpha-1 chain of human collagen is being tested in clinical trials to determine if it enhances bone formation in spinal fusions. We hypothesize that covalent attachment of P15 to titanium implants may also serve to promote osseointegration. To test this hypothesis, we measured osteoblast and mesenchymal cell adhesion, proliferation, and maturation on P15 tethered to a titanium (Ti-P15) surface. P15 peptide was covalently bonded to titanium alloy surfaces and incubated with osteoblast like cells. Cell toxicity, adhesion, spreading, and differentiation was then evaluated. Real-time quantitative PCR, Western blot analysis, and fluorescent immunohistochemistry was performed to measure osteoblast gene expression and differentiation. There was no evidence of toxicity. Significant increases in early cell attachment, spreading, and proliferation were observed on the Ti-P15 surface. Increased filapodial attachments, α(2) integrin expression, and phosphorylated focal adhesion kinase immunostaining indicated activation of integrin signaling pathways. qRT-PCR analysis indicated there was significant increase in osteogenic differentiation markers in cells grown on Ti-P15 compared to control-Ti. Western blotting confirmed these findings. Surface modification of titanium with P15 significantly increased cell attachment, spreading, osteogenic gene expression, and differentiation. Results of this study suggest that Ti-P15 has the potential to safely enhance bone formation and promote osseointegration of titanium implants.

Electrospun fibers from functional polyglycidol/poly(ε‐caprolactone) blends with defined surface properties

AbstractHighly hydrophilic but water insoluble fibers from star‐shaped poly(ethoxyethyl glycidyl ether) and poly(ε‐caprolactone) polymer blends in the submicron range were prepared using the electrospinning technique. The fibers were achieved in a smooth and homogeneous manner and do show tremendous decreased protein adsorption. Additionally, using alkyne‐ or vinyl sulfonate end capped polymer, fibers with the correspondent surface reactivity have been prepared. All fibers showed high biocompatibility and were highly hydrophilic but water stable. Furthermore, nonwovens based on functionalized poly(ethoxyethyl glycidyl ether) were equipped with small biofunctional molecules, e.g., the peptide sequence glycine‐arginine‐glycine‐aspartate‐serine (GRGDS). These fibers showed increased cell attachment compared with nonfunctionalized nonwovens. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Osteoblast activity on carbonated hydroxyapatite

Hydroxyapatite (HA), has been used commonly as a bone substitute and as a scaffold in bone tissue engineering. However it has certain drawbacks such as limited biodegradability and osteointegration properties. Other forms of HA, for example, carbonated hydroxyapatite (CHA) could prove to have enhanced bioactivity as they more closely mimic the chemical composition of the apatite found in human bone. The aim of this study was to test the efficacy of CHA in comparison to HA used as a control. The CHA (4.9 wt %) and the HA discs were seeded with MC3T3-E1 osteoblastic cells. Results revealed a trend of increased cell attachment on the HA discs at day 0, however, the cell proliferation on the CHA discs at 7 and 28 days showed no significant difference in comparison to the HA control. SEM of the CHA discs showed surface irregularities at 7 days indicating dissolution. Also at 7 days, SEM demonstrated cell attachment and extracellular matrix production on both the CHA and HA samples. There was no significant difference in the total amount of collagen produced in the CHA samples relative to the HA control samples at 28 days as evaluated by the hydroxyproline assay. Real time PCR revealed mRNA increase by 2.08, 7.62, and 9.86 fold for collagen I a1, collagen III a1, and osteocalcin respectively on the CHA as compared to the HA discs. This study demonstrates the use of CHA as a biocompatible material that has potentially increased biodegradation properties and osteogenic capability in comparison to HA.

EFFECTS OF GELATIN MODIFICATION ON RAPID PROTOTYPING PCL SCAFFOLDS FOR CARTILAGE ENGINEERING

The purpose of this study was to investigate the attachment and proliferation of cells on selective laser-sintered (SLS) polycaprolactone (PCL) scaffolds coated with gelatin for cartilage tissue engineering using chondrocytes isolated from the articular cartilage of swine. Scaffolds without modification were used as control groups. Cell proliferation was measured by cell count 1, 3 and 5 days after cell seeding into the scaffolds. The biocompatibility of the scaffold was examined by scanning electron microscopy (SEM). The PCL scaffolds coated with gelatin had higher hydrophilicity. The results provided a useful strategy for modifying the microenvironments to increase cell attachment, growth and the formation of extracellular matrix on scaffolds for cartilage tissue engineering.

Functionalization of reactive polymer multilayers with RGD and an antifouling motif: RGD density provides control over human corneal epithelial cell–substrate interactions

Our study demonstrates that substrates fabricated using a "reactive" layer-by-layer approach promote well-defined cell-substrate interactions of human corneal epithelial cells. Specifically, crosslinked and amine-reactive polymer multilayers were produced by alternating "reactive" deposition of an azlactone-functionalized polymer [poly(2-vinyl-4,4-dimethylazlactone)] (PVDMA) and a primary amine-containing polymer [branched poly(ethylene imine)] (PEI). Advantages of our system include a 5- to 30-fold decrease in deposition time compared to traditional polyelectrolyte films and direct modification of the films with peptides. Our films react with mixtures of an adhesion-promoting peptide containing Arg-Gly-Asp (RGD) and the small molecule D-glucamine, a chemical motif which is nonfouling. Resulting surfaces prevent protein adsorption and promote cell attachment through specific peptide interactions. The specificity of cell attachment via immobilized RGD sequences was verified using both a scrambled RDG peptide control as well as soluble-RGD competitive assays. Films were functionalized with monotonically increasing surface densities of RGD which resulted in both increased cell attachment and the promotion of a tri-phasic proliferative response of a human corneal epithelial cell line (hTCEpi). The ability to treat PEI/PVDMA films with peptides for controlled cell-substrate interactions enables the use of these films in a wide range of biological applications.

Graphene Oxide: A Nonspecific Enhancer of Cellular Growth

There have been multiple conflicting reports about the biocompatibility and antimicrobial activity of graphene oxide. To address this, we conducted a study to characterize the antimicrobial properties of graphene oxide (GO) and its biocompatibility with mammalian cells. When GO was added to a bacterial culture at 25 μg/mL, the results showed that bacteria grew faster and to a higher optical density than cultures without GO. Scanning electron microscopy indicated that bacteria formed dense biofilms in the presence of GO. This was shown by a large mass of aggregated cells and extracellular polymeric material. Bacterial growth on filters coated with 25 and 75 μg of GO grew 2 and 3 times better than on filters without GO. Closer analysis showed that bacteria were able to attach and proliferate preferentially in areas containing the highest GO levels. Graphene oxide films failed to produce growth inhibition zones around them, indicating a lack of antibacterial properties. Also, bacteria were able to grow on GO films to 9.5 × 10(9) cells from an initial inoculation of 1.0 × 10(6), indicating that it also lacks bacteriostatic activity. Thus, silver-coated GO films were able to produce clearing zones and cell death. Also, graphene oxide was shown to greatly enhance the attachment and proliferation of mammalian cells. This study conclusively demonstrates that graphene oxide does not have intrinsic antibacterial, bacteriostatic, and cytotoxic properties in both bacteria and mammalian cells. Furthermore, graphene oxide acts as a general enhancer of cellular growth by increasing cell attachment and proliferation.

Aqueous deposition of calcium phosphates and silicate substituted calcium phosphates on magnesium alloys

Abstract Attempts were made to deposit homogeneous films of calcium phosphates (CaPs) on two magnesium alloy systems, AZ31 and Mg–4Y, through an aqueous phosphating bath method. The deposition of silicate substituted CaPs by this aqueous method was also explored as silicate substitution is believed to increase the bioactivity of CaPs. The effect of doped and undoped coatings on the in vitro degradation and bioactivity of both alloy systems was studied. FTIR and EDX confirmed the deposition of Ca, P, and Si on both alloys and the coatings appeared to consist primarily biphasic mixtures of hydroxyapatite and β-TCP. These largely inhomogeneous coatings, as observed by SEM, were not shown to have any significant effect on maintaining the physiological pH of the culture medium in comparison to the uncoated samples, as the pH remained approximately in the 8.4–8.7 range. Interestingly, despite similar pH profiles between the coated and uncoated samples, CaP coatings affected the degradation of both alloys. These doped and undoped calcium phosphate coatings were observed to decrease the degradation of AZ31 whereas they increased the degradation of Mg–4Y. In vitro studies on cell attachment using MC3T3-E1 mouse osteoblasts showed that between the uncoated alloys, Mg–4Y appeared to be the more biocompatible of the two. Silicate substituted CaP coatings were observed to increase the cell attachment on AZ31 compared to bare and undoped CaPs coated samples, but did not have as great of an effect on increasing cell attachment on Mg–4Y.