Nano hydroxyapatite‐blasted titanium surface affects pre‐osteoblast morphology by modulating critical intracellular pathways

Abstract

Although, intracellular signaling pathways are proposed to predict the quality of cell-surface relationship, this study addressed pre-osteoblast behavior in response to nano hydroxyapatite (HA)-blasted titanium (Ti) surface by exploring critical intracellular pathways and pre-osteoblast morphological change. Physicochemical properties were evaluated by atomic force microscopy (AFM) and wettability considering water contact angle of three differently texturized Ti surfaces: Machined (Mac), Dual acid-etching (DAE), and nano hydroxyapatite-blasted (nHA). The results revealed critical differences in surface topography, impacting the water contact angle and later the osteoblast performance. In order to evaluate the effect of those topographical characteristics on biological responses, we have seeded pre-osteoblast cells on the Ti discs for up to 4 h and subjected the cultures to biological analysis. First, we have observed pre-osteoblasts morphological changes resulting from the interaction with the Ti texturized surfaces whereas the cells cultured on nHA presented a more advanced spreading process when compared with the cells cultured on the other surfaces. These results argued us for analyzing the molecular machinery and thus, we have shown that nHA promoted a lower Bax/Bcl2 ratio, suggesting an interesting anti-apoptotic effect, maybe explained by the fact that HA is a natural element present in bone composition. Thereafter, we investigated the potential effect of those surfaces on promoting pre-osteoblast adhesion and survival signaling by performing crystal violet and immunoblotting approaches, respectively. Our results showed that nHA promoted a higher pre-osteoblast adhesion supported by up-modulating FAK and Src activations, both signaling transducers involved during eukaryotic cell adhesion. Also, we have shown Ras-Erk stimulation by the all evaluated surfaces. Finally, we showed that all Ti-texturing surfaces were able to promote osteoblast differentiation up to 10 days, when alkaline phosphatase (ALP) activity and osteogenic transcription factors were up-modulated. Altogether, our results showed for the first time that nano hydroxyapatite-blasted titanium surface promotes crucial intracellular signaling network responsible for cell adapting on the Ti-surface.Biotechnol. Bioeng. 2017;114: 1888-1898. © 2017 Wiley Periodicals, Inc.

Nano hydroxyapatite-blasted titanium surface creates a biointerface able to govern Src-dependent osteoblast metabolism as prerequisite to ECM remodeling

“…As reported previously elsewhere [17], there are considerable differences in the topography of the titanium-based surfaces evaluated in this study, mainly documented by atomic force microscopy (insert in Fig. 1).…”

“…Three different titanium surfaces (discs) were investigated in this study, distinguishing by the surface properties, called Machined (Mc; control), Dual Acid-Etched (DAE), and acid-etched nanoHA-blasted (nHA). The nHA was obtained by using the Promimic HAnano-method, a detailed description can be found elsewhere [15][16][17]. Briefly, the samples were dipped into a stable particle suspension containing 10 nm in diameter HA particles followed by a heat treatment at 550 • C for 5 min in nitrogen atmosphere.…”

Mg–Al and Zn–Al Layered Double Hydroxides Promote Dynamic Expression of Marker Genes in Osteogenic Differentiation by Modulating Mitogen‐Activated Protein Kinases

“…Cell Adhesion Was Assayed by Violet Crystal : For the adhesion approach, MC3T3‐E1 cells were treated properly with the both LDH samples up to 24 h. Then, the cells were trypsinized, resuspended in conventional medium containing LDH, counted in hematological chamber, and plated (25 000 cells cm −2 ) in 96‐well plates . After plating, cell adhesion was estimated by incorporation of crystal violet dye at the 3 h and 24 h postplating times.…”

Layered Double Hydroxides Are Promising Nanomaterials for Tissue Bioengineering Application

“…Among the biomaterials, those devoted to bioengineering protocols need to recapitulate the wound‐healing steps triggering a moderate inflammatory cascade activating hemostasis and later requiring platelets and fibrinogen. In this context, advances in physics and chemistry area allowed the application of nanoscaled structures on modifying and functionalizing molecules/materials, ameliorating their performance in medical proposes . It is not new the purpose of layered double hydroxides (LDHs) as biomaterial .…”

“…Regrettably, serious preclinical concerns remain, the signaling cross talk with cells and biomaterials proposes are still the challenge for this promising therapeutic modality . Thus, we have explored and suggested in vitro approaches to predict the quality of the cell/biomaterial interaction, suggesting the intracellular signaling as a preliminary and reliable tool on this matter . Overall, we have shown that cells actively respond to biomaterials modifying their morphology as an immediate consequence of a well‐orchestrated cytoskeleton rearrangement upon integrin activation, where the phosphorylation of cofilin drives actin filaments rearrangement and governs cell adaptation on the substrate.…”