Modulation of RGD-Functionalized Polyelectrolyte Multilayer Membranes for Promoting Osteoblast Function

Abstract

Layer-by-layer deposition of polyelectrolyte multilayer (PEM) membranes has recently been applied successfully to a number of biomedical applications. This simple and versatile technique provides a broad surface modification platform, for example, for the display of biomolecules such as cell-adhesion peptides. In this work, we investigated the effects of PEM coatings on RGD-immobilization and osteoblast cell culture. RGD-containing peptides were conjugated to the amino groups of poly(allylamine hydrochloride) (PAH), and then adsorbed on top of 10-layer PAH/poly(acrylic acid) (PAA) multilayer membranes that were assembled at either pH 2.0 or pH 6.5. MG63 osteoblast-like cells were then seeded and cultured on the RGD-conjugated surfaces. We found that the cells adhered to and grew better on the RGD-conjugated PEM membranes. Furthermore, the cells grew better on the RGD-conjugated PEM coatings assembled at pH 6.5 than those assembled at pH 2.0. On the other hand, MG63 cells exhibited better differentiated phenotype on the pH 2.0 coatings compared to the pH 6.5 coatings with respect to alkaline phosphatase activity and calcium deposition, while cells did not express osteoblast phenotype on the PAH surfaces. These results clearly show that the base PEM membranes play an important role in RGD-immobilization and osteoblast functions.

Electrospun PLGA Fibers Incorporated with Functionalized Biomolecules for Cardiac Tissue Engineering

“…Peptide-conjugated polyelectrolytes have been previously used for surface modification to enhance cell affinity of biomaterials. 15,33,36,40 We suggest that ''isolation'' and reassociation'' of bioactive domains of a protein in polyelectrolytes such as PLL construct a biofunctional ''imitative-protein'' that can perform specific functions of a protein. Such ''imitative proteins'' have some advantages over native proteins.…”

“…The graft ratios of RGD and YIGSR to PLL were estimated as 15.0 and 13.9 mol%, respectively, with respect to the total moles of the amino groups of PLL, according to a previous procedure. 15 Preparation of PLGA ESFs PLGA solution (4-8 w/v% in 1.1.1.3.3.3-hexafluoro-2-propanol, HFP) with or without PLL-g-peptides was loaded into a 5 mL syringe fitted with a 21G blunted needle, which was connected to a high voltage (10-22 kV) power supplier for electrospinning. The polymer solution was fed at a rate of 4 mL/h using a syringe pump (Model 100 series; KDS).…”

“…[1][2][3][4][5] The types of cardiac patches include acellular xenogeneic extracellular matrix (ECM), [6][7][8] scaffolds 5,[9][10][11][12] , and cell sheets. [13][14][15] Morphological similarity between the native ECM and electrospun fibers (ESFs) provided great potential for the application of biologically modified ESFs in cardiac tissue engineering. Fibers fabricated by an electrically driven jet have been employed in various biomedical applications such as tissue engineered constructs and wound healing patches.…”

Modulation of the stemness and osteogenic differentiation of human mesenchymal stem cells by controlling RGD concentrations of poly(carboxybetaine) hydrogel

“…After cell culture, cell numbers were determined by a lactate dehydrogenase assay, according to a previously described procedure [22]. Intracellular alkaline phosphatase (ALP) activities were quantified by determining enzymatic conversion of 4-nitrophenyl phosphate disodium salt to p-nitrophenol, according to a published protocol [45].…”

Mussel-Inspired Multifunctional Hydrogel Coating for Prevention of Infections and Enhanced Osteogenesis

“…For example, the introduction of Arg-Gly-Asp (RGD) short peptide could enhance adhesion and function of osteoblasts. 17 Besides, the immobilization of inorganic nanoparticles, 15 growth factors, 15,18 and microRNA-loaded nanocapsules 19 has been shown to enhance bone regeneration.…”