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WIREs Nanomed Nanobiotechnol
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Advances in cellular and tissue engineering using layer‐by‐layer assembly

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Layer‐by‐layer (LbL) assembly is a self‐assembly technique used to develop multilayer films based on complementary interactions between film components. These multilayer films have had a significant impact on the fields of cellular and tissue engineering. The aim of cellular engineering is to understand and control cell behavior, which not only impacts applications in regenerative medicine but also other biomedical therapies that rely on cell interactions with biomaterials, including treatments for autoimmune disorders and cancer. Tissue engineering approaches to tissue repair and regeneration utilize three‐dimensional biomaterial scaffolds that interact favorably with cells. Cellular engineering studies can better inform the design of these scaffolds. The ease of tuning the chemical and mechanical properties of LbL films, the ability to coat a variety of medically relevant substrates (including cell culture surfaces and scaffolds), and the wide range of species that can be incorporated into these films (ranging from proteins to small molecules) have led to the successful use of LbL assembly for a variety of cellular and tissue engineering applications. The films used in these biomedical applications can be divided into those that release therapeutics, often with controlled stimuli‐responsive release behavior, and those that act without releasing these agents. This article is categorized under: Implantable Materials and Surgical Technologies > Nanomaterials and Implants Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement
Layer‐by‐layer (LbL) film assembly. A schematic of a (polymer 1/polymer 2)n bilayer film assembled using dip LbL assembly is shown. A negatively charged substrate is submerged in a solution containing a dissolved polycation, polymer 1. Following polymer 1 adsorption, a wash step removes weakly bound species. A polyanion, polymer 2, is then adsorbed to the growing film followed by a wash step, completing the deposition of the first bilayer. Repeating these steps n times allows the assembly of an LbL film containing n bilayers.
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Histological analysis of rat tissue at 9 weeks following implantation of bone morphogenetic protein‐2 (BMP‐2)‐releasing films (a) versus control scaffolds (b). Blue staining shows lamellar bone formation in (a) versus (b). Tissue exposed to BMP‐2‐releasing films showed active bone formation with osteoblasts (blue arrows), mature osteocytes (white asterisks), and cement line (white arrow) (c). Fatty marrow spaces were seen in mature lamellar bone (d and e). Woven bone formation occurred (f), while cartilaginous areas were remodeled into mature bone (g and h) at 4 weeks following implantation of the BMP‐2‐releasing scaffold. (Reprinted with permission from Ref . Copyright 2011 Elsevier)
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Examining biocompatibility of enzyme‐mediated, insulin‐releasing layer‐by‐layer (LbL) film‐coated implants. Hematoxylin and eosin staining of rat tissue in an untreated rat (a), tissue surrounding an uncoated control implant (b), and various formulations of LbL film‐coated implants (c–e). There are no signs of an adverse chronic inflammatory response; LbL film‐treated tissues look comparable to normal tissue (FB, fibroblast; L, lymphocyte; C, collagen). (Reprinted with permission from Ref . Copyright 2012 Elsevier)
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Human hepatocellular carcinoma cells (HepG2) cultured on various layer‐by‐layer (LbL) film architectures composed of poly(l‐lysine) (PLL), alginate (ALG), poly(allylamine hydrochloride) (PAH), and/or poly(sodium‐4‐styrene‐sulfonate) (PSS). Cell adhesion and proliferation varies with LbL film composition. (Reprinted with permission from Ref . Copyright 2008 Elsevier)
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Human bone marrow stromal cells (hMSCs) successfully adhere to and grow on photo‐cross‐linked layer‐by‐layer (LbL) films containing diphenylamine‐4‐diazoresin and pectin over time. HMSCs were cultured on LbL films for 1 day (a), 3 days (b), and 6 days (c). Quantification of cell density versus culture time is shown in (d). (Reprinted with permission from Ref . Copyright 2011 Elsevier)
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Implantable Materials and Surgical Technologies > Nanomaterials and Implants
Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement

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