Fabrication and Characterization of Recombinant Silk-Elastin-Like-Protein (SELP) Fiber.

Silk-elastin-like-protein polymers (SELPs) are genetically engineered recombinant protein sequences consisting of repeating units of silk-like and elastin-like blocks. By combining these entities, it is shown that both the characteristic strength of silk and the temperature-dependent responsiveness of elastin can be leveraged to create an enhanced stimuli-responsive material. It is hypothesized that SELP behavior can be influenced by varying the silk-to-elastin ratio.

Micropatterned cell sheets as structural building blocks for biomimetic vascular patches.

To successfully develop a functional tissue-engineered vascular patch, recapitulating the hierarchical structure of vessel is critical to mimic mechanical properties. Here, we use a cell sheet engineering strategy with micropatterning technique to control structural organization of bovine aortic vascular smooth muscle cell (VSMC) sheets. Actin filament staining and image analysis showed clear cellular alignment of VSMC sheets cultured on patterned substrates.

Predicting Silk Fiber Mechanical Properties through Multiscale Simulation and Protein Design.

Silk is a promising material for biomedical applications, and much research is focused on how application-specific, mechanical properties of silk can be designed synthetically through proper amino acid sequences and processing parameters. This protocol describes an iterative process between research disciplines that combines simulation, genetic synthesis, and fiber analysis to better design silk fibers with specific mechanical properties. Computational methods are used to assess the protein polymer structure as it forms an interconnected fiber network through shearing and how this process affects fiber mechanical properties.

Design Approaches to Myocardial and Vascular Tissue Engineering.

Engineered tissues represent an increasingly promising therapeutic approach for correcting structural defects and promoting tissue regeneration in cardiovascular diseases. One of the challenges associated with this approach has been the necessity for the replacement tissue to promote sufficient vascularization to maintain functionality after implantation. This review highlights a number of promising prevascularization design approaches for introducing vasculature into engineered tissues.

Introducing biomimetic shear and ion gradients to microfluidic spinning improves silk fiber strength.

Silkworm silk is an attractive biopolymer for biomedical applications due to its high mechanical strength and biocompatibility; as a result, there is increasing interest in scalable devices to spin silk and recombinant silk so as to improve and customize their properties for diverse biomedical purposes (Vepari and Kaplan 2007 Prog. Polym. Sci.

Silk-fibronectin protein alloy fibres support cell adhesion and viability as a high strength, matrix fibre analogue.

Silk is a natural polymer with broad utility in biomedical applications because it exhibits general biocompatibility and high tensile material properties. While mechanical integrity is important for most biomaterial applications, proper function and integration also requires biomaterial incorporation into complex surrounding tissues for many physiologically relevant processes such as wound healing. In this study, we spin silk fibroin into a protein alloy fibre with whole fibronectin using wet spinning approaches in order to synergize their respective strength and cell interaction capabilities.

Coordinated regulation of acid resistance in Escherichia coli.

Background: Enteric Escherichia coli survives the highly acidic environment of the stomach through multiple acid resistance (AR) mechanisms. The most effective system, AR2, decarboxylates externally-derived glutamate to remove cytoplasmic protons and excrete GABA. The first described system, AR1, does not require an external amino acid.

Silk-Its Mysteries, How It Is Made, and How It Is Used.

This article reviews fundamental and applied aspects of silk-one of Nature's most intriguing materials in terms of its strength, toughness, and biological role-in its various forms, from protein molecules to webs and cocoons, in the context of mechanical and biological properties. A central question that will be explored is how the bridging of scales and the emergence of hierarchical structures are critical elements in achieving novel material properties, and how this knowledge can be explored in the design of synthetic materials. We review how the function of a material system at the macroscale can be derived from the interplay of fundamental molecular building blocks.

Current approaches to electrospun nanofibers for tissue engineering.

The ultimate goal of tissue engineering is to replace damaged tissues by applying engineering technology and the principles of life sciences. To successfully engineer a desirable tissue, three main elements of cells, scaffolds and growth factors need to be harmonized. Biomaterial-based scaffolds serve as a critical platform both to support cell adhesion and to deliver growth factors.

Mussel-inspired surface modification of poly(L-lactide) electrospun fibers for modulation of osteogenic differentiation of human mesenchymal stem cells.

Development of biomaterials to control the fate of stem cells is important for stem cell based regeneration of bone tissue. The objective of this study is to develop functionalized electrospun fibers using a mussel-inspired surface coating to regulate adhesion, proliferation and differentiation of human mesenchymal stem cells (hMSCs). We prepared poly(L-lactide) (PLLA) fibers coated with polydopamine (PD-PLLA).

Development of functional fibrous matrices for the controlled release of basic fibroblast growth factor to improve therapeutic angiogenesis.

In this study, novel fibrous matrices were developed as a depot to store and liberate growth factors in a controlled manner. Specifically, heparin was covalently conjugated onto the surface of fibrous matrices (composites of poly[caprolactone] and gelatin crosslinked with genipin), and basic fibroblast growth factor (bFGF) was then reversibly immobilized. The immobilization of bFGF was controlled as a function of the amount of conjugated heparin.

Control of osteogenic differentiation and mineralization of human mesenchymal stem cells on composite nanofibers containing poly[lactic-co-(glycolic acid)] and hydroxyapatite.

We fabricated composite fibrous scaffolds from blends of poly(lactide-co-glycolide) (PLGA) and nano-sized hydroxyapatite (HA) via electrospinning. SEM-EDX and AFM analysis demonstrated that HA was homogeneously dispersed in the nanofibers, and the roughness increased along with the amount of incorporated HA. When hMSCs were cultured on these PLGA/HA composite nanofibers, we found that incorporation of HA on the nanofibers did not affect cell viability whereas increased ALP activity and expression of osteogenic genes as well as the calcium mineralization of hMSCs.

Modulation of osteogenic differentiation of human mesenchymal stem cells by poly[(L-lactide)-co-(epsilon-caprolactone)]/gelatin nanofibers.

Developing biomaterial scaffolds to elicit specific cell responses is important in many tissue engineering applications. We hypothesized that the chemical composition of the scaffold may be a key determinant for the effective induction of differentiation in human mesenchymal stem cells (hMSCs). In this study, electrospun nanofibers with different chemical compositions were fabricated using poly[(L-lactide)-co-(epsilon-caprolactone)] (PLCL) and gelatin.

In vitro osteogenic differentiation of human mesenchymal stem cells and in vivo bone formation in composite nanofiber meshes.

Tissue engineering has become an alternative method to traditional surgical treatments for the repair of bone defects, and an appropriate scaffold supporting bone formation is a key element in this approach. In the present study, nanofibrous organic and inorganic composite scaffolds containing nano-sized demineralized bone powders (DBPs) with biodegradable poly(L-lactide) (PLA) were developed using an electrospinning process for engineering bone. To assess their biocompatibility, in vitro osteogenic differentiation of human mandible-derived mesenchymal stem cells (hMSCs) cultured on PLA or PLA/DBP composite nanofiber scaffolds were examined.