Altered nanofeature size dictates stem cell differentiation.

The differentiation of stem cells can be modulated by physical factors such as the micro- and nano-topography of the extracellular matrix. One important goal in stem cell research is to understand the concept that directs differentiation into a specific cell lineage in the nanoscale environment. Here, we demonstrate that such paths exist by controlling only the micro- and nano-topography of polymer surfaces. Altering the depth (on a nanometric scale) of micro-patterned surface structures allowed increased adhesion of human mesenchymal stem cells (hMSCs) with specific differentiation into osteoblasts, in the absence of osteogenic medium. Small (10 nm) depth patterns promoted cell adhesion without noticeable differentiation, whereas larger depth patterns (100 nm) elicited a collective cell organization, which induced selective differentiation into osteoblast-like cells. This latter response was dictated by stress through focal-adhesion-induced reorganization of F-actin filaments. The results have significant implications for understanding the architectural effects of the in vivo microenvironment and also for the therapeutic use of stem cells.

Polysaccharide-coated thermosets for orthopedic applications: from material characterization to in vivo tests.

The long-term stability and success of orthopedic implants depend on the osseointegration process, which is strongly influenced by the biomaterial surface. A promising approach to enhance implant integration involves the modification of the surface of the implant by means of polymers that mimic the natural components of the extracellular matrix, for example, polysaccharides. In this study, methacrylate thermosets (bisphenol A glycidylmethacrylate/triethyleneglycol dimethacrylate), a widely used composition for orthopedic and dental applications, have been coated by electrostatic deposition of a bioactive chitosan-derivative. This polysaccharide was shown to induce osteoblasts aggregation in vitro, to stimulate cell proliferation and to enhance alkaline phosphatase activity. The coating deposition was studied by analyzing the effect of pH and ionic strength on the grafting of the polysaccharide. Contact angle studies show that the functionalized material displays a higher hydrophilic character owing to the increase of surface polar groups. The mechanical properties of the coating were evaluated by nanoindentation studies which point to higher values of indentation hardness and modulus (E) of the polysaccharide surface layer, while the influence of cyclic stress on the construct was assessed by fatigue tests. Finally, in vivo tests in minipigs showed that the polysaccharide-based implant showed a good biocompatibility and an ability for osseointegration at least similar to that of the titanium Ti6Al4V alloy with roughened surface.

Terminal sterilization of BisGMA-TEGDMA thermoset materials and their bioactive surfaces by supercritical CO2.

The development of biomaterials endowed with bioactive features relies on a simultaneous insight into a proper terminal sterilization process. FDA recommendations on sterility of biomaterials are very strict: a sterility assurance level (SAL) of 10(-6) must be guaranteed for biomaterials to be used in human implants. In the present work, we have explored the potential of supercritical CO(2) (scCO(2)) in the presence of H(2)O(2) as a low-temperature sterilization process for thermoset materials and their bioactive surfaces. Different conditions allowing for terminal sterilization have been screened and a treatment time-amount of H(2)O(2) relationship proposed. The selected terminal sterilization conditions did not notably modify the mechanical properties of the thermoset nor of their fiber-reinforced composites. This was confirmed by µCT analyses performed prior to and after the treatment. On the contrary, terminal sterilization in the presence of H(2)O(2) induced a slight decrease in the surface hardness. The treatment of the thermoset material with scCO(2) led to a reduction in the residual unreacted monomers content, as determined by means of high performance liquid chromatography (HPLC) analyses. Finally, it was found that a thermoset coated with a polysaccharide layer containing silver nanoparticles maintained a very high antimicrobial efficacy even after the scCO(2)-based terminal sterilization.

In vivo remodeling of human cell-assembled extracellular matrix yarns.

Cell-assembled extracellular matrix (CAM) has been used to produce vascular grafts. While these completely biological vascular grafts performed well in clinical trials, the in vivo remodeling and inflammatory response of this truly "bio" material has not yet been investigated. In this study, human CAM yarns were implanted subcutaneously in nude rats to investigate the innate immune response to this matrix. The impact of processing steps relevant to yarn manufacturing was evaluated (devitalization, decellularization, gamma sterilization, and twisting). We observed that yarns were still present after six months, and were integrated into a non-inflamed loose connective tissue. The CAM was repopulated by fibroblastic cells and blood vessels. While other yarns caused minor peripheral inflammation at an early stage (two weeks of implantation), gamma sterilization triggered a more intense host response dominated by the presence of M1 macrophages. The inflammatory response was resolved at six months. Yarn mechanical strength was decreased two weeks after implantation except for the more compact "twisted" yarn. While the strength of other yarns was stable after initial remodeling, the gamma-sterilized yarn continued to lose mechanical strength over time and was weaker than devitalized (control) yarns at six months. This is the first study to formally demonstrate that devitalized human CAM is very long-lived in vivo and does not trigger a degradative response, but rather is very slowly remodeled. This data supports a strategy to produce human textiles from CAM yarn for regenerative medicine applications where a scaffold with low inflammation and long-term mechanical properties are critical.

Human textiles: A cell-synthesized yarn as a truly "bio" material for tissue engineering applications.

In the field of tissue engineering, many groups have come to rely on the extracellular matrix produced by cells as the scaffold that provides structure and strength to the engineered tissue. We have previously shown that sheets of Cell-Assembled extracellular Matrix (CAM), which are entirely biological yet robust, can be mass-produced for clinical applications using normal, adult, human fibroblasts. In this article, we demonstrate that CAM yarns can be generated with a range of physical and mechanical properties. We show that this material can be used as a simple suture to close a wound or can be assembled into fully biological, human, tissue-engineered vascular grafts (TEVGs) that have high mechanical strength and are implantable. By combining this truly "bio" material with a textile-based assembly, this original tissue engineering approach is highly versatile and can produce a variety of strong human textiles that can be readily integrated in the body. STATEMENT OF SIGNIFICANCE: Yarn of synthetic biomaterials have been turned into textiles for decades because braiding, knitting and weaving machines can mass-produce medical devices with a wide range of shapes and mechanical properties. Here, we show that robust, completely biological, and human yarn can be produced by normal cells in vitro. This yarn can be used as a simple suture material or to produce the first human textiles. For example, we produced a woven tissue-engineered vascular grafts with burst pressure, suture retention strength and transmural permeability that surpassed clinical requirements. This novel strategy holds the promise of a next generation of medical textiles that will be mechanically strong without any foreign scaffolding, and will have the ability to truly integrate into the host's body.

In Vitro Mechanical Property Evaluation of Chitosan-Based Hydrogels Intended for Vascular Graft Development.

Vascular grafts made of synthetic polymers perform poorly in cardiac and peripheral bypass applications. In these applications, chitosan-based materials can be produced and shaped to provide a novel scaffold for vascular tissue engineering. The goal of this study was to evaluate in vitro the mechanical properties of a novel chitosan formulation to assess its potential for this scaffold. Two chitosan-based hydrogel tubes were produced by modulating chitosan concentration. Based on the standard ISO 7198:1998, the hydrogel tubes were characterized in vitro in terms of suture retention strength, tensile strength, compliance, and burst pressure. By increasing chitosan concentration, suture retention value increased to reach 1.1 N; average burst strength and elastic moduli also increased significantly. The compliance seemed to exhibit a low value for chitosan tubes of high concentration. By modulating chitosan concentration, we produced scaffolds with suitable mechanical properties to be implanted in vivo and withstand physiological blood pressures.