The future of carbon-based scaffolds in foot and ankle surgery.

Carbon may represent an alternative material suitable for future development as a soft-tissue substitute that potentially optimizes the biological and mechanical properties required for a graft product used in surgery. In addition, other modes of characterization such as 3-dimensional computational modeling may offer an insight into material performance in a biological environment. Further investigation is required to characterize and model the relationships between biological, mechanical, and design properties of this material to maximize its potential as a biomechanical scaffold and vehicle for delivering biologics that promote tissue repair and regeneration.

Cellular automata simulation of osteoblast growth on microfibrous-carbon-based scaffolds.

The objective of this study was to investigate the use of three fibrous carbon materials (T300, P25, and P120) for bone repair and develop and validate theoretical and computational methods in which bone tissue regeneration and repair could be accurately predicted. T300 was prepared from polyacrylonitrile precursor while P25 and P120 fibers were prepared from pitch, both common fiber precursors. Results showed that osteoblast growth on carbon scaffolds was enhanced with increased crystallinity, surface roughness, and material orientation. For unidirectional scaffolds at 120 h, there was 33% difference in cell growth between T300 and P25 fibers and 64% difference between P25 and P120 fibers. Moreover, for multidirectional fibers at 120 h, there was 35% difference in cell growth between T300 and P25 fibers and 43% difference between P25 and P120 fibers. Results showed that material alignment was integral to promoting cell growth with multidirectional scaffolds having the capacity for greater growth over unidirectional scaffolds. At 120 h there was 24% increase in cell growth between unidirectional alignment and multidirectional alignment on high-crystalline carbon fibers. Ultimately, data indicated that carbon scaffolds exhibited excellent bioactivity and may be tuned to stimulate unique reactions. Additionally, numerical and computational simulations provided evidence that corroborated experimental data with simulations. Results illustrated the capability of cellular automata models for assessing osteoblast cell response to biomaterials.

Hybrid carbon-based scaffolds for applications in soft tissue reconstruction.

Current biomedical scaffolds utilized in surgery to repair soft tissues commonly fail to meet the optimal combination of biomechanical and tissue regenerative properties. Carbon is a scaffold alternative that potentially optimizes the balance between mechanical strength, durability, and function as a cell and biologics delivery vehicle that is necessary to restore tissue function while promoting tissue repair. The goals of this study were to investigate the feasibility of fabricating hybrid fibrous carbon scaffolds modified with biopolymer, polycaprolactone and to analyze their mechanical properties and ability to support cell growth and proliferation. Environmental scanning electron microscopy, micro-computed tomography, and cell adhesion and cell proliferation studies were utilized to test scaffold suitability as a cell delivery vehicle. Mechanical properties were tested to examine load failure and elastic modulus. Results were compared to an acellular dermal matrix scaffold control (GraftJacket(®) [GJ] Matrix), selected for its common use in surgery for the repair of soft tissues. Results indicated that carbon scaffolds exhibited similar mechanical maximums and capacity to support fibroblast adhesion and proliferation in comparison with GJ. Fibroblast adhesion and proliferation was collinear with carbon fiber orientation in regions of sparsely distributed fibers and occurred in clusters in regions of higher fiber density and low porosity. Overall, fibroblast adhesion and proliferation was greatest in lower porosity carbon scaffolds with highly aligned fibers. Stepwise multivariate regression showed that the variability in maximum load of carbon scaffolds and controls were dependent on unique and separate sets of parameters. These finding suggested that there were significant differences in the functional implications of scaffold design and material properties between carbon and dermis derived scaffolds that affect scaffold utility as a tissue replacement construct.

A novel approach to control growth, orientation, and shape of human osteoblasts.

Carbon-based materials are considered to be promising as implants because of their unique mechanical and biocompatibility properties. This article investigates the use of carbon-based materials as a functional interface for tissue scaffolds and medical implants. Three basic parameters were explored: graphene orientation, crystallinity, and surface interaction. These parameters were measured using optical microscopy, x-ray diffraction, and atomic force microscopy. To explore the effect of the orientation, samples were made with and without a preferred carbon orientation. Conversely, the crystallinity was studied using graphitic and carbonaceous matrices. Finally, the surface interaction study was carried out using oxygen surface functionalized and non-functionalized carbon fibers. Fluorescent, confocal, and environmental scanning microscopy were used to visualize cell response. The cell attachment, proliferation, and elongation were prevalent on the unidirectional carbon preform. Cells tended to orient parallel to the fiber axis (parallel to the 002 graphene plane) and proliferate as a function of higher crystallinity, although the addition of oxygen or other functional groups disrupted the interaction between cells and graphene surface and inhibited the growth. In conclusion, osteoblast (bone-forming cells) attachment and overall growth is a function of crystallite size, graphene orientation, and carbon graphitization.

Differential binding of plasminogen, plasmin, and angiostatin4.5 to cell surface beta-actin: implications for cancer-mediated angiogenesis.

Angiostatin4.5 (AS4.5) is the product of plasmin autoproteolysis and consists of kringles 1 to 4 and approximately 85% of kringle 5. In culture, cancer cell surface globular beta-actin mediates plasmin autoproteolysis to AS4.5. We now show that plasminogen binds to prostate cancer cells and that the binding colocalizes with surface beta-actin, but AS4.5 does not bind to the cell surface. Plasminogen and plasmin bind to immobilized beta-actin similarly, with a Kd of approximately 140 nmol/L. The binding is inhibited by epsilon-aminocaproic acid (epsilonACA), indicating the requirement for a lysine-kringle domain interaction. Using a series of peptides derived from beta-actin in competitive binding studies, we show that the domain necessary for plasminogen binding is within amino acids 55 to 69 (GDEAQSKRGILTLKY). Substitution of Lys61 or Lys68 with arginine results in the loss of the ability of the peptide to block plasminogen binding, indicating that Lys61 and Lys68 are essential for plasminogen binding. Other actin peptides, including peptides with lysine, did not inhibit the plasminogen-actin interaction. AS4.5 did not bind actin at concentrations up to 40 micromol/L. Plasminogen, plasmin, and AS4.5 all contain kringles 1 to 4; however, kringle 5 is truncated in AS4.5. Isolated kringle 5 binds to actin, suggesting intact kringle 5 is necessary for plasminogen and plasmin to bind to cell surface beta-actin, and the truncated kringle 5 in AS4.5 results in its release from beta-actin. These data may explain the mechanism by which AS4.5 is formed locally on cancer cell surfaces and yet acts on distant sites.