Nanoindentation of soft hydrated materials for application to vascular tissues.

Soft hydrated materials, such as vascular tissues and other biomaterials, provide a number of challenges in the field of nanoindentation. However, the ability of nanoindentation to probe local, nanoscale mechanical properties of heterogeneous materials makes it desirable to adapt this technique for application to biologic tissues. To develop the field of nanoindentation for the analysis of soft hydrated materials, the goals of this study were fourfold: develop a sample hydration system, select an appropriate tip for soft material indentation, identify a substrate to be used for blunt tip alignment, and determine an appropriate control material for the development of future indentation protocols. A hydration system was developed that maintained sample hydration for over 8 h without completely submerging the sample. Further, a 100-microm radius of curvature conospherical tip was shown to be a suitable tip for indenting a variety of soft hydrated materials and back-illuminated agarose gel was found to be an effective material for use in tip alignment. Finally, agarose gel demonstrated similar qualitative and quantitative nanomechanical behavior to vascular tissue, suggesting that it will be an appropriate control material for the development of future indentation protocols for soft biologic tissues.

Mechanical and chemical characteristics of mineral produced by basic fibroblast growth factor-treated bone marrow stromal cells in vitro.

It has been shown that various organ and cell cultures exhibit increased mineral formation with the addition of basic fibroblast growth factor (bFGF) and phosphate ions in the medium. However, to date there has been no attempt to relate the chemical composition of mineral formed in vitro to a measure of its mechanical properties. This information is important for understanding the in vivo mineralization process, the development of in vitro models, and the design of tissue-engineered bone substitutes. In this study we examined the reduced modulus; hardness; and mineral-to-matrix, crystallinity, carbonate-to-mineral, and calcium-to-phosphorus ratios of mineral formed by bFGF-treated rat-derived bone marrow stromal cells in vitro. The cells were treated with 1 or 3 mM beta-glycerophosphate for 3 and 4 weeks. Both mechanical parameters, reduced modulus and hardness, increased with increasing beta-glycerophosphate concentration. The only chemical measure of the mineral composition that exhibited the same dependency was the mineral-to-matrix ratio. The values of crystallinity and carbonate fraction were similar to those for intact cortical bone, but the calcium-to-phosphorus ratio was substantially lower than that of normal bone. These data indicate that the mineral formed by bFGF-treated bone cells is mechanically and chemically different from naturally formed lamellar bone tissue after 4 weeks in culture. These results can be used to improve in vitro models of mineral formation as well as enhance the design of tissue-engineered bone substitutes.