Crosslinking of collagen gels by transglutaminase.

Collagen is commonly used as a tissue-engineering scaffold, yet its in vivo applications are limited by a deficiency in mechanical strength. The purpose of this work was to explore the utilization of a unique enzymatic crosslinking procedure aimed at improving the mechanical properties of collagen-based scaffold materials. Type I bovine collagen gel was crosslinked by transglutaminase, which selectively mediates the chemical reaction between glutamine and lysine residues on adjacent protein fibers, thus providing covalent amide bonds that serve to reinforce the three-dimensional matrix. The degree of crosslinking was verified by thermal analysis and amine group content. The denaturation temperature of crosslinked collagen reached a maximum of 66 +/- 1 degrees C. The chemical reaction was confirmed to be noncytotoxic with respect to bone marrow stromal cells acquired from New Zealand White rabbits. Tube-shaped cellular constructs fashioned from crosslinked collagen and bone marrow stromal cells were found to have burst pressures significantly higher than their noncrosslinked analogs (71 +/- 4 mmHg vs. 46 +/- 3 mmHg; p < 0.01). Thus, the transglutaminase mediated reaction served to successfully strengthen collagen gels while remaining benign toward cells.

Composition options for tissue-engineered bone.

The logical assembly of tissue-engineered bone is ultimately directed by the clinical status of the patient. The basic elements for tissue-engineered bone should include signaling molecules, cells, and extracellular matrix. The assembly of these basic elements may need to be modified by tissue engineers to account for patient variables of age, gender, health, systemic conditions, habits, and anatomical implant. Moreover, different regions of the body will have different functional loads and vascularity. This review discusses several basic options that may be necessary to engineer bone, including spatial and temporal assembly of signaling factors, cells, and biomimetic extracellular matrices. Moreover, the importance of the health care status of the patient who may be receiving the tissue-engineered composition is emphasized.