The vacuum-bioreactor technique allowed seeded chondrocytes to attach to PGA scaffolds within 48 hours and improved uniformity of cell distribution, compared with the spinner technique. Although formation of extracellular matrix may be stimulated by seeding chitosan scaffolds in the bioreactor, further evaluations of the seeding technique and characteristics of chitosan scaffolds are warranted.

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A significant amount of the data collected by cell biologists and tissue engineers relies on invasive imaging techniques to visualize dynamic structural and functional properties in engineered tissues. We report the use of optical coherence tomography and the comparative use of confocal microscopy to nondestructively and noninvasively monitor the structural and functional characteristics of three-dimensional engineered tissues over time. The engineered tissue model is composed of chitosan scaffolds and fibroblasts transfected with vinculin fused to green fluorescent protein. We image the developmental process of engineered tissues from changes of tissue microarchitecture to cell-matrix adhesions in three dimensions. These findings demonstrate the potential for optical coherence tomography in applications in cell and tissue biology, tissue engineering, and drug discovery.

Uniform distribution of cells and their extracellular matrix is essential for the in vivo success of bone tissue engineering constructs produced in vitro. In this study, the effects of biphasic calcium phosphate (BCP) granules embedded into chitosan scaffolds on the distribution, morphology, and phenotypic expression of osteoblastic cells were investigated. Mesenchymal stem cells (MSCs) and preosteoblasts were cultured on chitosan scaffolds with and without BCP under osteoblastic differentiation/maturation conditions for periods up to 4 weeks. The addition of 25 wt % BCP to chitosan created a uniform layer of calcium phosphate (CaP) precipitation similar to bone mineral on the scaffold surfaces as determined by scanning electron microscopy and X-ray spectroscopy. Scaffolds with this CaP layer yielded more uniform and complete cell and ECM distribution than chitosan scaffolds without BCP. The suggestion of chemotaxis in the appearance of this response was confirmed by successive experiments in a Boyden chamber. The CaP layer also altered morphology of cells initially attached to the scaffold surfaces, leading to higher expression of marker proteins of osteoblastic phenotype including alkaline phosphatase and osteocalcin. The use of chitosan/BCP scaffolds for culture of MSCs and preosteoblasts enhances bone tissue development in vitro.

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