Abstract
A challenge in engineering tissues is to supply parenchymal cells with suitable scaffolds which ideally reproduce the extracellular matrix (ECM). This paper tested the hypothesis of preserving the 'residual connective tissue' remaining after mechanical and enzymatic release of cells from human submandibular gland biopsies (that we named "natural ExtraCellular Matrix scaffolds", nECMsc) to be used as recycled natural scaffolds. The objective was to test if nECMsc and native salivary tissue were comparable morphologically, in ECM proteins composition, and in cell seeding efficiency. Following cell isolation procedures, nECMsc was kept, either fresh or frozen (sectioned into 12μm-thick slices), and examined with high-resolution electron microscopy (HRSEM) for its three-dimensional structure, and with picro-sirius-red staining and immunogold staining for ECM protein composition and distribution, respectively. nECMsc were seeded with human epithelial cells and fibroblasts to assess cell attachment and proliferation in short-term experiments. Under HRSEM, nECMsc had comparable fiber arrangement to original glands. Histochemical and immunogold-labeling examinations revealed the presence of collagen type I, III, and IV. Seeded epitheli...Continue Reading
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