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Natural extracellular matrix scaffolds recycled from human salivary digests: a morphometric study
Author(s) -
Lilliu MA,
Seo YJ,
Isola M,
Charbonneau AM,
Zeitouni A,
ElHakim M,
Tran SD
Publication year - 2016
Publication title -
oral diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.953
H-Index - 87
eISSN - 1601-0825
pISSN - 1354-523X
DOI - 10.1111/odi.12444
Subject(s) - extracellular matrix , decellularization , immunogold labelling , tissue engineering , staining , chemistry , extracellular , parenchyma , matrix (chemical analysis) , cell culture , pathology , microbiology and biotechnology , regeneration (biology) , biology , anatomy , biomedical engineering , ultrastructure , biochemistry , medicine , genetics , chromatography
Objective A challenge in engineering tissues is to supply parenchymal cells with suitable scaffolds which ideally reproduce the extracellular matrix ( ECM ). This study 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’, nECM sc) to be used as recycled natural scaffolds. The objective was to test whether nECM sc and native salivary tissue were comparable morphologically, in ECM proteins composition, and in cell seeding efficiency. Methods Following cell isolation procedures, nECM sc were kept, either fresh or frozen (sectioned into 12‐ μ m‐thick slices), and examined with high‐resolution electron microscopy ( HRSEM ) for its three‐dimensional structure, and with picrosirius red staining and immunogold staining for ECM protein composition and distribution, respectively. nECM sc were seeded with human epithelial cells and fibroblasts to assess cell attachment and proliferation in short‐term experiments. Results Under HRSEM , nECM sc had comparable fiber arrangement to original glands. Histochemical and immunogold‐labeling examinations revealed the presence of collagen types I, III , and IV . Seeded epithelial cells and fibroblasts attached, proliferated (14–55%), and were alive (86–99%) after 4–8 days of culture. Conclusions nECM sc retained native ECM proteins and maintained their distribution. Seeded cells remained viable on nECM sc.