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Biomedical subjects

J Glowacki

Publications and source records attributed to J Glowacki.

At least 109 records · Page 6Linked to original sources
PubMedDate not supplied

Perfusion of medium improves growth of human oral neomucosal tissue constructs.

Tissue engineering of the oral mucosa may be useful in congenital cleft palate repairs, defects following extirpative oncologic surgery, and periodontal disease. One of the limitations of in vitro growth of oral mucosal constructs is central necrosis of 3-dimensional tissues. We tested the hypothesis that medium perfusion would enhance oral mucosal histogenesis in vitro. Normal human oral keratinocytes were obtained from young to middle-aged adults. Porous 3-dimensional matrices were prepared from collagen and chondroitin sulfate with some crosslinked with glutaraldehyde. Each device was seeded with 5.0 x 10(5) human oral keratinocytes. The seeded matrices were cultured with or without perfusion of medium at 1.3 ml/min. Histologic analysis of samples cultured for 3, 7, or 14 days showed superior viability and proliferation when perfused. At day 7, the average number of cell layers of the neoepithelium of sponges in the perfused culture system (9.4 +/- 1.0) was 88% greater than for the nonperfused culture system (5.0 +/- 0.9, p<0.005). Glutaraldehyde crosslinking did not influence cellular proliferation or the extent of matrix's shrinkage in either culture system. This study shows that medium perfusion enhanced cell viability and proliferation of human oral keratinocytes cultured in porous 3-dimensional matrices.

Cell Division↗
PubMedDate not supplied

Perfusion enhances functions of bone marrow stromal cells in three-dimensional culture.

Perfusion of medium through three-dimensional (3D) collagen sponges enhanced viability and function of cocultivated marrow stromal and hematopoietic cell lines. Cells of the murine bone marrow stromal cell line GPIa were cultured in novel 3D collagen sponges, made from pepsin-digested bovine skin. Static cultures of sponges were maintained in dishes with media changes every other day. Perfused sponges were contained in a glass column with medium flow set at 1.3 mL/min. In some sponges, the 32D cl3 c-fms(m) (CRX-1) hematopoietic progenitor cell line was added 7 days after GPIa cells. At 7 and 16 days, light microscopic evaluation showed poor viability of cells in static sponge cultures. In perfused sponge cultures, there was greater cellularity throughout the sponge and abundant accumulation of metachromatic extracellular matrix surrounding GPIa cells. Chondroitin 6-sulfate and heparan sulfate were identified as components of the matrix by immunohistochemical methods. DNA synthesis was evaluated by 15-h exposure of cultures to bromodeoxyuridine (BrdU), with subsequent immunohistochemical localization with monoclonal anti-BrdU antibody. Cells positive for BrdU were identified at the outer surfaces of both static and perfused sponges; however, positive cells were also seen throughout the internal areas of the sponges that were perfused. These results suggest that better nutrient exchange occurred in perfused sponges. In static cocultures of GPIa and CRX-1 cells, there was no detectable viability of the IL-3-dependent CRX-1 cells; however, under perfused conditions, CRX-1 cells flourished within the sponges as documented by BrdU incorporation. Thus, medium perfusion enhanced GPIa stromal cell line viability and function in 3D collagen sponge cultures, as demonstrated by BrdU incorporation, matrix production, and support of CRX-1 cells. This novel culture system may be useful for examining the interactions of bone marrow stromal cells with extracellular matrix molecules, soluble and matrix-bound factors, and with other cell types.

Animals↗
PubMedDate not supplied

In vitro engineering of cartilage.

Because adult human cartilage shows poor capacity for repair and regeneration, innovative solutions are required for congenital and acquired degenerative cartilage lesions. Acquired lesions occur in young and old alike, the former being more at risk for sports-related injuries and the latter for age-related degenerative changes. Because cartilage is a relatively simple tissue with respect to its cellular homogeneity and avascularity, it has been a model for research of in vitro engineered tissues. Progress has been slow and obstructed on several levels. The adult chondrocyte has limited capacity for proliferation and has both catabolic and anabolic functions. These metabolic features must be controlled in order for engineered tissue to endure. Use of three-dimensional scaffolds can be combined with regulatory factors (cytokine, extracellular matrix [ECM], and mechanical) to optimize conditions for in vitro engineered cartilage. Cross-disciplinary interactions are likely to accelerate progress and to mediate application of advances made in other fields for consistently successful in vitro engineering of cartilage for all clinical needs.

Adult↗