Human gingival fibroblasts were cultured in vitro using as substrates an extracellular matrix (matrix) and polytetrafluoride (PTFE) membranes, which are used in guided tissue regeneration. To test the degree of biocompatibility of these membranes, the cellular proliferation and the accumulation of extracellular matrix (ECM) macromolecules were considered as parameters. The fibroblasts were cultured in vitro for 24 and 48 hours without serum on plastic, matrix, and PTFE membranes in the presence of 3H-thymidine, 3H-glucosamine, and 3H-proline to study the neo-synthesis of DNA, glycosaminoglycans (GAG), and collagen proteins, respectively. Studies on cell proliferation showed that fibroblasts grown on matrix membrane significantly increased 3H-thymidine incorporation, while fibroblasts grown on PTFE membrane decreased 3H-thymidine incorporation, compared to plastic used as a control. Moreover, the PTFE membrane induced a marked decrease of collagen and GAG accumulation both in the cellular and extracellular pool, while the matrix membrane provoked a decrease of the two macromolecules in the cellular pool and an increase in the extracellular one, compared to the control. The data we obtained demonstrate that matrix membranes are the most suitable to stimulate both cellular proliferation and ECM macromolecule accumulation.

Bone cells derived from the human jaw were cultured on titanium, titanium coated with hydroxyapatite (THA) or with plasma spray (TPS) to study the behaviour of the cells anchored to implant substrates. Bone cells were cultured in MEM with the addition of [3H]-thymidine to evaluate cellular proliferation, and [3H]-glucosamine to evaluate GAG synthesis and accumulation in the extra-cellular matrix (ECM). Moreover, to study the degradation of GAG bone cells were cultured in the presence of NH4Cl, an amine known to inhibit lysosomal activity. Our results show that TPS is the substrate that favours both cellular proliferation and the accumulation of GAG in the ECM.

Bone cells derived from human jaw were isolated from explants and grown in vitro. Subcultures were cultured on plastic (control) and metal substrates for 24 and 48 hours in medium containing 3H-glucosamine and labeled glycosaminoglycan (GAG) accumulation was measured. In bone cells cultured on metal substrates there was an evident reduction in the synthesis and secretion of radiolabeled macromolecules compared to bone cells cultured on plastic. Moreover, the accumulation of single GAG classes was specific for each substrate tested. The results showed that titanium was the only metal substrate studied in which the percentage of individual GAG classes remained the same as control cultures. GAG reduction was due to a decreased synthesis and not to an increased degradation as shown by the decrement of exoglycosidase activity. The metals also reduced the activity of transforming growth factor beta (TGF beta), measured using interleukin-1 assay method, a factor involved in the various phases of bone remodeling; in this case, too, cells grown on titanium showed the highest TGF beta activity compared to the other metal substrates studied. The results indicate that the substrate to which the cells adhere do exhibit specific differences in GAG composition and TGF beta activity. The differences observed may be important during in vivo events such as guided tissue regeneration and bone deposition.

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