Lieb, E. and Vogel, T. and Milz, S. and Dauner, M. and Schulz, Michaela B. (2004) Effects of transforming growth factor beta(1) on bonelike tissue formation in three-dimensional cell culture - II: Osteoblastic differentiation. TISSUE ENGINEERING, 10 (9-10). pp. 1414-1425. ISSN 2152-4947,
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We supplemented rat marrow stromal cells (rMSCs) seeded on poly(L-lactic- co-glycolic acid) fiber meshes with transforming growth factor beta(1) (TGF-beta(1)) to improve bone tissue formation for tissue engineering. Whereas our first study (Lieb, E., et al. Tissue Eng. 10, 1399 - 1413, 2004) investigated the effects of TGF-beta1 on matrix formation and mineralization, this second study focused on the differentiation of rMSCs to the osteoblastic phenotype in dynamic cell culture ( orbital shaker). We assessed a series of bone markers to determine a dosing regimen for TGF-beta(1) that enhances collagenous matrix formation and preserves or increases osteoblastic differentiation. Bone sialoprotein and osteonectin formation were investigated immunohistochemically and by RT-PCR. For alkaline phosphatase activity ( ALP), we employed an enzyme assay. Osteocalcin was examined by RT-PCR as well as by an immunoassay. Whereas bone sialoprotein appeared to be dose-dependently increased in the immunochemistical stainings after supplementation with TGF-beta(1), osteonectin remained unchanged. Both ALP activity and osteocalcin were suppressed by high doses of TGF-beta(1), such as single doses of 10 ng/mL or four doses of 1 ng/mL added once a week. Considering the effects of TGF-beta(1) 1 both on differentiation and on matrix formation and mineralization, TGF-beta(1) at 1 ng/mL, added once a week in the first 1 to 2 weeks, was selected as an effective dose to improve bonelike tissue formation in vitro.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ALKALINE-PHOSPHATASE; GENE-EXPRESSION; IN-VITRO; |
| Subjects: | 600 Technology > 615 Pharmacy |
| Divisions: | Chemistry and Pharmacy > Institute of Pharmacy > Pharmaceutical Technology (Prof. Göpferich) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 30 Jun 2021 10:09 |
| Last Modified: | 30 Jun 2021 10:09 |
| URI: | https://pred.uni-regensburg.de/id/eprint/37309 |
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