Osteogenic differentiation and proliferation of bone marrow-derived mesenchymal stromal cells on PDLLA + BMP-2-coated titanium alloy surfaces.
RhBMP-2 is clinically applied to enhance bone healing and used in combination with titanium fixation implants. The purpose of this in vitro study was to compare the osteogenic differentiation and proliferation of hMSC on native polished versus sandblasted titanium surfaces (TS) and to test their behavior on pure poly-D,L-lactide (PDLLA) coated as well as PDLLA + rhBMP-2 coated TS. Furthermore, the release kinetics of PDLLA + rhBMP-2-coated TS was investigated. Human bone marrow cells were obtained from three different donors (A: male, 16 yrs; B: male, 27 yrs, C: male, 49 yrs) followed by density gradient centrifugation and flow cytometry with defined antigens. The cells were seeded on native polished and sandblasted TS, PDLLA-coated TS and PDLLA + rhBMP-2-coated TS. Osteogenic differentiation (ALP specific activity via ALP and BCA assay) and proliferation (LDH cytotoxicity assay) was examined on day 7 and 14 and release kinetics of rhBMP-2 was investigated on day 3, 7, 10, and 14. We found significant higher ALP specific activity and LDH activity on native polished compared to native sandblasted surfaces. PDLLA led to decreased ALP specific and LDH activity on both surface finishes. Additional rhBMP-2 slightly diminished this effect. RhBMP-2-release from coated TS decreased nearly exponentially with highest concentrations at the beginning of the cultivation period. The results of this in vitro study suggest that native TS stimulate hMSC significantly stronger toward osteogenic differentiation and proliferation than rhBMP-2 + PDLLA-layered TS in the first 14 days of cultivation. The PDLLA-layer seems to inhibit local hMSC differentiation and proliferation.
Department of Orthopaedics and Trauma Surgery, University Duisburg-Essen, Essen, Germany.
Marcel
M
Haversath
Department of Orthopaedics, Medical Faculty, University Düsseldorf, Düsseldorf, Germany.
Tobias
T
Hülsen
Department of Orthopaedics, Medical Faculty, University Düsseldorf, Düsseldorf, Germany.
Carolin
C
Böge
Department of Orthopaedics and Trauma Surgery, University Duisburg-Essen, Essen, Germany.
Tjark
T
Tassemeier
Department of Orthopaedics and Trauma Surgery, University Duisburg-Essen, Essen, Germany.
Stefan
S
Landgraeber
Department of Orthopaedics, Medical Faculty, University Düsseldorf, Düsseldorf, Germany.
Monika
M
Herten
Department of Orthopaedics and Trauma Surgery, University Duisburg-Essen, Essen, Germany.
Sebastian
S
Warwas
Department of Orthopaedics, Medical Faculty, University Düsseldorf, Düsseldorf, Germany.
Rüdiger
R
Krauspe
Department of Orthopaedics and Trauma Surgery, University Duisburg-Essen, Essen, Germany.
Marcus
M
Jäger
D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D000497:Alloys / Q000494:pharmacology*; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D020099:Coated Materials, Biocompatible / Q000494:pharmacology*; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D051379:Mice; D008875:Middle Aged; D010012:Osteogenesis / Q000187:drug effects*; D011091:Polyesters / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology; D013499:Surface Properties; D014025:Titanium / Q000494:pharmacology*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D000497:Alloys; D055396:Bone Morphogenetic Protein 2; D020099:Coated Materials, Biocompatible; D011091:Polyesters; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; C033616:poly(lactide); D014025:Titanium; D000469:Alkaline Phosphatase
BMP-2; differentiation; human mesenchymal stromal cells; poly-D,L-lactide; proliferation