Mesenchymal stem cells cultured on a collagen scaffold: In vitro osteogenic differentiation.
OBJECTIVE
Management of periodontal defects has always been a challenge in clinical periodontics. Recently mesenchymal stem cells (MSC) have been proposed for tissue regeneration in periodontal disease and repair of large bone defects. Bone regeneration has to be supported by a scaffold which has to be biocompatible, biodegradable, and able to support cell growth and differentiation. The aim of this study was to evaluate osteogenic differentiation of MSC seeded on a collagen scaffold.


METHODS
MSC were obtained from adult rat bone marrow, expanded and cultured in plastic dishes or seeded in a collagen scaffold (Gingistat). MSC were induced towards osteogenic differentiation using osteogenic supplements. Cell differentiation and calcium deposits were evaluated by immunoblotting, immunohistochemistry, histochemical techniques, enzymatic activity assay, and SEM-EDX analysis. Biomaterial in vitro degradation was evaluated by measuring mass reduction after incubation in culture medium.


RESULTS
Rat MSC osteogenic differentiation was demonstrated by osteopontin and osteocalcin expression and an increase in alkaline phosphatase activity. MSC were distributed homogeneously in the collagen scaffold. Nodular aggregates and alizarin red stained calcium deposits were observed in MSC induced towards osteogenic differentiation cultured in dishes or seeded in the collagen scaffold. SEM-EDX analysis demonstrated that calcium co-localized with phosphorous. The biomaterial in vitro degraded in 4-5 weeks.


CONCLUSIONS
MSC from bone marrow differentiate towards osteogenic lineage, representing a suitable cell source for bone formation in periodontal regeneration. Gingistat collagen scaffold supports MSC distribution and differentiation, but its short degradation time may be a limitation for a future application in bone tissue regeneration.
Donzelli
E
E
Dipartimento di Neuroscienze e Tecnologie Biomediche, Università degli Studi di Milano-Bicocca, Via Cadore 48, 20052 Monza, MI, Italy. elisabetta.donzelli@unimib.it
Salvadè
A
A
Mimo
P
P
Viganò
M
M
Morrone
M
M
Papagna
R
R
Carini
F
F
Zaopo
A
A
Miloso
M
M
Baldoni
M
M
Tredici
G
G
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D000880:Anthraquinones / Q000032:analysis; D001854:Bone Marrow Cells / Q000502:physiology; D002118:Calcium / Q000032:analysis; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000502:physiology*; D004396:Coloring Agents / Q000032:analysis; D005260:Female; D007150:Immunohistochemistry / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000201:enzymology / Q000502:physiology* / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis / Q000502:physiology*; D053495:Osteopontin / Q000032:analysis; D010758:Phosphorus / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley
D000880:Anthraquinones; D004396:Coloring Agents; D015675:Osteocalcin; D053495:Osteopontin; D010758:Phosphorus; C010078:alizarin; D003094:Collagen; D000469:Alkaline Phosphatase; D002118:Calcium