GMP-grade preparation of biomimetic scaffolds with osteo-differentiated autologous mesenchymal stromal cells for the treatment of alveolar bone resorption in periodontal disease.
BACKGROUND
Periodontal disease is a degenerative illness that leads to resorption of the alveolar bone. Mesenchymal stromal cells (MSC) represent a novel tool for the production of biologic constructs for the treatment of degenerative bone diseases. The preparation of MSC differentiated into osteogenic lineage for clinical use requires the fulfillment of strict good manufacturing practice (GMP) procedures.


METHODS
MSC were isolated from BM samples and then cultured under GMP conditions. MSC were characterized phenotypically and for their differentiative potential. Cells were seeded onto collagen scaffolds (Gingistat) and induced to differentiate into osteogenic lineages using clinical grade drugs compared with standard osteogenic supplements. Alizarin Red S stain was used to test the deposition of the mineral matrix. Standard microbiologic analysis was performed to verify the product sterility.


RESULTS
The resulting MSC were negative for CD33, CD34 and HLA-DR but showed high expression of CD90, CD105 and HLA-ABC (average expressions of 94.3%, 75.8% and 94.2%, respectively). Chondrogenic, osteogenic and adipogenic differentiation potential was demonstrated. The MSC retained their ability to differentiate into osteogenic lineage when seeded onto collagen scaffolds after exposure to a clinical grade medium. Cell numbers and cell viability were adequate for clinical use, and microbiologic assays demonstrated the absence of any contamination.


CONCLUSIONS
In the specific context of a degenerative bone disease with limited involvement of skeletal tissue, the combined use of MSC, exposed to an osteogenic clinical grade medium, and biomimetic biodegradable scaffolds offers the possibility of producing adequate numbers of biologic tissue-engineered cell-based constructs for use in clinical trials.
Salvadè
A
A
Department of Neuroscience and Biomedical Technologies, University of Milano-Bicocca, Monza, Italy.
Belotti
D
D
Donzelli
E
E
D'Amico
G
G
Gaipa
G
G
Renoldi
G
G
Carini
F
F
Baldoni
M
M
Pogliani
E  M
E
Tredici
G
G
Biondi
A
A
Biagi
E
E
D020341:Absorbable Implants; D001672:Biocompatible Materials / Q000494:pharmacology*; D050071:Bone Density Conservation Agents / Q000494:pharmacology; D001857:Bone Matrix / Q000187:drug effects / Q000378:metabolism; D001861:Bone Regeneration / Q000187:drug effects / Q000502:physiology*; D001862:Bone Resorption / Q000209:etiology / Q000503:physiopathology / Q000628:therapy*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D048091:Guided Tissue Regeneration / Q000379:methods; D006801:Humans; D007568:Jaw / Q000473:pathology / Q000503:physiopathology; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology; D010510:Periodontal Diseases / Q000503:physiopathology / Q000628:therapy*; D017154:Stromal Cells / Q000187:drug effects / Q000502:physiology*