Bone repair using periodontal ligament progenitor cell-seeded constructs.
The success of tissue-engineering therapies is dependent on the ability of scaffolds to guide differentiation of progenitor cells. Here we present a new approach using a biomimetic construct composed of hydroxyapatite modified with an in vitro-derived extracellular matrix (HA-ECM) and seeded with periodontal ligament progenitor cells (PDLCs). The study aimed to investigate the effect of HA-ECM on osteogenic differentiation of PDLCs and in vivo evaluation of the PDLC-seeded HA-ECM constructs using a rat calvarial critical-sized defect model. After flow-cytometric phenotyping of PDLCs for typical mesenchymal stem cell markers, the PDLCs were cultured on HA-ECM or HA alone in osteogenic media and assessed by MTT, alkaline phosphatase (ALP) assays, and real-time qPCR at different time intervals after seeding. New bone formation induced by PDLC-seeded constructs was assessed by histomorphometric analysis at 12 weeks post-operatively. The PDLCs seeded on HA-ECM showed significantly higher ALP activity and up-regulation of bone-related genes. The treatment with PDLC-seeded HA-ECM significantly improved calvarial bone repair, with the highest amount of newly formed bone elicited by cell-seeded constructs cultured for 14 days. Our results highlight the PDLC-seeded HA-ECM constructs as a promising tool for craniofacial bone regeneration.
Department of Dental Medicine, Karolinska Institutet, Huddinge, Sweden.
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D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D040761:Biomimetic Materials / Q000737:chemistry; D001861:Bone Regeneration / Q000502:physiology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D004396:Coloring Agents; D003470:Culture Media; D017886:Durapatite / Q000737:chemistry; D005109:Extracellular Matrix / Q000502:physiology; D005434:Flow Cytometry; D008297:Male; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D010006:Osteoblasts / Q000502:physiology; D010012:Osteogenesis / Q000235:genetics / Q000502:physiology; D010513:Periodontal Ligament / Q000166:cytology*; D010641:Phenotype; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D012886:Skull / Q000601:surgery; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D015854:Up-Regulation
D004396:Coloring Agents; D003470:Culture Media; D013778:Tetrazolium Salts; D013844:Thiazoles; D017886:Durapatite; D000469:Alkaline Phosphatase; C022616:thiazolyl blue