beta-Tricalcium phosphate promotes cell proliferation, osteogenesis and bone regeneration in intrabony defects in dogs.
OBJECTIVE
This study investigates the effect of the new synthetic bone grafting material, high pure-phase beta-tricalcium phosphate (Cerasorb(1) M, granule size 500-1000microm), on the osteogenesis process and proliferation marker in bone marrow stromal cells (BMSCs) and its regenerative effect in the periodontal intrabony defects in dogs.


METHODS
The effect of Cerasorb(1) M (20 and 40mgml(-1) for 1 and 2 weeks) on the proliferation rate of BMSCs was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay on the proliferating cell nuclear antigen (PCNA) by immunoblotting and on alkaline phosphatase level by colourimetric assay. The regenerative effect of Cerasorb(1) M in the periodontal intrabony defects in dogs was investigated by histological and immunohistochemical analysis after 3 and 6 months of grafting.


RESULTS
Incubation of BMSCs with Cerasorb(1) M for 2 weeks led to significant increase in cell proliferation rate, which was associated with increased PCNA. Cerasorb(1) M significantly increased the production of alkaline phosphatase as a marker for the osteogenic stromal lineage and for differentiation and bone formation in BMSCs after 2 weeks. In the histological features and immunohistochemical analysis of PCNA of the intrabony defects in dogs augmented with Cerasorb(1) M, osteoid tissue with a plate-like structure and cellular mesenchymal proliferation besides osteoid islands joined by bridges were observed after 3 months. Six months after the implantation, the Cerasorb(1) M granules were replaced by abundant new plate-like bone besides PCNA-enriched, small, oval-shaped mononuclear cells and multinucleated-giant cells that were attached to newly formed bones. No remains of the Cerasorb(1) M granules could be seen after 3 and 6 months with the newly formed plate-like bones and no histological sign of inflammatory reaction or formation of foreign-body granulomas.


CONCLUSIONS
Cerasorb(1) M may induce cell proliferation via induction of PCNA that may induce early osteogenesis and bone formation. Cerasorb(1) M regenerated the bone completely in intrabony defects and that this regeneration was highly associated with PCNA expression in different cell lineage.
Neamat
Amany
A
Department of Surgery and Oral Medicine Researches, National Research Center, Cairo, Egypt.
Gawish
Abeer
A
Gamal-Eldeen
Amira M
AM
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000539:Alveolar Process / Q000601:surgery; D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000378:metabolism; D001861:Bone Regeneration / Q000187:drug effects*; D018786:Bone Substitutes / Q000494:pharmacology*; D002130:Calcium Phosphates / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004285:Dogs; D008297:Male; D008334:Mandible / Q000601:surgery; D051379:Mice; D010012:Osteogenesis / Q000187:drug effects; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
D018786:Bone Substitutes; D002130:Calcium Phosphates; D018809:Proliferating Cell Nuclear Antigen; C485817:beta-tricalcium phosphate; D000469:Alkaline Phosphatase