IADR Abstract Archives
Characterisation of Alternative Bone Graft Materials for Cleft Palate Repair
Objectives: Cleft lip/palate (CL(P)) is a common congenital craniofacial deformity with an average worldwide prevalence of 1 per 1000 live births, and requires lifelong surgical and non-surgical interventions. Alveolar bone grafts (ABG) at the mixed dentition stage are used in cleft repair and autologous bone is still regarded as the gold standard. However, donor-site morbidity is the major drawback. Therefore, synthetic substitutes are emerging as possible alternative ABG materials.
The purpose of this in vitro study was to develop a novel in vitro osteogenic progenitor cell-based model and to investigate the role of various calcium phosphate-based (CaP) materials in osteogenic differentiation.
Methods: Human embryonic palatal mesenchymal cells (HEPM) and normal gingival fibroblasts (MM1) were seeded on four (A, B, C, D) CaP scaffolds (Kuros Biosciences BV, The Netherlands), in α-Minimal Essential Medium (α-MEM) supplemented with 10% foetal calf serum (FCS) with/without DAG (dexamethasone, ascorbic acid and β-glycerophosphate) and/or bone morphogenic protein-2(BMP2) osteogenic reagents, for 21 days and analysed for biocompatibility and osteodifferentiation at different time points. Cell adhesion was investigated microscopically after staining the cells with methylene blue. Viability/proliferation was assessed by MTT assay, while expression of the osteogenic differentiation marker RUNX2 was visualised by immunofluorescence assay and quantified by using Image-J software.
Results: HEPM and MM1 cells were well attached to the materials. Material B stimulated cell proliferation more than other materials. HEPM expressed RUNX2 when cultured with the materials at different levels in the presence/absence of DAG and BMP2. In general, RUNX2 was expressed within 3 days and peaked on day 7. Material-wise, material D stimulated a higher expression level of RUNX2 than the other materials. However, in the presence of DAG, RUNX2 expression was significantly enhanced.
Conclusions: These data suggest that CaP scaffolds were suitable for expansion and osteogenic differentiation of HEPM and were therefore promising candidates as ABG alternative.
The purpose of this in vitro study was to develop a novel in vitro osteogenic progenitor cell-based model and to investigate the role of various calcium phosphate-based (CaP) materials in osteogenic differentiation.
Methods: Human embryonic palatal mesenchymal cells (HEPM) and normal gingival fibroblasts (MM1) were seeded on four (A, B, C, D) CaP scaffolds (Kuros Biosciences BV, The Netherlands), in α-Minimal Essential Medium (α-MEM) supplemented with 10% foetal calf serum (FCS) with/without DAG (dexamethasone, ascorbic acid and β-glycerophosphate) and/or bone morphogenic protein-2(BMP2) osteogenic reagents, for 21 days and analysed for biocompatibility and osteodifferentiation at different time points. Cell adhesion was investigated microscopically after staining the cells with methylene blue. Viability/proliferation was assessed by MTT assay, while expression of the osteogenic differentiation marker RUNX2 was visualised by immunofluorescence assay and quantified by using Image-J software.
Results: HEPM and MM1 cells were well attached to the materials. Material B stimulated cell proliferation more than other materials. HEPM expressed RUNX2 when cultured with the materials at different levels in the presence/absence of DAG and BMP2. In general, RUNX2 was expressed within 3 days and peaked on day 7. Material-wise, material D stimulated a higher expression level of RUNX2 than the other materials. However, in the presence of DAG, RUNX2 expression was significantly enhanced.
Conclusions: These data suggest that CaP scaffolds were suitable for expansion and osteogenic differentiation of HEPM and were therefore promising candidates as ABG alternative.