Growth and differentiation of alveolar bone cells in tissue-engineered constructs and monolayer cultures
| Autor: | Matej Drobnič, Elvira Malicev, N. Kregar Velikonja, M.E. Kreft, Darja Marolt, M. Rode |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Osteocalcin
Cell Culture Techniques Gene Expression Bioengineering Applied Microbiology and Biotechnology Tissue engineering Osteogenesis Alveolar Process medicine Humans Osteopontin Bone regeneration Cell Proliferation Tissue Engineering biology Chemistry Cell growth Cell Differentiation Osteoblast Alkaline Phosphatase Culture Media Cell biology Durapatite medicine.anatomical_structure Biochemistry Bone Substitutes biology.protein Alkaline phosphatase Biotechnology Explant culture |
| Zdroj: | Biotechnology and Bioengineering. 100:773-781 |
| ISSN: | 1097-0290 0006-3592 |
| DOI: | 10.1002/bit.21815 |
| Popis: | The ability to enhance bone regeneration by implanting autologous osteoblasts in combination with an appropriate scaffold would be of great clinical interest. The aim of our study was to compare the growth and differentiation of alveolar bone cells in tissue-engineered constructs and in monolayer cultures, as the basis for developing procedures for routine preparation of bone-like tissue constructs. Alveolar bone tissue was obtained from four human donors and explant cultures of the cells were established. Expanded cells were seeded on macroporous hydroxyapatite granules, and cultured in medium supplemented with osteogenic differentiation factors for up to 3 weeks. Control monolayer cultures were established in parallel, and cultured in media with or without osteogenic supplements. Cell proliferation, alkaline phosphatase (AP) activity and gene expression of AP, osteopontin and osteocalcin were determined under different culture conditions at weekly intervals. Cells in tissue constructs exhibited growth patterns similar to those in control monolayer cultures: enhanced proliferation was noted during the first 2 weeks of cultivation, followed by a decrease in cell numbers. AP activity at 3 weeks was higher in all cultures in osteogenic medium than in control medium. Gene expression levels were stable in monolayer cultures in both types of media whereas, in tissue constructs, they exhibited patterns of osteogenic differentiation. Light and scanning electron microscopy examination of the cell-seeded constructs showed uniform cell distribution, as well as cell attachment and growth into the interior region of the hydroxyapatite granules. Our results show that bone-like constructs with viable cells exhibiting differentiated phenotype can be prepared by cultivation of alveolar-bone cells on the tested hydroxyapatite granules. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text