Analysis of cell-seeded 3-dimensional bone constructs manufactured in vitro with hydroxyapatite granules obtained from red algae
| Autor: | Rolf Ewers, Dritan Turhani, Elisabeth Watzinger, Gert Wittwer, Martina Weiβenböck, Barbara Cvikl, Dietmar Thurnher, Kaan Yerit |
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| Rok vydání: | 2005 |
| Předmět: |
Bone Regeneration
Sialoglycoproteins Blotting Western Osteocalcin Cell Culture Techniques Tissue engineering Materials Testing Medicine Osteonectin Osteopontin Bone regeneration biology Tissue Engineering business.industry Reverse Transcriptase Polymerase Chain Reaction Mesenchymal stem cell Mesenchymal Stem Cells Anatomy Cell biology Durapatite Otorhinolaryngology Cell culture Bone Substitutes Rhodophyta biology.protein Microscopy Electron Scanning Alkaline phosphatase Surgery Electrophoresis Polyacrylamide Gel Oral Surgery business |
| Zdroj: | Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons. 63(5) |
| ISSN: | 0278-2391 |
| Popis: | Purpose Bone tissue engineering is a promising approach for the treatment of defective or lost bone in the maxillofacial region. Biocompatible and biodegradable scaffolds seeded with living cells are used to create functional tissue for load-bearing bone reconstruction. The aim of this study was to manufacture cell-seeded 3-dimensional bone constructs based on hydroxyapatite ceramic granule calcified from red algae and mesenchymal cambial-layer precursor cells. The ability of these cells to grow on hydroxyapatite ceramic was quantitatively investigated to evaluate 3-dimensional bone constructs for their potential use in bone tissue engineering. Materials and methods Mesenchymal cambial-layer precursor cells were isolated from mandibular periosteum biopsy samples of 3 patients. To manufacture 72 bone constructs, these cells and hydroxyapatite ceramic granules (C GRAFT/Algipore; Clinician’s Preference LLC, Golden, CO) were cultivated under osteogenic differentiation conditions in a rotating wall vessel system. After 6 and 21 days, histologic examination and scanning electron microscopy were performed. The absolute DNA content, protein synthesis, and alkaline phosphatase activity were also quantified. The osteoblastic phenotype of the constructs was confirmed by the expression of bone-specific genes (osteocalcin, osteonectin, osteopontin, and core binding factor α1) using semiquantitative reverse transcription-polymerase chain reaction and Western blot analysis. Results Cells within the constructs showed good viability, which was evidenced by an increase in DNA content over the culture period. The decrease in alkaline phosphatase-specific activity could be an indicator of the maturation of cells and the induction of mineralization. The osteoblastic phenotype of the constructs was demonstrated on protein and at the RNA level over the entire culture period. Conclusion We observed a positive effect of hydroxyapatite ceramic granules on mesenchymal cambial-layer precursor cell behavior in cell-seeded 3-dimensional bone constructs, indicating the potential applicability of C GRAFT/Algipore composites in bone tissue engineering. |
| Databáze: | OpenAIRE |
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