Cell sheet-engineered bones used for the reconstruction of mandibular defects in an animal model

Abstract

Abstract. The aim of the present study was to investigate the generation of cell sheet-engineered bones used for the reconstruction of mandibular defects. Bone marrow stem cells (BMSCs) were cultured and induced to generate osteoblasts. Poly(lactic-co-glycolic acid) (PLGA) scaffolds were wrapped with or without cell sheets and then implanted into dogs with mandibular defects in the right side (experimental group) or the left side (control group), respectively. Subsequently, X-ray analyses, and hematoxylin and eosin staining were performed at various time points (at 4, 8, 12 or 16 weeks post-implantation; n=4 at each time point). The osteogenesis in the experimental group was significantly improved compared with that in the control group. At 16 weeks after implantation, numerous Haversian systems and a few lamellar bones were observed at the periphery. In the control group, the engineered bone (without BMSC sheets) presented fewer Haversian systems and no lamellar bones. The optical density of the fresh bone in the experimental group was significantly higher compared with that in the control group (P<0.05). In conclusion, tissue-engineered bone with the structure of lamellar bones can be generated using BMSC sheets and implantation of these bones had an improved effects compared with the control group. Cell sheet transplantation was found to enhance bone formation at the reconstruction site of the mandibular defects. IntroductionBone loss is caused by various congenital and degenerative diseases, traumas or incorrect surgical procedures, leading to severe problems, particularly in elderly individuals (1).Tissue engineering is frequently used for the reconstruction of mandible defects. Yamato et al (2) have demonstrated a novel cell sheet engineering method for tissue regeneration, which uses temperature-responsive culture dishes (TRCDs). This technique allows for different types of cultured cells to be noninvasively harvested as intact sheets through simple temperature reduction, without the use of proteolytic enzymes (3). Using this method, the noninvasive transfer of these cell sheets can be achieved, while retaining the typical distributions of Na + /K + -ATPase, glucose transporter-1, sodium-glucose linked transporter-1, aquaporin-1, neutral endopeptidase and dipeptidylendopeptidase IV (4).In the present study, scaffolds of poly(lactic-co-glycolic acid) (PLGA) were produced, and composited with recombination human bone morphogenetic protein-2 (rhBMP-2) and vascular endothelial growth factor (VEGF). In addition, bone marrow stem cells (BMSCs) were cultured in TRCDs to form BMSCs sheets. PLGA/BMP-2/VEGF wrapped with BMSCs sheets were implanted into dogs with mandibular defects. The aim of the present study was to investigate the effects of tissue-engineered bones that have the same structure as normal bones and can be used for the reconstruction of bones with mandible defects. Materials and methodsAnimals. In this study, 16 healthy, adult, male mongrel dogs (age, 14 months; weight, 18-23 kg) were used. All the...

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