A biaxial rotating bioreactor for the culture of fetal mesenchymal stem cells for bone tissue engineering |
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| Authors: | Zhi-Yong Zhang Swee Hin Teoh Woon-Shin Chong Toon-Tien Foo Yhee-Cheng Chng Mahesh Choolani Jerry Chan |
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| Affiliation: | 1. Graduate Program in Bioengineering (GPBE), National University of Singapore, Singapore;2. Centre for Biomedical Materials Applications and Technology (BIOMAT), Department of Mechanical Engineering, Faculty of Engineering, National University of Singapore, Singapore;3. National University of Singapore Tissue Engineering Programme (NUSTEP), National University of Singapore, Singapore;4. Bioengineering Laboratory, Technology Centre for Life Sciences, Singapore Polytechnic, Singapore;5. Experimental Fetal Medicine Group, Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore and National University Hospital, Singapore;1. Department of Obstetrics and Gynaecology; VU University Medical Centre (VUmc), Amsterdam, The Netherlands;2. Department of Epidemiology and Biostatistics, VU University Medical Centre (VUmc), Amsterdam, The Netherlands;3. Department of Obstetrics and Gynaecology, Flevoziekenhuis, Almere, The Netherlands;1. Laboratory of Nuclear Energy Chemistry, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049, China;2. School of Radiological & Interdisciplinary Sciences and Collaborative Innovation Center of Radiation Medicine of Jiangsu Higher Education Institutions, Soochow University, 215123, Suzhou, China;1. Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Cologne, Germany;2. Fraunhofer Institute for Biomedical Engineering IBMT, St. Ingbert, Germany;3. Department of Chemistry, University of Cologne, Germany;4. Institute of Complex Systems, ICS-7: Biomechanics, Forschungszentrum Jülich GmbH, Jülich, Germany;5. Department of Pediatrics, Cairo University, Cairo, Egypt;6. Department of Internal Medicine III, University Clinics of Cologne, Cologne, Germany;7. Department of Paediatric Cardiology, University Clinics of Cologne, Cologne, Germany;8. Division of Pediatric Cardiology, University Children''s Hospital, Giessen, Germany;1. Department of Orthopaedics, Xijing Hospital, Fourth Military Medical University, Xi''an, Shaanxi, China;2. Department of Plastic and Reconstructive Surgery, Shanghai 9th People''s Hospital, Shanghai Key Laboratory of Tissue Engineering, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China;3. National Tissue Engineering Center of China, Shanghai 200241, China;4. No. 89 Hospital of PLA, China;5. Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457, Singapore;1. Department of Orthopedics, Xijing Hospital, The Fourth Military Medical University, Xi’an 710032, China;2. School of Mechanical Engineering, Shanghai Jiao Tong University, State Key Laboratory of Mechanical System and Vibration, Shanghai 200240, China;3. Department of Plastic and Reconstructive Surgery, Shanghai 9th People''s Hospital, Shanghai Key Laboratory of Tissue Engineering, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China;4. National Tissue Engineering Center of China, Shanghai 200241, China |
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| Abstract: | The generation of effective tissue engineered bone grafts requires efficient exchange of nutrients and mechanical stimulus. Bioreactors provide a manner in which this can be achieved. We have recently developed a biaxial rotating bioreactor with efficient fluidics through in-silico modeling. Here we investigated its performance for generation of highly osteogenic bone graft using polycaprolactone–tricalcium phosphate (PCL–TCP) scaffolds seeded with human fetal mesenchymal stem cell (hfMSC). hfMSC scaffolds were cultured in either bioreactor or static cultures, with assessment of cellular viability, proliferation and osteogenic differentiation in vitro and also after transplantation into immunodeficient mice. Compared to static culture, bioreactor-cultured hfMSC scaffolds reached cellular confluence earlier (day 7 vs. day 28), with greater cellularity (2×, p < 0.01), and maintained high cellular viability in the core, which was 2000 μm from the surface. In addition, bioreactor culture was associated with greater osteogenic induction, ALP expression (1.5× p < 0.01), calcium deposition (5.5×, p < 0.001) and bony nodule formation on SEM, and in-vivo ectopic bone formation in immunodeficient mice (3.2×, p < 0.001) compared with static-cultured scaffolds. The use of biaxial bioreactor here allowed the maintenance of cellular viability beyond the limits of conventional diffusion, with increased proliferation and osteogenic differentiation both in vitro and in vivo, suggesting its utility for bone tissue engineering applications. |
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